Objective: To elucidate the possible mechanism responsible for the improved protection of terminal warm blood cardioplegia (TWBC) after hypothermic cardiopulmonary bypass (CPB) through analysis of tubulin (TB) components changes in myocardial cells exposed to TWBC. Methods: Stable animal models of CPB were established in cats, which were then randomly divided into 2 groups. Group Ⅰ was subjected to intermittent cold blood cardioplegia (ICBC) whereas group Ⅱ to ICBC followed by TWBC before uncross-clamping. Left ventricular performance was then monitored and evaluated by LVSP, LVEDP, ±dp/dtmax and t-dp/dtmax in both groups and semi-quantitive analysis was conducted with Western blot method as to the content and constitution of TB in myocardial cells at 15 min, 120 min after aortic crossclamping (ACC) and 5 min,15 min, 60 min,120 min after reperfusion. Results: Within 120 min after reperfusion, systolic and diastolic functions decreased significantly in group Ⅰ as compared with group Ⅱ(P<0.05). At 115 min after ACC and 15 min after reperfusion, the content of free and polymerized TB in both groups had no difference (P>0.05). At 120 min after ACC and 5 minutes after reperfusion, there was a significant difference between groupⅠ andⅡ (P<0.01). Conclusion: TWBC accelerates the repolymerization of myocardial TB during hypothermic CPB, which may mediate the improved cardiac performance in the early stage of myocardial reperfusion.

Objective:To study the role of TNF-?in the mechanism of hepatocyte apoptosis induced by intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were divided randomly into 7 groups: control group and wounded 1, 2 , 4, 8, 12, 24 hour groups.The model of intestinal perforations due to abdominal firearm wound were established in wounded groups. Levels of plasma endotoxin were measured using ehromogenic limulus amehoeyte lysate test.Hepatic TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum TNF-? levels were determined at the same time. Results: Levels of plasma endotoxin, serum TNF-?, hepatic TNF-? content and hepatocyte apoptosis indexes in wounded groups were all significantly elevated compared with control group(P

Objective:To study the role of NF- ?B in the mechanism of liver injury following intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were assigned randomly into 7 groups: control group and wounded 1, 2, 4, 8, 12, 24 hours group.The model of intestinal perforations due to abdominal firearm wound was established in wounded groups. Hepatic NF-?B and TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum ALT levels were also determined at the same time. The alterations of hepatic tissue were observed under light microscope. Results: Levels of hepatic NF-?B activity in wounded groups were significantly elevated compared with control group, and two peaks appeared in 1 h group and 8 h group, respectively (P

The seasonal dynamics of the soil microbial growth and soil net respir ation rate were studied in the imminent Heptacodium miconioides community T he results indicate that the numbers of bacteria and actynomices of rhizosphere soil or that of bacteria ,fungi and actynomices of non rhizosphere soil as well as net respiration rate of soil maintain similar seasonal dynamics in which dis play mono peak curves and their biggest values occure in September but the bi ggest nu mber of fungi of rhizosphere soil is in October The microbial numbers of soil,e s pecially that of soil fungi ,are greatly affected by the water content and tempe rature of soil The net respiration rate of soil closely relate not only with t h e water content and temperature of soil,but also with the soil microbial numbers of non rhizosphere soil which are mostly responsible for the net respiration r ate of soil

BACKGROUND: The surgical resection rate of pdmary hepatic carcinoma is low, thus, the local treatment arose more attention. Microwave coagulation therapy can inactivate rather than kill the hepatic carcinoma ceils. Slow-release chemotherapy has been used in treating primary hepatic carcinoma because it can form local high concentrations and last for a long time. OBJECTIVE: To observe the therapeutic effect of epirubicin-loaded chitosan microspheres combined with microwave coagulation on treating hepatocellular carcinoma in mica. METHODS: Epirubicin-loaded chitosan microspheres were prepared by using emulsion-chemical cross linking technique. The surface morphology and particles size of chitosan microspheres were observed by scanning electron microscope. Ultraviolet speotrophotometer was used to analyze the entrapment efficiency, entrapment efficiency and cumulative release rates of epirubicin-loaded chitosan microspheres. Totally 24 mice with transplanted subcutaneous H22 HCC were divided into 4 groups, which were respectively treated by microwave coagulation therapy, intratumoral injected with physiological saline after microwave coagulation therapy, intratumoral injected with epirubicin after microwave coagulation therapy, intratumoral injected with epirubicin-ioaded chitosan microspheres after microwave coagulation therapy. The tumor inhibitory rate was calculated. RESULTS AND CONCLUSION: The average diameter of chitosan microsphere was 105 μm, with uniformed particle diameter. The ratio of drug loading was 11% and the entrapment was 80%. The in vitro drug cumulative release rate was 84% after 2 weeks. The tumor inhibitory rates of the microwave coagulation combined with physiological saline, epirubicin, and drug carried microspheres groups were 8%, 20%, and 47%. It suggested that chitosan microsphere is a safe and effective slow-release dosage form, which exhibits strong anti-tumor effect when combined with microwave treatment.

