OBJECTIVETo explore the inhibitory effects of endoplasmic reticulum-retained intrabody on the secretion of type IV collagenase and the invasion of human pulmonary giant cell carcinoma PG cells in vitro.

METHODSTwo expression plasmids were constructed, pcDNA3.1-CP.scFv and pcDNA3.1-ER.scFv encoding cytoplasm-retained and endoplasmic reticulum-retained single chain antibodies against the type IV collagenase, respectively. The intracellular antibody genes were transfected into the human pulmonary giant cell carcinoma PG cells. Western blot was performed to detect the expression of pcDNA3.1-CP.scFv and pcDNA3.1-ER.scFv. Gelatin zymography was performed to detect seretion of type IV collagenase in PG cells and Matrigel assay was employed for determination of the cell invasiveness.

RESULTSBoth of cytoplasm-retained and endoplasmic reticulum-retained introbodies, CP.scFv and ER.scFv, were expressed in PG cells. ER.scFv, significantly inhibited the secretion of type IV collegenase. As shown, matrix metalloproteinase 9 and matrix metalloproteinase 2 were inhibited by 85.7% and by 51.2%, respectively. However, CP.scFv did not show such inhibitory effect. The ER.scFv encoding gene-transfected PG cells were much less invasive than parental or vector control cells, the inhibition rate was 76.3% (P < 0.05), whereas CP.scFv encoding gene-transfected PG cells showed no reduction in invasiveness.

CONCLUSIONThose findings demonstrate that endoplasmic reticulum (ER)-retained intracellular antibody technology may selectively abrogate the activity of type IV collagenase in the protein trafficking and secretory pathway and effectively inhibit tumor cell invasion in vitro. Anti-type IV collagenase intrabody may be further used in cancer gene therapy.

Carcinoma, Giant Cell ; metabolism ; pathology ; Cell Line, Tumor ; Cytoplasm ; immunology ; Endoplasmic Reticulum ; immunology ; Genetic Vectors ; Humans ; Immunoglobulin Variable Region ; metabolism ; physiology ; Lung Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; immunology ; metabolism ; Matrix Metalloproteinase 9 ; immunology ; metabolism ; Neoplasm Invasiveness ; Plasmids ; Transfection

OBJECTIVETo compare the clinical benefits of rapid Rhino Stammberger sinus dressing (RR), a new nasal packing material for functional endoscopic sinus surgery (ESS), with traditional packing materials sorbalgon plus Vaseline gauze (SV) prospectively.

METHODSTwenty-four patients with chronic sinusitis of the similar grade were enrolled in the study. After ESS, the nasal cavities of each patient were packed with RR in the right side as observing group and SV in the left side as control group. SV in the left nasal cavity was removed in the first day after operation and RR remained in the right nasal cavity until the first endoscopic follow-up 1 week postoperatively. The same perioperative treatments such as irrigation and local steroid were given in both sides of nasal cavities. The grade of rhinalgia, nasal obstruction, discharge, bleeding and epiphora in each side of nasal cavities were assessed on a visual analogue scale (VAS) by the patients at the operation day (packing period), the first day and the second day after operation respectively. Postoperative endoscopic assessment of nasal cavities including crust, discharge, pseudomembrane, edema of mucosa, bubble and ostium obstruction were carried out by a rhinologist on VAS at 1, 2, 3, 4, 6, 8 weeks postoperatively. The follow-up continued until the complete mucosal healing.

RESULTSThree patients were dropped from statistical analysis. One of them with severe hypertension bleeding when he awoke from anesthesia, the RR was run out from the nasal cavity. The other two were lost in the follow-up. Twenty-one patients completed the follow -up ranging between 2-20 months. The scores of rhinalgia, epiphora at the operation day and the rhinalgia at the first day after operation in RR side were lower than SV side (P<0.05). The scores of the amount of crust at the first week after operation in RR side were lower than SV side (P<0.05). Statistically, no significant deference was found between RR and SV in the length of mucosal healing period.

CONCLUSIONSPatients felt more comfortable with the package of RR than traditional material. No pain and secondary bleeding happened. The efficacy of RR on mucosal healing was similar with SV. RR showed some advantages and could be used as packing material following ESS.

