Biosynthesis of spherical selenium nanoparticles with halophilic Bacillus subtilis subspecies stercoris strain XP for inhibition of strawberry pathogens / 生物工程学报

Biosynthesis of nanomaterials has attracted much attention for its excellent characteristics such as low energy consumption, high safety, and environmental friendliness. As we all know, the toxic selenite can be transformed into higher-value nanomaterials by using bacteria. In this study, nano-selenium was synthesized by halophilic Bacillus subtilis subspecies stercoris strain XP in LB medium supplemented with selenite (electron acceptor). The physicochemical characteristics of nano-selenium were analyzed by scanning electron microscope (SEM), X-ray energy dispersive spectral analysis (EDAX), X-ray diffraction (XRD), and fourier transform infrared spectroscopy (FTIR). Meanwhile, the antifungal activity of nano-selenium to strawberry pathogens (fusarium wilt, erythema, and purple spot fungi) was determined. The products from reduction of selenite by strain XP was amorphous spherical selenium nanoparticles (SeNPs) with a diameter range of 135-165 nm. The production of SeNPs was positively correlated with time (0-48 h) and no changes were observed on cell morphology. Selenium was dominant in the surface of SeNPs where the organic elements (C, O, N, and S) existed at the same time. SeNPs were coated with biomolecules containing functional groups (such as -OH, C=O, N-H, and C-H) which were associated with the stability and bioactivity of particles. Although the highest concentration of SeNPs had significant (P<0.05) inhibitory effects on three strains of strawberry pathogens, antifungal activity to erythema and fusarium wilt pathogenic fungi was higher than that to purple spot pathogenic fungi from strawberry. In conclusion, strain XP not only has strong tolerance to high salt stress, but can be also used to synthesize biological SeNPs with good stability and biological activity. Thus, the strain XP has bright perspectives and great potential advantage in pathogens control and green selenium-rich strawberry planting as well as other fields.

Analysis of hepatitis B virus-related indexes in the urine of HBV transgenic mice / 中国比较医学杂志

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.

To observe the NTBC dependence of Fah-knockout mice / 中国比较医学杂志

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.

Effects of chemically modified sugarcane bagasse on butanol production by immobilized Clostridium acetobutylicum XY16 / 生物工程学报

Sugarcane bagasse modified by polyethylenimine (PEI) and glutaraldehyde (GA) was used as a carrier to immobilize Clostridium acetobutylicum XY16 in the process of butanol production. The effects of chemically modified sugarcane bagasse on batch and repeat-batch fermentations were investigated. Batch fermentation was conducted with an addition of 10 g/L modified sugarcane bagasse and 60 g/L glucose, resulting in a high solvent concentration of 21.67 g/L and productivity of 0.60 g/(L x h) with the treatment of 4 g/L PEI and 1 g/L GA. Compared to the fermentations by free cells and immobilized cells on unmodified sugarcane bagasse, the productivity increased 130.8% and 66.7%, respectively. The fibrous-bed bioreactor also maintained a stable butanol production during repeat-batch fermentations, achieving a maximum productivity of 0.83 g/(L x h) with a high yield of 0.42 g/g.

Hepatic arterial transplantation of autologous bone marrow mesenchymal stem cells in treatment of decompensated liver cirrhosis / 中国组织工程研究

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.

Influences of D-galactosamine and lipopolysaccharide on liver tissue regeneration and repair in mice with partial hepatectomy / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of D-galactosamine (D-GaIN) and lipopolysaccharide (LPS) on liver tissue regeneration and repair in mice following liver injury induced by partial hepatectomy.</p><p><b>METHODS</b>A total of 40 male BALB/c mice were randomly assigned into 2 equal groups to receive intraperitoneal injections of D-GaIN (500 mg/kg) plus LPS (50 µg/kg, given 1 h later) or two doses of saline 24 h prior to 1/3 hepatectomy. The liver weight/body weight (LW/BW) ratio and liver regeneration rate were observed at different time points after partial hepatectomy. Liver cell injury was assessed using HE staining, hepatocyte proliferation evaluated with BrdU staining, and the oval cell proliferation observed with immunohistochemistry.</p><p><b>RESULTS</b>In mice receiving saline injection, the liver volume was nearly restored 9 days after partial hepatectomy, while in mice with D-GaIN and LPS injections, the liver failed to recover the normal volume even at 14 days, showing a significant difference in the liver regeneration rate between them [(22.6∓105.93)% vs (9.49∓32.55)%, P<0.001]. Significant degenerative changes of the hepatic cells were found in D-GaIN/LPS-treated group, while only mild inflammatory reaction was observed in saline-treated group after partial hepatectomy. Obvious hepatocyte proliferation was observed at day 7 in saline-treated group but not in D-GaIN/LPS-treated group. Oval cell proliferation in the portal area occurred 3 days after partial hepatectomy in D-GaIN/LPS-treated group.</p><p><b>CONCLUSION</b>D-GaIN and LPS can obviously inhibit hepatocyte regeneration after liver injury in mice. D-GaIN and LPS combined with partial hepatectomy can induce oval cell proliferation.</p>

Study on the immune tolerance status and blood biochemical index of hepatitis B virus transgenic mice / 中华传染病杂志

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.

