Objective To investigate the relationship of smoking, the gene polymorphisms of heine oxygenase-1, tumor necrosis factor-α, and human β-defensin-1 with chronic obstructive pulmonary disease (COPD). Methods A case-control study was held to compare the genotype frequency distribution of heine oxygenase-1, tumor necrosis factor-α, human β-defensin-1 by PCR -RFLP method and to analyze the relationship among them. Results The L allele gene frequency of heine oxygenase-1 was 23.44% in COPD group and 10.71% in control group with significant difference(P＜0.05). The heterogenesis gene frequency of tumor necrosis factor-α in COPD group and control group were 18. 24% and 7.53% (P＜0.01). The heterogenesis gene frequency of human β-defensin-1 was 14.28% in COPD group and 5.00% in control group with significant difference(P＜0. 01). Conclusions Smoking is a risk factor for COPD. Genetic polymorphisms of heme oxygenase-1, tumor necrosis factor-α, human β-defensin-1 are related with the development of COPD.

Objective To explore the bacteriostatic efficacy of snake venom antibacterial peptide OH-CATH and chitosan as ar-tificial implant coating on escherichia coli (E .coli) .Methods The catheters and dacron patches (blank group ,chitosan group ,cef-operazone group and antibacterial peptide group) were performed the pretreatment .Then the in vitro bacteriostatic test was conduc-ted to evaluate the antibacterial activity U value on E .coli ATCC 25922 and cephalosporin-resistant E .coli on 1 d and on 7 d after putting it into plasma .Results The cefoperazone group and the antibiotic peptide group had the very powerful antibacterial activity on E .coli ATCC 25922 on 1 ,7 d ,but the antibacterial activity of the cefoperazone group was stronger than that of the antibiotic peptide group(P<0 .05);the cefoperazone group had no antibacterial activity on cephalosporin-resistant E .coli ,but the antibiotic peptide group had significantly antibacterial activity on 1 ,7 d(P<0 .01) .Conclusion Snake venom antibacterial peptide OH-CATH has very obvious antibacterial effect on the standard strain and drug-resistant strain of E .coli and its value applied in artificial im-plant coating is affirmed .

Objective To investigate the efficacy of twice irradiating with focused ultrasound on recurrent and metastasized U14 cervical cancer implanted in the legs of mice. Methods Seventy-two mice with U14 cervical cancer cells implanted in their legs were divided into three groups randomly,with 24 rats in each group.One group received a single dose of focused ultrasound,while mice in the second group were irradiated twice and surgery resection was administered to the third group 7 or 8 days after the tumor was implanted.After 23 days post implantation of the tumor,local tumor recurrence and metastasis to the lungs and lymph nodes were evaluated. Results The inhibition rate after double irradiation was 61.70％ for local recurrence and 68.18％ for metastasis,significantly higher than in the other two groups. Conclusions Both single and double irradiation with focused ultrasound are effective for inhibiting local recurrence and metastasis,but double irradiation is more effective.

Objective To establish nucleic acid testing techniques for detecting Nipah virus (NiV) and Hendra virus (HeV), and to test the NiV and HeV in peripheral blood collected from domestic pigs, cows and goats in Chongqing. Methods Peripheral blood samples of 580 domestic pigs, 250 cows, 180 goats were collected from Chongqing since June 2007 to June 2008. The lymphocytes were separated by density gradient centrifugation and total RNA was extracted using Trizol method for detection of NiV and HeV with one-step real-time RT-PCR. Sequence identification and analysis were performed for positive PCR prod-ucts. Virus isolation and culture were adopted for positive samples, and epidemiologic reports were submit-ted. Results Nucleic acid detections searching for NiV and HeV were successfully performed in animal blood samples collected from Chongqing. "Takeoff points" were not found in fluorescence amplification curves of all samples. Curves kept the same slope, and assays were judged as negative. Conclusion Until now, Neither NiV or HeV infection has been found in domestic animals blood samples collected from Chongqing, which suggest a lower possibility of outbreaks of Nipah disease and Hendra disease in Chongqing in the near future.

