Objective:To investigate the association of tumour necrosis factor- ? (TNFs-?) gene polymorphism with Graves ophthalmopathy (GO)Chinese in Shandong . Methods: Allele and genotype frequencies were determined by polymerase chain reaction with sequence specific primers(PCR-SSP). Results: Allele and genotype frequencies of TNF-? +488、-308 were not different significantly between Graves′ disease(GD) and control, or GD subgroups with and without GO. There were also no significant differences between GO subgroups with different severity. Conclusion: There are no associations of TNF-? +488、–308 polymorphisms with GO susceptibility in Shandong Chinese.

Objective To investigate the relationship between acute biliary pancreatitis (ABP) and anomalous pancreaticobiliary duetal union (APBDU). Methods 131 patients with ABP were enrolled to test the serum total bilirubin (TB), alanine amintransferase (ALT), aspartate amintransferase (AST), alkaline phosphates (ALP), γ-glutamyl transferase (γ-GT). All the patients received medical treatment, and then these tests were performed again. Thereafter, all the patients underwent selective surgery and intra-operative cholangiography was performed to observe the pancreaticobiliary duetal union. Results 27 patients (20.6%) with APBDU were found in 131 patients. Among them, 8 cases (29.6%) was B-P subtype (TypeⅠ), 16 cases (59.3%) was P-B subtype (TypeⅡ) , and the remaining 3 cases was mixed subtype (TypeⅢ). A significant decrease of ALT, AST, ALP, γ-GT after non-surgical treatment in both group of APBDU and NAPBDU was noted (P＜0.05). The serum levels of ALT, AST,γ-GT in APBDU patients were (71.81± 23.19) U/L, (47.85±27.87) U/L, (52.86±31.49) U/L, respectively; and in NAPBDU patients were (51.96±15.40) U/L, (40.77±16.58) U/L, (34.86±26.47) U/L. The difference was statistically significant (P＜0.05). Condusions APBDU is an important etiology of ABP.

AIM:To investigate the effects of peroxiredoxin 4 ( Prdx4) protein expression levels on the migra-tion and invasion of human cervical cancer HeLa cells.METHODS:The plasmid pcDNA3.0-HA-Prdx4 was transfected into HeLa cells.The HeLa cells were infected with LV-Prdx4 RNAi vector to establish stable Prdx4 shRNA HeLa cells. The change in the expression of Prdx4 protein was validated by Western blotting.The wound-healing assay, and Transwell migration and invasion assays were performed to detect the migration and invasion of HeLa cells, respectively.RESULTS:The expression of Prdx4 protein was up-regulated in the HeLa cells after transfection with pcDNA3.0-HA-Prdx4 plasmid ( P<0.05), whereas it was down-regulated in the Prdx4 shRNA HeLa cells (P<0.05).The abilities of migration and inva-sion were significantly increased in Prdx4-overexpressing HeLa cells compared with non-transfected and mock plasmid trans-fected control groups ( P<0.01) .When Prdx4 was knocked down by shRNA, the migration and invasion of the HeLa cells were remarkably repressed compared with blank control group and negative control group ( P<0.01 ) .CONCLUSION:The up-regulation of Prdx4 expression facilitates the migration and invasion of HeLa cells, and the down-regulation of Prdx4 expression inhibits the migration and invasion of HeLa cells, indicating that Prdx4 may be a potential molecular target for cervical cancer therapy.

Objective To explore the value of cardiac troponin-I (cTnI), B-type natriuretic peptide (BNP) and blood lactic acid (Lac) on evaluation of severity and prognosis in patients with septic myocardial dysfunction (SMD). Methods According to retrospective analysis of clinical data,161 cases with sepsis were divided in to SMD group and non-SMD group. And the SMD group was further divided in to death group and survival group. Blood cTnI, BNP and Lac value in each group were detected respectively. The ROC curve was used to evaluate the forecast value of cTnI, BNP and Lac on prognosis for patients. Results The value of cTnI, BNP and Lac in SMD group were significantly higher than those in non-SMD group(P<0.05);The value of cTnI, BNP and Lac in death group among the SMD patients were significantly higher than those in survival group(P<0.05);cTnI, BNP and Lac contribute to predict the 28 day mortality rate of SMD. Conclusions Blood cTnI, BNP and Lac contributes to the assessment of the severity and the prognosis of septic patients with myocardial dysfunction.

