OBJECTIVETo investigate the relationship between hepatitis B virus (HBV) genotype, the mutation in basic core promoter (BCP) region/pre-core (Pre-C) region and the incidence of hepatocellular carcinoma (HCC) in Fusui county of Guangxi Zhuang Autonomous Region (Guangxi), a area with high incidence of HCC.

METHODSIn this case-control study, 53 HCC patients and 70 asymptomatic HBV carriers were enrolled. Blood samples were collected from them for serum separation and HBV DNA extraction. The DNA sequences of the S region and BCP/Pre-C region of HBV was determined by direct sequencing following nested-PCR amplification. The relationship between the genotype, gene mutation of HBV and the incidence of HCC was analyzed.

RESULTSThe mutation rates of the A1762T/G1764A in the BCP region and the T1858C in the Pre-C region of HBV were significantly higher in HCC group than in control group (94.3% vs. 75.7%, P = 0.006; 50.9% vs. 31.4%, P = 0.029). The mutation rate of A1775G was significantly higher in control group (28.6%) than in HCC group (13.2%) (P = 0.041). Multiple logistic regression analysis indicated that A1762T/G1764A and T1858C mutations are the risk factors for the development of HCC (OR = 5.459, 95% CI: 1.397-21.332, P = 0.015; OR = 3.881, 95% CI: 1.462-10.305, P = 0.006). A1775G is the protective factor in the development of HCC (OR = 0.192, 95% CI: 0.059-0.622, P = 0.006).

CONCLUSIONThe present investigation showed that BCP A1762T/G1764A, A1775G and Pre-C T1858C mutations are correlated with the incidence of HCC in Fusui county of Guangxi.

Carcinoma, Hepatocellular ; epidemiology ; virology ; Case-Control Studies ; China ; epidemiology ; DNA, Viral ; Genotype ; Hepatitis B virus ; genetics ; Humans ; Incidence ; Liver Neoplasms ; epidemiology ; virology ; Mutation ; Polymerase Chain Reaction ; Risk Factors ; Sequence Analysis, DNA

Objective:To study the improving effect of curcumin on cardiac function and its influence on myocardial collagen concentration in rats with myocardial infarction (MI).Methods:A total of 60 healthy adult male SD rats were randomly divided into sham operation group (n=10,survived rats n=9)and operation group [n=50,left an-terior descending artery was ligated,they were randomly divided into following four groups,on 28 d the treatment and survived rats in every group were:myocardial infarction (MI)group (n=9),control group (received intraper-itoneal injection of mixed solution,n= 10)and curcumin group (received intraperitoneal injection of curcumin, 100mg·kg-1 · d-1 n= 12)].After four weeks,M-mode echocardiography was used to measure left ventricular shortening fraction (LVFS),left ventricular ejection fraction (LVEF)and heart rate (HR).Masson staining was used to detect the myocardial fibrosis alteration after MI.Immunohistochemistry was used to measure expressions of myocardial collagen type Ⅰ and Ⅲ,and ratio of collagen Ⅰ/Ⅲ.Results:Compared with MI group and control group,after four weeks,there were significant rise in LVFS [(17.23±1.97)%,(19.34±0.83)% vs.(26.70± 1.15)%]and LVEF [(42.08±5.50)%,(41.63± 1.81)% vs.(56.76±2.49)%],significant reductions in HR [(433.16±20.05)beats/min,(433.04±24.17)beats/min vs.(403.96±7.08)beats/min],concentrations of myo-cardial collagen Ⅰ and Ⅲ,and ratio of collagen Ⅰ/Ⅲ [(13.5±0.9),(13.9±1.0)vs.(10.3±1.6)]in curcumin group,P <0.01 all.Masson staining indicated that acute myocardial infarction can cause significant left ventricular interstitial fibrosis.Conclusion:Curcumin could significantly improve cardiac function and inhibit myocardial fibro-sis level in rats with acute myocardial infarction.