Adult ; Aged ; Embolization, Therapeutic ; instrumentation ; methods ; Endoscopy ; Female ; Gels ; Humans ; Male ; Middle Aged ; Postoperative Hemorrhage ; therapy ; Prospective Studies ; Sinusitis ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the reliability and validity of the Chinese version of addiction severity index (ASI)-5th version (ASI-C-5), in illegal drug users receiving methadone maintenance treatment (MMT) in China.

METHODSOne hundred and eighty-six heroin addicts (144 men and 42 women) receiving MMT at three clinics in Guizhou province, southwest China, were recruited. They were all interviewed with a questionnaire of ASI-C-5 and 35 were re-interviewed at an interval of seven days to assess its test-retest reliability.

RESULTSCronbach's alpha for internal consistency of CSs varied from 0.60 to 0.81 in all domains. Test-retest reliability of composite scores (CSs) of ASI-C-5 were satisfactory (r=0.38-0.97). Based on item analysis and expert's suggestions, five items were deleted and one item was modified in ASI-C-5. Criterion validity of ASI-C-5 was found acceptable, as compared to addicts' self-rating anxiety scale (SAS) and self-rating depression scale (SDS) (r=0.59 and 0.45) except for social support rating scale (SSRS).

CONCLUSIONSASI-C-5 can be used for heroin addicts receiving MMT with acceptable reliability and validity.

Adult ; China ; Female ; Heroin Dependence ; drug therapy ; physiopathology ; Humans ; Male ; Methadone ; therapeutic use ; Reproducibility of Results ; Severity of Illness Index ; Surveys and Questionnaires

OBJECTIVETo investigate the role of Notch1 in the proliferation, migration and invasion of gastric cancer cells.

METHODSNotch1 expression was detected by RT-PCR and Western blot in four gastric cancer cell lines(SGC-7901, BGC-823, MKN-45 and MKN-28). The RNAi segments targeting Notch1 were designed and transfected into cell lines with the highest level of Notch1. The transcription and translation level of Notch1 expression was detected by real-time PCR and Western blot. The proliferation of transfected cells were determined by CCK8 test. Meanwhile, the migration and invasive ability of transfected tumor cells were detected by Transwell test.

RESULTSGastric cell line BGC-823 was selected with the highest expression of Notch1 at both mRNA level and protein level. The expression of Notch1 was significantly reduced after the transfection. The CCK8 test showed the growth rate was significantly reduced in RNAi group after two days(F=4.644, P=0.046). In migration test, the number of cells crossing through chambers in RNAi group(30.1±3.1) was significantly lower than that in the negative control group(44.5±3.2) and in the mock group(46.1±2.6)(P<0.05). In invasion test, the number of cells crossing through chambers in RNAi group(22.9±4.5) was also significantly lower than that in the negative control group (37.3±4.2) and in the mock group (37.8±3.7)(P<0.05).

CONCLUSIONDown-regulation of Notch1 inhibits the proliferation and reduces the ability of migration and invasion in gastric cell line BGC-823, suggesting that Notch1 may play an important role in the gastric carcinogenesis and metastasis.

Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Humans ; Neoplasm Invasiveness ; RNA Interference ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Receptor, Notch1 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; Transfection

Objective To understand the situation of client-retention to methadone maintenance treatment(MMT)program and related factors.Methods A Cohon study was adopted.In toml,1003 heroin addicts who were treated at 8 local MMT clinics with less than one month period.were recruited under nominal informed consent from Guizhou province.southwest part of China.during June to October 2006.Face-to-face interview and questionnaire administered to collect relevant information from the clients who were also followed nntil June 2007 to understand the situation on retention.Data were analyzed with Kaplan-Meier method to estimate the retention rate at different time spans while factors related to retention were analyzed with Cox proportional hazard regression model.Results All the clients were followed-up for 14 months,with an average retention of ten months.The retention rates of the clients were 68.8%and 57.4%at 6th-month and 12th-month of the treatment program.estimated by Kaplan-Meier method.Results of Cox regression analysis showed that factors influencing retention rate on MMT among the clients,including their awareness on MMT,dailv dose of methadone intake,and difierent MMT clinics which they were attached to.Risk influencing the withdrawal from MMT had a 20%decrease along with the increase when the daily dose of methadone intake reached 25 mg,with a hazard ratio of 0.80(P＜0.01).If the clients were aware that methadone was a life-time treatment when they began the MMT program.the risk for withdrawal would be lower than those who were not and the hazard ratio became 0.66(P＜0.05).Conclusion Our data showed that about half of the clients who were at the MMT program would still stick to it after 12-months.suggesting that the retention rate was not satisfactory and need to be improved.Factors as the MMT clinics themselves that the clients visited,(Laily methadone dosage they took,and the awareness on MMT etc.were important predictors to the rate of retention on MMT program.

