Objective To investigate the distribution of hepatitis delta virus (HDV) marker among hepatitis B virus (HBV) infected patients and to reveal its clinical significance.Method To collect the clinical data and sera samples of HBV infected patients and to detect HDAg,Anti-HDV as well as HBV infection markers by means of enzyme-linked immunosorbnent assay.These data combined with clinical diagnostic results and biochemical index were then analyzed.Result 462 samples of HBV infected patients were collected including 210 HBV carriers without symptom,175 chronic HBV infections,35 acute HBV infections and 42 liver fibrosis.The HDV infection rate was 4.8％ overall.The highest infection rate of 9.5％ was found in the group of liver fibrosis whereas the lower rate of 6.9％ was found in HBV chronic carriers.HDV infection rate was 7.8％ among the population of 40-60 years old,obviously higher than any other age groups.Conclusion HDV infection was significantly higher in the chronic HBV patients and liver fibrosis patients.Because HDV infection was highly associated with the progress of liver disease,we suggest the screen of HDV markers among hepatitis patients and discriminate whether the patient was co-infected with HDV.

OBJECTIVETo study mechanisms of reduction of the malignant activities of human naso-pharyngeal carcinoma cell CNE-2L2 induced by ectopic expression of BCSC-1 gene.

METHODSDNA was stained with propidium iodide and assayed upon a flow cytometer. Chromosomes were stained with Hoechest 33258. Adhesion of CNE-2L2 cells was detected by cell aggregation test. Protein expression on CNE-2L2 cells was examined by Western blot.

RESULTSCell cycle analysis showed that the percentage of CNE-2L2 cells was 55.1%, 43.4%, and 39.4% in G0/G1 phase, 25.2%, 28.7%, and 30.9% in S phase, and 19.7%, 27.9%, and 29.7% in G2/M phase for the cell with ectopic expression of BCSC-1 gene, wild type cell (W cells), and the cell transduced with the mock (M cell). Many mitotic cells were found in W cells and M cells. In contrast, almost no mitotic cell was observed in the cells with ectopic expression of BCSC-1 gene. Ectopic BCSC-1 expression resulted in cell aggregation, enhanced expression of E-cadherin, cx-catenin, and p53.

CONCLUSIONSEctopic BCSC-1 expression causes enhancement of adhesion of CNE-2L2 cells associated with enhanced expression of E-cadherin and alpha-catenin, arrest of cell in G1 phase, which may be associated with enhanced expression of p53. These alteration may play a role in the reduction of malignant activities of the cells with ectopic expression of BCSC-1 gene.

Cell Adhesion ; Cell Cycle ; physiology ; Cell Line, Tumor ; Humans ; Nasopharyngeal Neoplasms ; Neoplasm Proteins ; biosynthesis ; genetics

Objective To clone and express a high mobility group box 1(HMGB1)protein of Schistosoma japonicum(Main-land strain)and analyze its function. Methods The DNA fragment of open reading frame encoding Sj HMGB1 protein was ampli-fied by RT-PCR from the mRNA of S. japonicum worms,then it was subcloned into the expression vector pET28a(+)to form the recombinant expression plasmid SjHMGB1-pET28a. The recombinant expression plasmid was transformed into the component E. coli BL21(DE3),and the tranformant containing recombinant expression plasmid was induced with IPTG to express the recombi-nant protein SjHMGB1. The recombinant SjHMGB1 protein was purified by affinity chromatography with nickel chelating affinity chromatography agarose gel. The Gel retard experiment and animal immunization were performed to analyze the DNA binding ca- pacity and the immunologic property of recombinant SjHMGB1. The expression levels of HMGB1 in different life cycle stages of S. japonicum were analyzed by Western bloting and RT-PCR. Female ICR mice were immunized with the recombinant SjHMGB1 pro-tein and infected with 45±2 cercariae of S. japonicum after three immunizations. Forty-two days post-infection,the worms and eggs of S. japonicum were recovered from the portal vein and liver tissue,respectively. The worm and egg reduction rates were calculat-ed respectively. Results A 530 bp of specific DNA fragment was amplified from mRNA of S. japonicum by RT-PCR,which was the open reading frame(ORF)encoding SjHMGB1protein confirmed by DNA sequencing analysis. The recombinant expression plasmid SjHMGB1-pET28a was constructed by cloning the ORF of SjHMGB1 into a expression vector pET28a(+). The bacterium transformants containing the recombinant plasmid expressed a soluble recombinant protein about 28 kDa after induced by IPTG, and the recombinant SjHMGB1 protein was purified by nickel chelating affinity chromatography. The gel retard experiment showed that the recombinant SjHMGB1 protein could bind to both supercoiled DNA and linear DNA,and the recombinant protein immu-nized mice produced high titers of antiserum IgG. Western bloting indicated that the recombinant SjHMGB1 protein was recognized specifically by the S. japonicum-infected mice serum. Above results showed that the recombinant SjHMGB1 protein possessed both functional activity and immunogenicity as the natural protein. RT-PCR and Western blot results showed that SjHMGB1 was abun-dantly expressed in the adult and egg stages whereas barely detectable in the cercaria stage. The immune protection experiment showed that the recombinant SjHMGB1 induced mice to produce high titers of specific antibody IgG but failed to conduct an effec-tive immune protection against S. japonicum. Conclusion The gene encoding HMGB1 from S. japonicum and the soluble recombi-nant SjHMGB1 protein with natural functional activity are obtained,and the recombinant SjHMGB1 has a high immunogenicity but is not able to induce an effective immune protection against S. japonicum.

