The Hengduan Mountain region is located at the eastern end of the Himalayan range and in the sout... more The Hengduan Mountain region is located at the eastern end of the Himalayan range and in the southeastern corner of the Qinghai-Tibet Plateau in eastern Asia. It is probably the richest temperate region in the world biologically, with about 8000 species recorded from the 500,000 km 2 region. A high rate of polyploidy was expected in the Hengduan Mountains because of the unusually high species diversity and endemism, the widespread alpine environment in this region, and the importance of polyploidy in plant evolution. Chromosome numbers for 552 taxa of native Hengduan angiosperms were obtained from published literature. If the criterion of infrageneric polyploidy is used (i.e., the possession of three or more basic chromosome sets in a nucleus), the frequency of polyploidy is only 22%. It is 37% and 58% if taxa with haploid chromosome number n ϭ 14 or more, or n ϭ 11 or more are considered to be polyploids, respectively. Several highly diversified groups such as Aconitum and Delphinium also show a low proportion of polyploidy (18.6% and 0%, respectively) in the Hengduan Mountains. Examples from other large plant groups (e.g., Saxifraga, Ligularia, and Cremanthodium) and endemic genera (e.g., Solms-laubachia, Tibetia, and Nomocharis) again indicated that polyploidy, especially infrageneric polyploidy, may have played a minor role in the evolutionary diversification of these plants in the Hengduan Mountains.
ABSRACT. Two new species of Wikstroemia from Sichuan, China, are described and illustrated. Wikst... more ABSRACT. Two new species of Wikstroemia from Sichuan, China, are described and illustrated. Wikstroemia guanxianensis from Dujiangyan Xian is similar to W. dolichantha in the one-sided development of the disc and in the elongate inflorescence. It differs from W. dolichantha in the oval, wider leaves (to 3.5 cm wide) rounded at the base, longer pedicel (1.5 mm), and disc quadrate, apex truncate, undulate. Wikstroemia jiulongensis is similar to W. fargesii, but differs from it in young branches with scattered pubescence, the peduncles absent or very short, and a quadrate disc with undulate apex. A key to similar species of Wikstroemia in Sichuan is provided.
The karyotypes of five species of Solms-laubachia (Brassicaceae) from Hengduan Mountains (Sichuan... more The karyotypes of five species of Solms-laubachia (Brassicaceae) from Hengduan Mountains (Sichuan and Yunnan provinces) are investigated for the first time. The karyotype formulas of S. minor, S. eurycarpa, S. pulcherrima, and S. linearifolia are the same and all have 2n=14=6m+6sm+2st, whereas that of S. retropilosa is 2n=28=12m+12sm+4st. The first four species are diploids, the last is tetraploid, and the
A distinctive, previously undescribed species of Epimedium (Berberidaceae), which we propose nami... more A distinctive, previously undescribed species of Epimedium (Berberidaceae), which we propose naming E. tianmenshanensis, was found during a floristic survey in Hunan, China. It is similar to E. baojingense and E. franchetii, but has smaller flowers and petals with a short, blunt spur. We determined the phylogenetic position of the new species using DNA sequences of nuclear ITS and two chloroplast regions (atpB-rbcL intergenic spacer and matK). Molecular evidence indicates that E. tianmenshanensis occupies a distinct group related to E. baojingense.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To investigate the effects of metformin on fatty livers in insulin-resistant rats. Thirty-one mal... more To investigate the effects of metformin on fatty livers in insulin-resistant rats. Thirty-one male Wistar rats were randomly divided into a normal (n=8) and an experimental group (n=23). The normal group rats were fed a standard diet, and those of the experimental group with a high-fat diet. After 8 weeks, 7 rats from the experimental group were sacrificed to verify whether the model was established successfully. Then the experimental group was randomly subdivided into two groups: one with metformin treatment (n=8) and one without (n=8). After 6 weeks, insulin sensitivity was measured with glucose infusion rate (GIR) by euglycermic hyperinsulinemia clamp technique. Aminotransferase, triglyceride (TG) and free fatty acids (FFA) were measured by biochemical methods, insulin by radioimmunoassay and tumor necrosis factor-alpha (TNF alpha) by ELISA. The steatosis changes and inflammation activity of all rat livers were scored histologically. Compared with the model group, the insulin resistance, liver index, visceral adiposity and weight loss of the metformin group were dramatically ameliorated. The steatosis and the inflammatory activity in the livers and the level of serum aminotransferase of the metformin group were also significantly decreased. Furthermore, metformin treatment lowered serum TG, liver lipid accumulation and the production of FFA and TNF alpha. Metformin can significantly improved insulin resistance and fatty liver development in rats fed a high-fat diet. It may become a new choice for fatty liver treatment in the future.