Objective To investigate the epidemiological characteristics,clinical features,diagnosis,treatment and prognosis of benign tumor of the bile ducts.Methods The retrospective cross-sectional multicenter study was conducted.The clinical data of 136 patients with benign tumor of the bile ducts who were admitted to the eight hospitals between January 2007 and December 2016 were collected,including 70 in the Eastern Hepatobiliary Surgery Hospital of the Second Military Medical University,19 in the First Affiliated Hospital of Zhengzhou University,15 in the First Affiliated Hospital of Xi'an Jiaotong University,11 in the First Affiliated Hospital of Dalian Medical University,7 in the Affiliated Hospital of North Sichuan Medical College,6 in the Southwest Hospital of the Third Military Medical University,4 in the Cancer Center of Sun Yat-Sen University and 4 in the Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine.All the patients received laboratory and imaging examinations,and then underwent corresponding surgery when treatment planning was respectively determined by comprehensive hospitals according to clinical features and results of examinations.Surgical procedures were performed based on the results of intraoperative frozen section in rapid pathological diagnosis.Observation indicators:(1) epidemiological characteristics;(2) clinical features;(3) results of laboratory and imaging examinations;(4) treatment situations;(5) follow-up situations.Follow-up using outpatient examination and telephone interview was performed to detect the survival of patients up to March 2017.Measurement data with normal distribution were represented as-x±s.Results (1) Epidemiological characteristics:Of 136 patients,the male to female ratio was 1.78 ∶ 1.The incidence of whole bile duct tumors was from high to low,including 52 patients with duodenal papilla adenoma,32 with extrahepatic bile duct adenoma,24 with intrahepatic biliary cystadenoma,11 with intrahepatic papillary adenoma,9 with intrahepatic bile duct epithelial tumor,7 with epithelial tumor of duodenal papilla and 1 with neuroendocrine tumor of duodenal papilla.Among 136 patients,adenomas (including cystadenoma) was detected in 108 patients,papillomas in 11 patients,intraepithelial neoplasias in 16 patients,neuroendocrine tumor in 1 patient;intrahepatic bile duct benign tumors in 44 patients and extrahepatic bile duct (including duodenal papilla) benign tumors in 92 patients.(2) Clinical features:of 44 patients with intrahepatic bile duct benign tumors,29 had abdominal pain,fever and abdominal masses,4 had jaundice,11 had no obvious clinical symptoms and were diagnosed by physical examination.Among 92 patients with extrahepatic bile duct benign tumors,76 developed obstructive jaundice,68 were accompanied by abdominal pain or colicky pain,8 were combined with pancreatitis and 2 deveIoped hemobilia,some patients were combined with multiple clinical symptoms.(3) Results of laboratory and imaging examinations:82 patients received CA19-9 test,results of 22 patients were abnormal,with a level of (148-± 126)U/mL.Ninety-seven patients received carcinoembryonic antigen (CEA) test,with a level of test of (2.7±2.0) μg/L,and a level of CEA in 1 patient was slight abnormal,with a level of 11.2 pμg/L.One hundred and thirty-six patients underwent preoperative ultrasound examinations,showing unqualified hepatic and bile duct space occupying lesions and bile duct dilatation.Ninetyfive patients underwent preoperative computed tomography (CT),80 underwent preoperative magnetic resonance imaging (MRI) or magnetic resonanced cholangio-pancreatography (MRCP),and 13 underwent preoperative endoscopic ultrasonography (EUS).Twenty-nine patients were considered for intrahepatic bile duct benign tumors;76 were diagnosed with obstructive jaundice,with uncertain benign or malignant tumors;other patients had bile duct space occupying,considering bile duct tumor (including cancer).Twelve patients with bile duct obstruction underwent ERCP,showing obstruction site and morphology.(4) Treatment situations:among 136 patients,65 underwent pancreaticoduodenectomy,17 underwent right hemihepatectomy + cholecystectomy,16 underwent cholecystectomy + hepatic left lateral lobectomy,11 underwent left hemihepatectomy + cholecystectomy,11 underwent duodenal papillary local excision+papilla reconstruction,11 underwent Roux-en-Y choledochojejunostomy anastomosis,4 underwent cholecystectomy + extrahepatic bile duct local excision + end-to-end bile duct anastomosis and 1 underwent endoscopic mucosal resection of duodenal papillary adenomna.Of 136 patients with postoperative complications,25 were complicated with pancreatic leakage,11 with bile leakage,2 with postoperative hemorrhage and 1 with hepatic failure.Two patients with pancreatic leakage died of massive hemorrhage caused by abdominal infection,1 died of hepatic failure and other patients were discharged from hospital after symptomatic treatment.(5) Follow-up situations:47 of 136 patients were followed up for 3-123 months,with a follow-up rate of 34.6％.During follow-up,2 patients undergoing duodenal papillary local excision + papilla reconstruction had canceration,and other patients had good survival.Conclusions There is a low clinical incidence of benign tumor of the bile ducts,which is more common in male than in female,and in adenomas (including cystadenoma) and papillomas.The preoperative imaging examinations or ERCP biopsy pathological examination can increase an accuracy of preoperative diagnosis.Benign tumors with high canceration rates need positive surgical treatment.

