Objectiv To evaluate the usefulness of dot immuno-gold filtration assay(DIGFA) for the diagnosis of Paragonimus infection.Methods During 2003 to 2006,72 cases suspected of paragonimiasis in Zhejiang Province were examined with DIGFA for rapid detection of specific antibodies against Paragonimus(Pw-DIGFA).The diagnosis was primarily established with the presence of antibodies,experience of ingesting raw freshwater crabs or crayfishes and clinical presentations.The cases were treated with praziquantel and followed-up at 3 and/or 6 months post-treatment.Antibody level in patients(pre-and post-treatment) were detected in parallel and analyzed comparatively by Pw-DIGFA and ELISA.Results The result of detection by Pw-DIGFA was in agreement with that of ELISA.28 of 72 cases were antibody positive and 44 cases were negative.Among the 28 positives,26 cases had a history of eating raw freshwater crab or crayfishes and the other 2 cases drank freshwater from brook before.21 cases showed paragonimiasis-related clinical symptoms such as low-grade fever,cough,or changes in image examination,while the other 7 cases showed only eosinophilia in peripheral blood(15%-70%).The mean absorbance values(A450) of positive sera,negative sera and normal sera tested by ELISA were 1.7361,0.2973 and 0.2657 respectively.There was significant difference between the positive cases and the negative cases(t=12.047,P0.05).At 3 month post-treatment,serum antibody in 5 cases whose clinical symptoms and physical signs relieved or disappeared decreased 2-5 titers and that of one case who relapsed with new signs increased by one titer.In Pw-DIGFA,the dot color of 5 cured cases showed a little weaker than that of pre-treatment and the relapsed case displayed similar response.At 6 month post-treatment,7 sera of clinically cured cases showed significantly weaker response than that of pre-treatment.The antibodies of those sera dropped 3-6 titers.Conclusion Pw-DIGFA is of supplementary value for clinical diagnosis of paragonimiasis.Antibody detection by pre-and post-treatment using Pw-DIGFA shows potential for the evaluation of therapeutic effect.

Objective To investigate the different expression pattern of Src-homology2 domain phosphatase (SHP)-1 and SHP-2 in human papiilomavirus (HPV)6/11 infected condyloma acuminatum (CA) and the significance of the difference. Methods HPV6/11 related CA cases were diagnosed by in situ hybridization. The expression and distribution of SHP-1 and SHP-2 were examined by SP immunohistochemistry technique in skin samples from 40 HPV 6/11 positive CA cases and 20 healthy control (foreskins). Results The positive rates of SHP-1 and SHP-2 were 80% and 85% respectively in CA, which were significantly higher than those in healthy control cases (only 35% and 30%, respectively, X2=11.87,P<0.01; X2 =18. 15,P<0. 01) . The SHP-1 and/or SHP-2 positive cells in CA skin lesions were mainly distributed in prickle layer, showing as brown yellow, with the positive staining located in cytoplasm. Contrastively, the SHP-1 and/or SHP-2 positive cells in healthy controls were rare and mainly distributed in basal layer, showing as pale yellow with the positive staining located in cytoplasm. There was no significant correlation between the expression of SHP-1 and SHP-2 in CA( rs = 1.0, P>0.05 ). Conclusion The expressions of SHP-1 and SHP-2 increase in HPV6/11 positive CA, which suggest that with the infection of HPV6/11, SHP-1 and SHP-2 may play a regulatory role in the proliferation of keratinocytes.

To determine the diagnostic value as well as the evaluation value in therapy, the specific IgM and IgG anti-bodies of the infected and treated rabbits were detected using soluble egg antigen(SEA) and adult worm antigen(AWA) by ELISA. By using SEA to detect the IgM antibodies, the serum antibody level rapidly dropped 7 weeks after infection even without treatment, and those of IgM in sera detected by AWA rose early than IgG detected by SEA. After 5 month treatment, IgM detected by AWA and IgG detected by SEA were still positive. From these observations, it is evident that SEA is a better antigen to detect specific IgG in the diagnosis of schistosomiasis, AWA is a better one for early diagnosis. The use of AWA as antigen to detect IgM showed high diagnostic value both in acute and chronic schistosomiasis. However, detection of the IgM/ IgG in ELISA using SEA and AWA could not evaluate the therapeutic effect well.

