0.05).However,the percentage of patients with cholecystolithiassis was 86.5% in IGC group,and 50.6% in GC group(P=0.000).Besides the percentage of IA stage in IGC group(29.7%) was relatively higher than that(9.0%)in GC group(P=0.03);the surgical resection rate of tumor in IGC group was 56.8% and 32.6% in GC group(P=0.01).Nevertheless,the percentage of advance stage in IGC group(43.2%) was relatively lower than that in GC group(74.2%)(P=0.001).The overall 1,3,and 5-year survival rate of IGC group was 70.0%,31.2% and 26.8% repectively,and the mean survival time was17 months(51?13);which were significantly higher than those in GC group,in which the 1,3,5-year survival rate was 27.0%,17.7% and 15.1% repectively and the mean survival time was(25?8),5 months(all P=0.006).Single factor analysis showed that the survival time in IGC patients was influenced by the TNM stage(P=0.000),pT-category(P=0.000),operation-category(P=0.008);however,postoperative pathological grade(P=0.080),age(P=0.188) and sex(P=0.234) had no influence on survival rate.According to multivariate analysis,pT-category(P=0.000)was an independent factor for the survival time of IGC.Conclusions Comparing with GC group,IGC has a higher percentage of cholecystolithiassis,IA tumor stage and surgical resection rate,and thus,it has relatively better progonosis.pT-category is the vital independent prognostic factor in IGC.If a patient in ICG has been misdiagnosed during the primary operation,the patient should be re-operated for radical excision as soon as possible,except when the tumor is in stage Tis or T1a.

Laser techniques are widely applied in medical research and military affairs. The characters of laser make it the best way to evoke pain.Pain induced by laser stimuli is influenced by laser parameters such as wavelength, pulse duration and stimulus area in addition to the properties of skin such as the distance from the brain, type and color of skin. In this review,both laser evoked pain and factors influencing it are discussed.

Objective To investigate whether cell lines expressing apolipoprotein E-enhanced green fluorescence protein (apoE-EGFP) can support assembly of infectious hepatitis C virus particles. Methods apoE stably down-regulated Huh7. 5. 1 cell lines (sh-apoE cell lines) were established by shRNA gene silencing technique, and cell lines expressing apoE-EGFP fusion protein (apoE-EGFP cell lines) were established. The culture supernatants of wild-type Huh7. 5. 1 cells, control cells (sh-NT cells), sh-spoE cells and apoE-EGFP cells transfected with HCV RNA were collected and the HCV titer of supernatants was determined by TCID50. The interaction of apoE with HCV structure protein E2 was examined by immunofluorescence and confocal microscopy, and the expression of apoE-EGFP on the surface of HCV particles purified by FLAG-specific affinity gll was analyzed by Western blotting analysis. Results The apoE-EGFP fusion protein was highly expressed in sh-apoE cell lines. The infectivity of HCV from apoE-EGFP cell culture supernatant was not significantly different with those of HCV from Huh7. 5. 1 and sh-NT cells. apoE-EGFP infused protein had fluorescent co-location with HCV structural protein E2, and was detected on the surface of HCV particles purified by FLAG-specific affinity gll. Conclusion The apoE-EGFP fusion protein is an important component of HCV particles and apoE-EGFP cell lines can support the assembly of HCV particles.

To identify the prevalence and the distribution of ticks infected with spotted fever group rickettsiae (SFGR) in Xunke Area of Heilongjiang Province ,China ,partial outer membrane protein A gene (ompA) and citrate synthase gene (gltA) specific fragments were assessed using the PCR method .The positive products were sequenced .Result showed that the pres-ence of SFGR was 14 of 60 in detection Dermacentor silvarum cases ,while the overall positive rate was 23 .33% .Its nucleotide sequence of ompA showed 99 .3% and 99 .18% similarity with nucleotide sequence of Rickettsia sp .JL-02 and Rickettsia rao-ultii respectively .And the evolutionary positions of SFGR species were similar with Rickettsiamontana and Rickettsiamassili-ae .It's concluded that the nature focus of tick-borne spotted fever did exist in the area of Xunke Area of Heilongjiang Province , China .

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

The leakage of genetically engineered microorganism(GEM) at the beginning of operation was the important content for ecological risk assessment when GEM was inoculated in a membrane bioreactor(MBR) for bioaugmentation. The effects of different operating conditions on leaking density and intercepting efficiency of GEM were investigated in a submerged microfiltration MBR at the beginning of operation. The interception characteristics were also discussed. The results showed that different operating conditions had different influences on intercepting efficiency:higher sludge concentration was profit for interception,while higher aeration intensity and flux had opposite effects on interception. When the inoculated density of GEM was 1.0?1010 CFU/mL,the leaking densities varied from 1.0?102 CFU/mL to 2.5?102 CFU/mL under different operating conditions at the beginning of operation and the maximum intercepting efficiency could be higher than 8 lg. The main factors determining intercepting efficiency at the beginning of operation were membrane module interception,sludge adsorption as well as suspended GEM transfer inhibition and their contribution shares under certain conditions were 82.3%,14.9% and 2.8%,respectively. Gel layer formation during MBR stable operation was helpful to increase intercepting efficiency. The contribution shares for GEM interception of membrane module,sludge and gel layer were 75.3%,10.7% and 14.0%,respectively,under certain conditions.

