Objective To analyze the prognostic factors in patients with cervical squamous cell carcinoma of stage Ⅰb and Ⅱa treated by surgery,and to investigate their guid roles in available post-operation adjuvant therapy. Methods The clinicopathologic records of 306 patients with cervical squamous cell carcinoma of stage Ⅰb and Ⅱa who underwent radical hysterectomy and pelvic lymphadenectomy were retrospectively analyzed, and the prognostic factors were explored by univariate and multivariate methods. Independent prognostic factors were identified by COX proportional hazards regression model. Results The overall 5-year survival rate of these 306 patients was 78.1%. In univariate survival analysis, the poor prognostic factors included poor differentiation, positive pelvic lymph nodes, deep stromal invasion, parametrial extension, tumor size≥4 cm, and lymph vascular space involvement (P

On the basis of the Uncaria transcriptome, specific primers were designed for UrSTR. The full-length cDNA of UrSTR (GeneBank: OL310251) was 1 541 bp, encoding 345 amino acid residues, and the promoter region sequence of UrSTR (GeneBank: OL310252) was 1 179 bp. Phylogenetic tree is revealed that UrSTR had a closest relationship with STR from Ophiorrhiza pumila and Ophiorrhiza japonica. Localization of UrSTR protein is revealed located in the vacuole membrane. Plant-care analysis indicated that the promoter region sequence of UrSTR, covering multiple light, stress and hormone-response cis-regulatory elements, and verified transcriptional activity. The results of SDS-PAGE show that pET-28a-UrSTR recombinant protein was successfully expressed, and the size was anticipated. The UrSTR prokaryotic expression system needs to be optimized in the later stage. The research lays the foundation for further purification to study its structure and functional characterization of the UrSTR protein.

Objective To screen the differentially expressed genes in esophageal squamous cell cancer (ESCC) and normal tissue of esophageal mucosa in Kazakh. Methods RNA was extracted from the ESCC sections in Kazakh patients, and was amplified to obtain cRNA. The gene expression profiles in ESCC and normal tissue of esophageal mucosa were detected by HG-U133 Plus 2.0 gene chip. The results were analyzed by bioinfor-matics. Results One hundred and seventy differentially expressed genes in ESCC and normal tissue of esophageal mucosa were found, with a difference of more than 10 times in expression levels. Of the 170 genes, 39 were up-regulated (signal log ratio > 3 ) and 131 down-regulated (signal log ratio < - 3). These factors such as cell cycle regulation, apoptosis, cytoskeleton; extracellular matrix, intracellular signal transduction, protein translation and synthesis, and immunological functions were correlated with the genes with abnormal expression. Conclusion The use of oligonucleotide microarray accurately and efficiently screen the 170 target genes in ESCC in Kazakh. It is suggested that these genes may be related to the carcinogenesis and development of ESCC in Kazakh.

Objective To compare the effect of cimetidine associated with bletilla striata,rhubarb on peptic ulcer,cushingulcer,acute erosive gastritis,bleeding of esophagus and gastric funds varicosis of cirrhotic portal hyper- tension and gastri cancer.Methods 136 patients with upper gastrointestinal hemorrhage were randomly divided into 2 groups.The treated group(n=69)was given cimitidine associated with bletilla striata,rhubarb and the control group(n=67)was given cimitidine only.The effects of hemostasis in 2 groups were observed.Results The effect of hemostasis in the treated group was better than that in the control group on peptic ulcer,cushingulcer,acute ero- sive gastritis and gastric cancer(P＜0.05).There were no difference between 2 groups about bleeding of esophagus and gastric funds varicosis of cirrhotic portal hypertension(P＞0.05).Conclusion The effect of hemostasis on up- per gastrointestinal hemorrhage treated by integrated traditional Chinese medicine and western medicine might achieve better on cushingulcer,acute erosive gastritis,peptic ulcer and gastri cancer.It has less adverse effect and more safety.

In this article, the status of spindle assembly checkpoint and the alteration of its major component, Mad2 protein level were examined in A2780 and SKOV3 ovarian cancer cell lines. Recombinant eukaryotic expression plasmid pEGFP-Mad2 was transfected into paclitaxel-resistant SKOV3 cells and Mad2 protein was knocked down by Mad2-specific siRNA in paclitaxel-sensitive A2780 cells. Then the expression level of Mad2 gene was detected by Western blotting. Flow cytometry revealed that SKOV3 cells were not fully arrested in G(2)/M phase in contrast to A2780 cells in the presence of paclitaxel. However, paclitaxel sensitivity assay showed that sensitivity to paclitaxel was reversed after the transfection in both cell lines in terms of number of cells arrested at G(2)/M phase and the expression of Bcl-2 was significantly changed. These results suggest that weakened spindle checkpoint with reduced expression of Mad2 is associated with resistance to paclitaxel in ovarian cells and Bcl-2 may be involved in this process.

OBJECTIVEElectrical and structural remodeling are of importance for the occurrence and maintenance of atrial fibrillation. We observed association between atrial connexin protein expression and fibrosis in a canine model of prolonged rapid atrial pacing.

METHODS"J"-type electrodes were placed in the right atrial appendage under the guidance of X-ray in 16 dogs, Animals in model group (n = 8) received fast pacing (400 beats/min) for 10 weeks while animals in control group (n = 8) maintained at sinus rhythm. Limb-lead ECGs were recorded at 2, 4, 6, 8 weeks respectively. Burst stimulation was applied to induce atrial fibrillation in all animals after 10 weeks, animals were sacrificed thereafter and the left atrial tissues were taken for myocardial collagen measurement (Masson staining) and myocardial ultrastructure examination and detection of protein expression of connexin (Cx) 40 and 45 (immune staining). Procollagen type III N-terminal peptide and type IV collagen in serum were also detected by radioimmunoassay.