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2... more To detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2 cells. To develop a kind of RNAi that specifically targets human augmenter of liver regeneration by synthesizing small interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on hALR expression. The expression of hALR in HepG2 cells was observed with immunocytochemistry. The RNAi plasmid pSIALR-A and the unrelated control plasmid pSIALR-B were transfected into HepG2 cells. Forty-eight hours after transfection, the protein level of hALR was measured with immunocytochemistry; meanwhile, the reverse transcription PCR (RT-PCR) was performed to detect the expression of hALR mRNA. hALR was expressed by HepG2 cells. siRNA plasmid pSIALR-A, which targets the cDNA of hALR and the unrelated control plasmid pSIALR-B, was successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly, compared with that of pSIALR-B. The results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To screen the hALR-interacting protein by phage-displayed technique and identify its biological a... more To screen the hALR-interacting protein by phage-displayed technique and identify its biological activities. The specific phage clones that interacted with target protein hALR from a cDNA library of hepatocarcinoma cells were selected using the T7 phage-displayed technique. The acquired cDNA inserts were sequenced and analyzed by bioinformatic tools. The biological activities of the phage-displayed peptide affecting QGY hepatocarcinoma cells were studied using 3H-TdR method. The cDNA inserts with 212 bp were acquired after 4 rounds of biopanning. They showed 100% homology with citron kinase. The phage-displayed peptide and the peptide combined with hALR affected QGY cells proliferation. hALR-interacting peptide can be specifically screened by phage-displayed technique. Citron kinase that interacted with hALR potentially plays an important role in the proliferation of hepatocarcinoma cells.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To create a rat insulin resistant fatty liver model. 14 male Wistar rats were randomly divided in... more To create a rat insulin resistant fatty liver model. 14 male Wistar rats were randomly divided into a model and a control group. The model rats were fed a high-fat diet (45% of energy from fat) for 8 weeks, and the control group a standard diet (19% of energy from fat). Insulin sensitivity was measured with glucose infusion rate (GIR) by the euglycermic hyperinsulinemia clamp technique. The aminotransferase, triglyceride (TG), free fatty acid (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by biochemical methods, and insulin was measured by radioimmunoassay. The histological and ultrastructural changes of all rat livers were scored using light and electron microscopy. Rats fed the high-fat diet developed panlobular macrovesicular steatosis, lobular inflammatory cell infiltration and abnormal mitochondria, whereas those fed the standard diet had normal livers. All model group rats had elevated levels of aminotransferase, TG, FFA, insulin and liver index, and low GIR. In addition, the high-fat diet increased MDA and decreased SOD. A fatty liver and insulin resistance model was successfully developed in rats fed a high-fat diet for 8 weeks, which provided a useful experimental tool for elucidating pathogenesis and treatment of fatty liver.
To take advantage of the small interfering ribonucleic acid (siRNA) targeting the human augmenter... more To take advantage of the small interfering ribonucleic acid (siRNA) targeting the human augmenter of liver regeneration (hALR) and anti-hALR monoclonal antibody (McAb) to inhibit the function of hALR, and to demonstrate whether the growth of hepatoma is influenced by siRNA targeting hALR and anti-hALR McAb through inhibiting expression of hALR. This study was conducted in the Key Laboratory of Molecular Biology for Infectious Diseases, Ministry of Education, Institute for Viral Hepatitis, Chongqing Medical University, China, between January 2005 and May 2007. We transfected siRNA plasmid pSIALR-A, which targeted the complementary deoxyribonucleic acid (cDNA) of hALR and the unrelated control plasmid pSIALR-B into human hepatocellular liver carcinoma cell line (HepG2) cells. Then, the proliferation of HepG2 cells, after being treated with pSIALR-A and anti-hALR McAb was detected. The growth of the xenograft tumor was observed after being treated with pSIALR-A and anti-hALR McAb in nude mice. We successfully constructed expressing plasmid pSIALR-A and pSIALR-B. The pSIALR-A inhibited the expression of hALR in HepG2 cells significantly. The siRNA targeting hALR and anti-hALR McAb inhibited obviously the growth of HepG2 cells in vitro. siRNA targeting hALR and anti-hALR McAb significantly inhibited the growth of xenograft tumor in 5 nude mice. Anti-hALR McAb inhibited apparently the autonomous growth of HepG2 cells. Our results demonstrated that anti-hALR McAb inhibited the autonomous growth of hepatoma cells obviously, moreover, hALR maintained the autonomous growth of hepatoma cells in vitro through an autocrine mechanism.
To construct a yeast expression system of human augmenter of liver regeneration (hALR) and to exa... more To construct a yeast expression system of human augmenter of liver regeneration (hALR) and to examine its bioactivity in vitro. With PCR and gene recombination techniques, cDNA of open reading frame of hALR was obtained from recombinant plasmid pcDNA3.1-hALR and inserted into plasmid pPIC9. The cDNA of hALR from recombinant plasmid pPIC9-hALR demonstrated by sequencing was subcloned into plasmid pPIC9K. The recombinant plasmid pPIC9K-hALR was transformed into GS115 with electroporation. hALR was expressed by GS115 under the induction of 5 mL/L methanol and purified with ultrafiltration after it was analyzed by 15% SDS-PAGE and Western blot. The effects of hALR on in vitro proliferation of QGY and HepG(2) cells were evaluated by (3)H-TdR methods. The correctness and integrity of recombinant plasmids pPIC9-hALR and pPIC9K-hALR were identified by restriction digestion, PCR and sequencing methods, respectively. hALR as a secretive protein was successfully expressed by GS115. Its molecular weight was about 15 ku and the target protein was about 60% of the total protein in the supernatant from GS115 with plasmid pPIC9K-hALR. The results of Western blot of hALR showed the specific band. The high qualitative hALR was obtained through ultrafiltration. hALR could stimulate in vitro proliferation of QGY and HepG(2) cells in a dose-dependent manner, but there was a difference in reactivity to hALR between QGY and HepG(2). The hALR as a secretive protein can be successfully expressed by GS115. It may stimulate in vitro proliferation of QGY and HepG(2) cells at a dose-dependent manner. But QGY and HepG(2) cells have different reactivities to hALR.
