Objective To explore different pathogenesis of chronic hepatitis B(CHB)and asymptomatic H BV carriers(ASCs)by identifying differentially expressed genes.Methods Subtracted library was constructed by suppression subtraetive hybridization(SSH),and α-defensin was identified by dot blot hybridization.Peripheral blood was collected from 46 CHB patients and 11 ASCs.and the expressions of α-defensin mRNA in peripheral blood mononuclear cell and protein in plasma were determined by the real time RT-PCR and enzyme-linked immunosorbent assay(ELISA).Results Real time RT-PCR showed that the expression of α-defensin mRNA in blood samples of CHB was 1.4-fold higher than that of ASCs.As shown by ELISA,the plasma level of α-defensin in CHB was higher than that of ASCs [(216.40±81.25)μg/L vs.(156.00±57.26)μg/L,t=2.23,P<0.05].Conclusion α-defensin may involve in the pathogenesis of CHB,for it iS over-expressed in CHB patients.

Calculi ; Humans ; Lung Diseases ; Male ; Middle Aged ; Pulmonary Alveoli ; pathology

Objective To explore the difference of pathogenesis between chronic hepatitis B (CHB) and asymptomatic hepatitis B virus(HBV) carriers(ASCs) in children.Methods Subtracted libraries was constructed by suppression subtractive hybridization (SSH) from peripheral blood mononuclear cell (PBMC) of two groups, beta-2-microglobulin (?_2-MG) was screened as one of differentially expressed genes. Serum was collected from children with CHB and ASCs, the differential expression of ?_2-MG was confirmed by enzyme immunoassay.Results Subtractive libraries were constructed successfully, the differentially expression of ?_2-MG and lactroferrin were up-regulated in CHB children.And the differentially expression of ?_2-MG on the level of protein was confirmed.Conclusion The up-regulation of ?_2-MG in CHB children is involved in the pathogenesis mechanisms.

Objective To use gene engineering technique to express the humanin.Methods Humanin segment was synthesized and cloned into the prokaryocyte expression vector pGEMEX-1 and pet44a. The plasmid was transformed to E. coli BL21(DE3) by screening and identifying to obtain the high expressing positive clone.Results The humanin segment was cloned into vector successfully,and expressed efficiently with the inducement of IPTG. The molecular weight of the expressed recombinant protein of pGEMEX-1-humanin was about 28 kd while it was 63 kd in pet44a-humanin,and the expressed quantity of fusion protein was 40 percentage of the total bacterial protein while it was 20 percent in pet44a-humanin.Conclusion The humanin has been cloned and expressed successfully and the high efficiently expressed recombinant protein might contain the intact important structure domain of the humanin.

OBJECTIVETo construct a eukaryotic expression plasmid pcDNA3.1(-)-Humanin.

METHODSThe recombinant plasmid pGEMEX-1-Humanin was digested with restriction endonucleases BamH I and Hind III and the Humanin gene fragments, about 100 bp length, were obtained. Then the Humanin gene fragments were inserted into eukaryotic expression vector pcDNA3.1(-) and the recombinant plasmids pcDNA3.1(-)-Humanin were identified by sequencing.

RESULTSRecombinant plasmid DNA successfully produced a band which had the same size as that of the Humanin positive control. The sequence of recombinant plasmids accorded with the Humnain gene sequence.

CONCLUSIONSA eukaryotic expression plasmid of Humanin was successfully constructed.

Base Sequence ; Cloning, Molecular ; methods ; Escherichia coli ; genetics ; metabolism ; Humans ; Intracellular Signaling Peptides and Proteins ; Molecular Sequence Data ; Molecular Weight ; Plasmids ; genetics ; Protein Engineering ; methods ; Proteins ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry

Objective To investigate the effects of heparin coating on intimal hyperplasia in implanted decellularized xenografts.Methods The resected canie carotid arteries were decellularized,and heparin coating was partially performed.Eighteen rabbits were divided into non-heparin-coated group(n=9)and heparin-coated group(n=9).During implantation,only the left carotid between the anastomotic stoma was ligated.Doppler ultrasonography was performed 1,3 and 12 weeks post-implantation to measure the luminal diameter,and the hemodynamic parameters such as PSV,RI and PI were calculated.All animals were sacrificed,histological observations were conducted at 12 weeks post-implantation,and I/(I+ M)was calculated.Results Except for 1 week post-implantation in the ligated side,the luminal diameters in non-heparin -coated group were significantly smaller than that of pre-implantation.Besides,those of the non-ligated side at each time points were significantly smaller than the ligated side(P