RESULTSTwo dogs died in model group due to atrial rupture induced cardiac tamponade or lung emboli. Spontaneously atrial fibrillation was not observed in all animals, but two dogs developed atrial flutter and atrial premature beats. Atrial fibrillation was induced by burst stimulation in 4 out of 6 dogs in model group and in none of the dogs in control group. Atrial myocardial collagen volume fraction was significantly increased in model group compared with the control group (P < 0.05). Ultrastructure examination in atrial tissue evidenced disorder, fracture, collagen fiber proliferation, mitochondrial swelling, blurred cristae, and intercalated disc distortion, expansion, part of gap junction disappears in model group. The serum levels of procollagen type III N-terminal peptide and type IV collagen in model group were significantly higher than in the control group (P < 0.05). The protein expression of Cx 40 in atrial myocardium in model group was significantly higher than in control group (P < 0.05), while Cx 45 protein expression was similar between two groups (P > 0.05). The left atrial CVF was positively correlated with Cx 40 (r = 0.671, P < 0.01).

CONCLUSIONIncreased myocardial fibrosis is positively correlated with upregulation of myocardial Cx 40 protein expression in left atrium in rapid atrial paced canine.

Animals ; Atrial Fibrillation ; metabolism ; pathology ; Cardiac Pacing, Artificial ; Connexins ; metabolism ; Disease Models, Animal ; Dogs ; Fibrosis ; Heart Atria ; Myocardium ; metabolism ; pathology

Objective To perform molecular cloning and sequencing, bioinformatics analysis,protein expression and function of extracellular signal regulated kinase (EgERK1) of Echinococcus granulosus in Xinjiang. Methods The specific primers of EgERK1 were designed and total RNA was extracted from Echinococcus granulosus in Xinjiang. EgERK1 gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and prokaryotic expression plasmid pET28a-EgERK1 was constructed and sequenced. The sequences were analyzed by DNA sequencing and bioinformatics technology. The recombinant EgERK1 protein was induced and expressed. The biological function was detected using sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot. Results The sequence of RT-PCR product was 1125 bp, encoding 374 amino acids with isoelectric point of 6.34.This gene was a new ERK-homologues gene indicated by BLAST, named EgERK1(EU701008).Homology comparisons indicated that the homology of EgERK1 and EmMPK1from Echinococcus multilocularis was 95.45%, and was 43.04%-61.88% to ERK from Caenorhabditis elegans, S. cerevisiae, D. melanogaster and human. Phylogenetic analysis showed that EgERK1 clustered with EmMPK1. Bioinformatics analysis predicted that EgERK1 contained a highly conserved T-X-Y motif and activation loop segment of ERK-like kinase.Western blot results showed the EgERK1 recombinant protein could reacted specifically with anti-human ERK monoclonal antibody. Conclusion A new EgERK1 gene of Echinococcus granulosus is successfully cloned and its recombinant protein could reacted specifically with ERK1/2 antibody, which provides the basis for further study of EgERK1 function in the host-parasite interaction.

Objective To establish a real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) for detection of human Herpesvirus-8 (HHV-8) viral load. Methods pMD19-T recombinant vectors inserted with an open reading frame (ORF) 26 of HHV-8 or β-actin gene were constructed respectively. A sensitive RT-qPCR method was established and optimized. The effectivity of the method was evaluated by determining the HHV-8 viral loads in 30 (formalin fixed, paraffinised)biopsy samples of Kaposi's sarcoma. Results The key factors for optimizing the method included anneal temperature and extension. The standard curve showed that the Ct value of ORF26 and β-actin had a good linear relationship (r2 ＞0.990) with the standard samples. The melt curve and electrophoresis showed the specificity of our study. The sensitivity of this method was very high and the detection rate could reach 100%. The viral loads were significantly higher in patients with classic Kaposi's sarcoma compared to patients with acquired immunodeficiency syndrome-associated Kaposi's sarcoma(69.18 va 8. 63, x2 =7.950,P=0.005).Conclusions The established RT-qPCR method is highly sensitive, which can be used as a routine assay for detecting HHV-8.This system offers a good platform for diagnosing other causative organism.

Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

Objective To establish serological detection methods of human herpesvirus 8 (HHV-8).Methnds Three potent antigenic fusion proteins.K8.1,ORF65 and ORF73 C of HHV- 8 were synthesized using E.coli system.The sera were detected using lhese antigenic proteins.The positive sera were from 12 patients with Kaposi's sarcoma and 32 patients with acquired immunodeficiency syndrome-related Kaposi's sarcoma.The negative sera were from 20 patients with cutaneous tumors and children under 15 years old.Western blot and enzyme-linked immunosorbent assay (EI.ISA)were employed to determine the immunogenicity of each recombinant protein and the sensitivity and specificity of ELISA using the complex antigens.Results Three types highly purified HHV 8 specific recombinant pro teins with potent antigenicity were successfully synthesized.The sensitivity of ELISA using the above complex antigens was significantly higher than traditional immuno-flurescent assay (IFA)detecting the positive and negative sera,whieh were 81.8%,34.4%,respectively.And the specificity of ELISA was 97.9%.Conclusion K8.1,ORF65 and ORE73 C are good candidate antigens for establishing HHV-8 serological detection methods,which have better sensitivity and specificity.