Human augmenter of liver regeneration (hALR) is highly expressed in hepatocellular carcinoma (HCC... more Human augmenter of liver regeneration (hALR) is highly expressed in hepatocellular carcinoma (HCC), but rarely expressed in normal liver cells. ALR could stimulate the proliferation of HCC cell line in vitro, but has no effect on normal hepatocytes. This study was to investigate the inhibitory effect of blocking the expression of hALR on the proliferation of HCC cell line HepG2 using small interfering RNA (siRNA) targeting ALR and anti-ALR monoclonal antibody (McAb). RNA interference (RNAi) plasmid pSIALR-A targeting hALR cDNA and control plasmid pSIALR-B were constructed and transfected into HepG2 cells, respectively. After transfection, the expression of green fluorescent protein was observed under a fluorescent microscope to calculate transfection efficiency, the protein level of hALR was measured by immunocytochemistry, meanwhile, the expression of hALR mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). (3)H-TdR incorporation approach was performed to detect the proliferation of HepG2 cells after transfection and after neutralization with anti-hALR McAb. hALR was expressed in HepG2 cells. Plasmids pSIALR-A and pSIALR-B were successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly by 83% as compared with pSIALR-B. pSIALR-A specifically inhibited the growth of HepG2 cells after transfection. (3)H-TdR incorporation of pSIALR-A/HepG2 cells was significantly lower than that of pSIALR-B/HepG2 cells (67 687+/-6 548 vs. 104 807+/-5 713, P<0.05). Anti-hALR McAb markedly inhibited the autonomous growth of HepG2 cells (P<0.05). hALR is highly expressed in HepG2 cells. The short interfering RNA targeting hALR gene could specifically suppress the expression of hALR, and subsequently inhibit the growth of HepG2 cells. Anti-hALR McAb could partially inhibit the autonomous growth of HepG2 cells.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat ... more To evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat augmenter of liver regeneration (rALR) injection in treating rats with acute hepatic failure. Acute hepatic failure (AHF) was induced in rats using D-gal (1.2 g/kg). The rats were then randomly divided into 6 groups. Group I received no further treatment and served as blank controls; group II received 1 ml buffered saline once through intrasplenic injection; group III received 1 ml rALR; group IV received 2 x 10(7)/ml hepatocytes; group V received 2 x 10(7) hepatocytes suspended in 1 ml rALR (50 microg/kg) and group VI received 2 x 10(7) hepatocytes in 1 ml cyclosporine A (10 mg/kg). The rats of the various treated groups were sacrificed at day 1, 5 and at week 2 and their livers and spleens were examined histopathologically. Blood samples of the rats were also obtained to determine the levels of TNF alpha and IL-1 beta. There were no significant differences in survival between group I, II and III rats. 33.3% of the group IV rats survived for 2 weeks. At week 2, the survival rate of group V rats was significantly higher than that of group IV, but there was of no statistical significant increase when compared to that of group VI rats. Hepatocytes transplanted into spleens survived for 5-7 days in the spleens of group IV and VI rats, but they survived at least 2 weeks in group V. The average serum TNF alpha level in group IV was significantly higher than that in groups V and VI on the first postoperative day, but after four days, only the difference between group IV and group VI was significant (P < 0.05). The average serum level of TNF alpha in group II was higher than that in groups IV, V and VI on the first postoperative day (P < 0.05), but there were no significant differences between those in groups IV, V and VI on the 1st and the 5th postoperative days. Newborn rat hepatocyte intrasplenic transplantation combined with rALR is effective in treating acute hepatic function failure induced by D-gal in rats. The transplanted hepatocytes can survive for at least 2 weeks in the spleens. The rALR mixed with the hepatocytes injected into the spleens may be able to facilitate the hepatocyte regeneration, to inhibit liver cell apoptosis and to suppress the cellular immunity.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)e... more To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)expression and releasing in macrophage induced by lipopolysaccharide(LPS). The murine macrophage-like cell line RAW264.7 cultured in vitro divided into LPS group and LPS+EP group. The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR. The cytoplasmic and nuclear HMGB1 levels were detected by Western blot. The contents of HMGB1 and TNF-α and IL-6 protein in cultured cells supernatant were detected by ELISA. Immunocytochemistry and confocal laser-scanning microscopy were used to confirm the relocation and distribution of intracellular HMGB1 protein in RAW264.7 cells. HMGB1 mRNA expression in the LPS+EP group was significantly lower than in LPS alone, at 24, 36 and 48 hours. In the LPS+EP stimulation group, the cytoplasm stained weakly while the nuclear stain was stronger than that of the LPS group at the same time points. Both TNF-α and IL-6 levels in LPS+EP group were sign...
