Objective:To construct the Resuscitation-promoting factor A(RpfA) expression plasmid of Mycobacterium tuberculosis, and express and purify it in E.coli.Methods:RpfAgene of Mycobaeterium tuberculosis was amplified with PCR method from the H37Rv,cloned into pET32a(+) vector,and then transformed into E.coli Top10.After identifying by restriction digestion and DNA sequencing,the constructed recombinant plasmid was transformed into E.coli BL21,and expressed the recombinant protein under the control initiated by T7 after induction with IPTG.The target protein was identified by His-tag in-gel Stain and purified by using affinity chromatograghy.Results:The size of the RpfA gene digested by restricted enzymes accorded with the theoretical size and the DNA sequence with the reported one in literatures.66ku protein was found by SDS-PAGE and His-tag in-gel Stain.Conclusion:The recombinant expression plasmid pET32a(+)-RpfA was successfully cloned and constructed, and the recombinant protein with high purity was obteined;which laid the basis for further study.

Objective:To construct the Resuscitation-promoting factor B(RpfB)expression plasmid of Mycobacterium tuberculosis, and express and purify it in E.coli.Methods:RpfB gene of Mycobacterium tuberculosis was amplified with PCR method from the H37Rv,cloned into pET32a(+)vector,and then transformed into E.coli Top10.After identifying by restriction digestion and DNA sequencing,the constructed recombinant plasmid was transformed into E.coli BL21,and expressed the recombinant protein under the control initiated by T7 after induction with IPTG.The target protein was identified by Western-blot and purified by using affinity chromatograghy.Results:The size of the RpfB gene digestedby restricted enzymes accorded with the theoretical size and the DNA sequence with the reported one in literatures.61KDa protein was found by SDS-PAGE and Western-blot.Conclusion:The recombinant expression plasmid pET32a(+)-RpfBv of RpfB gene was successfully cloned and constructed,and the recombinant protein with high purity was obteined;which laid the basis for further study.

ObjectiveTo explore the causation of nosocomial infection in disabled children.MethodsThe causation of nosocomial infection in disabled children (n=1014) was analyzed retrospectively. The relationship between nosocomial infection and the factors, such as constituent ration in the infections parts, time in hospital, age and contact history separately were analyzed.ResultsOut of the 1014 patients, 291 cases were infected (the rate was 29%). The main site was in the respiratory system (276 cases, 94.8%). Among that, 73.2 % in upper respiratory tract and 21.6% in lower respiratory tract. The infection of digestive canal was only 7.6 %. The nosocomial infection rate in children under 1, 1～3, and above 3 years old were 44.6%, 37.8% and 27.5% respectively. The rate of those with contagious contact history was 31.6%, significantly higher than the ones without contact history (9.1%) (P≤0.05). There was no significant difference between infection rate and the time in hospital.ConclusionThe nosocomial infection is related to the age, contagious contact history, hospital environment and body nutritional situation, but not with time in hospital.

Objective To assess the microscopic features of lentigo by using in vivo confocal laser scanning microscopy (CLSM).Methods This study included 30 patients clinically diagnosed with solar lentigo and 10 patients with lentigo simplex.Lentigo lesions and perilesional normal skin were examined by in vivo CLSM.Tissue specimens were also obtained from lesional and perilesional normal skin and subjected to routine histopathologic examination.Results The CLSM features of lentigo were mainly observed at the dermo-epidermal junction.In all of these cases of lentigo,there was an increase in the length and number of rete ridges,with a marked hyperpigmentation of the basal layer.Several distinct microscopic patterns were observed,such as extended,irregularly shaped papillary dermis surrounded by highly refractile cells (presumed to be basal keratinocytes).Histopathologically,there was an infiltrate of a small quantity of melanophages and lymphocytes in superficial dermis.Conclusion As far as lentigo lesions are concerned,CLSM images are consistent with histopathological findings.

Reflux is a common post-gastrectomy complication due to the destruction of physiological antireflux mechanism,which affects patients' post-surgery life quality greatly.It's one of the basic requirements of digestive tract reconstruction to reduce the incidence of post-surgery reflux,and nowadays there are many kinds of anti-reflux surgical methods,which are also hot topics both at home and abroad.The non-surgical anti-reflux treatments are mainly focused on gastrointestinal motility promotion,mucosa protection,acid suppression,and so on.There are still some controversies concerned with which surgical or non-surgical way to choose.It's of great significance to have a deep insight into the associated mechanisms and make a wise decision on the surgical management with proper non-surgical assistance.

OBJECTIVE:To compare the efficacy and safety between caspofungin and fluconazole in the treatment of severe pulmonary fungal infection in intensive care unit. METHODS:36 patients with systemic fungal infections in intensive care unit were randomly assigned to receive caspofungin 50 mg?d-1 which was added into 250 mL normal saline and infused in no less than 1h for 14 days after initial dose of 70 mg(caspofungin group,n=18) or fluconazole 400 mg?d-1 for 14 days by iv gtt (fluconazole group,n=18). The curative efficacy and adverse events in the two groups were observed. RESULTS:The effective rate was 100.0% in caspofungin group versus 66.7% in fluconazole group. The incidence rates were 22.2% and 72.2%,respectively. There were significant differences in the overall effective rate between two groups(P

OBJECTIVE:To compare the efficacy and safety of linezolid vs.vancomycin in the treatment of MRSA-inducing ventilator-associated pneumonia in ICU.METHODS:In randomized controlled trials,54 patients with MRSA-inducing ventilator-associated pneumonia in ICU were randomly divided into 2 groups.Linezolid group (n=24) were administered with linezolid 600 mg every 12 h for 10 days,ivgtt.Vancomycin group (n=30) were given vancomycin 500 mg every 8h for 10 days ivgtt.Adverse events and the efficacy of two groups were investigated.RESULTS:Comparison results of linezolid group vs.vancomycin group were as follows:effective rates:100% vs.66.7%,MRSA eradication rates:87.5% vs.43.3%,adverse reactions rates:12.5% vs.13.3%.There were significant differences in the effective rates and MRSA eradication rates between two groups.CONCLUSION:The efficacy and safety of linezolid in the treatment of MRSA-inducing ventilator-associated pneumonia is superior to vancomycin.

In order to understand the vibration regularity and morphologic alteration of vocal folds objectively and directly without invading and disturbing the normal pronunciation, the voice acoustic multilateral analysis was conducted and electroglottograph was defecated for 59 cases of normal voice and 102 cases (136 times) of pathological voice by Dr. Speech software through computer' s multimedia technique. The results showed that except F0 there was no sexual difference in the parameters such as jitter, shimmer, NNE and SDF0; there was obvious difference in the parameters between normal and pathological voice. There was a falling tendency for the parameters between preoperation and postoperation. The Egg curve's variation can well reflect the morphologic alteration of vocal folds in voice diseases, and the abnormal ratio is 83.7 % and obvious lower in postoperation reexamined cases.

Objective To evaluate the clinical usefulness of manometry of esophgeal varices with endoscopic balloon technique. Methods The variceal pressure was measured by endoscopic balloon technique in 26 patients with portal hypertension, at the same time , laboratory parameters of liver function were determined. ResultsDetermination of variceal pressure was successful in 23/26 patients, while no side effects was observed. The intraobserver coefficient of variation was 7.18% ?2.64%,and there was a significant(P