OBJECTIVETo study effects of ectopic expression of BCSC-1 gene on the malignant activi-BCSC-1 cDNA was isolated by RT-PCR ties of human nasopharyngeal carcinoma cell CNE-2L2.

METHODSand inserted into pMAL-c2X and pcDNA4/myc-His A. BCSC-1 protein was expressed in prokaryocytes. Rabbit antiserum to BCSC-1 was developed by means of immunization of rabbit with the BCSC-1 protein. Expression of BCSC-1 gene in wild type CNE-2L2 cell (W cell) was examined by real-time RT-PCR and immunofluorescence staining with the antiserum as a probe. pcDNA4/myc-His A-BCSC-1 was transfected into W cell at the presence of LipofectAmine. The cells were selected by G418 and cloned. Ectopic expression of BCSC-1 gene in W cell was examined by Western blot. Cell growth was detected by drawing of growth curves and colony formation tests. Cells were inoculated into nude mice. Size of tumors was assayed once a week. Lungs of the mice were sectioned continuously and metastatic loci in lungs were examined upon a microscope.

RESULTSRabbit BCSC-1 antiserum was prepared. Expression of BCSC-1 gene in W cell was found to be very low. CNE-2L2 cell with ectopic expression of BCSC-1 gene was developed. Growth in vitro, colony formation, tumorigenesis in nude mice, and lung metastasis of the tumor were profoundly inhibited of the cell with ectopic expression of BCSC-1 gene in comparison with controls, wild type cell and the cell transfected with mock. Conclusion Ectopic expression of BCSC-1 gene exerts profound inhibitive effect on the malignant activities of CNE-2L2 cell.

Animals ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Lung Neoplasms ; secondary ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Proteins ; biosynthesis ; Neoplasm Transplantation ; Rabbits ; Transplantation, Heterologous

Objective To evaluate the corrosion resistance of a new titanium alloy for dental restoration in artificial saliva. Methods In simulated oral environment, the electrochemical behavior of a new titanium alloy(Ti-12. 5Zr-3Nb-2. 5Sn) for dental restoration based on the d-electron alloy design method with high elastic modulus, high mechanical and good biological safety properties was investigated together with that of Ti-6Al-4V and TA2 as control groups. The anodic polarization curve and polarization resistance of these alloys were analyzed and the element release of Ti-12.5Zr-3Nb-2.5Sn and Ti-6Al-4V alloy after immersion in artificial saliva for 1,2,3,5,7,15 d were measured. Results Polarization curve indicated that Ti-6Al-4V alloy had lower breakdown potential(0. 8 V) than Ti-12. 5Zr-3Nb-2. 5Sn alloy did (>2. 5 V) . Ti-6A1-4V alloy showed higher passivation current density (1.45 × 10-4 -1.09 × 10-3 A/cm2) than Ti-12. 5Zr-3Nb-2. 5Sn alloy(3. 32 × 10-6-3.46 × 10-5 A/cm2) and TA2(5. 03 × 10-6-2. 65 × 10-5 A/cm2) did. Polarization resistance results showed that polarization resistance volume of TA2, Ti-12. 5Zr-3Nb-2. 5Sn alloy and Ti-6Al-4V alloy were 371.0, 252.0, and 60. 1 kΩ · cm2 respectively. With the increasing of dipping time in artificial saliva, the ion release of Ti-12. 5Zr-3Nb-2. 5Sn alloy and Ti-6Al-4V alloy increased to different degrees. Ti-6Al-4V alloy always showed more ion release than Ti-12. 5Zr-3Nb-2. 5Sn alloy did in the experiment. Conclusions Data from this study indicated that Ti-12. 5Zr-3Nb-2. 5Sn alloy, as a dental restoration material, had good corrosion resistance in artificial saliva.

BACKGROUNDType 2 diabetes mellitus (T2DM) has traditionally been considered to affect mainly the elderly; however, the age at diagnosis has gradually reduced in recent years. Although the incidence of young-onset T2DM is increasing, it is still not fully clear the onset characteristics and risk factors of early-onset T2DM. The aim of this study was to describe the initiating characteristics of early-onset T2DM in Chinese patients and evaluate the risk factors for diabetes mellitus.

METHODSThis cross-sectional controlled study was performed using a questionnaire survey method in outpatients of multiple centers in China. A total of 1545 patients with T2DM with an age at onset of <40 years were included, and the control group consisted of subjects aged <40 years with normal blood glucose level.

RESULTSIn patients with young-onset T2DM, the mean age and initial hemoglobin 1Ac at diagnosis were 32.96 ± 5.40 years and 9.59 ± 2.71%, respectively. Most of the patients were obese, followed irregular diet pattern and sedentary lifestyle, had life or work pressure, and had a family history of diabetes mellitus. Compared with subjects with normal blood glucose level, logistic regression analysis showed that waist-to-hip ratio (odds ratio [OR] 446.99, 95% confidence interval [CI] 42.37-4714.87), family history of diabetes mellitus (OR 23.46, CI 14.47-38.03), dyslipidemia (OR 2.65, CI 1.54-4.56), diastolic blood pressure (OR 1.02, CI 1.00-1.04), and body mass index (OR 0.95, CI 0.92-0.99) are independent factors for early-onset T2DM.

CONCLUSIONSWe observed that abdominal obesity, family history of diabetes mellitus, and medical history of hypertension and dyslipidemia are independent risk factors for early-onset T2DM. It is, therefore, necessary to apply early lifestyle intervention in young people with risk of diabetes mellitus.

Adult ; Blood Glucose ; analysis ; Cross-Sectional Studies ; Diabetes Mellitus, Type 2 ; blood ; etiology ; Female ; Glycated Hemoglobin A ; analysis ; Humans ; Male ; Risk Factors ; Waist-Hip Ratio

OBJECTIVETo investigate the effect of valproic acid on the expression of P27(Kip1) and P170 and drug resistance of leukemia HL60/HT cell line and explore its possible mechanisms.

METHODSHL-60/HT cells were derived from HL-60 cells induced by harringtonine (HT) in gradient concentrations. The inhibitory effect of valproic acid on the proliferation of HL-60 and HL-60/HT cells was evaluated by MTT assay, and the P27(Kip1) expression, P170 expression and cell cycle of the cells were analyzed with flow cytometry.

RESULTSThe multidrug-resistant HL-60/HT was acquired, which showed a stable drug-resistant index with increased IC(50) of HT, VCR, DNR and Ara-c by 9.30, 5.20, 4.91 and 3.65 folds, respectively, as compared with those of HL60 cells. The expression of P27(Kip1) in HL-60/HT cells was significantly lower but P170 expression significantly higher than that of HL-60 cells and normal mononuclear cells (P<0.05). The expressions of P27(Kip1) and P170 showed no significant difference between normal mononuclear cells and HL-60 cells. The growth inhibition rate of VPA combined with Ara-C was significantly higher than that of valproic acid or Ara-C alone in HL-60/HT cells and HL-60 cells (q=1.37 and 1.51, respectively). HL-60/HT and HL-60 cells cultured in the presence of VPA resulted in a significant increase in the expression of P27(Kip1) and the G(1)-phase cells (P<0.05), but the expression of P170 underwent no significant changes (P>0.05).

CONCLUSIONHL-60/HT cells have lower P27(Kip1) expression compared with HL-60 cells. Valproic acid can inhibit the growth of HL-60/HT cells and enhance their Ara-C sensitivity possibly by increasing P27(Kip1) expression and causing cell cycle arrest in G(1) phase.

Antineoplastic Agents ; pharmacology ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Cytarabine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Glycoproteins ; genetics ; metabolism ; HL-60 Cells ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; metabolism ; Valproic Acid ; pharmacology