Objective To investigate the relationship between hepatitis B virus (HBV) genotype,the mutation in basic core promoter(BCP)region/pre-core(Pre-C)region and the incidence of hepatocellular carcinoma(HCC) in Fusui county of Guangxi Zhuang Autonomous Region (Guangxi),a area with high incidence of HCC. Methods In this case-control study,53 HCC patients and 70 asymptomatic HBV carriers were enrolled. Blood samples were collected from them for serum separation and HBV DNA extraction. The DNA sequences of the S region and BCP/Pre-C region of HBV was determined by direct sequencing following nested-PCR amplification. The relationship between the genotype,gene mutation of HBV and the incidence of HCC was analyzed. Results The mutation rates of the A1762T/G1764A in the BCP region and the T1858C in the Pre-C region of HBV were significantly higher in HCC group than in control group(94.3%vs. 75.7%,P=0.006;50.9%vs. 31.4%,P=0.029). The mutation rate of A1775G was significantly higher in control group (28.6%) than in HCC group (13.2%)(P=0.041). Multiple logistic regression analysis indicated that A1762T/G1764A and T1858C mutations are the risk factors for the development of HCC (OR=5.459,95%CI:1.397- 21.332,P=0.015;OR=3.881,95%CI:1.462-10.305,P=0.006). A1775G is the protective factor in the development of HCC(OR=0.192,95%CI:0.059-0.622,P=0.006). Conclusion The present investigation showed that BCP A1762T/G1764A,A1775G and Pre-C T1858C mutations are correlated with the incidence of HCC in Fusui county of Guangxi.

Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.

he aim of this study was to obtain a cell-penetrating cytoglobin (Cygb), which combines the transmembrane function of cell-penetrating peptides TAT with the anti-aging and anti-fibrotic role of cytoglobin. The Cygb gene was complexed with TAT gene by overlapping PCR, inserted into the vector pET22b to construct the recombinant expression plasmid (pET22b-TAT-Cygb) and then transformed into Escherichia coli BL21 (DE3). The fusion protein TAT-Cygb, whose expression was induced by lactose, was purified by CM Sepharose Fast Flow Protocol and verified by Western blotting. The final TAT-Cygb had a molecular weight of 23 kDa with 95% purity, as shown by SDS-PAGE. As demonstrated by bioactivity experiments, TAT-Cygb exhibited a high specific peroxidase activity up to (422.30 ± 0.36) U/mg. Both TAT-Cygb and Cygb pretreatment group could protect Hacat cells against oxidation of H2O2, but only TAT-Cygb treatment group could remedy cells injuried by H2O2 (RGR = 98%), which was significantly different from Cygb treatment group (RGR = 79%). We successfully obtained the bioactive and cell-penetrating fusion protein TAT-Cygb that has the potential application in anti-aging, anti-fibrotic and anti-cancer.
Blotting, Western ; Cell Line ; Cell-Penetrating Peptides ; biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; metabolism ; Gene Products, tat ; Genetic Vectors ; Globins ; biosynthesis ; Humans ; Hydrogen Peroxide ; Recombinant Fusion Proteins ; biosynthesis

In this paper, the preliminary study on antioxidant, enhancement of antioxidant enzymes activity, reducing the content of oxygen free radicals, delaying skin aging of the recombination cytoglobin (rCygb) purified by our lab were investigated through human keratinocyte cell line (HaCAT) H2O2 oxidative stress model, mouse skin aging model caused by continuous subcutaneous injection D-gal, rat acute liver injury model induced by CCl4 and rat skin wound healing model. The results showed that rCygb improved the activities of total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px) and catalase (CAT), reduced the activities of lactate dehydrogenase (LDH) and alanine aminotransferase (ALT) as well as decreased the content of malondialdehyde (MDA). Skin biopsy showed that rCygb promoted angiogenesis, increased expression of collagen and improved the anti-inflammatory ability. All results displayed that rCygb improved the oxygen free radical scavenging ability, delayed skin aging and promoted wound healing.

To investigate the protective effect of polyethylene glycol (PEG) modified recombinant cytoglobin (PEG-rCygb) on acute liver damage in mice. The acute liver injury model of KM mice was induced by CCl4 and then treated with PEG-rCygb, The liver and blood samples were collected for biochemical and histopathological analysis. The results showed that PEG-rCygb reduced the liver mass index and decreased significantly the levels of alanine amiotransferase (AST) and aspartate transaminase (ALT) in mouse serum. In liver tissues, the content of malondialdehyde (MDA) was decreased, whereas the content of glutathione (GSH) was increased in PEG-rCygb treated group. PEG-rCygb also elevated the activities of total super oxidedismutase (T-SOD) and catalase (CAT) in liver tissues. HE staining of liver tissue slices revealed that PEG-rCygb relieved fatty degeneration of liver, decreased inflammatory factors and reduced liver cell injury. Further in vitro experiments indicated that the protective effects of PEG-rCygb on hepatic stellate cell (HSC) against H2O2 were enhanced compared with that of rCygb. All results indicated that the PEG-rCygb promoted oxygen free radical scavenging ability and prevented acute liver injury in KM mice induced by CCl4.
Animals ; Carbon Tetrachloride ; Chemical and Drug Induced Liver Injury ; prevention & control ; Free Radical Scavengers ; metabolism ; Globins ; biosynthesis ; genetics ; therapeutic use ; Liver ; enzymology ; Male ; Mice ; Polyethylene Glycols ; chemistry ; Protective Agents ; therapeutic use ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use

