Reliability and Validity of the Chinese Version of the Lymphedema Quality of Life Questionnaire / 中国医学科学杂志(英文版)

Objective To study the reliability and validity of the Chinese version of the Lymphedema Quality of Life Questionnaire (LYMQOL) in lymphedema patients. Methods LYMQOL was translated into Chinese. The Chinese version of the LYMQOL was distributed with the official Wechat account "Lymphedema Channel" to lymphedema patients who were recruited from October 28

[Cloning and function identification of gene 'admA' and up-stream regulatory sequence related to antagonistic activity of Enterobacter cloacae B8].

To reveal the antagonistic mechanism of B8 strain to Xanthomonas oryzae pv. oryzae, transposon tagging method and chromosome walking were deployed to clone antagonistic related fragments around Tn5 insertion site in the mutant strain B8B. The function of up-stream regulatory sequence of gene 'admA' involved in the antagonistic activity was further identified by gene knocking out technique. An antagonistic related left fragment of Tn5 insertion site, 2 608 bp in length, was obtained by tagging with Kan resistance gene of Tn5. A 2 354 bp right fragment of Tn5 insertion site was amplified with 2 rounds of chromosome walking. The length of the B contig around the Tn5 insertion site was 4 611 bp, containing 7 open reading frames (ORFs). Bioinformatic analysis revealed that these ORFs corresponded to the partial coding regions of glyceraldehyde-3-phosphate dehydrogenase, two LysR family transcriptional regulators, hypothetical protein VSWAT3-20465 of Vibrionales and admA, admB, and partial sequence of admC gene of Pantoea agglomerans biosynthetic gene cluster, respectively. Tn5 was inserted in the up-stream of 200 bp or 894 bp of the sequence corresponding to anrP ORF or admA gene on B8B, respectively. The B-1 and B-2 mutants that lost antagonistic activity were selected by homeologuous recombination technology in association with knocking out plasmid pMB-BG. These results suggested that the transcription and expression of anrP gene might be disrupted as a result of the knocking out of up-stream regulatory sequence by Tn5 in B8B strain, further causing biosythesis regulation of the antagonistic related gene cluster. Thus, the antagonistic related genes in B8 strain is a gene family similar as andrimid biosynthetic gene cluster, and the upstream regulatory region appears to be critical for the antibiotics biosynthesis.

Tea polyphenols inhibit Pseudomonas aeruginosa through damage to the cell membrane.

This paper aims to delineate the inhibition mechanism of tea polyphenols (TP) toward Pseudomonas aeruginosa by cell membrane damage. Morphological changes in bacteria treated with TP were investigated by transmission electron microscopy, with results indicating that the primary inhibitory action of TP is to damage bacterial cell membranes. TP also increased the permeability of the outer and inner membranes of P. aeruginosa and disrupted the cell membrane with the release of small cellular molecules. A proteomics approach based on two-dimensional gel electrophoresis and MALDI-TOF/TOF MS analysis was used to study the differences in the membrane proteins of TP-treated P. aeruginosa and those of control samples. Twenty-seven differentially expressed proteins were observed in the treated and the control groups. Most of the proteins identified by MALDI-TOF/TOF MS were enzymes (dihdrollpoamide dehydrogenase 50s ribosomal protein, and so on), which may have induced the metabolic disorder of the bacteria and resulted in their death.

[Intervention trial on HIV/AIDS among men who have sex with men based on venues and peer network].

