Objective: To clone the full-length cDNA encoding 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphatereductase (HDR) gene from Dendrobium officinale (DoHDR), then to analyze the expression difference in different tissues and expression patterns of DoHDR induced by signal molecule. Methods: RT-PCR and RACE technologies were used to clone the full length cDNA of DoHDR. The analyses of homologous comparison and phylogenetic tree were performed using DNAMAN and MEGA6.0 softwares, then the expression patterns of DoHDR were studied by real-time PCR. Results: The DoHDR gene was successfully obtained (GenBank accession number KC344827), and the full-length cDNA was 1 658 bp, coding the protein containing 460 amino acids. DoHDR had high homology (≥ 80%) with HDR proteins from other plants. Tissue expression analysis showed that DoHDR had the highest expression in the leaves, followed by roots, stems, and protocorm. Quantitative PCR results showed that DoHDR could be induced by signal molecule such as abscisic acid (ABA) and salicylic acid (SA). Conclusion: The cDNA encoding DoHDR is cloned. It is helpful for the future research on the mechanism of terpenoid biosynthesis in D. officinale.

The desired DNA product of KGPcd and KGP-hag was obtained from the total DNA of Porphyromonas gingivalis by PCR with two pairs of gene specific primers. The segment of KGPcd and KGP-hag (about 1.5kb and 1.6kb) was inserted into pGEM-T easy Vector. The double-stranded DNA of the postitive clone was analyzed by restriction endonuclease mapping and DNA sequenceing. The sequences of KGPcd and KGP-hag were consistent with those of the references appeared. The proteins of KGPcd and KGP-hag will be obtained for further study.

Female ; Humans ; Hysterectomy ; Laparoscopy ; Natural Orifice Endoscopic Surgery ; Singapore ; Vagina/surgery*

ObjectiveTo explore the effects of sacral neuromodulation using a new tined-lead electrode on neurogenic bladder.MethodsThe use of a new tined-lead electrode for sacral neuromodulation was evaluated in a study including 5 consecutive patients with neurogenic bladder.The tined leads were implanted at the S3 foramen under the X-ray screening.Subjects completed the recording of detailed voiding diary pre-and post-operation including fluid intake,voided volume,leaked volume,catheterized volume,frequency,accompanying symptoms and sensation.Vesicourethral function was assessed by video-urodynamics.ResultsUrinary frequency and voided volume were improved 22% and 49% respectively in one patient with spinal bifida.Urinary frequency,voided volume and residual volume were improved 0.7%,11% and 46% respectively in another one.Urinary frequency,voided volume and residual volume were improved 0.4%,18% and 44% respectively in the third one.Frequency of leakage and leaked volume were improved 36% and 54% respectively in the patient with brain trauma.Frequency of CIC and catheterized volume were improved 42% and 54% respectively,and indexes of urodynamics were improved 37%～45% in the patient with spinal cord injury.ConclusionA new tined-lead electrode for sacral neuromodulation provide a new alterative and minimally invasive procedure to treat neurogenic bladder.

Objective To discuss the impact of psychological behavior interventions on psychologic status and self-efficacy of perioporafive people with breast cancer. Methods 37 patients with breast cancer were randomly divided into conventional care group and psychological support group. General duty nursing and health education were adopted to conventional care group. Both psychological support interventions and self-regulation training were adopted to psychological support group. Both groups were evaluated by Hamilton Anxiety Scale (HAMA), Hamilton Depression Rating Scale(HAMD) and General Perceived Self-Efficacy Scale (GSES) after been hospitalized and operated. Results There was significant difference in the scores of conventional care group and psychological support group evaluated by HAMA and HAMD before and after intervention(P <0.01). The score of psychological support group evaluated by GSES is higher than conventional care group(P <0.05). Conclusions Psychological behavior interventions can ameliorate psychological anxiety and depression of porioporative people with breast cancer, and can improve patients' self-efficacy.