ABSTRACT Rubia L. is the type genus of the coffee family Rubiaceae and the third largest genus in... more ABSTRACT Rubia L. is the type genus of the coffee family Rubiaceae and the third largest genus in the tribe Rubieae, comprising ca. 80 species restricted to the Old World. China is an important diversity center for Rubia where about half of its species occur. However, its internal phylogenetic relationships are still poorly understood. The objective of the present study is to contribute to the phylogenetic relationships within Rubia, using nuclear ITS and six plastid markers and focusing on species from China. Twenty-seven species of Rubia were sampled to infer their phylogeny using Maximum Parsimony, Maximum likelihood and Bayesian analyses. The monophyly of Rubia is supported, provided that R. rezniczenkoana is excluded from Rubia and transferred to Galium as a new combination: G. rezniczenkoanum (Litv.) L. E. Yang & Z. L. Nie. Within Rubia two clades are clearly supported. They correspond to the traditional sect. Rubia s.l. (A) and sect. Oligoneura (B), and are morphologically mainly separable by their pinnate (A) versus palmate (B) leaf venation. Plesiomorphic features are the pinnate leaf veining in sect. Rubia s.l. and the occurrence of some species with opposite leaves and true stipules in sect. Oligoneura. In sect. Oligoneura one can assume an evolution from species with opposite leaves and true stipules (as in the R. siamensis group) to those with whorls of two leaves and two leaf-like stipules (as in ser. Chinenses and the R. mandersii group) and finally with whorls of 6 or even more elements (as in ser. Cordifoliae). The correlation between clades recognized by DNA analyses and available differential morphological features is partly only loose, particularly in the group of R. cordifolia with 2x, 4x and 6x cytotypes. This may be due to rapid evolutionary divergence and/or hybridization and allopolyploidy.
Although little morphological differentiation is detected between intercontinental disjunct popul... more Although little morphological differentiation is detected between intercontinental disjunct populations of Phryma , this monotypic genus shows distinct molecular divergence corresponding to its distribution in eastern Asia and eastern North America . This study further employs amplified fragment length polymorphism ( AFLP) analyses using two selective primer pairs to quantify genetic diversity and evaluate the morphological stasis . Most of the molecular variance is accounted for by variance among populations between regions . Cluster and PCA analyses revealed that Phryma constitutes two major groups in line with their geographic distribution , with one genetically distinct group from the eastern Asia and the other with accessions from eastern North America . The results robustly supported the distinct genetic divergence and morphological stasis in Phryma .
Three new phenolic biphenyl derivatives (1-3) and one new lignan glycoside (4) were isolated from... more Three new phenolic biphenyl derivatives (1-3) and one new lignan glycoside (4) were isolated from the leaves and twigs of Osteomeles schwerinae. The structures of the new compounds were established by spectroscopic data interpretation. The inhibitory effects of 1-4 on rat lens aldose reductase in vitro were examined, and compounds 1-3 markedly inhibited the enzyme with IC50 values of 3.8 to 13.8 μM. In addition, the effects of these isolates on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in zebrafish larvae were investigated. Compound 1 was the most effective in reducing HG-induced dilation of hyaloid-retinal vessels.
It has been suggested that many plants now found in the arctic originated from ancestors that occ... more It has been suggested that many plants now found in the arctic originated from ancestors that occurred at high altitudes in the southern mountains of the Northern Hemisphere during the Tertiary. However, this hypothesis has rarely been tested using a molecular phylogenetic approach. Here, we present a fossil-calibrated molecular phylogeny of Lagotis, an arctic-alpine genus with the greatest diversity in the Qinghai-Tibetan Plateau (QTP) and Central Asian mountains, based on five chloroplast (matK, psbA-trnH, rps16, trnG-S, trnL-F) and nuclear ribosomal ITS DNA markers. Within this framework, we infer the ancestral area and biogeographic history of the genus. Four major clades (A-D) within Lagotis were recovered with strong support, which largely correspond to the previous classification of the genus. Within clade A, Lagotis species from QTP were distributed among several subclades, and L. integrifolia from Central Asia was sister to L. glauca and L. minor from the arctic and subarctic region. The Bayesian molecular dating and the ancestral area reconstruction analyses suggested that Lagotis could have originated in the QTP in the Miocene (Tertiary), and that the genus radiated from the Miocene to Pleistocene. The diversification of Lagotis probably took place predominantly in the QTP and it subsequently spread to the Central Asian highlands, followed by northward migration into the arctic. Our results support the hypothesis that the "Central Asiatic Highland Corridor" as an important route for the migration of the flora between the arctic and QTP. ) and alignment are available in the Supplementary Data section of the online version of this article at
The Hengduan Mountain region is located at the eastern end of the Himalayan range and in the sout... more The Hengduan Mountain region is located at the eastern end of the Himalayan range and in the southeastern corner of the Qinghai-Tibet Plateau in eastern Asia. It is probably the richest temperate region in the world biologically, with about 8000 species recorded from the 500,000 km 2 region. A high rate of polyploidy was expected in the Hengduan Mountains because of the unusually high species diversity and endemism, the widespread alpine environment in this region, and the importance of polyploidy in plant evolution. Chromosome numbers for 552 taxa of native Hengduan angiosperms were obtained from published literature. If the criterion of infrageneric polyploidy is used (i.e., the possession of three or more basic chromosome sets in a nucleus), the frequency of polyploidy is only 22%. It is 37% and 58% if taxa with haploid chromosome number n ϭ 14 or more, or n ϭ 11 or more are considered to be polyploids, respectively. Several highly diversified groups such as Aconitum and Delphinium also show a low proportion of polyploidy (18.6% and 0%, respectively) in the Hengduan Mountains. Examples from other large plant groups (e.g., Saxifraga, Ligularia, and Cremanthodium) and endemic genera (e.g., Solms-laubachia, Tibetia, and Nomocharis) again indicated that polyploidy, especially infrageneric polyploidy, may have played a minor role in the evolutionary diversification of these plants in the Hengduan Mountains.
