AIM: To study the protective effect of cardiomyopeptidin (CMP) attributed to polypeptide on cultured myocardial cells injured by anoxia-reoxygenation.METHODS: The anoxia-reoxygenation injury model were developed, anoxia for 60 min and reoxygenation for 30 min. The effect of CMP on myocardial ultrastructure was observed. ［Ca2＋］i was estimated with adherent cell analysis and sorting 570(ACAS 570) laser cytometer and measured with fluorescent dye Fura-2-AM, the lipid fluidity of cellular membrane was determined by fluorescence polarization technique. RESULTS: CMP could obviously improve the ultrastructure of myocardial cells and dose-dependently decrease ［Ca2＋］i and increase the lipid fluidity of cellular membrane, CMP also could markedly reduce the chromaticity value of pseudo-colour graphic model of Ca2＋. CONCLUSION: Cardiomyopeptidin has an obvious protective effect on cultured myocardial cells injured by anoxia-reoxygenation, this may be related to its effect of decreasing ［Ca2＋］i and increasing lipid fluidity of cellular membrane.

ObjectiveTo research the hepatitis B virus (HBV) replication and immune tolerance status of transgenic mice for elucidating the pathogenesis of hepatitis B and evaluating new drugs against HBV.Methods SPE grade HBsAg negative nontransgenic and transgenic mice with the same genetic background were recruited in this study.HBsAg,HBeAg and HBV DNA were detected by chemiluminescent method.Pre-S1 and HBcAg were detected by enzyme linked immunosorbont assay (ELISA).Liver pathology was examined and HBsAg expressions at different stages were determined by immunohistochemical staining.The lymphocyte proliferation of mice was detected by flow cytometry and interferon (IFN)γ-producing T lymphocytes was determined by enzyme linked immunospot (ELISPOT).The expressions of Toll-like receptor (TLR)2 and TLR9 in splenocyte suspension and splenic dendrite cells (DC) were determined by double-labeling immunofluorescence.The data were analyzed by t test and F test.ResultsHBsAg,preS1,HBeAg,HBcAg were expressed and HBV DNA was replicated in HBV transgenic mice,while anti-HBs,anti-HBc,and anti-HBe were all negative.There were no obvious pathological changes in liver tissues.HBsAg was expressed in cytoplasm and HBcAg in nucleus of hepatocytes.After stimulated with HBsAg,T lymphocyte proliferation capacity of HBV transgenic mice was (697.6±67.3) cpm,which was much lower than that of nontransgenic mice [( 1315.5 ±191.6) cpm].The number of spot forming cells of IFNγ-producing splenocytes from transgenic mice after HBsAg stimulation was 8.25 ± 1.10,which was obviously lower than that of nontransgenic mice (28.50±4.21) (F=155.967,P=0.000).The expressions of CD11c+,TLR2 and TLR9 on DC from both HBV transgenic and nontransgenic mice were not different significantly (all P＞0.05).The HBsAg expressions in liver tissues were observed in 18-day-old fetal mice and 1-day-old newborn mice.ConclusionsThe HBV transgenic mice can express HBV-related antigens,and are immune tolerant to the antigens.The innate and acquired immunity of the HBV transgenic mice are normal,which is similar to chronic asymptomatic HBV carriers of human.Therefore,HBV transgenic mouse is an ideal animal model.

Objective To explore clinical treatment of postoperative hyponatremia of pituitary tumor. Meth-ods 32 cases of patients with postoperative hyponatremia of pituitary tumor were randomly divided into control group and treatment group. The control group(n = 15) was treated with routine western medicine, and the treatment group(n = 17) was treated with water limited and sodium supplemented based on the control group. The clinical effect was observed between control group and treatment group. Results The clinical effect in the treatment group was signifi-cantly higher than that of the control group(P <0.05). Conclusion The treatment of water limited and sodium sup-plemented on postoperative hyponatremia of pituitary tumor is better.

In our laboratory, injection of HGF, prepared by the modified Labrecque's method, was used for the treatment of experimental hepatic injury in rats. It was found that the serum estradiol level was significantly higher in the group of HGF treatment than that in the control group (P

AIM: To study the protective effect of cardiomyopeptidin (CMP) attributed to polypeptide on cultured myocardial cells injured by anoxia-reoxygenation.METHODS: The anoxia-reoxygenation injury model were developed, anoxia for 60 min and reoxygenation for 30 min. The effect of CMP on myocardial ultrastructure was observed. [Ca 2+ ] i was estimated with adherent cell analysis and sorting 570(ACAS 570) laser cytometer and measured with fluorescent dye Fura-2-AM, the lipid fluidity of cellular membrane was determined by fluorescence polarization technique. RESULTS: CMP could obviously improve the ultrastructure of myocardial cells and dose-dependently decrease [Ca 2+ ] i and increase the lipid fluidity of cellular membrane, CMP also could markedly reduce the chromaticity value of pseudo-colour graphic model of Ca 2+ . CONCLUSION: Cardiomyopeptidin has an obvious protective effect on cultured myocardial cells injured by anoxia-reoxygenation, this may be related to its effect of decreasing [Ca 2+ ] i and increasing lipid fluidity of cellular membrane. [

Objective To observe the NTBC dependence of Fah-knockout mice and study the biological characteristics in order to use the model more effectively.Methods Examine the progressive changes in body weight, survival time, liver pathology and serological markers after the NTBC withdrawal.Results After removing of NTBC, Fah-knockout mice lost their body weight gradually, and finally died in 5 to 7 weeks, along with increased serum ALT, AST levels and deformation of the hepatocytes.Conclusions Fah-knockout mice have a strong drug dependence of NTBC and could be the ideal model to hereditary tyrosinemia type I and other liver injury.