In traditional oral practice, the presystemic interactions with gut microbiota is an important mechanism underlying the holistic health benefits of Chinese herbal medicines (CHMs), making the study of CHMs distinct from the research of Western medicines of which the systemic exposure (level in blood) is the starting point and the core. Gut microbial metabolism complements host metabolism in maintaining metabolic homeostasis of many biologically important endogenous molecules and the disposition of numerous exogenous compounds. Among them, the widely distributed gut bacterial β-glucuronidases (BGUSs) coordinate with host UDP-glucuronosyltransferases (UGTs) to play a role in the occurrence and intervention of diseases by affecting the glucuronidation homeostasis and altering the intestinal local and/or systemic exposure of endogenous compounds and xenobiotics. On one hand, many ingredients of CHMs undergo enterohepatic circulation; On the other hand, CHMs can act on BGUSs directly or indirectly change the distribution and function of BGUSs through reprogramming gut microbiome. The multiple interactions between BGUSs and CHMs may play an important role in the overall therapeutic benefits of CHMs. This work firstly summarizes the latest research progress on BGUSs; then the physiological, pathological and pharmacological significance of BGUSs are exemplified with representative endogenous and exogenous compounds from the aspects of nutrient utilization, metabolic homeostasis, and therapeutic response based on the varied substrate spectra of BGUSs; finally, the scattered data in literature were integrated to summarize the multiple interactions between BGUSs and CHMs, highlighting the important role of BGUSs in the holistic actions of CHMs.

Objective To investigate the relationship between single nucleotide polymorphism (SNP) of Fas ligand (FasL) and fulminant hepatitis B in Han Chinese. Methods HBV infected subjects were enrolled in this case-control study, including 233 cases of inactive HBsAg carrier, 68 patients with fulminant hepatitis B,100 cases of spontaneous hepatitis B clearance, 102 patients with hepatitis B virus (HBV) related cirrhosis and 112 patients with HBV related primary hepatocellular carcinoma. The blood samples and clinical data were collected. FasL-844T/C polymorphisms of enrolled subjects were examined by TaqMan real time fluorescent genotyping polymerase chain reaction (RT-PCR). A adjusted odds ratios (OR)and 95% confidence intervals (CI)were calculated using the Logistic regression model. Results After adjusting the factors of gender and age, binary Logistic regression analyses indicated that the genotype frequencies of FasL-844 CC,CT,TT in inactive HBsAg carriers were 50. 64% ,39. 91% and 9. 44% respectively, and those in cases of fulminant hepatitis B were 79. 41%, 17. 65% and 2. 94%, respectively. The analysis also revealed that FasL-844CC genotype in inactive HBsAg carriers was high risk factor of developing fulminant hepatitis B (OR =4. 729,95%CI:0. 510 - 21. 282,P = 0. 043), while there were no statistic significances in other cases (P>0. 05). Conclusion The inactive HBsAg carriers harboring FasL-844CC may have greater susceptibility to fulminant hepatitis B, which need arouse high attention.

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. We have previously identified the benzoyl sulfathiazole derivative II as a non-competitive PTP1B inhibitor with in vivo insulin sensitizing effects. Preliminary SAR study on this compound series has been carried out herein, and thirteen new compounds have been designed and synthesized. Among them, compound 10 exhibited potent inhibition against human recombinant PTP1B with the IC50 value of 3.97 micromol x L(-1), and is comparable to that of compound II.
Humans ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; Structure-Activity Relationship ; Sulfathiazoles ; chemistry ; pharmacology

OBJECTIVETo investigate the protective effect of melatonin on residual hair follicle cells of scald rats at early stage.

METHODSEighteen male Sprague-Dawley rats were randomly divided into scald group, treatment group, sham group , with 6 rats in each group. The rats in scald group and treatment group were subjected to 30% TBSA partial thickness scald on the back, and were resuscitated with balanced solution after 1 hour, while those in sham group were immersed in water at 37 degrees C for 25 s to simulate scald, and did not receive fluid replacement. Rats in treatment group were intraperitoneally injected with 10 mg/kg melatonin solution at 1 minute, 8 hours and 12 hours after scald, while those in sham group and scald group were given equal volume of 1% alcohol sodium-isotonic saline instead. Tissue samples were harvested at 6, 12 and 24 post scald hours (PSH) for determination of MDA and GSH levels. Apoptosis of residul hair follicle was detected by TUNEL method and immunohistochemistry of caspase-3.

RESULTSThe level of MDA in scald group at each time point was much higher than that in sham group (P < 0.01) and treatment group (P < 0.05), and it peaked at 12 PSH. The changes in GSH were just opposite to that of MDA. Under fluorescence microscope, the residual hair follicle cells were blue, and the apoptotic cells appeared green. The apoptosis rate in scald group at 6, 12, 24 PSH was obviously higher than that in sham (P < 0.01) and treatment groups (P < 0.05), which was (20.2 +/- 3.4)% vs (4.3 +/- 2.3)% vs (10.9 +/- 3.2)%, (31.2 +/- 3.6)% vs (5.1 +/- 2.5)% vs (19.1 +/- 3.7)%, (22.4 +/- 2.7)% vs (4.1 +/- 2.4)% vs (13.1 +/- 3.4)%, respectively. The score of caspase-3 positive cell in scald group was higher than those in sham group (P < 0.01) and treatment group (P < 0.05).

CONCLUSIONSThere is obvious correlation between oxidative stress and apoptosis rate of hair follicle cells in rats with partial thickness scald. Early administration of melatonin may have anti-apoptosis ability for residual hair follicle cells by attenuation of oxidative stress.

Animals ; Apoptosis ; Burns ; drug therapy ; metabolism ; Hair Follicle ; cytology ; metabolism ; Male ; Melatonin ; therapeutic use ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley