Background and purpose:Runt-related transcription factor 3 (RUNX3) has a signiifcant relation with gastric cancer. Many researches have confirmed that rs760805 AA can increase the risk of gastric cancer. The purpose of the study was to investigate the relationship between the rs760805 polymorphism of the RUNX3 gene and gastric cancer. Methods: The rs760805 genotypes were determined by PCR-based DNA sequence measuring analysis and direct DNA sequencing in 310 incident cases with gastric cancer and 327 controls recruited in Shandong. Results:The frequency of TT genotype was 15.16%in gastric cancer patients and 20.49%in normal controls, and the corresponding percentages for AT and AA genotypes were 48.39%and 36.45%, and 52.60%and 26.91%, respectively. Compared to TT genotype, AT genotypes were associated with an increased risk of gastric cancer (OR=1.24, 95%CI`:0.81-1.92;OR=1.83, 95%CI:1.15-2.92). Conclusion:The rs760805 polymorphism of the RUNX3 gene is associated with increased susceptibility to gastric cancer.

To explore the cause and pathology of hemifacial spasm and evaluate the therapeutic efficacy of butulin A, 240 HFS patients were studied for therapeutic efficacy of multifocal facial injections with botulin A. Symptomatic alleviations after the injection were graded in comparison with the symptoms in the pre-treatment stage. Electromyography (EMG) was performed in 68 patients at random. Following the treatment, 96% of the patients were relieved of symptomes, but in 80% of the patients the symptom relapsed about one half a year later. The results showed that the latent period of EMG was longer than normal facial muscle, and the amplitude of evoked movement potential of EMG of orbiticularis occuli muscle and orbiticularis oris muscle was decreased. The cause of hemifacial spasm was resulted from pathological stimulation of the facial nerve, producing abnormal nervous conduction. The therapeutic hemifacial spasm with botulin A is an easy and effective method.

Objective To construct a prokaryotic expression vector of Salmonella paratyphi A ompA gene, and to determine immunogenieity and immonuprotection of the recombinant expressed product rOmpA and carrying frequency of ompA gene in S. paratyphi A isolates. Methods ompA gene was amplified from a clinical S. paratyphi A strain JH01 by PCR, cloned into T-A cloning vector, and then the prokaryotic expression vector was constructed. The rOmpA expression was determined by SDS-PAGE. Antigenicity and immunoreactivity of rOmpA were determined by immunodiffusion test, Micro-Widal's test and Western blot assay, ompA gene exiting in 98 S. paratyphi A isolates was detected by PCR. By using immunoprotective effect of rOmpA against the challenge of S. paratyphi A strain 50001 in mice were determinded. Results Both the nucleotide and putative amino acid sequence identities of the cloned ompA gene were 100% com-pared to the reported corresponding sequences. The expression yield of rOmpA was approximately 65% of the total bacterial proteins, rOmpA was able to induce specific antibody in rabbits, and reacted efficiently with rabbit antisortm or the antiserum against whole cell of S. paratyphi A detected by Western blot. 94.9% (93/98) of the S. paratyphi A isolates had ompA gene. 41.7% (5/12) and 58.3% (7/12) of the mice im-murtized with 100 and 200 μg rOmpA were survival after lethal challenge with S. paratyphi A strain 50001. The titer of antibody to the H antigens of S. paratyphi spp in the sera from rOmpA immunized mice and sur-viral mice in the S. paratyphi A challenge detected by Micro-Widal's test was 1:5-1:40. Conclusion ompA gene has an extensive distribution in clinical isolates of S. paratyphi A. rOmpA possesses an immunogenicity and a certain immunoprotective effect, and can be used as the candidate antigen in genetic engineering vaccine.

Objective To analyze the correlation between covalently closed circular DNA (cccDNA) in the peripheral blood mononuclear cells (PBMC) of hepatitis B virus (HBV)-infected patients and serum HBV DNA,hepatitis B surface antigen (HBsAg),hepatitis B e antigen (HBeAg) and liver histology of hepatitis B patients,and to explore the clinical significance of HBV cccDNA detection in PBMC.Methods One hundred and eight patients with chronic HBV infection were involved in this study.PBMC were extracted using density gradient centrifugation.HBV cccDNA in PBMC and serum HBV DNA were detected by real-time fluorescence quantitative polymerase chain reaction.HBsAg and HBeAg were detected by chemiluminescence immunoassay.Liver biopsy was conducted in 59 out of the 108 patients.Chi-square test was used to compare the categorical variables.Correlation analysis was used to compare quantitative variables.Nonparametric test was used to compare the non-normal distribution parameters.Results In the overall population,HBV cccDNA in PBMC was positive in 59 patients (54.6％).Eleven of the 15 patients with liver failure were found to be HBV cccDNA positive,which was significantly higher than that in the acute hepatitis B group (only 2 of the 8 patients were HBV cccDNA positive; x2 =4.960,P＜0.05).One hundred and eight patients were categorized into three groups according to their serum HBV DNA levels,with group A:＞5 lg copy/mL,group B:3-5 lg copy/mL and group C:＜3 lg copy/mL.The proportions of HBV cccDNA positivity in PBMC in three groups were 76.1％ (51/67),5/18 and 13.0％ (3/23),respectively.Comparing with patients with lower HBV DNA (group B and C),the proportion of HBV cccDNA positivity was higher in patients with higher HBV DNA (group A; x2=14.751,P＜0.05 and x2 =28.384,P＜0.05,resepectively).The HBV cccDNA quantitation in PBMC was positively correlated with the serum HBV DNA level and HBsAg quantification (r=0.554,P＜0.05 and r=0.497,P＜0.05,respectively).The proportion of HBV cccDNA positivity in PBMC of patients with liver histology ≥G2 and/or ≥S2 was significantly higher than that in patients with liver histology ＜ G2/S2 (x2 =9.159,P＜0.05).Conclusions HBV cccDNA exists in PBMC of hepatitis B patients.The HBV cccDNA quantitation in PBMC is positively correlated with the serum level of HBV DNA and HBsAg quantification,and is also associated with liver histology injury.

Objective To synthesize 131I labeled anti-neuropilin-1 monoclonal antibody A6 (131IA6) and evaluate its biodistribution and imaging in malignant glioma xenografts.Methods (1) A6 was labeled with 131I by Iodogen method under the optimum labeling conditions,then the labeling efficiency,radiochemical purity and stability were measured in vitro.(2) In vitro bioactivity,cellular uptake and receptor affinity of 131I-A6 with U87MG cells were measured.(3) The nude mice bearing human U87MG cells were randomly divided into 5 groups with 5 in each group.The nude mice were sacrificed by cervical dislocation and dissected at 24,48,72,96,and 120 h,respectively,after intravenous injection of 1.2 MBq 131I-A6.The biodistribution of the agent was measured as ％ID/g,and the ratios of tumor/blood (T/B) and tumor/muscle (T/M) were calculated.(4) SPECT/CT imaging was performed in 6 mice including 3 in the competitive inhibition control group at 24,48,72,96,and 120 h post injection.Two-sample t test was used for data analysis.Results (1) The labeling yield of 131I-A6 was (95.46±3.34)％,and the radiochemical purity was more than 95％.At 96 h of incubation in PBS,the radiochemical purity was more than 85％.(2)131I-A6 had rapid accumulation in U87MG cells and reached the peak of (15.80±1.30)％ at 1 h.When the probe was incubated with large excesses of non-radioactive A6,the uptake level of 131I-A6 in U87MG cells was significantly inhibited (t=2.862,P＜0.05).Kd of 131I-A6 binding to NRP-1 was (1.67±0.14) nmol/L in U87MG cells.(3) Biodistribution study showed that the uptake in blood,liver and tumor was (8.00±1.42),(7.68±1.56) and (6.00±1.24) ％ID/g at 24 h,respectively.The uptake in muscle,brain and bone was lower.The T/B and T/M were 0.78±0.10 and 3.20±0.30 at 24 h,and they reached the highest level of 1.87±0.50 and 7.13±0.24 at 120 h.(4) The SPECT imaging showed that the tumors could be visualized at 24 h and delineated more clearly at 120 h post injection of 131I-A6.Conclusions 131I-A6 can be easily synthesized by Iodogen method with high radiochemical purity.The specific tumor uptake of 131I-A6,which correlates with NRP-1 expression in gliomas,suggests that it may be a new promising tumor targeting radiotracer.

[Summary] To investigate the effect of microRNA126 on glucose metabolism in the normal liver cell lines. In vitro, the chang liver cell lines were cultured. Under the most effective transfection conditions ascertained above, microRNA126 mimic, microRNA126 inhibitor, and relative negative control were transfected into the cultured normal liver cells. And the transfection efficiency was tested by realtime fluorescent quantitative PCR. After 48 hours, the cells were stimulated with synthetic insulin ( 100 nmol/L ) and respective substrates for 2 hours. Then the glycogenesis, gluconeogenesis, and glycolysis in cells were measured. The level of microRNA126 of the microRNA126 mimic group was higher than the other groups, and the difference was statistically significant ( P<0. 05 ). MicroRNA126 mimic group significantly decreased glucose utilization, reduced glycogen synthesis, effectively increased the account of gluconeogenesis, reduced lactate production, and pyruvate kinase activity ( all P<0. 05). The over-expressing microRNA126 in hepatocytes may reverse the function of glucose metabolism, and enhance output of hepatic glucose.

ObjectiveTocomparethedifferencesoftwostocksofguineapigs,thealbinoguineapigsandpigment guinea pigs , in establishing dyslipidemic model , to evaluate their lipid-lowering action , and to compare their properties for development of hyperlipidemia .Methods Two stocks of the 5-week-old guinea pigs were randomly divided into two groups, normal group (NC) and model group (Model).For the NC group, 12 guinea pigs were fed with normal chew .For the model group , after fed with high-fat diet for four weeks , 24 male guinea pigs were randomly grouped and treated with vehicle (VC group) and pitavastatin (Pit group) calcium, respectively, by gavage as well as received high-fat diet.Before and after modeling and pitavastatin treatment , blood samples were collected and subjected to analysis of plasma TC , TG, HDL-C and LDL-C, respectively .Results In the normal group , the blood lipid levels of albino guinea pigs were more stable than that of the pigmented pigs with the increase of age .After fed with high-fat diet , the plasma lipid levels of TC , TG and LDL-C were significantly increased in the two strains of guinea pigs , while HDL-C showed a decrease to varying degrees .Interestingly , the lipid level in the albino guinea pigs was significantly higher than that of pigment guinea pigs . And also, after drug administration for four weeks , pitavastatin treatment significantly decreased the elevated lipid level of TC, TG and LDL-C in the albino guinea pigs compared with that in the pigment guinea pigs .Conclusions The albino guinea pigs and pigment guinea pigs demonstrate certain differences in establishing dyslipidemic model and evaluating lipid -lowering pharmacodynamics .However , compared with the pigment guinea pigs , the albino guinea pigs have obvious superiority because of easy establishment of hyperlipidemia model and are more sensitive to lipid -lowering drugs .

Objective To investigate the function and mechanism of miR-149 in nasopharyngeal carcinoma (NPC).Methods The expression of miR-149 was examined by real-time PCR and calculated by 2-△△Ct method. The cell proliferation was analyzed by MTT assay. The cell migration and invasion were shown by the wound healing assay and transwell migration assay, and the expression of E-cadherin was detected by Western blot. Results The expression of miR-149 was higher in NPC cell lines 5-8F and 6-10B than that in normal immortalized nasopharyngeal epithelial NP69. MTT assay showed that miR-149 promoted the proliferation of NPC cell lines. The wound healing assay showed miR-149 promoted the mobility and invasion of NPC cell lines. Inhibition of miR-149 reduced the ability of NPC cell lines to proliferate and invade. miR-149 downregulated the expression of E-cadherin, whereas antagomir which mediated knockdown of miR-149 significantly upregulated the expression of E-cadherin. Conclusion miR-149 might be involved in the invasion and metastasis of NPC through regulation of epithelial-mesenchymal transition (EMT).

