Objective To express Japanese encephalitis virus (JEV) E protein in Escherichia coli. Methods The gene coding for JEV E protein was amplified by using RT-PCR, and cloned into plasmid pET-28a. The constructed plasmid was transformed into E.coli BL21 (DE3). Expression of E protein by the transformants was induced by IPTG and analyzed by SDS-PAGE and Western blot. Results Five nucleotide changes leading to amino substitutions were identify in the E protein gene of our laboratory strain of JEV when compared to a published gene sequence of JEV E protein. The yield of expressed JEV E protein with relative molecular mass approximately 52?10 3 was 35% of total bacterial proteins. Conclusions JEV E protein was expressed successfully in E.coli, which should be useful for the production of diagnostic reagents and the analysis of gene structure/function of the E protein.

In order to control the quality of all parts of the process for culturing,preparing and using clinically the human embryo olfactory ensheathing cells (OECs),the conduct standard of culturing human OECs from olfactory bulb is formulated. Because the strict conduct process can reduce the human factor as much as possible and ensure the stability of the cell quality. Therefore,during the culturing process of human embryo OECs, as for the embryo sample of OECs from olfactory bulb,the standard of lab condition and cell culture must be set strictly. The key steps of conduct process must be regulated,the freezing and resuscitating process of OECs must be controlled,the test method for microbial contamination during cell culture must be proposed and the risk must be reduced,then the quality of the human OECs from olfactory bulb can be guaranteed during the clinical application.

ObjectiveTo explore the feasibility and potential benefit of olfactory ensheathing cell (OEC) intraspinal transplantation in the treatment of intractable chronic neuropathic pain after spinal cord injury (SCI).Methods17 patients, 15 male and 2 female, with intractable chronic neuropathic pain after spinal cord injury was treated by OEC implant from November, 2004 to November, 2007. The age ranged from 18 to 68 (mean 40.4) years. The etiology of cord impairment included car accidents, falls, radiation damage, machine extrusion, gun-shot, and diving. The patients suffered severe persistent pain for 6 to 309 (mean 102.2) months, and the time points when cell therapy were administrated in the patients ranged from 6 to 312 (mean 105.9 months) after their injuries. Olfactory bulbs were harvested and trypsinized down to single fetal OECs. They were cultured for 12～14 days before implant. The fetal OECs were transplanted by injection into spinal cord at opposing ends of the injury site. The degree of pain was assessed and compared before operation and long-term follow-up according to the International Association of Neurorestoratology Spinal Cord Injury Functional Rating Scale (IANR-SCIFRS), i.e., 0 point means extreme pain, uncontrolled; 1 point, severe pain, narcotics required; 2 points, mild pain, ordinary pain killer effective; 3 points, no pain.ResultsThe follow-up and pain reevaluation were performed at 0.5 to 88 months with an average of 17.5 months after cell transplantation. The mean score of pain amelioration is 1.2 points.ConclusionThe OEC intraspinal transplantation appears to have a promising role in treatment of intractable chronic neuropathic pain after SCI.

Objective To analyze the effects of Hual qi huang granules on children with mycoplasma pneumoniae pneumonia.Methods A randomized,multicenter parallel controlled clinical trial was carried out.A total of 3 000 cases of hospitalized children with mycoplasma pneumoniae pneumonia were selected.All of them were given treatment for mycoplasma pneumoniae pneumonia with macrolide antibiotics and symptomatic treatment.They were randomly divided into 2 groups:research group and control group.The children of research group were give oral Huai qi huang granules for three months.According to the classification of pneumonia,these two groups were divided into:lobar pneumonia research group,lobar pneumonia control group,lobular pneumonia research group,lobular pneumonia control group.The hospitalization duration of fever,length of hospital stay,the absorption area of lung inflammation and pneumonia severity sores were observed.The frequency of upper respiratory infections,bronchitis,pneumonia were observed in 3 months after discharge.Results 2 378 cases were investigated.The hospitalization duration of fever,length of hospital stay of research group were significantly shorter than that of in control group (P ＜ 0.001).The children with lobar pneumonia,2 weeks after treatment,the absorption of consolidation of the lobar pneumonia research group is significantly better than lobar pneumonia control group (P ＜0.001).After two weeks treatment,the pneumonia scores of lobar pneumonia research group is lower than lobar pneumonia control group (P ＜ 0.05).Followup of 3 months after hospital discharge,frequency of upper respiratory infection and bronchitis of research group,were significantly lower than that of control.In addition,appetite increased significantly in research group than control (P ＜ 0.001).There are 21 cases with drug associated adverse reactions (mild diarrhea),including 12 cases of research group,9 cases of control group,and there was no statistical significance (P ＞0.05).Conclusion Standard treatment combined with oral Huai qi huang granules in the treatment of mycoplasma pneumoniae pneumonia,can significantly shorten hospitalization duration of fever,length of hospital stay and reduce the severity score of pneumonia.Three months oral Huai qi huang granules can significant reduce the frequency of respiratory infections and bronchitis,also can increase patients appetite,and be safe.