This study was aimed to investigate the effect of MK886, a mPGES-1 inhibitor, on apoptosis and drug resistance of leukemia HL-60/A cell line. Expression of mPGES-1 was assayed by QT-PCR and Western blot. The effect of MK886 on HL-60/A cell proliferation was assayed by CCK-8 method, and flow cytometry was used to detect cell apoptosis. The expression of Akt and P-Akt was detected by Western blot. PGE2 was measured by ELISA. Effect of MK886 (10 µmol/L) on the chemotherapeutic sensitivity of HL-60/A cells and expression of mdr-1 mRNA and P170 protein were investigated too. The results indicated the expression of mPGES-1 was higher in HL-60/A cells. MK886 inhibited HL-60/A cell proliferation and induced apoptosis in a time- and concentration-dependent manner. Expression of mPGES-1 and P-Akt and synthesis of PGE2 decreased significantly. MK886 reduced expression of mdr-1 and P170 protein and enhanced the sensitivity of HL-60/A cells to chemotherapeutic drugs. It is concluded that MK886 can inhibit HL-60/A cell proliferation, induce apoptosis and enhance sensitivity to chemotherapeutic drugs, the mechanism of which possibly associates to down-regulation of mPGES-1/PGE2 synthesis, reduction P-Akt expression and decreasing mdr-1 and P170 protein expression.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Indoles ; pharmacology

OBJECTIVETo observe the effects of pravastatin on platelet-derived nitric oxide system in hypercholesterolemia (HC) and atherosclerosis (AS) in rabbits, and the relationship between these changes and atherosclerosis courses.

METHODSThirty male New Zealand white rabbits were randomly divided into three groups, 12 in group A, 12 in group B, and 6 in group C. All of them were fed daily with cholesterol-rich food during the first 12 weeks. In addition, in group A, pravastatin (10 mg) was orally administered daily. At the end of the 12th week, 6 in group A and B were killed randomly and their aortas were removed and the pathologic changes were observed. In the following 12 weeks, food enriched with cholesterol was substituted with normal food in all three groups. Pravastatin treatment was continued or started in the remaining members of group A and group B, but not in group C. At the end 24th week, all rabbits were killed and their aortas were examined for the fatty-streaks or atherosclerotic plaques. The expressions of endothelial NOS (eNOS) mRNA and inducible NOS (iNOS ) mRNA, NOS activity, NO production and the level of the serum lipids were measured at 0, 6th, 12th, 18th and 24th week.

RESULTSThe expression levels of platelet-derived NOS mRNA, eNOS mRNA ratio in group A had no difference at above time points, while in group B were reduced significantly at 6th week and 12th week compared with at 0 week (P <0.01), and increased at 18th week and 24th week compared with 12th week (P <0.05). The expression levels of eNOS mRNA in group C were reduced at 6th, 12th and 18th, 24th week compared with 0 week (P <0.05 and P <0.01, respectively), and were reduced in groups B and C compared with group A at 6th ,12th week (P < 0.05) and increased in group A and B compared with group C at 18th, 24th week (P <0.01). The expression levels of iNOS/mRNA among the three groups had no difference. Pathologic finding of the arteries: AS was not found in group A from the 12th to 24th week. While in group B, there were a lot of fatty-streaks on the entire intima of all large arteries at the 12th week. There were also fatty-streaks in the ascending aorta, but were improved at the 24th week. In group C, there were marked plaques in the entire aorta at the 24th week.

CONCLUSIONSThe expressions of platelet-derived eNOS mRNA, NOS activity, NO production are decreased in HC or AS rabbits. Pravastatin can up-regulate expressions of platelet-derived eNOS mRNA, NOS activity, leading to preventing or improving the pathological courses of AS.

Animals ; Atherosclerosis ; blood ; pathology ; Blood Platelets ; metabolism ; Disease Models, Animal ; Male ; Nitric Oxide ; blood ; genetics ; Nitric Oxide Synthase ; blood ; genetics ; Pravastatin ; pharmacology ; RNA, Messenger ; genetics ; Rabbits

To investigate the effect of a microsomal prostaglandin E synthase-1 (mPGES-1) inhibitor MK886 on cell cycle of the human acute myeloid leukemia HL-60 cells. HL-60 cells were treated with different concentration of MK886 (10, 25, 50 µmol/L) for 24 h. Flow cytometry, Western blot and ELISA were used to measure cell cycle, cyclin D1, mPGES-1, PGE(2), Akt, P-Akt and C-MYC. The results indicated that after treated with MK886, the percentage of HL-60 cells decreased in G(0)/G(1) phase and increased in S phase, and expressions of mPGES-1, cyclin D1, P-Akt and C-MYC and synthesis of PGE(2) decreased significantly. It is concluded that MK886 can arrest HL-60 cells in G(0)/G(1) phase, the mechanism of which is possibly associated to inhibition of mPGES-1 expression, reduction of PGE(2) synthesis, suppression of Akt phosphorylation and C-MYC expression, down-regulation of cyclin D1 expression.
Cell Cycle ; drug effects ; HL-60 Cells ; Humans ; Indoles ; pharmacology ; Intramolecular Oxidoreductases ; antagonists & inhibitors ; Leukemia ; metabolism ; pathology ; Prostaglandin-E Synthases