AIM:To check the physical interaction between GST-Na+-K+-ATPase domain and recombinant human augmenter of liver regeneration (rhALR) by GST pull down assay. METHODS: With PCR and genetic recombinant techniques, the coding region of ? subunit of Na+-K+-ATPase was cloned into expressing plasmid pGEX-4T and identified by endonuclease digestion and sequencing methods. Under the inducing of 0.1 mmol/L IPTG, the fusion protein GST-Na+-K+-ATPase domain was highly expressed by E.coli DH-5?. After hypersound quassating, the GST-Na+-K+-ATPase domain was purified by glutathione agarose beads and the physical interaction with rhALR was checked by GST pull down assay. RESULTS: Analysis by SDS-PAGE showed the rhALRs of monomer and dimmer in GST-Na+-K+-ATPase domain lane. The Western blotting of the GST-pull down assay showed the same results as well. CONCLUSION: The Na+-K+-ATPase domain is associated with rhALR specifically in vitro.

Objective To detect and analyze the HBV?related indexes in the urine of HBV transgenic mice and further understand the biological characteristics of transgenic mice, and to clarify the tissue sources of HBV?related indexes. Methods HBV?related indexes in the urine of transgenic mice were tested using enzyme?linked immune sorbent assay ( ELISA ) and fluorescence quantitative PCR ( real?time RCR ) . The tissue sources were confirmed by several experiments, i. e. hydrodynamic transfection of mice, RNA interference to inhibit HBV?expression in the transgenic mice, and to infect normal mice with HBV?positive serum from patients. Results Expression of HBsAg, HBeAg and HBV?DNA was present in the urine of transgenic mice, of which the HBsAg expression level was high (6674 ± 619?8 IU/mL), but lower than that in the serum (16470 ± 2704 IU/mL). The level of HBsAg expression in the urine of male mice was higher than that in female mice. The level of HBeAg expression in the urine was lower and the HBeAg positive rate of urine was higher than that of blood, and the levels of HBeAg expression showed significant inter?individual and inter?sexual differences. HBV?DNA level reached 103 -105 copy/mL in the urine, but no related antibody expression was detected. The experiments such as hydrodynamic infection test indicated that the HBV?related indexes in the urine are derived from replication in the kidneys rather than secreted from the liver, entered into the blood circulation, and discharged from the urine. The kidneys are an independent expression site of HBV. Conclusions The expression of HBV?related indexes is present in the urine of transgenic mice and it is a long?term expression along with the age in months, of which the expression levels of HBsAg and HBV?DNA are rather high and stable. HBsAg titer in the urine of the male mice is higher than that of female mice. HBeAg expression level in the male mice is more stable compared with that in female mice. No expressions of various kinds of antibodies have been found in the urine. The kidneys are an independent expression site of HBV.

AIM:To generate monoclonal antibodies against human augmenter of liver regeneration (rhALR). METHODS:After BALB/C mice were immunized by the purified rhALR, the cells of spleen were fused with the cells of SP2/0; The titer and speciality were respectively fathomed from ascites or foster fluid by ELISA and Western-blot test. RESULTS:2 hybridoma cell lines were successfully obtained. The McAbs titer from ascites and foster fluid are respectively about 10-3-10-5 and 10-2-10-3. It is evident that the two McAbs were directed at different epitopes. CONCLUSIONS:The McAbs have higher speciality. It is significantly useful of the value that how hALR distribute in tissue organs, how the hALR signals the metabolism in the body and the control distribution of the hALR on cell growth on the translational level and so on is researched.

BACKGROUND:Autologous bone marrow mesenchymal stem cel s can treat decompensated liver cirrhosis, however, little evidence has addressed the control ed clinical research in hepatitis B patients with decompensated live cirrhosis.
OBJECTIVE:To evaluate the clinical efficacy and safety of autologous bone marrow mesenchymal stem cel s in the treatment of hepatitis B with decompensated live cirrhosis.
METHODS:A total of 67 hepatitis B patients with decompensated live cirrhosis were divided into two groups according to their wishes to receive stem cel transplantation. The control group (34 patients) only received oral administration of nucleoside analog antivirus and supportive treatment. The treatment group (33 patients) received autologous bone marrow mesenchymal stem cel s transplantation via hepatic artery plus antivirus and supportive treatment. The liver functional index, clinical signs and symptoms, adverse reactions were observed and compared at 4, 12, 24 weeks after treatment.
RESULTS AND CONCLUSION:After treatment, al patients’ symptoms were improved to varying degrees. After 4 weeks of treatment, the liver functional indexes were al significantly improved compared with before treatment, the levels of alanme aminotransferase, cholinesterase and prothrombin activity in treatment group were significantly ameliorated compared with control group (P<0.05). At 12 and 24 weeks of treatment, the alanme aminotransferase, albumin, total bilirubin, cholinesterase and prothrombin activity in control group and treatment group showed statistical y significant differences compared with before treatment (P<0.05). At the same time point, al the indicators in the treatment group were significantly ameliorated compared with control group (P<0.05). The Child-pugh score and model for end-stage liver disease score declined at 4, 12, 24 weeks after treatment, showing significant differences compared with before treatment. The difference was also significant at the same time point between two groups. The treatment of nucleoside analogue antivirus combined with autologous bone marrow mesenchymal stem cel s transplantation on hepatitis B patients with decompensated liver cirrhosis is an effective method to improve liver function and blood coagulation function, with symptom improvement, safety and low risk.