ABSRACT. Two new species of Wikstroemia from Sichuan, China, are described and illustrated. Wikst... more ABSRACT. Two new species of Wikstroemia from Sichuan, China, are described and illustrated. Wikstroemia guanxianensis from Dujiangyan Xian is similar to W. dolichantha in the one-sided development of the disc and in the elongate inflorescence. It differs from W. dolichantha in the oval, wider leaves (to 3.5 cm wide) rounded at the base, longer pedicel (1.5 mm), and disc quadrate, apex truncate, undulate. Wikstroemia jiulongensis is similar to W. fargesii, but differs from it in young branches with scattered pubescence, the peduncles absent or very short, and a quadrate disc with undulate apex. A key to similar species of Wikstroemia in Sichuan is provided.
The karyotypes of five species of Solms-laubachia (Brassicaceae) from Hengduan Mountains (Sichuan... more The karyotypes of five species of Solms-laubachia (Brassicaceae) from Hengduan Mountains (Sichuan and Yunnan provinces) are investigated for the first time. The karyotype formulas of S. minor, S. eurycarpa, S. pulcherrima, and S. linearifolia are the same and all have 2n=14=6m+6sm+2st, whereas that of S. retropilosa is 2n=28=12m+12sm+4st. The first four species are diploids, the last is tetraploid, and the
A distinctive, previously undescribed species of Epimedium (Berberidaceae), which we propose nami... more A distinctive, previously undescribed species of Epimedium (Berberidaceae), which we propose naming E. tianmenshanensis, was found during a floristic survey in Hunan, China. It is similar to E. baojingense and E. franchetii, but has smaller flowers and petals with a short, blunt spur. We determined the phylogenetic position of the new species using DNA sequences of nuclear ITS and two chloroplast regions (atpB-rbcL intergenic spacer and matK). Molecular evidence indicates that E. tianmenshanensis occupies a distinct group related to E. baojingense.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To investigate the effects of metformin on fatty livers in insulin-resistant rats. Thirty-one mal... more To investigate the effects of metformin on fatty livers in insulin-resistant rats. Thirty-one male Wistar rats were randomly divided into a normal (n=8) and an experimental group (n=23). The normal group rats were fed a standard diet, and those of the experimental group with a high-fat diet. After 8 weeks, 7 rats from the experimental group were sacrificed to verify whether the model was established successfully. Then the experimental group was randomly subdivided into two groups: one with metformin treatment (n=8) and one without (n=8). After 6 weeks, insulin sensitivity was measured with glucose infusion rate (GIR) by euglycermic hyperinsulinemia clamp technique. Aminotransferase, triglyceride (TG) and free fatty acids (FFA) were measured by biochemical methods, insulin by radioimmunoassay and tumor necrosis factor-alpha (TNF alpha) by ELISA. The steatosis changes and inflammation activity of all rat livers were scored histologically. Compared with the model group, the insulin resistance, liver index, visceral adiposity and weight loss of the metformin group were dramatically ameliorated. The steatosis and the inflammatory activity in the livers and the level of serum aminotransferase of the metformin group were also significantly decreased. Furthermore, metformin treatment lowered serum TG, liver lipid accumulation and the production of FFA and TNF alpha. Metformin can significantly improved insulin resistance and fatty liver development in rats fed a high-fat diet. It may become a new choice for fatty liver treatment in the future.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2... more To detect whether there is an expression of human augmenter of liver regeneration (hALR) in HepG2 cells. To develop a kind of RNAi that specifically targets human augmenter of liver regeneration by synthesizing small interfering RNA (siRNA) in vivo, and to assess the inhibitory effect of this siRNA on hALR expression. The expression of hALR in HepG2 cells was observed with immunocytochemistry. The RNAi plasmid pSIALR-A and the unrelated control plasmid pSIALR-B were transfected into HepG2 cells. Forty-eight hours after transfection, the protein level of hALR was measured with immunocytochemistry; meanwhile, the reverse transcription PCR (RT-PCR) was performed to detect the expression of hALR mRNA. hALR was expressed by HepG2 cells. siRNA plasmid pSIALR-A, which targets the cDNA of hALR and the unrelated control plasmid pSIALR-B, was successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly, compared with that of pSIALR-B. The results showed that the small interfering RNA targeting hALR suppresses the expression of hALR in a sequence-specific manner
