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.

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.

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.

Objective To investigate the dermoscopic and confocal microscopic features of Riehl's melanosis,as well as their association with histopathological findings.Methods Ten patients with a previously established diagnosis of Riehl's melanosis were recruited.The lesions of the patients were observed using dermoscopy and in vivo confocal laser scanning microscopy (CLSM),followed by histopathologic analysis.Results On dermoscopy and CLSM,all the lesions showed the following features:pseudonetwork,liquefaction degeneration of the basal cell layer,and incontinence of pigment.Conclusion Both dermoscopy and in vivo CLSM can serve as noninvasive auxiliary diagnostic tools for Riehl's melanosis.

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 investigate the effect of methylprednisolone (MP) combined with puerarin in preventing the spinal cord ischemia-reperfusion injury (SCII).Methods The model of ischemia-repeffusion injury was established by clipping the abdominal aorta of rats for 30 min.Eighty male SD rats were randomly and equally divided into 4 groups: the blank group in which the abdominal aorta was only exposed,the positive drug group in which intravenous MP injection (30 mg/kg) was conducted 30 min before the establishment of the ischemia-reperfusion injury model,the therapy group in which intravenous MP (10 mg/kg)and puerarin (30 mg/kg) injection was conducted 30 min before the establishment of the model,and the model group in which the physiological saline equivalent to the volume of MP in the positive drug group were injected into the caudal vein of rats 30 min before the establishment of the model.Ten rats were taken from each group and the homogenation of the spinal cord was used to test expression level of superoxide dismutase (SOD) and sodium-potassium adenosinetriphosphatase (Na-K ATPase).For another 10 rats in each group,the BBB scoring system was used to evaluate the neural function of the hind limb immediately after recovery from anesthesia,12 h and 24 h after reperfusion,and the spinal cord was obtained 24 h after reperfusion to observe the morphous of the spinal cord neurons.Results The expression levels of SOD and Na-K ATPase in the model group,positive drug group and therapy group 3 h after reperfusion were significantly lower than that in the blank group; the expression levels of SOD and Na-K ATPase in positive drug group and therapy group were significantly higher than that in the model group,and there was no significant difference between the positive drug group and the therapy group with regard to the levels of SOD and Na-K ATPase.The BBB score was better in the positive drug group and the therapy group than those in the blank group and model group.Microscope observation showed that the degree of injury between positive drug group and therapy group was equivalent,while each of them was lower than that in the model group.Conclusion MP combined with puerarin plays a great role in the prevention of the SCII.

Basement Membrane ; pathology ; ultrastructure ; Biopsy ; Glomerulonephritis, IGA ; pathology ; Humans ; Kidney ; pathology ; ultrastructure ; Kidney Glomerulus ; pathology ; ultrastructure ; Lupus Nephritis ; pathology ; Microscopy, Electron, Transmission ; Paraffin Embedding ; Specimen Handling ; methods

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