OBJECTIVE: To determine feasibility and effectiveness of the intervention on HIV/AIDS among MSM based on venues and peer network. METHODS: The intervention trial was conducted in Mianyang and Yibin in Sichuan province, where the cultural and social environment were similar. These two cities have no HIV/AIDS intervention conducted yet before this study. The intervention was conducted in Mianyang, while Yibin was regarded as control, in which education materials related HIV/AIDS and VCT service were available. Intervention in Mianyang included MSM venue intervention distributing the education materials, condom and promoting HIV-test and STIs clinic referral by 40 MSM as Popular Opinion Leaders who received the knowledge and intervention skill training.Meanwhile, Popular Opinion Leader intervention was implemented in MSM peer network to advocate safe sex. After 6-month intervention the survey was conducted to assess the effectiveness of intervention. RESULTS: The scores of knowledge related HIV/STDs and self-efficacy of condom use was 1.293 (95%CI: 0.657 - 1.292, P < 0.05) and 1.556 (95%CI: 0.656 - 2.456, P < 0.05) higher in post-intervention than in pre-intervention which was (12.42 +/- 0.232) and (10.25 +/- 0.327) respectively in Mianyang, while no significant changing in Yibin during the time. Score of knowledge related HIV/STDs increase 0.577 (95%CI: -0.173 - 1.327, P > 0.05) in post-intervention compared with (10.40 +/- 0.412) in pre-intervention and score of self-efficacy of condom use decreased 0.362 from 9.86 +/- 0.547 in pre-intervention (95%CI: -1.458 - 0.534, P > 0.05). In the six months prior to survey, the rate of unprotected sexual intercourse with male casual sexual partners in last 3 times decreased to 11.0% (22/200) (OR(adjusted) = 0.472, 95%CI: 0.265 - 0.841, P < 0.05) from 19.5% (39/200) baseline in Manyang, while in Yibin that increased to 19.0% (38/200) from 17.5% (35/200) in baseline (OR(adjusted) = 1.153, 95%CI: 0.660 - 2.014, P > 0.05). The rate of HIV-test increased significantly from 9.0% (18/200) to 22.0% (44/200) (OR = 2.852, 95%CI: 1.583 - 5.138, P < 0.05) in intervention city and Accordingly in the control, that was 24.5% (29/200) in baseline and 24.0% (28/200) in post-intervention (OR = 0.960, 95%CI: 0.548 - 1.682, P > 0.05). No difference was found in number of male sexual partner pre- and post-intervention both in intervention and control city. CONCLUSION: The intervention based on MSM venues and peer network among MSM is feasible and can increase knowledge related HIV/STDs and self-efficacy and as well as condom use and HIV testing.

[HIV risk behavior based on intervention among men who have sex with men peer groups in Anhui province].

OBJECTIVE: To determine the feasibility and the effectiveness of HIV risk behavior intervention characterized by initiator taking the lead combined with peer's participation, as to preventing HIV epidemic through promoting condom use and reducing the number of sexual partners among men who have sex with men (MSM) groups. METHODS: Subjects were recruited via peer referral chain. Twelve key MSM were recruited as initiators in bars or other MSM venues in 3 cities of Hefei, Wuhu and Fuyang. Then, each initiator recruited up to 3 MSM to participate and also each of them continued recruiting others. A total of 218 eligible MSM were recruited, and there were four intervention activities conducted. Firstly, twelve initiators were trained according to intervention manual and then intervention activities were implemented by initiators based on their referral chain. Participants were required to complete self-administrated questionnaire at baseline and the third month after intervention finished. The comparison of the results before and after intervention was conducted two months later to see any improvement in HIV/AIDS knowledge, and condom use. RESULTS: Of 218 participants, 170 (77.9%) were followed up in assessment three months later. The results from paired t-Test and Chi Square Test showed that intervention increased HIV/STDs related knowledge (baseline, 14.71 +/- 2.59; follow-up, 16.95 +/- 1.81; t = -10.647, P < 0.01) and the rate of having female sexual partner during previous 2 months (baseline, 17.6%; follow-up, 11.2%; P < 0.01) were of significant differences. Meanwhile, the intervention increased rate of condom use in the last three times of anal intercourse with homosexual partners, casual homosexual partners and primary homosexual partners (baseline, 55.3%, 43.2%, 49.1%; follow-up, 65.2%, 52.2%, 60.9%; chi(2) = 9.979, P < 0.01; chi(2) = 5.797, P < 0.05; chi(2) = 13.082, P < 0.01; respectively) and decreased rate of non-condom use in the last anal intercourse with homosexual partners, casual homosexual partners and primary homosexual partners (baseline, 41.2%, 35.3%, 45.3%; follow-up, 25.3%, 27.1%, 31.2%; P < 0.01, P < 0.05, P < 0.01; respectively) were all of some improvement. Other relevant indicators of 218 participants with 170 followed were compared, excepting the above similar findings, there were no differences in rate of number of female sexual partner during previous 2 months and rate of condom use in the last three anal intercourse with casual homosexual partners and rate of non-condom use in the last anal intercourse with casual homosexual partners. CONCLUSION: HIV risk behavior intervention based on MSM peer groups is feasible and might increase the condom use among MSM.

Identification of anrF gene, a homology of admM of andrimid biosynthetic gene cluster related to the antagonistic activity of Enterobacter cloacae B8.