ObjectiveTo identify the pathogen of an aseptic meningitis outbreak which occurred in Linyi City of Shandong Province during the summer of 2009,and to analyze the genetic variations of Coxsackicvirus B5 (CVB5) isolates.MethodsForty-two cerebrospinal fluids (CSF) specimens were collected from aseptic meningitis cases and virus isolation was performed. The viral RNA was extracted and amplified from the positive specimens using reverse transcription polymerase chain reaction (RT-PCR).The partial VP1 coding region was purified and sequenced. The phylogenetic trees based on VP1 sequences were constructed among CVB5 isolates and those in GenBank.ResultsSeventeen enteroviruse strains were isolated from 42 CSF samples with 40.5％ isolation positive rate. All these strains were identified as CVB5 using both microneutralization test and molecular typing methods. Homology comparisons indicated that the nucleotide acid identities and amino acid sequence identities were 92.3 ％- 100.0％ and 98.7 ％- 100.0％,respectively among these CVB5 isolate.s,and compared with the Faulkner prototype strain,which were 81.0％-82.4％ and 96.6％97.0％,respectively.Phylogenetic analysis on VP1 sequences showed that all CVB5 could be separated into four genogroups of A,B,C and D.Isolates of this outbreak belonged to genogroup D.Interestingly,two distinct genogroups in the phylogenetic tree were observed among the 17 isolates.Conclusions CVB5 is responsible for the outbreak of aseptic meningitis in Linyi City of Shandong Province,China. The genetic diversity is high among the isolates and all belong to genogroup D.

Objective To analyze the expression characteristics of excision repair cross-complementing 1 (ERCC1), topoisomeraseⅡ (TOPOⅡ), ribonucleotide reductase M1 (RRM1), β3-tubulin and thymidylate synthase (TS) in lung cancer and their associations with the pathological types. Methods The immunohistochemical EnVision method was used to determine the expression of ERCC1, TOPOⅡ, RRM1,β3-tubulin and TS in 548 patients who were diagnosed as lung cancer from January 2011 to December 2014. Variance analysis was performed to analyze their expression characteristics among different pathological types and correlation. Results The expression positive rates of ERCC1, TOPOⅡ, RRM1, β3-tubulin and TS were 61.86 % (339/548), 91.06 % (499/548), 62.59 % (343/548), 73.18 % (401/548) and 70.44 % (386/548), respectively. The expression of ERCC1 was weak positive mostly (P<0.05), meanwhile the expression of TOPOⅡ was medium-strong positive mostly (P<0.05). In ERCC1 group, the positive rate of squamous cell carcinoma was higher than that of adenocarcinoma [57.39 % (167/291) vs. 42.61 % (124/291), P=0.000]. In weak positive of TOPOⅡ group, the proportion of adenocarcinoma was higher than that of squamous cell carcinoma [23.58 % (100/137) vs. 8.73 % (37/137), P=0.000]. In medium-strong positive of TOPOⅡ group, the proportion of squamous cell carcinoma was higher than that of adenocarcinoma [47.41 % (201/287/) vs. 20.28%(86/287), P=0.000]. The expressions of ERCC1, TOPOⅡ, RRM1,β3-tubulin and TS were irrelevant (r=0.4, P=0.397). Conclusions The expressions of ERCC1 and TOPOⅡ are higher in squamous cell carcinoma than those in adenocarcinoma. The expression of ERCC1 is weak positive mostly, meanwhile the expression of TOPOⅡis medium-strong positive mostly. There is no correlation between them.