0.05). Among 36 sera collected at 12 months post-treatment, the antibody negative conversion rates were 80.6% (29/36) and 58.3% (21/36), there was a significant difference between the two assays (P

Objective To analyse the effectiveness of the preoperative 3-dimensional conformal radiotherapy for large nephroblastoma up to 10 centimeters in diameter,and to investigate more effective preoperative therapies for intermediate and advanced nephroblastoma.Methods 32 cases of nephroblastoma were treated with preoperative radiotherapy with a dose fractionation as follows:150-200 cGy/fraction,5 fraction/week,the total dose of 1 000-2 000cGy,the mean dose of 1 600 cGy.During the radiotherapy,the B ultrasonic examination and CT were performed weekly to measure the variation of tumor volumes.The time of operation were determined based on the overall health status of patients,the shrinkage of tumor,and adhesions between tumor and adjacent vital organs.Radiontherapy was terminated one week before operation.4 patients who were found tumor adhered to normal tissues around kidney during operation were placed silver clip,and were given postoperative radiotherapy with additional dose of 1 000-2 000 cGy and the mean dose of 1 200 cGy.Results The median tumor reduction rate was 37 ％.The effective rate of preoperative radiotherapy was 100 ％.The complete resection rate was 87.5 ％.2-years tumor-free survival rates was 84.4 ％ and 5-years was 78.1％.There was no surgery-related death.Conclusion Preoperative 3-dimensional conformal radiotherapy reduces tumor volume,and raises resection and survival rate.

Objective To investigate the role of CD4 ~+ CD25~+ regulatory T lymphocytes (Treg)in modulating the cellular immune response and pathogenesis of murine pulmonary tuberculosis.Methods Inactivation of Treg was achieved by intraperitoneal injection anti-CD25 (clone PC61,50 μ/mouse) in PC61 group, and rat-IgG (50 μ/mouse) was injected intraperitoneally in control group. All the mice were inoculated intravenously with H37Rv 0. 1 mL (1 × 10~6 CFU) 3 days after Treg inactivation. The effects of Treg inactivation in different tissues were analyzed by flow cytometry. The cellular immune response, pulmonary histopathology and bacterial load were determined in vitro at different time points. The data were compared using homogeneity of variance F test and non-paired t test. Results In spleen, the percentages of Treg/CD4 T lymphocytes in PC61 group and control group were (21. 13± 3. 58)% and (30. 42± 4. 20)%, respectively at day 10 of inoculation (t = 2. 38, P < 0. 05), and those were (16. 12 ± 1. 26)% and ( 17. 34± 1. 62)%,respectively at day 30 of inoculation (t = 0. 84,P>0. 05). The percentages of Foxp3~+/CD4~+ T lymphocytes in PC61 group and control group were (32. 07 ± 3. 95)% and (60. 55 ± 5. 48)%,respectively at day 10 of inoculation (t = 5. 96, P<0. 05). Similar results were achieved in the peripheral blood. Bacillus calmette-guerin (BCG)-specific 1L-17 (ng/L) secreted by murine spleen cells in PC61 group and control group at day 10, 30 and 60 of inoculation were 5. 1± 0.9 vs 0, 43. 1± 10.0 vs5. 9± 2. 8 and 124.8 ± 5.8 vs 102. 5±8. 1, respectively (t = 7. 90, t=5. 10,t = 3. 19; all P<0.05); those of BCG-specific IFN-γ (ng/L) were 28. 4 ± 8. 2 vs 4. 0±1. 3, 685. 9± 128. 6 vs418. 7±20.4 and 310.9±119. 7 vs 32. 8±7. 5, respectively(tO = 4. 21,t = 8. 43, t = 3. 27; all P<0.05);those of TNF-a (ng/L) were 38. 6±5.0 vs 16. 3±4. 0, 112. 9 ±12. 3 vs 71. 5±12. 6 and 86. 2±8. 2vs0, respectively(t = 4. 95, t=3. 33,t/=14.8; all P<0. 05). The lung bacterial load at day 10 of inoculation in PC61 group was lower than that in control group (t = 4. 63, P < 0. 01), but the differences were not significant thereafter. The changes of lung histopathology at late stage of infection (day 120) in PC61 group were less severe than those in control group. Conclusions Murine Tregs increase dramatically after Mycobacterium tuberculosis infection. Treg could inhibit the specific cellular immunity against Mycobacterium tuberculosis, and therefore, may facilitate the persistent infection of Mycobacterium tuberculosis and development of tuberculosis.