Objective: To explore independent risk factors of coronary artery calcification (CAC) and analyze correlation among risk factors of CAC and serum osteopontin (OPN) level. Methods: According to results of 64-slice spiral computed tomography (MSCT) coronary angiography, a total of 65 patients were continuously enrolled and divided into CAC group (n=37) and non-CAC control group (n=28). Enzyme linked immunosorbent assay (ELISA) was used to measure serum level of OPN. Single factor and multiple factor Logistic regression analysis’s were used to analyze risk factors of CAC. Spearman’s straight line analysis was used to analyze correlation between risk factors of CAC and serum OPN. Results: 1、 The age, hypertension, diabetes, poor eating habits，lack of exercise, overweight, etc., which were independent risk factors of CAC （OR=3.47～12.96, P=0.018～0.003）by single factor Logistic regression analysis, were inducted to multiple factor Logistic regression analysis, its result showed that age, overweight, poor sleep quality, poor eating habits were independent risk factors of CAC, OR=35.31～5.17, P＜0.01～0.05; 2、Serum level of OPN in CAC group was significant higher than that of non-CAC control group ［(39.919±11.879) μg /L vs. (24.000±6.000) μg /L，P<0.01］; 3、The Spearman straight line correlation analysis indicated that serum level of OPN was correlated with risk factors of CAC : positively correlated with LDL-C, overweight, age, TC（r=0.487～0.286,P＜0.001～＜0.05）, and positively correlated with poor sleep quality, diabetes, poor eating habits, lack of exercise（r=4.10～2.24， P<0.01～0.05）; negatively correlated with HDL-C（r=-0.250,P＜0.05）. Conclusion: Correlation analysis indicates that age, overweight, poor sleep quality, poor eating habits etc. are independent risk factors of CAC；Serum OPN level is correlated with LDL-C, overweight，age, diabetes, lack of exercise etc., so these indicate that must decrease OPN level and risk factors of CAC to relieve CAC and slow down its development.

The aim of this study was to reveal the protection role and the related mechanism of cytoglobin on the oxidation induced hepatic stellate cell damage. We applied siRNA to interfere the endogenous cytoglobin gene, used recombinant cytoglobin protein to treat the completely activated human hepatic stellate cell line LX-2 and the incompletely activated primary rat hepatic stellate cells, or over-expressed cytoglobin protein in LX-2 cells. We used two different oxidative-stress related models, the hydrogen peroxide model and the iron-overload model in our experiments and investigated the proliferation status and the intracellular superoxide level of the cells. The results showed that endogenous cytoglobin exerted significant protective effects on hydrogen peroxide or iron-overload induced LX-2 cell damage, confirming that upregulation of cytoglobin was the protective response of activated hepatic stellate cells to oxidative stress. Recombinant cytoglobin protein could protect LX-2 cells from oxidation induced damage, and prevent primary rat hepatic stellate cells from excessive proliferation and injury. The cytoplasmic reactive oxygen species (ROS) scavenging capacity of the recombinant cytoglobin protein was not as good as its capacity in scavenging ROS outside the cells, likely owing to the lack of active transporting mechanisms. Intracellular over-expression of cytoglobin protein could exert significant protective effect on LX-2 cells treated with hydrogen peroxide or iron-overload. Our results would accelerate the exploitation of new anti-fibrotic targets.
Animals ; Cell Line ; Globins ; genetics ; pharmacology ; Hepatic Stellate Cells ; cytology ; pathology ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidative Stress ; drug effects ; Protective Agents ; pharmacology ; RNA, Small Interfering ; genetics ; Rats ; Reactive Oxygen Species ; metabolism

Recombinant adeno-associated viral vectors (rAAV) have been widely used as gene therapy vectors in clinical trials. Here, we reviewed the genomic structures and replication mechanisms of wt-AAV. Then, the assembly of capsid and the encapsidation of genomic DNA, two major events during AAV pakaging, was discussed in detail. Although the overall pattern of virus assembly and encapsidation is known, the molecular mechanisms and the structure-function relationship involved in these processes are not well understood. Further elucidatation of these processes may improve the production technology of rAAV and develop gene drug based on rAAV.
Capsid ; physiology ; Capsid Proteins ; genetics ; DNA, Viral ; genetics ; Dependovirus ; genetics ; physiology ; Genetic Vectors ; Genome, Viral ; Virus Assembly ; genetics ; physiology