Objective To construct the bicistronic retroviral vectors containing GFP gene and human survivin gene(wildtype and T34A mutant). Methods Survivin fragments were amplified from eukaryotic expression vectors by PCR, then cloned into pMIG retroviral vectors. The constructed recombinant was identified by double digestion with restriction enzymes. The constructed retroviral vectors was transfected into Phoenix E package cells under mediation of liposome. The expression of mRNA was examined by RT-PCR and the transcription of gene was determined by FACS analysis of GFP. Results The target fragments were successfully bound to pMIG vector. After the vectors were transfected into NIH3T3 cells, the expression of mRNA was confirmed by RT-PCR and the expression of GFP was confirmed by FACS analysis. Conclusion Human survivin gene wildtype and T34A mutant retroviral vectors have been constructed successfully, which can provide strong molecular tools for further studies and a novel approach for selective cancer gene therapy.

Objective To study the changes of serum high-sensitivity C-reactive protein (ha-CRP) and depression in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS),and investigate the relationship between them.Methods Thirty healthy volunteers (control group) and 54 OSAHS patients (OSAHS group) were recruited for the study.The level of serum hs-CRP was determined by scatter rate nephelometry,and the state of depression was assessed by self-rating depression scale (SDS).Results The level of serum hs-CRP and SDS score were higher in OSAHS group than those in control group [(48.8 ± 12.7) scores vs.(36.3 ± 6.3) scores,(3.3 ±0.7) mg/L vs.(1.4 ± 0.4) mg/L](P＜0.01).SDS score and the level of serum hs-CRPwere positively correlated to apnea-hypopnea index(AHI) (r = 0.636,0.628 ;P<0.01) and negatively related to the MSaO2 (r =-0.509,-0.614;P <0.01) and LSaO2 (r =-0.607,-0.512;P <0.01).The level of serum hs-CRP was positive correlation to SDS score (r = 0.536,P<0.01).SDS score was related to the AHI,the level of serum hs-CRP and LSaO2 in multiple linear regression(F= 33.31,P = 0.002).Conclusion Depression is correlated to AHI and the level of serum hs-CRP in patients with OSAHS.

Objective To evaluate the effect and safety of tiotropium plus sustained-release theophylline in treating stable moderate chronic obstructive pulmonary diseases (COPD) . Methods A randomized clinical trail was conducted in 80 patients with stable moderate COPD, who were admitted to our hospital from March 2008 to December 2009. All patients were divided into two groups randomly and accepted tiotropium (groupⅠ ,n =40) or tiotropium plus sustained-release theophylline (group Ⅱ ,n =40) treatment for 12 weeks. Results Seventy patients completed the study,in which 36 cases had tiotropium alone and 34 cases had combination therapy. At the end of treatment, no statistically significant difference was found between the two groups in the comparisons of FEV1, FEV1%, FVC, FEV1/FVC, activity score and impacts score of the St George's Respiratory Questionnaire (SGRQ) (Ps > 0. 05). While the symptoms score (53.08 ± 12.25 vs.59.39±9.74,t=2.39,P<0.05) and total score (23. 91 ±2. 57 vs. 25. 27 ±2.14, t =2.40, P <0.05) of SGRQ were significantly lower in group Ⅱ compared to group Ⅰ. Six adverse events,including dry mouth (n =4), palpitation (re = 1) , constipation (n = 1), were observed in tiotropium alone group during the treatment period. Eleven adverse events were observed in combination treatment group, including dry mouth (re = 3) ,stomach discomfort or nausea (n = 3), diarrhea (re = 2) , palpitation (n = 2), constipation (n = 1) . The incidences of adverse reactions of two groups had no significant difference (P > 0. 05). Conclusion Compared with the single application of tiotropium, tiotropium plus sustained-release theophylline therapy can alleviate symptoms and improve life quality in patients with stable moderate COPD,without significantly increasing adverse reactions.

