Objective To establish an ELISA method for detection of the concentration of TAP in human urine samples.Methods TAP-BSA was used as coating antigen. The TAP was a competitor to TAP-BSA. They reacted to the limited amount of monoclonal antibody against TAP.Results The optimal concentration of the coating antigen was 250ng/ml.The dilutions of monolclonal antibody against TAP and sheep anti-mouse IgG were 25?g/ml and 1∶4 000 respectively. The optimal range was from 0.69 to 1 000ng/ml.The assay provided a sensitivity of 0.69ng/ml.The coefficients of variation of intra-assay and inter-assay were 9.10% and 10.33% respectively. The average recovery rate of TAP was 97.70%.Conclusion The indirect competitive enzyme-linked immunosorbent assay is established.

Objective To evaluate the therapeutic effects of combining acupuncture and physical therapy with drugs on treating Duchenne muscular dystrophy (DMD).Methods Sixty infants with DMD were randomly divided into a treatment group and a control group (n =30 in each).Both groups were injected with Bozhi glycopeptide and given levocamitine and fructose sodium diphosphate orally.Moreover,the treatment group was additionally given acupuncture and physical therapy.Before and after 90 days of treatment,the time to walk 10 metres and to climb stairs of 2 metres high was measured,as were the serum creatine phosphate kinase (CPK),lactate dehydrogenase (LDH) and aspertate aminotransferase (AST) levels of all participants.Results After treatment,the average walking and stair climbing times of both groups were significantly shorter,and those of the treatment group were significantly shorter than those of the control group.Moreover,the average CPK,LDH and AST levels in the treatment group were significantly lower than before treatment and than those of the control group after treatment.There was a significant difference in the overall response rates,with 93％ observed in the treatment group and 73％ in the control group.Conclusion Combining acupuncture and physical therapy with drugs can significantly increase the mobility and improve the serum CPK,LDH and AST levels of children with DMD.

Objective To study the relationship between the expression of EBV lytic genes and systemic lupus erythematosus (SLE). Methods The blood samples from 44 SLE patients and 43 matched normal controls were tested for BamHⅠ-W, the specific DNA fragment of EBV, by polymerase chain reaction(PCR)-Southern analysis. RT-PCR and Southern blotting were used to detect the EBV lytic genes (including immediate early genes BZLF1 and BRLF1, early genes BARF1, late genes BcLF1 and BLLF1) in EBV DNA-positive SLE patients. Results The EBV DNA was positive in 32 SLE patients and 3 controls. There was significant difference between the SLE patients and controls in the EBV DNA expression ( ?2 = 39.18, P 0.05). In the EBV DNA-positive SLE patients, 2 (both with active SLE) were positive for BARF1 mRNA; 14 (11 with active SLE and 3 with inactive SLE) were positive for BcLF1 mRNA; none was positive for BZLF1,BRLF1 or BLLF1 mRNA. No lytic genes were expressed in any of the 3 EBV DNA-positive controls. Conclusion EBV infection may be related to the development of SLE. EBV lytic infection exists in some SLE patients. EBV is at a low level of lytic activity in patients with SLE.

Objective To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 anti-gen. Methods The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction en-zymes,and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both frag-ments were modified by Klenow fragment to form blunt end,then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1-Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction,a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band,which sug-gested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis,the recombi-nant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombi-nant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.

Objective To introduce and evaluate a new surgical technique about concealed penis.Methods The incision was chosen on the outer lamina of the bilateral sides of penis.The excision of fibrous cord of penis dartos and suprapubic fat pad and skin fixation on the penis root were done to the patients.For the severe patients,the superficial layer of the suspensory ligament of penis must be excised.Thirty-nine cases were treated with this new technique,22 cases with Devine technique,and 23 cases with modified Shiraki technique and clinical comparative research was carried out.Results Penis of 39 cases were found 2 to 3 centimeters pro-pubic projection.Within 3 to 36 months of follow-up,no obvious reduction of penis was found and the erection function was normal.By new evaluat standard of surgical efficacy,the clinical efficacy of new technique,Devine′s and modified Shiraki′s technique were 92.31%(36/39 cases),77.27%(17/22 cases) and 78.26%(18/23 cases),respectively and the efficacy rate in new technigue was significcmthy higher than that of Devines and modified Shirak′s groups(Pa

Objective To observe the treatment of ~(99)Te-MDP on typeⅡcollagen induced arthritis (CIA)in rat,and the effect on the expression of synovial MMP-3 and TIMP-1 mRNA.To explore the mech anisms of the ~(99)Te-MDP in the treatment of rheumatoid arthritis.Methods The rats in which C1A(n=24)were divided into three group:the control group(n=8),~(99)Tc-MDP group(n=8)and Methotrexate group(n=8). Arthritis were evaluated by arthritis index and histopathological index and the expressions of MMP-3 and TIMP-1 mRNA in synovium were detected by RT-PCR.Results①The arthritis indexs of the control group, the methotrexate group,the ~(99)Tc-MDP group were increased with time.②The histopathological scnres of the control group were significantly higher than those of methotrexate group and ~(99)Tc-MDP group(P＜0.01).The histopathological scores of cartilage destruction and bone erosion of ~(99)Tc-MDP group were lower than those of methotrexate group(P＜0.05).③The levels of MMP-3 mRNA of the control group,~(99)Tc-MDP group, methotrexate group were notably higher than those of the control group(P＜0.01).The levels of control group was notably higher than that of the ~(99)Tc-MDP group(P＜0.05).There was not significant difference in all groups on the levels of TIMP-1 mRNA(P＞0.05).Conclusions ~(99)Tc-MDP can notably relieve the arthritis symdrome and retard the catilage damage and bone erosion of CIA in rats,and could significantly decrease the MMP-3 mRNA in the synovium.Which may be one of the therapeutic mechanism.~(99)Tc-MDP is better than methotrexate in retarding catilage and bone erosion and decreasing MMP-3 mRNA in CIA rats in a 3-week therapeutic intervention.