Objective To investigate the source and genetic background of methicillin-resistant Staphylococcus aureus in the year of 2006,in China. Methods From January to December 2006,a total number of 302 consecutive and non-repetitive methicillin-resistant Staphylococcus aureus were collected from 17 Teaching hospitals in 15 areas. Genotypes of SCCmec were determined by multiplex PCR and multilocus sequence typing (MLST) was used to type the house-keeping genes. The implementation of the spa typing method was straightforward,and the results obtained were reproducible,unambiguous,and easily interpreted. Results All areas but Dalian harbored SCCmec Ⅲ while Dalian harbored SCCmec Ⅱ most. There were two strains in Guangzhou,harboring SCCmec Ⅳ. There were four strains of sequence type(ST),with ST239 accounted for 46.7% and ST5 accounted for 44.4%. ST59 accounted for 6.7% and ST88 accounted for 2.2%. There were fourteen strains of Spa typing,with t30 accounted for 52.6% ; t37 accounted for 27.2% ; t2 accounted for 12.9% ; t632 accounted for 2.3% ; t437 accounted for 1.3% ; t570,t601 accounted for 0.7% ; t377,t459,t796,t899,t1152,t2649 accounted for 0.3% ; no-typing accounted for 0.3%,respectively,pvl gene was not detected. Conclusion The main clone strains were ST239-MRSA-SCCmec Ⅲ-t30,ST5-MRSA-SCCmec Ⅱ-t2,with unique geographic distributions across the whole nation.

OBJECTIVETo evaluate HIV risk behaviors among heroin drug users who were treated in methadone maintenance treatment (MMT) clinics.

METHODSA prospective cohort study recruited and followed up clients of eight MMT clinics treated no more than two and half months in Guizhou province, China. Through face-to-face interviews, the baseline and following up informations were collected. And the baseline information included both demographic information and HIV risk behaviors, the following up informations included only HIV risk behaviors. The baseline investigation started in June, 2006 and the following up investigation finished in June, 2007. A total of 1003 heroin drug users were recruited at baseline, among them 666 (66.4%) were still at treatment by the end of follow up and 469 (70.4%) clients participated in the follow up interview. Wilcoxon two sample test and McNemar test were used to test for changes in HIV risk behaviors between baseline and following up investigation.

RESULTSAmong the 469 MMT clients who were followed up, the average days of reported heroin use decreased from 26.4 to 0.9 in the past 30 days (Z = 27.21, P < 0.05). Average days of alcohol use at baseline were 3.3 but 3.7 at follow up (Z = 0.45, P = 0.96). Needle-sharing behavior reported in the past 30 days decreased from 1.3% at baseline to 0.2% at follow up (χ(2) = 5.00, P = 0.025). At baseline, 5.5% (26/469) subjects reported having multiple sex partners in the past 30 days compared to 3.4% (16/469) at following up (χ(2) = 3.18, P = 0.08).6.4% (30/469) subjects reported casual sex with non-regular sex partners in the past 30 days at baseline compared to 5.1% (24/469) at following up (χ(2) = 0.95, P = 0.33). Of those who reported having casual sex relationship in the past 30 days 56.7% (17/30) reported using condoms at baseline but 58.3% (14/24) reported using condoms at follow up (χ(2) = 1.96, P = 0.16).

CONCLUSIONMMT was observed to decrease needle-sharing HIV risk behavior. However, decreased HIV sexual risk behaviors were not observed at statistical significant level.

Adult ; Female ; HIV Infections ; psychology ; Heroin Dependence ; drug therapy ; psychology ; Humans ; Male ; Methadone ; administration & dosage ; therapeutic use ; Opiate Substitution Treatment ; Prospective Studies ; Risk-Taking ; Sexual Behavior

OBJECTIVETo investigate the proportion of methadone maintenance treatment (MMT) clients continuing to use heroin and to explore its associated risk factors.

METHODSAll 1003 heroin addictive patients were recruited from eight MMT clinics in Guizhou province and followed up for 14 months. During the 14 months, each MMT client received random urine tests. MMT clinics, gender, marital status, employment, ethnicity, religion, previous drug using method, having a history being in detoxification program, age, methadone dosage, education level and length of drug use were treated as potential predicting variables into the GEE model.