Objective To analyze the genetic characteristics of echovirus 6 ( E-6) strains isolated from patients with acute meningitis/encephalitis syndrome ( AMES) in 2014 and sewage samples in 2013—2014 in Shandong province and to investigate their correlations.Methods Enterovirus strains were isolated from cerebrospinal fluid, stool and throat swab samples collected from 940 cases of AMES and 96 sewage samples used for environmental surveillance.The positive isolates were identified by molecular typing meth-od.Homologous and phylogenetic analyses based on the VP1 sequences of E-6 isolates were performed.Re-sults Altogether 47 E-6 strains were isolated from patients with AMES in 2014, accounting for 29.56%of all isolated enteroviruses ( EVs) strains.No E-6 strains were isolated from AMES cases in 2013.Data of the environmental surveillance showed that E-6 virus strains had been frequently detected in sewage samples since the summer of 2013 to the end of 2014.In total, 40 E-6 virus strains were isolated (7.87% of total isolated EVs strains) in 2013 and 139 E-6 virus strains (26.18%) in 2014.Phylogenetic analysis indicated that the E-6 isolates recruited in this study belonged to clusters A and C with high intracluster sequence iden-tities between AMES and environmental isolates.The nucleotide identities were 98.3%-100% among cluster A E-6 virus strains isolated from AMES cases in 2014 and 96.6%-100% among cluster A E-6 virus environ-mental isolates during the surveillance year 2013—2014.The cluster A E-6 virus strains shared 97.1%-100% nucleotide identities between the AMES and environmental isolates.For cluster C E-6 virus strains, the nucleotide identities were 100%, 98.7%-100% and 99.1%-100%, respectively.Conclusion The epidemic of viral encephalitis in Shandong province in 2014 was associated the transmission of two lineages of E-6 virus.Environmental surveillance revealed the potential epidemic of E-6 virus even before the epidemic of viral encephalitis in Shandong province, indicating the possibility of using environmental surveillance for early warning of related diseases.

Objective To analyze the genetic characterization of VP1 region of Coxsackie virus A10(CVA10)isolated from clinical specimens of hand, foot and mouth disease(HFMD) patients in Shandong Province. Methods Clinical specimens were collected from some of HFMD patients from 2008 to 2009. The virus was isolated by cell culture. Total RNA was extracted, and the VP1 genes of the isolates were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. The genotypes were identified by molecular typing method and bioinformatics analysis.Homologous comparison and phylogenetic analysis of representative CVA10 strains were performed.Homologous comparison between the Shandong isolates and strains obtained from GenBank were performed and phylogenetic analysis of some representative CVA10 strains were performed. Results Three hundred and thirty viruses strains were isolated from 760 clinical specimens collected from HFMD patients, and 17 of them were identified as CVA10. The homologies of nucleotide and amino acid of the 17 CVA10 strains were 82.3%-100.0% and 94.2%-100.0%, respectively. Compared with the prototype strain of CVA10 (Kowalik/USA/2003), the homologies of nucleotide and amino acid were 75.6%-76.8% and 90.2%-93.2%, respectively. Interestingly, Shandong CVA10 strains were clustered into two distinct subgroups in the phylogenetic tree. Conclusions CVA10 is one of the causative agents of HFMD. Two independently circulating subgroups of CVA10 exist in Shandong province.

ObjectiveTo investigate the effect of targeted regulation of the Wnt2 gene by microRNA(miR-21) on the proliferation and migration of HepG2 hepatoma cells. MethodsQuantitative real-time PCR was used to measure the mRNA expression of miR-21 in HepG2 hepatoma cells and normal liver cell line LO2. HepG2 cells were transfected with miR-21 inhibitor, and then the expression of miR-21 and cell proliferation, migration, and apoptosis were analyzed for the inhibitor group and the control group. The protein expression of Wnt2 was measured for the two groups, and dual-luciferase reporter assay was used to verify the association between miR-21 and the Wnt2 gene. The t-test was used for comparison of continuous data between groups. ResultsThe relative expression of miR-21 in HepG2 cells was significantly higher than that in LO2 cells (1.978±0.035 vs 1.586±0.022, t=16.424, P＜0.05). After the transfection of miR-21 inhibitor, the inhibitor group had significantly lower expression of miR-21 than the control group (0.857±0.017 vs 1.684±0.039, t=33669, P＜0.05). Compared with the control group after the transfection of miR-21 inhibitor, the inhibitor group had a significant reduction in the proliferation ability of HepG2 cells (P＜0.05), a significantly lower number of cells passing through the Transwell chamber (83.72±15.06 vs 147.85±20.64, t=4.347, P＜0.05), and a significantly higher cell apoptosis rate (25.67%±3.95% vs 10.27%±2.14%, t=5937, P＜0.05). The inhibitor group had significantly lower relative expression of Wnt2 in HepG2 cells than the control group (0.862±0.127 vs 1.306±0.218, t=3.048, P＜0.05). TargetScan software showed that miR-21 inhibitor significantly inhibited the luciferase activity of the cells transfected with wild-type Wnt2-3′UTR plasmid (0.972±0.102 vs 0.612±0.092, t=4.219, P＜005), while there was no effect on the luciferase activity of the cells transfected with mutant Wnt2-3′UTR plasmid (0.982±0.093 vs 0911±0.128, t=0.972, P＞0.05). ConclusionInhibition of miR-21 expression can effectively inhibit the proliferation and migration of HepG2 cells, promote the apoptosis of HepG2 cells, and inhibit the over-activation of the Wnt signaling pathway, and therefore, it may become one of the potential target genes for liver cancer treatment.