According to the characteristics of sludge samples from full-scale wastewater treatment bioreac-tors, the essential total DNA extraction method for most environmental samples, lysozyme-SDS-phenol/ chloroform method, was modified to improve sample pretreatment, intensify cell lysis and enhance the effi-ciency of impurity removal. Obtain a general total DNA extraction method for industrial sludge samples. Such a method was applied for total DNA extraction of sludge samples from several running full-scale an- aerobic or aerobic bioreactors in Shijiazhuang, China. The results indicated that the modified method was suitable for all the sludge samples in this study, showing the satisfying generality. The extracted total DNA of all sludge samples were pure, with about 1.8 of A260/ A280 ratio. The method was also efficient; with average total DNA yield of over 0.7 mg/g and maximum yield of 0.85 mg/g. Moreover, all the extracted to- tal DNA samples could serve as templates directly to amplify 16S rDNA by PCR. The PCR products could be separated well by denaturing gradient gel electrophoresis (DGGE) and the DGGE band patterns were clear enough to be used for further analysis. All these facts indicated that the total DNA extraction method provided in this study could meet the requirements of sludge samples research, from full-scale wastewater treatment bioreactors, using molecular biology technologies.

Objective:To analyze the role of preoperative circulating tumor cell(CTC) and circulating tumor vascular endothelial cells (CTEC) in the diagnosis of gastric cancer and its correlation with the clinicopathological characteristics of gastric cancer.Methods:Sixty-two gastric cancer patients and 11 patients of benign gastric diseases were enrolled. Subtraction enrichment (SE) and immunofluorescence staining-chromosome fluorescence in situ hybridization (i·FISH) were used to integrate the unique SE-i ·FISH technology platform detecting patients′ CTC and CTEC.Results:The number of CTC in the gastric cancer group was significantly higher than that in the control group ( t=2.693, P=0.009); the number of CTEC in the gastric cancer group was higher than the control group ( t=2.015, P=0.048). With the cut-off value being set at 9 cells/6 ml in blood, the sensitivity of CTC in the diagnosis of gastric cancer is 84%, and the specificity is 82% (AUC=0.876, 95% CI, 0.792-0.963, P<0.01); When set at 6 cells/6 ml, the sensitivity of CTEC in the diagnosis of gastric cancer is 50%, and the specificity is 100%(AUC=0.727, 95% CI, 0.603-0.851, P=0.02). CTC positive is closely related to tumor location(χ 2=4.292, P=0.038 ) and TNM stage(CTC≥10, χ 2=4.848, P=0.028; CTC≥11, χ 2=6.234, P=0.013). CTEC positive is closely related to serum CA19-9(χ 2=4.858, P=0.028) and serum CA724 (χ 2=4.108, P=0.043 ) . Conclusion:SE-i·FISH technology has high sensitivity and specificity in the detection of CTC and CTEC of gastric cancer.

The study was conducted to explore the response of growth and yield of Tulipa edulis to flower bud removal and artificial pollination. And flower bud removal and artificial pollination were carried out in the squaring period and bloom stage respectively. The morphological index and biomass indicators were determined and the yield was counted in harvest time. Result showed that flower bud removal was beneficial to the growth of T. edulis, resulting in increasing growth index, biomass as well as the yield of bulb. The diameter and dry weight of T. edulis fruit by artificial pollination were increased significantly compared with the control. Seed setting percentage increased to 100%, and the number of seed as well as the single grain weight increased by 69.03% and 16.48%, respectively, which did not significantly affect the bulb production. In conclusion, Flower bud removal treatment accelerates bulb biomass increase, so as to improve its yield. Artificial pollination raised significantly seed setting percentage, seed number as well as the single grain weight.
Biomass ; Botany ; methods ; Flowers ; growth & development ; physiology ; Pollen ; growth & development ; physiology ; Pollination ; Tulipa ; growth & development ; physiology

OBJECTIVETo evaluate whether garlicin can prevent reperfusion no-reflow in a catheter-based porcine model of acute myocardial infarction (AMI).

METHODSTwenty-two male Chinese mini swines were randomized into 3 groups: sham-operation group (n=6), control group (n=8), and garlicin group (n=8). The distal part of left anterior descending coronary artery (LAD) in swines of the latter two groups was completely occluded by dilated balloon for 2 h and a successful AMI model was confirmed by coronary angiography (CAG) and electrocardiograph (ECG), which was then reperfused for 3 h. In the sham-operation group, balloon was placed in LAD without dilatation. Garlicin at a dosage of 1.88 mg/kg was injected 10 min before LAD occlusion until reperfusion for 1 h in the garlicin group. To assess serial cardiac function, hemodynamic data were examined by catheter method before AMI, 2 h after occlusion and 1, 2, and 3 h after reperfusion. Myocardial contrast echocardiography (MCE) and double staining with Evans blue and thioflavin-S were performed to evaluate myocardial no-reflow area (NRA) and risk area (RA).

RESULTSLeft ventricular systolic pressure and left ventricular end-diastolic pressure significantly improved in the garlicin group after reperfusion compared with the control group P<0.05) and 2 h after AMI (P<0.05). MCE showed garlicin decreased reperfusion NRA after AMI compared with the control group (P <0.05). In double staining, NRA/RA in the garlicin group was 18.78%, significantly lower than that of the control group (49.84%, P<0.01).

CONCLUSIONSGarlicin has a preventive effect on the porcine model of myocardial infarction reperfusion no-reflow by improving hemodynamics and decreasing NRA.

Allyl Compounds ; pharmacology ; therapeutic use ; Animals ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Contrast Media ; Disease Models, Animal ; Disulfides ; pharmacology ; therapeutic use ; Hemodynamics ; drug effects ; Male ; Myocardial Infarction ; complications ; diagnostic imaging ; drug therapy ; pathology ; Myocardial Reperfusion ; No-Reflow Phenomenon ; complications ; diagnostic imaging ; drug therapy ; pathology ; Swine ; Swine, Miniature ; Thiazoles ; metabolism ; Ultrasonography