AIM: To identify the gene (s) related to the antagonistic activity of Enterobacter cloacae B8 and to elucidate its antagonistic mechanism. METHODS: Transposon-mediated mutagenesis and tagging method and cassette PCR-based chromosomal walking method were adopted to isolate the mutant strain(s) of B8 that lost the antagonistic activity and to clone DNA fragments around Tn5 insertion site. Sequence compiling and open reading frame (ORF) finding were done with DNAStar program and homologous sequence and conserved domain searches were performed with BlastN or BlastP programs at www.ncbi.nlm.nih.gov. To verify the gene involved in the antagonistic activity, complementation of a full-length clone of the anrF gene to the mutant B8F strain was used. RESULTS: A 3 321 bp contig around the Tn5 insertion site was obtained and an ORF of 2 634 bp in length designated as anrF gene encoding for a 877 aa polyketide synthase-like protein was identified. It had a homology of 83% at the nucleotide level and 79% ID/87% SIM at the protein level, to the admM gene of Pantoea agglomerans andrimid biosynthetic gene cluster (AY192157). The Tn5 was inserted at 2 420 bp of the gene corresponding to the COG3319 (the thioesterase domain of type I polyketide synthase) coding region on B8F. The antagonistic activity against Xanthomonas oryzae pv. oryzae was resumed with complementation of the full-length anrF gene to the mutant B8F. CONCLUSION: The anrF gene obtained is related to the antagonistic activity of B8, and the antagonistic substances produced by B8 are andrimid and/or its analogs.

[Cloning and analysis of the complete genome of a Goose circovirus from Yongkang Zhejiang].

To investigate the mechanism of avian influenza outbreak wildly in water fowls, the co-pathogens, especially that caused immuno-depression were studied. A pair of degenerated primers, which amplified a fragment of 552bp in length, was designed and synthesized based on the published goose and other Circovirus sequences. The specific PCR product was amplified from the goose sample of Yongkang avian influenza case of Zhejiang Province. The fragment was then sequenced and the result showed the existence of the GoCV. The opposite part of the genome was further amplified using inverse primers designed based on the 552bp sequence obtained and the 1821bp full length genomic sequence of GoCV-yk01 was compiled using Seqman program of DNAStar. Sequence analysis showed that the GoCV-yk01 possessed common features of circovirus included potential replication associated stem-loop structure and the Rep protein motifs. Homology analysis showed that the sequence of GoCV-yk01 had 91% approximately 93% similarity to that of Taiwan and Germany strains. Phylogenetic analysis with ClustalW, however, showed that the GoCV-yk01 was on a different branch away from Taiwan and Germany strains. Circovirus usually causes immuno-depression and results in secondary infection, as it infects rapid growing cells as lymphocyte. It is speculated that the infection of GoCV may be a part of the reason of the avian influenza outbreak of the Yongkang case. The GoCV-yk01 is the first GoCV strain reported in mainland of China.

[Construction and verification of the effectiveness of pMBL: a cloning vector of exported proteins encoding genes].

The beta-lactamase was used as the reporter of expression and transmembrane secretion in this paper. A fragment of Amp resistance gene encoding the mature part of beta-lactamase (delta P delta SP Amp, i.e. without promoter and signal peptide coding sequences) was amplified from pUC18 vector. The upstream primer has BglII, BclI, BamHI in three reading frame respectively, in order to in frame fuse and express target genes together with the downstream reporter in finally constructed vector. Meanwhile, pET-28 was digested with the restriction enzymes BglII and Bst1107 I. The 2.8 kb fragment with replication origin, Kan resistance gene and MCS was recovered, filled, self-ligated and resulted in a plasmid pKan-B. The Bgl II site on pKan-B was then filled and the plasmid pKan was obtained. The delta P delta SP Amp gene, which was first cloned into pGEM-T-EASY vector, was inserted into pKan between EcoR I and XbaI sites. A plasmid pMBL-E was selected, with which the bacteria host could grew on Kan plate but not on plate with both Amp and Kan. An EcoRI site beside HindIII on the plasmid pMBL-E was then filled, and the plasmid pMBL, a cloning vector of the exported proteins encoding genes was finally obtained. Both results of the restriction enzyme digestion and sequencing demonstrated the correctness of the construction. The Tet resistance gene, a transmemebrane protein encoding gene, was applied to verify the effectiveness of the reporter in the vector. Cut with EcoRI and BamHI, a 375 bp fragment including promoter and 96 animo acids coding sequence (including signal peptide) of Tet was obtained from pBR322 vector. The fragment was then ligated to the vector pMBL which had been cut with both enzymes of EcoRI and BglI, or EcoRI and BclI, or EcoRI and BamHI (as 0, +1, +2 respectively of the beta-lactamase gene reading frame). Kan and Amp double resistant colonies only grew with the EcoRI and BglII combination (0 position). Restriction enzyme digestion and sequencing results of the recombinant plasmid showed that Tet resistance gene, which promoted the expression and transmembrane secretion of downstream beta-lactamase, was inserted in a correct reading frame into the vector. Thus, the results verified the effectiveness of the constructed vector pMBL, which may be used effectively to clone genes encoding exported proteins with promoters and signal peptide sequences.