Objective:In this study,a prokaryotic expression of the 3-ketosteroid-Delta (1)-dehydrogenase (KSDD) which came from Arthrobacter simplex was built.Moreover,in order to investigate the catalytic mechanism of KSDD and improve its stability,the structure of KSDD was predicted by computer and the critical sites were confirmed by site-directed mutations.Methods:The recombinant plasmid was constructed by eukaryotic expression vector pET-22b and the recombinant strain was constructed and expressed in Escherichia coli BL21 (DE3).High-performance liquid chromatography was used to determine the transformation rate of 4-AD to ADD.The KSDD structure and key sites were predicted by SWISS-MODEL.Site-directed mutations for the amino acid residues of key sites were constructed and activities of the mutations were detected.Results:The recombinant strain E.coli pET-22-ksdd was successfully constructed.It was induced to express the dehydrogenase by IPTG and the conversion rate of 4-AD to ADD was 27％ at 21 ℃.The structure of 3-ketosteroid-Delta (1)-dehydrogenase and the four key sites was analyzed by SWISS-MODEL.Four mutants,Y120R,Y320L,Y488F and G492Y were constructed.Mutants Y120R and Y488F were inactivated,so they were proved to be the key active sites of KSDD.The conversion rate of mutant Y320L was consistent with that of wild type,but the stability at 37 ℃ was improved.The conversion rate of mutant G492Y was 1.2 times of the wild type and the stability has been improved at 37 ℃.Conclusions:At present,there are few studies about the structure and catalytic mechanism of dehydrogenase.The active sites of the enzyme were verified by this study,which laid the foundation for the further study of the properties of the enzyme KSDD.

Objective To study the sex hormone levels and erectile dysfunction(ED)in patients with obstructive sleep apnea hypopnea syndrome(OSAHS)and to investigate the relationship between them.To assess the efficacy of continuous positive airway pressure(CPAP)in the treatment of ED with OSAHS.Methods The 46 OSAHS patients(OSAHS group)and 30 healthy voluntrers(control group)were recruited in the study.The states of ED were evaluated by 5-item version of the International Index of Erectile Function(IIEF-5)and the levels of serum sex hormone were tested by chemiluminescence immune assay in both group.Erectile function were reassessed and the levels of FSH,LH,T were retested in 22 ED with OSAHS patients after 3 months of CPAP treatment.Results Compared to the control group,the levels of serum FSH[(9.65?1.08)U/L vs(10.90?1.12)U/L],LH[(9.97?0.98)U/L vs(11.98?1.52)U/L]and T[(8.96?0.89)nmol/L vs(10.67?1.01)nmol/L]were significantly decreased in patients with OSAHS(all P

Obiective To study the characteristics and the correlation factors of ADR induced by Ginkgo Leaf Extract and Dipyridamole Injection on the purpose of providing reference for clinical rational administration.Methods All ADR cases that induced by Ginkgo Leaf Extract and Dipyridamole Injection reported between January 2006 and August 2009 were retrieved and statistically analyzed.Results Of all 24 ADR patients induced by Ginkgo Leaf Extract and Dipyridamole Injection,the female patients were more than the male,and the senile patients had highest incidence 41.67%.Most ADR events happened within 30min(70.83%).ADR mainly manifested as adverse reactions of skin and appendants(33.33%),followed by central nervous system adverse reaction.Conclusion Great importance should be attached to ADR induced by Ginkgo Leaf Extract and Dipyridamole Injection.Medication monitoring should be enhanced to avoid occurrence of ADR clinically.

Objective To study the polysomnography(PSG) indexes which can reflex the severity of anoxia in obstructive sleep apnea-hypopnea syndrome(OSAHS) more comprehensively. Methods Sixtythree OSAHS patients' PSG indexes, including the apnea-hypopnea index (AHI), apnea time/total sleeping time (AM), hypoventilation time/total sleeping time (HM), mean oxygen saturation (MSaO2),the lowest oxygen saturation (LSaO2) and the longest apnea time (LAT) were analyzed retrospectively. Results In patients with OSAHS,the AHI was positively correlated with AM, HM and LAT(P ＜ 0.01 or ＜ 0.05 ), and was negatively correlated with MSaO2 and LSaO2(P ＜ 0.01 ). The AHI had linear regression relationship with AM,HM and LSaO2 (F =202.264,P ＜0.01). Conclusions Among the PSG indexes,AM is intimately associated with AHI. Therefore,compared with LSaO2,AM and HM can reflex the severity of anoxia in patients with OSAHS more accurately.

