Objective: To modify the mono-antigen DNA vaccine of Mycobacterium tuberculosis with ubiqutin, so as to obtain more potent immune response. Methods: We constructed Mycobacterium tuberculosis MPT64 antigen DNA vaccine (pM) and ubiquitin-MPT64 fusion gene DNA vaccine (pUM). The constructed DNA vaccines were intramuscularly inoculated into female BALB/c mice separately. The serum antibodies(including IgG, IgG1,and IgG2a), cytokines (IFN-γ, IL-4) and cytotoxic T lymphocyte(CTL) response were determined in immunized mice. Results: The IgG level in the pM group was higher than that in the pUM group(P<0.01) and the ratio of IgG2a/IgG1 in the pM group was lower than that in the pUM group([2.16±0.3] vs [4.48±0.4],P<0.05). The IFN-γ level was higher (P<0.01) and the IL-4 level was lower (P<0.01) in the pUM group than those in the pM group. Furthermore, the pUM group had higher CTL activity than the pM group. It was indicated that the fusion gene DNA vaccine induced weaker humoral immune response but stronger cellular immune response compared to single gene DNA vaccine. Conclusion: The fusion gene DNA vaccine constructed in the present study might be more effective for prevention against tuberculosis than the single gene DNA vaccine.

Objective: To clone the Ag85 A antigen gene of Mycobacterium tuberculosis and express it in E. coli k802. Methods: The Ag85 A gene was amplified from the genome of Mycobacterium tuberculosis by PCR and was inserted into prokaryotic expressing vector pGEX5T to construct pGEX5T-Ag85A recombinant plasmid. The expression of Ag85A protein in E. coli k802 was induced by IPTG. The expressed protein was purified by affinity chromatography and the antigenicity of the expressed protein was confirmed by Western blotting assay. Results: A band about 0.9 kb in length was obtained by PCR and the recombinant plasmid pGEX5T-Ag85A was successfully constructed. A new band about 58 000 in length was observed after IPTG induction in E. coli. The relative molecular weight of the expressed protein was consistent with that expected. A single protein band of 58 000 in length was obtained after purification. The expressed protein could be specifically recognized by the sera of patients with tuberculosis patients. Conclusion: The Ag85A gene of Mycobacterium tuberculosis has been successfully cloned and expressed in E. coli k802, which paves a way for further studies on diagnosis and therapy of tuberculosis.

Objective To evaluate advantages and safety of bipolar electrodes for the treatment of submucosal myoma of uterus.Methods Three hundred patients with submucous myoma of uterus were performed with bipolar electrodes under hysteroscope,including 212 cases of type 0 submucous myoma,54 cases of type ?,and 34 cases of type Ⅱ.Results Total volumes of uterine distention fluid used during operation were 500-2000 ml(mean,626 ml);operation time was 20-50 min(mean,25 min);intraoperative blood loss was 10-20 ml.Cardio-cerebral syndrome occurred in one case and was relieved by timely treatment.There was no operation discontinuation as a consequence of pain.The diameters of resected myomas were 3-5 cm,with an average of 3.8 cm.The postoperative diagnosis showed leiomyosarcoma in 1 case and was performed by radical hysterectomy resection combined with a pelvic lymphadenectomy at 10 day postoperatively.During the follow-up period of 1-24 months in 277 cases,40 cases(type I in 12 cases;type II in 28 cases) needed hysteroscope operation again.Conclusions Hysteroscopic resection for submucosal myoma of uterus with dipolar electrodes has advantages of safety,minimal invasion,quicker recovery,higher efficiency,so it is worthy of being recommended.

Objective To explore the clinical effects of hysteroscopy combined with mifepristone on the diagnosis and treatment of abortive remnants.Methods A total of 56 patients with abortive remnants,who had undergone uterine curettage after the abortion,were treated with mifepristone combined with hysteroscopy.Before hysteroscopy,mifepristone was taken orally at a dose of 100 mg daily(50 mg,Bid) for 7 days.After the surgery,those who still had remnants in the uterus were given mifepristone at the same dose for 14 days.Results No patient had complication during and after the hysteroscopy except for one,who had abortive remnants at the uterine horn,experienced blood loss of 200 ml.In 40 patients,the abortive remnants were removed completely by hysteroscopy.Two weeks after the surgery,laboratory examination of the patients showed normal blood ?-hCG,and B-ultrasonography revealed no space-occupying mass in the uterus.In the other 16 patients,remnants were detected in the uterine horns after the hysteroscopy,and mifepristone was prescribed.These patients received reexaminations 2 months after the operation,and no abnormal blood ?-hCG or remnants in the uterus was found.Among the 56 patients,33 were followed up for 1-6 months(mean,3 months).During the follow-up,their symptoms including vaginal bleeding or drainage,amenorrhea,and abdominal pain disappeared,and the menstruation became normal.Conclusions Hysteroscopy combined with mifepristone is an effective measure to diagnose and treat abortive remnants.

We took such measures in internal medicine practice teaching in five-year program as actualizing special teachers’supervision system,adopting multiple teaching methods (enlightening,case-based,question-based,multimedia),taking interview of teachers and students,and enhancing doctor-patient communication.