RESULTSThe average age of the 1003 clients was (33.3 +/- 6.1) years old, the average length of drug use was (8.0 +/- 4.0) years, and the average daily methadone dosage was (38.0 +/- 16.6) mg. Among the 1003 clients, 26.0% were female drug users, 15.5% were divorced and 35.8% had a full time job. The rate of positive urine samples was approximately 30% for the first 10 months of follow-up, after which the positive rate decreased. The model found that different MMT clinics, the average daily methadone dosage (RR = 0.98, P = 0.003), treatment duration (RR = 0.95, P = 0.029) and years of education (RR = 0.94, P = 0.014) were associated with patients positive urine test.

CONCLUSIONContinued heroin use was common in MMT clinics in Guizhou province; increasing the dosage of methadone and the duration in the treatment program might decrease the continued use of heroin among clients in MMT clinics.

Adolescent ; Adult ; Ambulatory Care ; China ; epidemiology ; Female ; Heroin Dependence ; drug therapy ; epidemiology ; Humans ; Male ; Methadone ; therapeutic use ; Risk Factors ; Risk-Taking ; Young Adult

Objective To study the identification characteristics of rRNA genes on Yersinia (Y.)pestis.Methods By means of comparative genomics,we compared the rRNA genome sequences of nine completely sequenced strains of Y. pestis isolated from China and other countries by Clustal W software.we also compared the 2000 bp sequence adjacent to the rRNA genes,rRNA genes and 16S-23S rRNA spacer region respectively to determine the identification features of rRNA genes for Y. pestis.Results There were 6 rRNA gene clusters in the strains of D182038,D106004,Z176003 and CO92 respectively(6 copies strain).There were 7 rRNA gene clusters in the strains of 91001,KIM,Nepa1516,Antiqua and Pestoides F(7 copies strain).According to the 2000 bp sequence,13 types of rRNA gene clusters could classify the strains between the 6 copies and 7 copies.There were 4 types of tRNA gene among the 16S-23S rRNA spacer region that could classify the strains among the 6 copies and 7 copies strains respectively.The number of point mutation among the 23S rRNA gene was statistically different in some copies under ANOVA analysis(F=0.548,P=0.815>0.05 among the strains and F=5.228,P<0.01 among the copies).Conclusion The 2000 bp sequence adjacent to the rRNA genes,tRNA gene and 23S rRNA gene sequence could serve as the identification sign of rRNA genes for classifing the strains of Y. pestis.

Objective Measurement and analysis of the complete genome sequences of Yersinia Pestis from a new plague natural foci and adjacent foci in China, to know the genetic relationship among the epidemic strain isolated in Yulong (D 106004) and Jianchuan strains (D 182038) and the Tibetan strain ( Z 176003 ). Methods Three complete genome sequences were sequenced using the whole-genome shotgun and Solexa method and comparative genomics analysis was done among the three sequences. Genome comparative analysis among the coding sequences was done by BLAST software, SNPs finding was done by the program, genome rearrangements were analyzed using MAUVE software. Results All of the genomes of Yersinia pestis strains D182038, D106004 and Z176003 consist of a single circular chromosome and three virulence plasmids, pMT1, pCD1 and pPCP1. They had similar characteristics in chromosome and plasmid features, and there were no significant difference in coming sequence (CDS) of the cluster of orthologous groups of proteins (COG) functional classification and the number of insertion sequence in the three strains (x2 =3.03, 0.257, all P ＞ 0.05). The comparative genomics results showed that the three bacteria had 2882 genes with 100％ homology, of 3636 genes predicted in D106004, 2994 were identical with D182038's and 3113 with Z176003's, and of which 240 had 90％ homology with D182038's and 200 with Z176003 's. Synonymous single nucleotide polymorphisms(sSNPs) were 59 and 68, and non-synonymous SNPs(nsSNPs) were 104 and 203 between strains D106004 and Z176003/D182038. There were 11 segments rearrangements between D106004 and Z176003, which was less than 16 segments rearrangements between D106004 and D182038. ConclusionsThe three strains are highly homologous, the Yulong strain has more similarity with Tibet strain than with Jianchuan strain, the strain from Yulong foci may be evolved from Tibet foci.