Objective To explore the association between polymorphisms of interferon-gamma gene intron 1 at position+874 (IFN-γ+874) gene and the susceptibility of HBV and/or HCV infection with different clinical outcomes. Methods IFN-γ+874 gene SNP were detected in 277 subjects including 79 chronic HBV/HCV coinfections,69 individuals only with HBV infection,55 individuals only with HCV infection and 74 controls,by sequence specific primers-PCR (SSP-PCR). Hepatocellular injury as suggested by alanine aminotransferase (ALT) was detected by Beckman LX-20. The status of viral particles in serum was determined by RT-nPCR. The possible association of the polymorphism of IFN-γ+874 with the susceptibility of HBV and/or HCV infection and the outcome of these infections were analyzed. Results (1) IFN-γ+874 AA frequency in individuals with chronic HBV,HCV,HBV/HCV coinfections were significant higher than that in controls (X~2=16.15,P=0.01); OR (95% CI) of IFNγ+874 AA in chronic infection with HBV,HCV,HBV/HCV coinfections appeared to be 2.70 (1.24-5.92),3.22 (1.43-7.25) and 4.02 (1.88-8.55) compared with + 874 TA. No significant differences were found among HBV,HCV,HBV/HCV coinfections (X~2=1.97,P=0.73). There were no significant association of IFN-γ +874 A/T allele frequency with HBV and/or with HCV infection (X~2=4.87,P=0.18). (2)The clinical outcomes of mild chronic hepatitis (CH),moderate/severe CH and cirrhosis with HBV and/or HCV infection were associated with IFN-γ+874 AA [X~2＝14.17,P＝0.03;OR＝3.09(1.51-6.33),3.85 (1.70-8.70),3.14 (1.08-9.17)]. No significant relationships were found between IFN-γ+874 A/T allele frequency and the clinical outcome of HBV/HCV infection (X~2＝6.07,P＝0.11). (3)There were no significant associations of IFN-γ+874 genotype/allele frequency with HCV duplication (X~2=2.36,P=0.31). (4) There were no significant associations of IFN-γ+874 genotype/allele frequency with abnormal ALT (X~2=0.15,P=0.93). Conclusion These results suggested that polymorphisms in the IFN-γ +874 had some influence on chronic HCV and/or HBV infection,and on the outcome of HCV and/or HBV infections. IFN-γ+874 AA genotype and T allele were possible risk to chronic HBV and/or HCV infections and to the outcomes of HBV and/or HCV infection. However,IFN-γ+874 TA genotype might serve as possible protective factors to them.

Objective To study the relationship between polymorphisms in interleukin-2gene at position-330 (IL-2-330) and the clinical outcome of hepatitis B virus (HBV) and/or hepatitis C virus (HCV) infection. Methods 277 subjects were recruited including 79 chronic HCV co-HBV infection, 55 chronic HCV infection, 69 chronic HBV infection and 74 controls. Single nucleotide polymorphisms of IL-2-330 was investigated by restricted fragment long polymorphism-PCR (RFLP-PCR). Hepatocellular injury, as revealed by alanine aminotransferase (ALT) was detected by Beckman LX-20 analyzer. The presence of hepatitis C viral particles in serum was determined by RT-nPCR. Results ( 1 ) IL-2-330 polymorphisms showed close association with persistent HBV and/or HCV infection. IL-2-330 TT was associated with an increased risk, but IL-2-330 GG with a reduced risk of persistent HBV and/or HCV infection (χ2=14.24, P=0.03 ) with ORs (95%CI) as 7.14(2.13-23.81 ), 3.46 (1.17-10.02) and 2.93 (1.15-7.46) respectively. However,IL-2-330 TT/GG did not significantly differ between patients with HBV and/or HCV infection (χ2=2.09, P=0.72). IL-2-330 T allele was associated with an increased risk, but the -330G allele was associated with a reduced risk of chronic HBV/HCV infection (χ2=12.33,P=0.01),with ORs (95% CI) as 2.26 (1.39-3.69) , 1.82 ( 1.09-3.03 ) and 1.73 ( 1.10-2.73 ) respectively. (2) IL-2-330polymorphisms showed significant association with the outcome of HBV and HCV infection ( χ2=13.52, P=0.04). IL-2-330 TT was associated with an increased risk, but-330 GG with a reduced risk of mild CH, moderate/severe CH, and cirrhosis. The ORs (95%CI) appeared to be 3.33(1.75-6.32), 3.31 (1.75-6.26), 11.23 (3.09-40.76) respectively. IL-2-330 T allele was associated with an increased risk, but the -330 G allele was associated with a reduced risk of mild CH, moderate/severe CH and cirrhosis (χ2= 12.32, P=0.01 ), with ORs as 1.86(1.32-2.63), 1.71 (1.27-2.31) and 2.77(1.57-4.89) respectively. (3) The polymorphisms of IL-2-330 showed no association with HCV RNA replication (χ2=0.83, P=0.66; χ2=0.20, P=0.66). The polymorphisms of IL-2-330 were not significantly associated with abnormal ALT ( χ2= 1.10, P=0.58; χ2=0.08, P=0.78). Conclusion These results suggested that IL-2-330 TT/T was associated with an increased risk, but IL-2-330GG/G was associated with reduced risk of persistent HBV and/or HCV infection, and with the development of mild CH,moderated/severe CH,and cirrhosis.

AIMNucleoside analogues have become the most promising candidates of anti-HBV drugs. In this study, beta-L-D4A was synthesized and explored its inhibitiory action against hepatitis B virus (HBV) in 2. 2. 15 cells derived from HepG2 cells transfected with HBV genome.