OBJECTIVE To study infection and drug resistance of Mycoplasma from female genital tract.METHODS Mycoplasma in cervical secretion from inflammed female genital tract were identified by cultivation,and the sensitivities to antibiotics were also performed.RESULTS Mycoplasma were detected in 318 specimens among 588 patients with genital tract inflammation(54.1%),among which Ureaplasma urealyticum(Uu) accounted for 312(98.1%),and Mycoplasma hominis was 6(1.9%).The resistance to 12 kinds of antibiotics indicated that the sensitivities to 8 kinds of antibiotics were higher than 80% and the sensitivities to erythromycin,spiramycin and tetracycline were the lowest.CONCLUSIONS The infectious rates of Uu are on big rise in female genital tract with inflammation and the resistant strains increase continuously.

Objective To determine whether it is necessary to execute pre-labor pubic shaving.Methods Women(336 cases)who received pubic shaving through vaginal delivery from 2003 to 2006 were set as the control group.Women(103 cases)who were waiting for delivery currently in our hospital were set as the test group.The test group only received perineal cleaning without pubic shaving.If episiotomy was needed during the operation,the perineal hair Was cut by scissors.The perineum infectious rate and postpartal infection Was compared in the two groups.We interviewed the patients attitude for pubic shaving and counted the numing time used for pubic shaving.Results The perineal infection in the test group was 5.8%,which was no statistically different from that of the control group(2.4%),U=0.33,P＞0.05.Totally 95% women in the test group support this method demonstrated humanization.In the telephone folow-up visit for patients in the control group they comphined itch and discomfort during the regeneration of pubes.The average numing time for pubic shaving Was(4.6±0.6)minutes.Conclusions No pubic shaving for pre-labor skin preparation was feasible.If operation was disturbed by pubes we could use scissors to cut it short.This method could reduce the skin infection and propagation of blood-derived infectious disease.It increased the comfort of parturient,saved time and proved to be a nursing method of humanization.

Objective To explore the effect of MPT64 antigen from Mycobacterium tuberculosis on RAW264. 7 macrophages and the related mechanism. Methods MPT64 was purified after expression in E. crSi and verified by Western blotting analysis. The RAW264. 7 differentiation was induced by phorbol myristate acetate (PMA) and the resultant cells were divided into three groups according to different treatments: negative control, purified protein derivative of BCG(BCG-PPD) and BCG-PPD+MPT64 treatment groups. After 16 h incubation, flow cytometry was used to examine apoptosis of macrophages, and the levels of TNF-α and IL-10 in the supernatants were determined by ELISA. Results BCG-PPD treatment induced apoptosis of RAW264. 7 macrophages, and compared with BCG-PPD group, the apoptotic level of macrophages was significantly lower in BCG-PPD+MPT64 group (P<0. 05). We also found that the supernatant TNF-a level in the BCG-PPD group was significantly higher than that in negative group (P<0. 01) and the IL-10 levels were not significantly defferent between the two groups. Compared with BCG-PPD group, the IL-10 levll was significantly increased in BCG-PPD + MPT64 group (P<0. 01) and the TNF-a levels were not significantly different between the two groups. Conclusion MPT64 may act as a virulence factor and can inhibit the apoptosis of macrophages induced by BCG-PPD, which is probably through increasing IL-10 level.

Objective: To observe the clinical efficacy of heat-sensitive moxibustion for vascular dementia and explore its mechanism. Methods: A total of 70 patients with vascular dementia were randomized into an observation group and a control group, with 35 cases in each group. The control group was treated with piracetam, and the observation group was treated with heat-sensitive moxibustion on the basis of the treatment of the control group. The treatment lasted for 8 weeks. Before and after the treatment, the mini-mental state examination (MMSE), activity of daily living (ADL) and traditional Chinese medicine (TCM) symptom scores were assessed, and the levels of acetyl choline (Ach) and homocysteine (Hcy) were measured. The efficacy was evaluated after treatment. Results: The total effective rate of the observation group was significantly higher than that of the control group (P<0.05). After treatment, the MMSE and ADL scores in the observation group decreased significantly, and were lower than those in the control group (all P<0.05); the TCM symptom score of the observation group decreased significantly (P<0.05), while that of the control group had no significant change (P>0.05); the plasma Ach level in the observation group increased significantly (P<0.05), and the Hcy level decreased significantly (P<0.05), which were statistically different from those in the control group (both P<0.05). Conclusion: Heat-sensitive moxibustion plus piracetam is effective in treating vascular dementia. It can significantly improve dementia symptoms and ADL, which may be related to the correction of plasma Ach and Hcy levels.

OBJECTIVE:To observe the curative effect of Danhong injection in treating cerebral infarction.METHODS:30 patients(trial group) were assigned to receive Danhong injection,while 26(control group) to receive Ligustrazine injection.The neurologic impairment score,clinical efficacy and ADRs during medication were compared between the two groups.RESULTS:The total effective rates in trial group vs.in control group were 86.67% vs.65.38%,showing significant differences between the two groups(P