Objective Lipocalin-2(LCN2) can promote the M1 approach of microglia.The study was to explore the effect of activated microglia induced by LCN 2 on depression pathogenesis in rats and its mechanism . Methods According to the chronic stress depression model created by Willner , 40 adult male SD rats were divided into 4 groups(n=10): normal control group;UCMS group;UCMS+LCN2 siRNA group;LCN2 siRNA control group .A series of stress stimulation was given on UCMS group and UCMS +LCN2 siRNA group for 21 days to create depression model .At 7 days after the stress stimulation , the rats in UCMS+LCN2 siRNA group and LCN2 siRNA control group were anaesthetized by 0.4mg intraperitoneal injection of 10% chloral hydrate , followed by intrathecal injection of LCN2 siRNA(0.015 μL/g, 3 times a week) to the rats till the end of the stress (21 days).At the same time, the same volume of isotonic saline was given to normal control group and UCMS group .The weight of rats was measured every week and the sucrose preference and the forced swimming test were applied to measure behavior of the rats after the experiment .The hippocampus of the rats were extracted and immunofluorescence and western blot were applied to detect the expressions of microglia specific markers:LCN2 and the Iba. Results At 3 week, the weight of rats in UCMS+LCN2siRNA group was higher than that of UCMS group ([262.82 ±0.01]g vs [179.98 ±0.08]g, P<0.05).The weight of rats in normal control group and LCN2 SiRNA control group increased significantly higher than the other two groups (P<0.05).The sucrose preference values of normal control group (0.82 ±0.01),UCMS+LCN2 siRNA group(0.81 ±0.01) and LCN2 siRNA control group(0.82 ±0.01) were higher than that of UCMS group (0.25 ±0.04) (P<0.05).The fixed time of the forced swimming test of UCMS+LCN2siRNA group decreased significantly compared with UCMS group ([4.64 ±0.8]s vs [23.11 ±2.63]s, P<0.05).The LCN2 expression of UCMS group was significantly greater than the other groups (P<0.05).The Iba1 expression in the hippocampus of the UCMS group increased significantly compared with other groups . Conclusion LCN2 is associated with the pathogenesis of de-pression induced by chronic stress reaction and is mechanism may be related to the activation of microglia in the central nervous system of rats.

ObjectiveTo investigate the prevalence and distribution of 16S rRNA methylase gene and research the relationship with drug resistant spectrum.And preliminary explore its role in molecular epidemiology analysis.MethodsCollected 69 clinical isolates of non repetitive ESBL-producing Klebsiella pneumoniae in our hospital from Mar to Sep 2010.Detection 16S rRNA methylation enzyme gene by PCR,and analyze ESBL genetype and integron gene of the positive strains.All PCR products were sequenced for determination.Plasmid conjugation test and plasmid elimination method to determine dissemination of 16S rRNA methylase gene.Then we used ERIC-PCR genotyping technology for the establishment of DNA fingerprinting.ResultsIn sixty-nine strains,twenty isolates were rmtB positive (28.9％),two isolates were armA positive,and two strains coproduce rmtB and armA.All positive isolates carried the CTX-M gene,detemined by sequencing,14 strains of CTX-M-14 gene,6 strains of CTX-M-15 gene,14 strains carried TEM1 gene,8 strains carried SHY gene,sequencing showed that 5 strains of SHV-12 gene,3 strains of SHV-11 gene,3 strains carried OXA-10 gene,3 strains carried VBE-1 gene.In addition,the intl gene was found in 12 isolates of 20 rmtB positive strains.All the intl gene positive strains were divided into five kinds gene cassettes,which contained drfA25,drfA1,drfA12,aadA1,aadA2,sat and blaVEB-1 genes.Respectivily,16S rRNA methylase gene positive strains were divided into five genetypes using ERIC-PCR technology.A genetype was the advantage popular clones.Conjugative plasmid and elimination test found that rmtB gene was located in a plasmid in KP5 and KP16 isolates with A genetype,and can disseminate by conjugation.ConclusionA high prevalence of 16S rRNA methylase gene-rmtB was found among clinical ESBL-producing K.pneumoniae isolates in our hospital,which could lead to resistant to almost all aminoglycoside at a high level.Both horizontal gene transfer and clonal spread were responsible for the dissemination of the rmtB gene.In addition,K.pneumoniae co-producing ESBLs,16S rRNA methylation enzymes and class Ⅰ integron existed and were spreading.