Objective To investigate the feasibility of congenital cytomegalovirus (CMV)infection screening by saliva polymerase chain reaction.Methods From November 1,2010 to February 29,2012,6733 newborns born in Changzhou Maternal and Child Health Care Hospital were enrolled.Saliva samples (0.2 ml) were collected within 3 days after birth,CMV-DNA was detected by real time-polymerase chain reaction and hearing screening was done with EroScan transient-evoked otoacoustic emissions at the same time.The positive rate of congenital CMV infection screening was calculated and clinical manifestations were analyzed.Chi square test was applied to statistical analysis.Results Totally 6733 newborns were screened and 107 of them were found to be positive with CMV DNA,the positive rate was 1.59％ (107/6733),among which 88 were asymptomatic (82.2％) and 19 were symptomatic (17.8 ％).The major clinical manifestations of the neonates with positive CMVDNA were pathological jaundice (13 cases),hepatomegaly (5 cases),granulocytopenia,thrombocytopenic purpura,anemia and small for gestational age (two cases each).Fourteen newborns had only one major clinical manifestation,three newborns had two major clinical manifestations and two newborns had three major clinical manifestations.There was no statistical difference between newborns with positive and negative CMV DNA on hearing screening [hearing loss in one ear:8.4％ (9/107) vs 5.8％ (382/6626); hearing loss in two ears:3.7 ％ (4/107) vs 2.4 ％ (159/6626),x2 =2.776,P=0.241].Conclusion It is feasible to screen congenital CMV infection with saliva sample.

Objective To investigate the factors associated with vision and hole closure for idiopathic macular hole (IMH) after vitrectomy surgery.Methods Eighty-nine eyes of 89 patients with IMH were enrolled in this retrospective study.There were 15 males and 74 females.The patients aged from 42 to 82 years,with the mean age of (64.13 ± 7.20) years.All subjects underwent best corrected visual acuity (BCVA) and optical coherence tomography (OCT) examinations.The BCVA ranged from 0.01 to 0.4,with the mean BCVA of 0.12 ± 0.09.The MH stages was ranged from 2 to 4,with the mean stages of 3.56 ± 0.77.The basal diameter ranged from 182 μm to 1569 μm,with the mean basal diameter of (782.52± 339.17) μm.The treatment was conventional 25G pars plana vitrectomy combined with phacoemulsification and intraocular implantation.Fortyone eyes received internal limiting membrane peeling and 48 eyes received internal limiting membrane grafting.The follow-up ranged from 28 to 720 days,with the mean follow-up of (153.73 ± 160.95) days.The visual acuity and hole closure were evaluated on the last visit and the possible related factors were analyzed.Results On the last visit,the BCVA ranged from 0.02 to 0.8,with the mean BCVA of 0.26±0.18.Among 89 eyes,vision improved in 45 eyes (50.56％) and stabled in 44 eyes (49.44％).Eighty-six eyes (96.63％) gained MH closure but 3 eyes (3.37％) failed.By analysis,patients of early stages of MH and smaller basal diameter of MH will gain better vision outcome (t=2.092,2.569;P＜ 0.05) and patients of early stage MH will gain high hole closure rate after surgery for IMH (t=-5.413,P＜0.05).However,gender,age,duration,preoperative BCVA,surgery technique,gas types and follow-up time had no relationship with the effect after surgery for IMH (P＞0.05).Conclusions Stages of MH and basal diameter of MH may be the factors associated with the visual outcome for idiopathic macular hole after surgery.However,age,gender,duration,surgery patterns,gas types and followup time showed no effects on operational outcomes.

Objective:To discuss the characteristics of auditory brainstem response in preterm very low birth weight (VLBW) babies and to investigate the correlations between the ABR and clinical characteristics. Method: Fifty-nine VLBW babies (118 ears) were enrolled in the study and 30 term normal babies as the control group. Tympanometry, acoustic reflex, DPOAE, ABR were obtained in all the babies. Result:The prevalence of hearing loss in VLBW babies was higher than normal term babies and babies with perinatal complications higher than those without perinatal complications. There was no correlations between ABR threshold and gestational age, birth weight, postconceptional age, negative correlations between wave Ⅰ,Ⅲ and Ⅴ latencies Ⅰ-Ⅲ ,Ⅲ-Ⅴ and Ⅰ-Ⅴ intervals and postconceptional age. Wave Ⅰ and Ⅴ latencies, Ⅰ-Ⅲ and Ⅲ-Ⅴ intervals differed significantly between the two groups. Conclusion:The perinatal complications were the most important causes of the hearing loss in preterm VLBW babies than the gestational age and birth weight. There was a high prevalence of peripheral hearing loss in the preterm VLBW babies. Combining OAE and automated ABR should be applied for hearing screening. Regular follow-up was very important in all the preterm VLBW neonatal.