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To screen the hALR-interacting protein by phage-displayed technique and identify its biological a... more To screen the hALR-interacting protein by phage-displayed technique and identify its biological activities. The specific phage clones that interacted with target protein hALR from a cDNA library of hepatocarcinoma cells were selected using the T7 phage-displayed technique. The acquired cDNA inserts were sequenced and analyzed by bioinformatic tools. The biological activities of the phage-displayed peptide affecting QGY hepatocarcinoma cells were studied using 3H-TdR method. The cDNA inserts with 212 bp were acquired after 4 rounds of biopanning. They showed 100% homology with citron kinase. The phage-displayed peptide and the peptide combined with hALR affected QGY cells proliferation. hALR-interacting peptide can be specifically screened by phage-displayed technique. Citron kinase that interacted with hALR potentially plays an important role in the proliferation of hepatocarcinoma cells.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To create a rat insulin resistant fatty liver model. 14 male Wistar rats were randomly divided in... more To create a rat insulin resistant fatty liver model. 14 male Wistar rats were randomly divided into a model and a control group. The model rats were fed a high-fat diet (45% of energy from fat) for 8 weeks, and the control group a standard diet (19% of energy from fat). Insulin sensitivity was measured with glucose infusion rate (GIR) by the euglycermic hyperinsulinemia clamp technique. The aminotransferase, triglyceride (TG), free fatty acid (FFA), superoxide dismutase (SOD) and malondialdehyde (MDA) were measured by biochemical methods, and insulin was measured by radioimmunoassay. The histological and ultrastructural changes of all rat livers were scored using light and electron microscopy. Rats fed the high-fat diet developed panlobular macrovesicular steatosis, lobular inflammatory cell infiltration and abnormal mitochondria, whereas those fed the standard diet had normal livers. All model group rats had elevated levels of aminotransferase, TG, FFA, insulin and liver index, and low GIR. In addition, the high-fat diet increased MDA and decreased SOD. A fatty liver and insulin resistance model was successfully developed in rats fed a high-fat diet for 8 weeks, which provided a useful experimental tool for elucidating pathogenesis and treatment of fatty liver.
To take advantage of the small interfering ribonucleic acid (siRNA) targeting the human augmenter... more To take advantage of the small interfering ribonucleic acid (siRNA) targeting the human augmenter of liver regeneration (hALR) and anti-hALR monoclonal antibody (McAb) to inhibit the function of hALR, and to demonstrate whether the growth of hepatoma is influenced by siRNA targeting hALR and anti-hALR McAb through inhibiting expression of hALR. This study was conducted in the Key Laboratory of Molecular Biology for Infectious Diseases, Ministry of Education, Institute for Viral Hepatitis, Chongqing Medical University, China, between January 2005 and May 2007. We transfected siRNA plasmid pSIALR-A, which targeted the complementary deoxyribonucleic acid (cDNA) of hALR and the unrelated control plasmid pSIALR-B into human hepatocellular liver carcinoma cell line (HepG2) cells. Then, the proliferation of HepG2 cells, after being treated with pSIALR-A and anti-hALR McAb was detected. The growth of the xenograft tumor was observed after being treated with pSIALR-A and anti-hALR McAb in nude mice. We successfully constructed expressing plasmid pSIALR-A and pSIALR-B. The pSIALR-A inhibited the expression of hALR in HepG2 cells significantly. The siRNA targeting hALR and anti-hALR McAb inhibited obviously the growth of HepG2 cells in vitro. siRNA targeting hALR and anti-hALR McAb significantly inhibited the growth of xenograft tumor in 5 nude mice. Anti-hALR McAb inhibited apparently the autonomous growth of HepG2 cells. Our results demonstrated that anti-hALR McAb inhibited the autonomous growth of hepatoma cells obviously, moreover, hALR maintained the autonomous growth of hepatoma cells in vitro through an autocrine mechanism.
To construct a yeast expression system of human augmenter of liver regeneration (hALR) and to exa... more To construct a yeast expression system of human augmenter of liver regeneration (hALR) and to examine its bioactivity in vitro. With PCR and gene recombination techniques, cDNA of open reading frame of hALR was obtained from recombinant plasmid pcDNA3.1-hALR and inserted into plasmid pPIC9. The cDNA of hALR from recombinant plasmid pPIC9-hALR demonstrated by sequencing was subcloned into plasmid pPIC9K. The recombinant plasmid pPIC9K-hALR was transformed into GS115 with electroporation. hALR was expressed by GS115 under the induction of 5 mL/L methanol and purified with ultrafiltration after it was analyzed by 15% SDS-PAGE and Western blot. The effects of hALR on in vitro proliferation of QGY and HepG(2) cells were evaluated by (3)H-TdR methods. The correctness and integrity of recombinant plasmids pPIC9-hALR and pPIC9K-hALR were identified by restriction digestion, PCR and sequencing methods, respectively. hALR as a secretive protein was successfully expressed by GS115. Its molecular weight was about 15 ku and the target protein was about 60% of the total protein in the supernatant from GS115 with plasmid pPIC9K-hALR. The results of Western blot of hALR showed the specific band. The high qualitative hALR was obtained through ultrafiltration. hALR could stimulate in vitro proliferation of QGY and HepG(2) cells in a dose-dependent manner, but there was a difference in reactivity to hALR between QGY and HepG(2). The hALR as a secretive protein can be successfully expressed by GS115. It may stimulate in vitro proliferation of QGY and HepG(2) cells at a dose-dependent manner. But QGY and HepG(2) cells have different reactivities to hALR.