Objective To investigate molecular epidemiology and antimicrobial susceptibility of carbapenem-resistant strains of Klebsiella pneumoniae isolated from children.Methods From July 2010 to June 2011,twelve non-replicate clinical isolates of carbapenem-resistant Klebsiella pneumoniae were consecutively collected from children inpatients in the Second Hospital of Wenzhou Medical Colloge.All of the isolates were identified by the automated microbiology systems.Modified Hodge test was used to screen strains producing carbapenemases.Pulsed field gel electrophoresis(PFGE) was performed to analyze the homogeneity of genomic DNA of Klebsiella pneumoniae.KPC,IMP,GIM,SPM,SME,OXA-10,bla(s),VIM gene and integrase gene were amplified by PCR and then sequenced to cofirm the genotypes;Plasmid conjugation experiment was used to study the transfer method of bacterial resistance.Plasmid-curing test were used to initally locate the resistant genes.Results One(8.3％),5(41.7％),7(58.3％),1(8.3％),1(8.3％) and4(33.3％) of12isolates were susceptible to gentamicin,tobramycin,amikacin,ciprofloxacin,levofloxacin and trimoxazole,respectively.All isolates carried KPC-2,TEM-1 and SHV genes(six for SHV-11-like,six for SHV-12-like).Eleven of twelve isolates with KPC-2 gene carried CTX-M genes(4 for CTX-M-14-like,6 for CTX-M-15-like).Two isolates carried OXA-10 genes,and one isolates carried PER-1 gene.None of NDM-1,GIM,SPM,SIM and VIM carbapenemase genes was detected in 12 isolates.All of 12 isolates carried Int 1 genes.The plasmids of 2 isolates were transgerred into the recipients E.coli EC600.PCR and sequence analysis revealed that blaTEM-1 and blaCTX-M-15-like were co-transferred with the KPC-2 gene to the recipients.Elimination of KPC-2-encoding plasmid from Kp7 and Kp12 resulted in imipenem susceptibility in the two isolates.Amplification revealed that KPC-2 gene was lost by the plasmid-curing test.Of the 12 isolates,5 patterns were obtained by PFGE.Pattern B and C were the main drug resistant clones.Conclusion KPC-2 gene are the major carbapenemase genes in Klebsiella pneumoniae isolated from children,including ESBLs and integrase.Some resistance genes can be disseminated by plasmids.

Objective:To probe the method of isolating outer hair cells (OHC) from each of four turns of the guinea-pig cochlea. Method:From eight guinea pigs the organ of Corti from each of four turns of the cochlea were dissected, and then treated using enzyme. Result:A fair amount of living OHCs from each of four turns were obtained. The length of OHCs from each of four turns were 23.81,34.50,60.48 and 71.37 μm. Conclusion:The key to success in isolating OHCs from each of four turns of the cochlea is to know very well the anatomical characteristics of each of four turns of the cochlea and be operated in accordance with normal rules.

Objective:To investigate the relationship between herpesviridae and malignant or benign laryngeal diseases.Method:128 paraffin-embedded laryngeal squamous cell carcinoma and laryngeal epithelium hyperplastic lesions were detected by polymerase chain reaction (PCR) and PCR-ISH for herpesviridae. Result:HSV-1 was detected in 10 cases by PCR,among them 3 were laryngeal squamous cell carcinoma (LSCC),1 was carcinoma in situ(CIS),4 were laryngeal polyps and 2 were laryngeal keratosis. Except 1 LSCC and 1 CIS, 8 of 10 cases were positive while detected by PCR-ISH. In benign diseases, signals were shown from basal layer to superficial cell; in malignant lesions, the signals were scattered in the diseases.Conclusion:Most of laryngeal diseases were not related to herpesviridae, but HSV-1 may acts as initiator in the development of a few cases.

Objective To study the effect of Qizhen capsule combined with docetaxel injection on the neoadjuvant chemotherapy of breast cancer and its effects on cyclooxygenase (COX-2), cancer antigen 15-3 (CA15-3),and vascular endothelium growth factor (VEGF).Methods 84 patients with breast cancer were selected in our hospital from January 2015 to August 2016, those patients were divided into observation group and control group according to coin method.The control group received docetaxel injection, epirubicin injection, cyclophosphamide neoadjuvant chemotherapy, the observation group was treated with Qizhen capsule on the basis of the control group.The clinical efficacy, COX-2, CA15-3 and VEGF levels, quality of life and adverse reactions were compared between the two groups.Results After treatment, the total effective rate in the observation group (90.48%) was significantly higher than the control group (57.14%) (P<0.05).After treatment, the levels of COX-2, CA15-3 and VEGF in the observation group were significantly lower than the control group (P<0.05).The scores of cognitive function, emotional function, social function, role function and physical function of life quality in the observation group were significantly higher than the control group ( P<0.05 ) .Cardiac toxicity, nausea and vomiting, thrombocytopenia, alopecia, neutropenia and leukopenia were significantly lower in the observation group than the control group (P<0.05). Conclusion Qizhen capsule combined with docetaxel injection can significantly reduce the levels of COX-2, CA15-3 and VEGF in the neoadjuvant chemotherapy of breast cancer, and the clinical curative effect is good, which can effectively improve the quality of life of patients and reduce the adverse reaction rate.