Objective To examine the expression of toll-like receptor 2(TLR2)in peripheral blood monocytes and explore its association with disease stages and clinical manifestations and to explore the patho- genesis of rheumatoid arthritis(RA).Methods The expression of toll-like receptor 2 in peripheral blood monocytes from 47 RA patientis(27 in active stage and 20 in stable stage)and 18 normal individuals were de- tected by flowcytometry and RT-PCR.Results The expression of toll-like receptor 2 in peripheral blood monocytes in patients with active disease was significantly increased compared to non-active patients and nor- mal individuals,The expression was found to correlate with the Disease Active Score(DAS),serum C-reactive protein(CRP)level and the erythrocyte sedimentation rate(ESR),but not correlate with rheumatoid factor (RF)and the anti-cyclic citrullinated peptide(CCP)antibody.Conclusion The expression of toll-like recep- tor 2 in peripheral blood monocytes of patients with active RA is significantly increased.And the expression is correlated with disease activity index.The innate immune system is activated in patients with active disease. And the increased expression may promote the activities of monocytes.

Objective To determine the effects of~(99)Tc-MDP on joint inflammation and bone destruc- tion in collagen-induced arthritis(CIA)rats model and its effect on tumor necrosis factor alpha(TNF-?). Methods CIA was induced by immunization of male SD rats with an emulsion of collagen.~(99)Tc-MDP or placebo was intravenous infused to rats for 20 days.Joint inflammation was assessed by arthritis index.Lesions of bone were assessed based on the histological changes in ankle joints,radiographic analysis in hind paw with Larsen score.Systemic TNF-?level was measured by radioimmune assay.Results~(99)Tc-MDP suppressed joint swelling(P

Using uniform design to optimize some relative factors influencing the expressing of t-PA variant-rPA(K) in E.coli system, and found the best expressive conditions. They were(by using 300ml culture fask): the volume of media was 25ml, inducing time was 5.3 hours, pH was 6.0, the concentration of IPTG was 0.1mmol/L, inducing time was 25℃, and best culture media was HD. After being optimized, the yield of expression had been improved from 0.16 to 0.48, and it was as 3 times as before. The results above will offer the basement for purification and renaturation of rPA(K).

Objective To investigate the clinical effect of pulmonary rehabilitation therapy including respiratory exercise and vibration expectoration on patients with pulmonary infection after abdominal surgery.Methods A retrospective case control study was conducted.Seventy-six patients with pulmonary infection after abdominal surgery admitted to the First Affiliated Hospital of Hunan Normal University from September 2015 to September 2016 were enrolled.According to whether accept the pulmonary rehabilitation therapy or not,the patients were divided into two groups.In the control group (n =35),the convemional expectoration method was adopted.The patients in pulmonary rehabilitation group (n =41) received both methods of the control group and pulmonary rehabilitation treatment,including respiratory exercise (effective cough,lip reduction breathing),respiratory exercise device (respiratory exerciser tri-ball),and vibrated expectoration.The 24-hour sputum volume,degree of comfort,inflammatory and pulmonary function parameters,and recovery situation were recorded in the two groups.Results ① There were no significant differences in the parameters of inflammation and pulmonary function before treatment between the two groups.After treatment,the white blood cell (WBC) and C-reactive protein (CRP) in both groups were significantly decreased,and the forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) were significantly increased.The above changes in pulmonary rehabilitation group were more significant than those of the control group [WBC (× 109/L):11.12 ± 2.88 vs.13.42 ± 2.62 at 3 days,8.22 ± 1.48 vs.9.27 ± 1.92 at 5 days;CRP (mg/L):13.47 ± 4.77vs.16.03±4.94 at 3 days,9.69±1.56 vs.11.77±1.41 at 5 days;FEV1 (L):2.48±0.14 vs.2.29±0.16 at 3 days,FEV1/FVC:0.78±0.04 vs.0.75±0.04 at 3 days;all P ＜ 0.05].② The 24-hour sputum volume within 3 days of pulmonary rehabilitation group were significantly higher than that of the control group (mL:30.51 ± 4.15 vs.18.30 ± 3.64at 1 day,31.08±3.22 vs.20.37±3.20 at 2 days,29.03±2.55 vs.19.03±2.51 at 3 days,all P ＜ 0.01].③ In the pulmonary rehabilitation group,the recovery time of pulmonary infection symptoms (days:5.44 ± 1.45 vs.6.20 ± 1.55),the days of antibiotic use (days:12.61 ± 3.15 vs.15.03 ± 3.78),the time of getting out of the bed (days:4.05 ± 0.74vs.4.51±0.89),and the hospital days (days:19.95±3.90 vs.22.00±4.42) were significantly shorter than those of the control group (all P ＜ 0.05),and the degree of comfort was significantly better than that of the control group (comfort score:2.71 ±0.90 vs.2.14±0.91,P ＜ 0.01).Conclusion The application of pulmonary rehabilitation including respiratory exercise and vibration expectoration in abdominal surgery patients with pulmonary infection can promote recovery,and it has a good clinical and practical application value.