Human augmenter of liver regeneration (hALR) is highly expressed in hepatocellular carcinoma (HCC... more Human augmenter of liver regeneration (hALR) is highly expressed in hepatocellular carcinoma (HCC), but rarely expressed in normal liver cells. ALR could stimulate the proliferation of HCC cell line in vitro, but has no effect on normal hepatocytes. This study was to investigate the inhibitory effect of blocking the expression of hALR on the proliferation of HCC cell line HepG2 using small interfering RNA (siRNA) targeting ALR and anti-ALR monoclonal antibody (McAb). RNA interference (RNAi) plasmid pSIALR-A targeting hALR cDNA and control plasmid pSIALR-B were constructed and transfected into HepG2 cells, respectively. After transfection, the expression of green fluorescent protein was observed under a fluorescent microscope to calculate transfection efficiency, the protein level of hALR was measured by immunocytochemistry, meanwhile, the expression of hALR mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). (3)H-TdR incorporation approach was performed to detect the proliferation of HepG2 cells after transfection and after neutralization with anti-hALR McAb. hALR was expressed in HepG2 cells. Plasmids pSIALR-A and pSIALR-B were successfully constructed. Both immunocytochemistry and RT-PCR showed that pSIALR-A inhibited the expression of hALR in HepG2 cells significantly by 83% as compared with pSIALR-B. pSIALR-A specifically inhibited the growth of HepG2 cells after transfection. (3)H-TdR incorporation of pSIALR-A/HepG2 cells was significantly lower than that of pSIALR-B/HepG2 cells (67 687+/-6 548 vs. 104 807+/-5 713, P<0.05). Anti-hALR McAb markedly inhibited the autonomous growth of HepG2 cells (P<0.05). hALR is highly expressed in HepG2 cells. The short interfering RNA targeting hALR gene could specifically suppress the expression of hALR, and subsequently inhibit the growth of HepG2 cells. Anti-hALR McAb could partially inhibit the autonomous growth of HepG2 cells.
Zhonghua gan zang bing za zhi = Zhonghua ganzangbing zazhi = Chinese journal of hepatology
To evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat ... more To evaluate the effects of newborn rat hepatocyte intrasplenic transplantation combined with rat augmenter of liver regeneration (rALR) injection in treating rats with acute hepatic failure. Acute hepatic failure (AHF) was induced in rats using D-gal (1.2 g/kg). The rats were then randomly divided into 6 groups. Group I received no further treatment and served as blank controls; group II received 1 ml buffered saline once through intrasplenic injection; group III received 1 ml rALR; group IV received 2 x 10(7)/ml hepatocytes; group V received 2 x 10(7) hepatocytes suspended in 1 ml rALR (50 microg/kg) and group VI received 2 x 10(7) hepatocytes in 1 ml cyclosporine A (10 mg/kg). The rats of the various treated groups were sacrificed at day 1, 5 and at week 2 and their livers and spleens were examined histopathologically. Blood samples of the rats were also obtained to determine the levels of TNF alpha and IL-1 beta. There were no significant differences in survival between group I, II and III rats. 33.3% of the group IV rats survived for 2 weeks. At week 2, the survival rate of group V rats was significantly higher than that of group IV, but there was of no statistical significant increase when compared to that of group VI rats. Hepatocytes transplanted into spleens survived for 5-7 days in the spleens of group IV and VI rats, but they survived at least 2 weeks in group V. The average serum TNF alpha level in group IV was significantly higher than that in groups V and VI on the first postoperative day, but after four days, only the difference between group IV and group VI was significant (P < 0.05). The average serum level of TNF alpha in group II was higher than that in groups IV, V and VI on the first postoperative day (P < 0.05), but there were no significant differences between those in groups IV, V and VI on the 1st and the 5th postoperative days. Newborn rat hepatocyte intrasplenic transplantation combined with rALR is effective in treating acute hepatic function failure induced by D-gal in rats. The transplanted hepatocytes can survive for at least 2 weeks in the spleens. The rALR mixed with the hepatocytes injected into the spleens may be able to facilitate the hepatocyte regeneration, to inhibit liver cell apoptosis and to suppress the cellular immunity.
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)e... more To elucidate the mechanism of ethyl pyruvate (EP) inhabit high mobility group protein B1 (HMGB1)expression and releasing in macrophage induced by lipopolysaccharide(LPS). The murine macrophage-like cell line RAW264.7 cultured in vitro divided into LPS group and LPS+EP group. The expression of HMGB1 mRNA in cultured cell was determined by RT-PCR. The cytoplasmic and nuclear HMGB1 levels were detected by Western blot. The contents of HMGB1 and TNF-α and IL-6 protein in cultured cells supernatant were detected by ELISA. Immunocytochemistry and confocal laser-scanning microscopy were used to confirm the relocation and distribution of intracellular HMGB1 protein in RAW264.7 cells. HMGB1 mRNA expression in the LPS+EP group was significantly lower than in LPS alone, at 24, 36 and 48 hours. In the LPS+EP stimulation group, the cytoplasm stained weakly while the nuclear stain was stronger than that of the LPS group at the same time points. Both TNF-α and IL-6 levels in LPS+EP group were sign...