This paper summarized Professor PENG Peichu's experience in the differentiation and treatment of prostate cancer in three phases and four stages. It is considered that prostatic cancer is categorized into root deficiency and branch excess， with depletion of healthy qi as the root， and the accumulation of cancer toxin as the minifestation. Clinical diagnosis and treatment of prostatic cancer can be divided into three phases and four stages according to the exuberance and decline of pathogenic and healthy qi and the changes of deficiency and excess of yin and yang. In the initial accumulation phase of cancer toxin （yang excess stage）， the key pathogenesis is the accumulation of dampness， heat and static blood， and internal generation of cancer toxin， and the treatment should be resolving toxins， fighting cancer and dispelling yang excess. In the phase of healthy qi deficiency and toxin accumulation （yin deficiency stage）， with the lung and kidney yin deficiency， dampness， heat and static toxin accumulation as the key pathogenesis， the treatment should be centered on mutual generation between metal and water to nourish kidney yin， supplemented with the method of clearing heat and draining dampness， activating blood and resolving toxins， for which self-made Nanbei Formula（南北方）is usually used. In the phase of yang deficiency and cold stagnation （yang deficiency stage and yin excess stage）， with the spleen and kidney yang deficiency， cold dampness stagnation， static heat and toxin accumulation as the key pathogenesis， the treatment should be warming and tonifying spleen and kidney to dissipate cold accumulation； for deficiency of both yin and yang， and excess pathogen obstruction， modified Yanghe Decoction（阳和汤） is recommended， while for yang deficiency， cold congealing and blood stasis， self-made Wenshen Sanjie Formula（温肾散结方） can be used， and for cold dampness binding with cancer toxin， and cold complex with heat， self-made Quanan Formula （泉安方） is advised.

BACKGROUNDNowadays it is now a focus topic in schistosomiasis research to find ideal vaccine candidates and new drug targets for developing anti-schistosomiasis vaccine. We cloned a new gene, casein kinase II beta subunit, of Schistosoma japonicum (S. japonicum) and express it in Escherichia coli (E. coli).

METHODSThe ESTs obtained in our laboratory were analyzed by homologous searching, and a new gene was recognized. The full-length cDNA of the new gene was obtained by joining the 3'RACE PCR fragment and the EST clone. To express the new gene, the cDNA was cloned into pGEX-4T-1 vector and then transformed into E. coli JM109. The recombinant protein was analyzed by SDS-PAGE and Western-blot.

RESULTSA 908 bp cDNA was isolated from S. japonicum and identified to be casein kinase II beta subunit gene by sequence analysis. The open reading frame of the gene encodes a protein of 217 amino acids exhibiting 75.8%, 75.8%, 73.9%, 68.2%, 51.6% identity to the amino acids sequence of the corresponding genes of Homo sapiens (H. sapiens), Xenopus laevi (X. laevi), Drosophila melanogaster (D. melanogaster), Caenorhabditis elegan (C. elegan), and Schizosaccharomyces pombe (S. promber) respectively. The predicted molecular weight of the protein was 24.921 kDa. The new cDNA sequence had been submitted to GenBank, and its accession number is AY241391. This cDNA was subcloned into the pGEX-4T-1 vector and expressed in E. coli JM109. The recombinant protein could be recognized by the S. japonicum infected rabbit serum.

CONCLUSIONThe full-length cDNA sequences encoding S. japonicum casein kinase II beta subunit were firstly sequenced, cloned, and expressed in E. coli.

Amino Acid Sequence ; Animals ; Base Sequence ; Blotting, Western ; Casein Kinase II ; chemistry ; genetics ; Cloning, Molecular ; DNA, Complementary ; chemistry ; isolation & purification ; Escherichia coli ; genetics ; Molecular Sequence Data ; Rabbits ; Schistosoma japonicum ; enzymology ; genetics