Generation and characterization of a series of monoclonal antibodies against recombinant human augmenter of liver regeneration / 中国病理生理杂志

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.

Protective effect of cardiomyopeptidin on cultured myocardial cells injured by anoxia-reoxygenation / 中国病理生理杂志

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.

Effects of dendritic cells pulsed by hepatitis B surface antigen on the functions of cytokine-induced killer cells / 中国医师杂志

Objective To explore the effects of dendritic cells(DCs) pulsed by hepatitis B surface antigen(HbsAg) on the proliferation and function of cytokine-induced killer(CIK) cells.Methods The peripheral blood mononuclear cells(PBMCs) were isolated by the conventional method,pulsed by HbsAg,and added into the CIK cells.The ~3H-TdR incorporation was used to determine the proliferation of CIK cells and lactate dehydrogenase(LDH) release was used to measure the specific killer activity of CIK cells induced by HBsAg-pulsed DCs.Results The HBsAg-pulsed DCs could induce the proliferation of CIK cells and strengthen the killer activity of CIK cells induced by HBsAg-pulsed DCs(P

Protective effect of cardiomyopeptidin on cultured myocardial cells injured by anoxia-reoxygenation / 中国病理生理杂志

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. [

Interaction between human augmenter of liver regeneration and Na~+-K~+-ATPase in vitro / 中国病理生理杂志

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.

Use of high-level HBV replication transgenic mice for evaluating drugs treating hepatitis B virus / 中国病理生理杂志

AIM:To study the high-level HBV replication transgenic mice for evaluation of drugs treating hepatitis B virus.METHODS:The HBV transgenic mice were treated respectively with lamivudine,large dose recombinant hepatitis B protein vaccine,?-1b interferon,siRNA to evaluate their pharmacodynamics and mechanism of action.RESULTS:HBV DNA titre was reduced significantly in transgenic mice which were treated with lamivudine(100 mg?kg-1?d-1),recombinant hepatitis B protein vaccine(HBsAg 6 ?g/mouse),?-1b interferon(50 ?g /mouse),respectively.Recombinant hepatitis B protein vaccine and ?-1b interferon promoted the level of IL-2 and IFN-? and increased the Elispot number of spleen cells secreting IFN-? in the treated transgenic mice.HBV transgenic mice were treated with RNAi expression vector pU6-siHBV against HBV through vena caudalis by hydrodynamics technique.Five days later,the level of serum HBsAg was reduced by 56.7% and the inhibition lasted at least 14 days.The HbcAg(+)cells were decreased obviously by immunohistochemistry detection in liver tissue,but the RNAi did not reduce the serum HBV DNA titre.CONCLUSION:These inbreeding high-level HBV replication transgenic mice are reliable and feasible for evaluating the anti-HBV drugs and have its economical and convenient superiority.

The specific T cell response in mice inoculated with HBV vaccine / 中国病理生理杂志

AIM: To evaluate the specific cellular immune response in mice inoculated with the recombinant hepatitis B virus (HBV) surface vaccine (rHBs). METHODS: Spleen T lymphocyte reactivity to rHBs was assessed by a proliferation assay and cytokine secretion. BALB/c mouse were intraperitoneally inoculated with rHBs at doses of 0.65, 1.25, 2.5 or 5 ?g for once or twice. 4 weeks later, spleen lymphocytes were harvested and restimulated with rHBs in experimental group or with PBS as control. The spleen lymphocytes were labeled with [~3H]-thymidine for 3 days. The [~3H]-thymidine incorporation was measured, which expressed as the incorporation of [~3H] (counts?min~ -1 ) and stimulation index (SI) was calculated by the method of dividing the cpm obtained in the experimental group by that in control group. The content of IL-2 and IFN-? secreted from the spleen lymphocyte were measured by the method of ELISA. RESULTS: 2 weeks after primary vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.55, 1.93, 2.41, 2.81 ng/L, respectively. IL-2 was 5.48?8.88, 9.28?6.98, 28.53?14.32, 64.69?20.88 ng/L, respectively. IFN-? was 8.22?8.61, 9.89?9.34, 20.27?15.50, 30.77?22.12 ng/L, respectively. 2 weeks after boost vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.61, 2.05, 3.74, 3.62 ng/L, respectively. The IL-2 was 5.75?5.04, 102.53?67.52, 177.13?91.12, 332.10?124.31 ng/L, respectively. IFN-? was 3.63?4.42, 28.33?13.04, 59.66?25.75, 80.73?19.30 ng/L, respectively. CONCLUSION: Specific proliferation activity and IL-2, IFN-? secretion from the spleen lymphocytes of the mouse inoculated with rHBs are produced,that the strength is dependent on the dose of vaccination.

STUDY ON EFFECT OF HGF ON SERUM ESTRAD1OL LEVEL OF THE RATS WITH LIVER INJURY / 解放军医学杂志

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