ABSTRACT Rubia L. is the type genus of the coffee family Rubiaceae and the third largest genus in... more ABSTRACT Rubia L. is the type genus of the coffee family Rubiaceae and the third largest genus in the tribe Rubieae, comprising ca. 80 species restricted to the Old World. China is an important diversity center for Rubia where about half of its species occur. However, its internal phylogenetic relationships are still poorly understood. The objective of the present study is to contribute to the phylogenetic relationships within Rubia, using nuclear ITS and six plastid markers and focusing on species from China. Twenty-seven species of Rubia were sampled to infer their phylogeny using Maximum Parsimony, Maximum likelihood and Bayesian analyses. The monophyly of Rubia is supported, provided that R. rezniczenkoana is excluded from Rubia and transferred to Galium as a new combination: G. rezniczenkoanum (Litv.) L. E. Yang & Z. L. Nie. Within Rubia two clades are clearly supported. They correspond to the traditional sect. Rubia s.l. (A) and sect. Oligoneura (B), and are morphologically mainly separable by their pinnate (A) versus palmate (B) leaf venation. Plesiomorphic features are the pinnate leaf veining in sect. Rubia s.l. and the occurrence of some species with opposite leaves and true stipules in sect. Oligoneura. In sect. Oligoneura one can assume an evolution from species with opposite leaves and true stipules (as in the R. siamensis group) to those with whorls of two leaves and two leaf-like stipules (as in ser. Chinenses and the R. mandersii group) and finally with whorls of 6 or even more elements (as in ser. Cordifoliae). The correlation between clades recognized by DNA analyses and available differential morphological features is partly only loose, particularly in the group of R. cordifolia with 2x, 4x and 6x cytotypes. This may be due to rapid evolutionary divergence and/or hybridization and allopolyploidy.
Although little morphological differentiation is detected between intercontinental disjunct popul... more Although little morphological differentiation is detected between intercontinental disjunct populations of Phryma , this monotypic genus shows distinct molecular divergence corresponding to its distribution in eastern Asia and eastern North America . This study further employs amplified fragment length polymorphism ( AFLP) analyses using two selective primer pairs to quantify genetic diversity and evaluate the morphological stasis . Most of the molecular variance is accounted for by variance among populations between regions . Cluster and PCA analyses revealed that Phryma constitutes two major groups in line with their geographic distribution , with one genetically distinct group from the eastern Asia and the other with accessions from eastern North America . The results robustly supported the distinct genetic divergence and morphological stasis in Phryma .
Three new phenolic biphenyl derivatives (1-3) and one new lignan glycoside (4) were isolated from... more Three new phenolic biphenyl derivatives (1-3) and one new lignan glycoside (4) were isolated from the leaves and twigs of Osteomeles schwerinae. The structures of the new compounds were established by spectroscopic data interpretation. The inhibitory effects of 1-4 on rat lens aldose reductase in vitro were examined, and compounds 1-3 markedly inhibited the enzyme with IC50 values of 3.8 to 13.8 μM. In addition, the effects of these isolates on the dilation of hyaloid-retinal vessels induced by high glucose (HG) in zebrafish larvae were investigated. Compound 1 was the most effective in reducing HG-induced dilation of hyaloid-retinal vessels.
It has been suggested that many plants now found in the arctic originated from ancestors that occ... more It has been suggested that many plants now found in the arctic originated from ancestors that occurred at high altitudes in the southern mountains of the Northern Hemisphere during the Tertiary. However, this hypothesis has rarely been tested using a molecular phylogenetic approach. Here, we present a fossil-calibrated molecular phylogeny of Lagotis, an arctic-alpine genus with the greatest diversity in the Qinghai-Tibetan Plateau (QTP) and Central Asian mountains, based on five chloroplast (matK, psbA-trnH, rps16, trnG-S, trnL-F) and nuclear ribosomal ITS DNA markers. Within this framework, we infer the ancestral area and biogeographic history of the genus. Four major clades (A-D) within Lagotis were recovered with strong support, which largely correspond to the previous classification of the genus. Within clade A, Lagotis species from QTP were distributed among several subclades, and L. integrifolia from Central Asia was sister to L. glauca and L. minor from the arctic and subarctic region. The Bayesian molecular dating and the ancestral area reconstruction analyses suggested that Lagotis could have originated in the QTP in the Miocene (Tertiary), and that the genus radiated from the Miocene to Pleistocene. The diversification of Lagotis probably took place predominantly in the QTP and it subsequently spread to the Central Asian highlands, followed by northward migration into the arctic. Our results support the hypothesis that the "Central Asiatic Highland Corridor" as an important route for the migration of the flora between the arctic and QTP. ) and alignment are available in the Supplementary Data section of the online version of this article at
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