METHODSbeta-L-D4A was stereo-controlled synthesized from D-glutamic acid, and the structure was identified by IR, 1H NMR and MS. 2. 2. 15 Cells were placed at a density of 5 x 10(4) per well in 12-well tissue culture plates, and treated with various concentrations of beta-L-D4A for 6 days. At the end, medium was processed to obtain virions by a polyethlene glycol precipitation method. At the same time, intracellular DNA was also extracted and digested with Hind III. Both of the above DNA were subjected to Southern blot, hybridized with a 32P-labeled HBV probe and autoradiographed. The intensity of the autoradiographic bands was quantitated by densitometric scans of computer and EC50 was calculated. 2. 2. 15 cells were also seeded in 24-well tissue culture plates, and cytotoxicity with different concentrations was examined by MTT method. IC50 was calculated.

RESULTSThe synthesized compound structure conformed with beta-L-D4A; Autoradiographic bands showed similar for supernatant and intracellular HBV DNA. Episomal HBV DNA was inhibited in a dose-dependent manner. EC50 0.2 micromol x L(-1). The experiment of cytotoxicity gained IC50 200 micromol x L(-10.

CONCLUSIONbeta-L-D4A has been synthesized successfully. beta-L-D4A possessed potent inhibitory effect on replication of HBV in vitro with low cytotoxicity, TI value was 1 000. It is expected to be developed clinically into a new anti-HBV drug.

Antiviral Agents ; chemical synthesis ; chemistry ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Replication ; drug effects ; DNA, Viral ; drug effects ; Dideoxyadenosine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Genome, Viral ; Hepatitis B virus ; drug effects ; genetics ; physiology ; Humans ; Liver Neoplasms ; pathology ; Transfection ; Virus Replication ; drug effects

Objective To investigate the 16S rRNA methylase genes and Aminoglycoside modifying enzymes(AMEs)genes in Enterobacter cloacae isolated from the People's Liberation Army 98th Hospital,Huzhou district,Zhejiang province,China.Methods 40 strains of Enterobacter cloacae were isolated from the inpatients between September,2003 and November,2004.5 kinds of 16S rRNA methylase gene (including armA,rmtA,rmtB,rmtC and rmtD)and 9 kinds of AMEs gene[including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6')-Ⅰ b,aac(6')-Ⅱ,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ]were analyzed by PCR and verificated by DNA sequencing.Results In 40 strains of Enterobacter cloacae,the positive rates of genes of rmtB,aac(3)-Ⅱ,aac(6')-Ⅰ b,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ were 12.5%(5/40),27.5%(11/40),72.5%(29/40),32.5%(13/40),5.0%(2/40)and 5.0%(2/40),respectively.8 kinds of the rest of genes were all tested negative.The total positive rate of AMEs gene was 85.0%(34/40).Among 29 strains of Enterobacter cloacae that the aac(6')-Ⅰ b gene was positive,through PCR and verification by DNA sequencing,7 strains(24.1%)were confirmed to take the aac(6')-Ⅰ b-cr(the GenBank register number:EF375620,EU159121)alone,18 strains(62.1%)were confirmed to take the aac(6')-Ⅰ b-Suzhou(EU085533)alone,3 strains(10.3%)were confirmed to take both aac (6')-Ⅰ b-Suzhou and aac(6')-Ⅰ b-cr while only 1(3.4%)was aac(6')-Ⅰ b(the classical type).Conclusion There was lower positive rate of 16S rRNA methylase gene but very high AMEs genotypes in Enterobacter cloacae isolated from inpatients and the finding of rmtB gene was reported for the first time in the world.At least 5 kinds of AMEs gene existed in Enterobacter cloacae were isolated and they were the new host of both gene of aac(6')-Ⅰ b-cr and aac(6')-Ⅰ b-Suzhou,with aac(6')-Ⅰ b-Suzhou gene was the predominance subtype in aac(6')-Ⅰ b.

Objective To investigate the gene expression profiles of peri-implantation endometrium of high responders during controlled ovarian hyperstimulation. Methods High responders with cancelled embryo-transfer during controlled ovarian hyperstimulation (high responder group, n=4) and healthy fertile volunteers (control group, n=3) were performed endometrial biopsies during peri-implantation. Histologic changes of endometrium were observed by HE staining, genes of differential expression were screened with microarrays Affymetrix U133A 2.0 and identified by Real-time PCR. The biological process analysis was performed by online biological information analysis tool PANTHER. Results The ehdometrium was in mid-secretory phase in control group, while development delay was found in some glandular organs in endometrium of high responder group. Three hundred and sixty-four genes of differential expression were screened, among which 233 were up-regulated genes and 131 were down-regulated genes. OPN, PLA2G2, DPPIV, IGFBP5 and SSAT were identified as endometrial function-related genes, whose Real-time PCR findings were positively correlated to gene signal values detected by microarray(r=0.44, P<0.01). PANTHER analysis indicated that genes of differential expression participated in the biological processes of cytokine signal transduction and immunological regulation. Conclusion Ovarian high response affects the gene expression profiles of peri-implantation endometrium, which may be one of the causes of sub-optimal endometrial receptivity.