Objective To investigate reflectance confocal microscopy (RCM) features of irritant cutaneous reactions to sodium lauryl sulphate (SLS) in healthy persons aged from 18 to 60 years,to analyze effects of age and gender on cutaneous reactions,and to estimate the value of RCM in objective evaluation of cutaneous reactions.Methods An occlusive patch test was performed on the back of 120 healthy testees with 0.1％ and 0.5％ SLS solution (0.1％ and 0.5％ SLS groups) and distilled water (negative control group) for 48 hours.At different time points after the patch removal,clinical evaluation and RCM were performed.Results RCM imaging in the 0.1％ and 0.5％ SLS groups showed parakeratosis,indistinct structure of the stratum corneum,spongiosis and infiltration of inflammatory cells in the epidermis,and telangiectasia in the papillary dermis.The incidence of RCM features reached the peak until 24 hours after the removal of 0.1％ and 0.5％ SLS patches,and the incidence of telangiectasia in the dermis was up to 66.7％ and 95.0％ in the 0.1％ and 0.5％ SLS groups respectively.At 24 hours after the removal of 0.5％ SLS patch,the incidence of spongiosis was significantly lower in the males than in the females (68.9％ [42/61] vs.84.7％ [50/59],x2 =4.24,P ＜ 0.05).However,the incidence of spongiosis was significantly higher in testees aged 18-40 years than in those aged 41-60 years at 24 hours after the removal of 0.1％ SLS patch (53.3％[32/60] vs.35.0％[21/60],x2 =4.09,P ＜ 0.05).For the other RCM features,there were no significant differences in their incidence between different genders or age groups after the removal of 0.1％ and 0.5％ SLS patches (all P ＞ 0.05).Clinical evaluation showed that after the removal of 0.1％ and 0.5％ SLS patches,no significant difference in the incidence of irritant cutaneous reactions was observed between the males and the females or between the testees aged 18-40 years and those aged 41-60 years (all P ＞ 0.05).There were good correlations between the clinical evaluation results and RCM features.At 24 hours after the removal of 0.1％ SLS patch,the correlation coefficient between spongiosis and clinical evaluation results was up to 0.77,so was that between telangiectasia in the dermis and clinical evaluation results (both P ＜ 0.001).However,at 0.5 hour after the removal of SLS patches,clinical evaluation showed that the positive reaction rates were 2.5％ (3/120) and 12.5％ (15/120) in the 0.1％ and 0.5％ SLS groups respectively.In the meantime,there were 17.5 ％ (21 / 120) and 51.7％ (62/120) of testees manifesting more than 2 RCM features in the 0.1％ and 0.5％ SLS groups respectively,which were more similar to the clinical evaluation results at 24 hours after the removal of SLS patches (34.2％ [41/120] and 85.0％ [102/120] in the 0.1％ and 0.5％ SLS groups respectively) compared with the clinical evaluation results at 0.5 hour after the removal of SLS patches.Conclusions Neither gender nor age affects irritant cutaneous reactions to 0.1％ and 0.5％ SLS.Compared with clinical evaluation,RCM can evaluate cutaneous reactions more objectively and accurately in the early stage of irritant reactions.

OBJECTIVETo investigate the correlation between intrahepatic eovalently closed circular (ccc)DNA of hepatitis B virus (HBV) and pathogen-and patient-related parameters.

METHODSUltrasound-guided liver biopsies were obtained from 60 patients with chronic HBV infection. Levels of intrahepatic HBV cccDNA and serum HBV DNA were measured by quantitative fluorescence PCR. Level of serum hepatitis B surface antigen (HBsAg) was measured by chemiluminescence immunoassay. Clinical parameters, such as alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), alkaline phosphatase (AKP), albumin, globulin (GLO), white blood cell, platelet, prothrombin-international normalized ratio, were measured by standard assay. Demographic information was recorded.The correlation between intrahepatic HBV cccDNA and pathogen-and patient-related parameters was assessed.

RESULTSIntrahepatic HBV cccDNA level was negatively correlated with age, GLO, ALT and grade of necroinflammation. Patients with age of 30 years or more showed significantly higher level of HBV cccDNA level than patients under 30 years-old (7.44±0.58 and 5.66±1.35; t=7.157, P less than 0.001). Intrahepatic HBV cccDNA level was positively correlated with serum HBV DNA level (r=0.916, P less than 0.001) and serum HBsAg level (r=0.727, P less than 0.001). The median ratio of HBV cccDNA to HBV DNA was 1.18, and of HBV cccDNA to HBsAg was 1.67.

CONCLUSIONIntrahepatic HBV cccDNA levels decrease with age, level of ALT, level of GLO and grade of liver necroinflammation, but increase with level of serum HBV DNA and level of serum HBsAg. To a certain extent, serum HBV DNA and serum HBsAg levels may be a sufficient marker of intrahepatic HBV cccDNA levels.

Age Distribution ; Alanine Transaminase ; Aspartate Aminotransferases ; Biomarkers ; DNA, Circular ; DNA, Viral ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Real-Time Polymerase Chain Reaction ; Serologic Tests