Objective To explore the relationship between pulse blood pressure and cognitive function for the elders in Yuhuan County,Zhejiang Province.Methods Multi -stage cluster method was used to select 1 700 residents aged over 60 years in Yuhuan County,Zhejiang Province.Mini -Mental State Examination (MMSE)was used to screen cognitive impairment.Locally weighted scatter plot smoothing was used to explore the local linear trend between MMSE score and diastolic.The association between pulse pressure or blood pressure and cognitive function was analyzed by multivariate logistic regression analysis.Results The prevalence of cognitive impairments for the elders aged over 60 years in Yuhuan County was 15.74%.There was significant difference between the pulse blood pressure of cognitive impairment group (64.01 ±16.31)mmHg and none -cognitive impairment group (58.97 ±14.67)mmHg.The t value was 4.97.Pulse blood pressure and the elderly MMSE score showed a significant positive correlation.Multivariate logistic regression analysis showed that cognitive function related to diastolic blood pressure (OR =1.010,95% CI:1.001 -1.019,P =0.034)after adjusting several covariate.Conclusion Normal cognitive function was associated with decreased pulse blood pressure among the elders aged over 60 years in Yunhuan County,Zhejiang Province.

Objective To explore the association between different visual display terminal use and physical symptoms among adults.Methods Hangzhou,Jiaxing,Huzhou,Jinhua and Quzhou City were selected,where residents aged 18 years old and above were investigated using Mitofsky-Waksberg two-stage sampling.Information of demography,different visual display terminal use,eye and musculoskeletal symptoms,eye disease and eye relax were collected.Backward stepwise logistic regression was used to explore the association between different visual display terminal use and the symptoms (P value for addition ＜ 0.05,P value for removal ＞ 0.1).Results Totally 350 adults responded to the investigation,with a response rate of 48.54％.Finally,333 respondents were included in the analysis,among which 120 reported eye symptoms (36.04％) and 71reported musculoskeletal symptoms (21.32％).More time of computer use at work(0.5-4 h:OR=2.006,95％ CI:1.021-3.943;4-6 h:OR=3.578,95％ CI:1.751-7.315;＞6 h:OR=4.874,95％ CI:1.897-12.521) and eye disease (OR =4.361,95％ CI:2.465-7.714) were significandy associated with higher odds ratios in eye symptoms.Similar pattern was seen for the association on time of computer use at work with musculoskeletal symptoms (0.5-4 h:OR =3.815,95 ％ CI:1.809-8.045;4-6 h:OR =4.974,95 ％ CI:2.281-10.845;＞ 6 h:OR =5.934,95％ CI:2.347-15.005).More time of watching TV on television (＞ 2 h:OR =2.051,95％CI:1.068-3.939) and women (OR =1.884,95％ CI:1.052-3.372) were also observed to be statistically associated with musculoskeletal symptoms.However,no statistical association was observed between the two symptoms and entertainment purpose use of computer/cell phone/pad/kindle (P ＞ 0.05).Conclusion Computer use at work was significantly associated with eye symptoms,especially for those having eye diseases.It was also statistically associated with musculoskeletal symptoms,especially for women.

OBJECTIVE: To construct two types of Wnt-inducible secreted protein 3(WISP3) gene's mutants(1000T/C,840delT) found in spondyloepiphyseal dysplasia tarda with progressive anthopathy (SEDT-PA) patients, and to observe their expression in COS-7 cells. METHODS: Full-length cDNA of wild type WISP3 gene(WT-WISP3) was amplified from human chondrocytes by RT-PCR, and site-directed mutagenesis was used to obtain full-length cDNAs of the mutated WISP3 genes(MUT1000T/C and MUT840delT). The recombined plasmids WT-WISP3/pcDNA3.1(+), MUT1000T/C/pcDNA3.1(+) and MUT840delT/pcDNA3.1(+) were transfected transiently into COS-7 cells by liposome-mediated method, and pcDNA3.1(+) vector was used as a control. The total RNA and protein of the transfected COS-7 cells were extracted after 48 hours of transfection. The expression of WISP3 gene in the transfected COS-7 cells was detected by semi-quantitative RT-PCR and Western blot. RESULTS: By restriction endonuclease analysis and sequencing, the sequence of MUT1000T/C and MUT840delT were consistent with that mutated in SEDT-PA, and the open reading frames matched with the vector sequence. Semi-quantitative RT-PCR and Western blot showed that the recombined plasmids were highly expressed in COS-7 cells. CONCLUSION WISP3 gene's mutants of SEDT-PA are successfully constructed by genetic recombination, and expressed in COS-7 cells, which lays the foundation for the further study on its molecular functions in SEDT-PA.
Animals ; Base Sequence ; CCN Intercellular Signaling Proteins ; COS Cells ; metabolism ; Chlorocebus aethiops ; Humans ; Insulin-Like Growth Factor Binding Proteins ; biosynthesis ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Osteochondrodysplasias ; genetics ; metabolism ; Transfection