BACKGROUND: An ideal marked molecule has not been found to detarmine bone marrow mesenchymal stem cells (MSCs) so as to make sure the homogenicity.OBJECTIVE: To verify the in vitro differentiation from BrdU-treced MSCs into osteoblasts.DESIGN, TIME AND SETTING: A cytological observation/n vitro was performed in Shanxi Medical University from January to October 2008.MATERIALS: Wistar rats aging 4 weeks old were provided by Experimental Animal Center of Shanxi Medical University.METHODS: MSCs were isolated and cultured by using density gradient cantrifugation combined with attachment culture method.At about 80% confluence, trypsin was used for passage and amplification. MSCs at density of 5×1010/L were inoculated in a 25-mm culture dish with L-DMDM culture medium containing dexamethasone, β -phosphoglycarol, vitamin C, and 10% fetal bovine serum. The third-passaged MSCs were labeled in vitro with 10 μmol/L BrdU tracer. Thereafter, 10 visual fields were randomly selected to calculate numbers of positive and negative ceils so as to obtain BrdU tracing rate under a fluorescence microscope (×200).MAIN OUTCOME MEASURES: Inverted microscope was used to observe cell morphology; flow cytometry was used to detect cell surface antigen, differentiation into osteoblasts, and BrdU tracing rate in vitro.RESULTS: The purified MSCs which were like fibroblasts were adherent and fusiform. The third-passaged cells were changed equidirectionally and whirlpool-arranged, and the survival rate was more than 95%. The seventh-passaged cells still grew rapidly.CD44, CD71, and CD105 expressions were positive, but CD45 expression was negative. Black particles were visualized in MSCs after Von kossa staining. BrdU tracing rate was more than 90%.CONCLUSION: Density gradient centrifugation combined with attachment culture method can effectively isolate and purify rat MSCs which are cultured in vitro for a long period and differentiated into osteoblasts. BrdU tracer is safe, effective, and convenient to successfully label MSCs.

Objective To investigate the prevalence and associated factors of fecal incontinence (FI) and urinary incontinence (UI) in postpartum Chinese women.Methods Questionnaires about FI and UI symptoms were completed via telephone interviews conducted within six months after delivery.Multi-variant Logistic analysis was applied for relation between delivery mode and FI or UI.Results (1) Two thousand and twelve postpartum women were admitted into this study,among which 14 (0.7％) had FI within 6 months after delivery.Logistic regression analysis showed that FI was significantly associated with forceps delivery (OR =20.09,95 ％ CI:3.64-110.90,P =0.000),and mediolateral episiotomy (OR=6.11,95％ CI:1.29-28.80,P=0.024).(2) Among the 2012 women,the prevalence of UI,stress urinary incontinence (SUI),urgent urinary incontinence (UUI)and mixed urinary incontinence (MUD was 10.04％ (n=202),8.15％ (n=164),0.94％ (n=19)and 0.94 ％ (n =19),respectively.Logistic regression analysis found that SUI prevalence was related to maternal age (OR =1.07,95％ CI:1.04-1.11,P =0.000),maternal weight before delivery (OR=1.04,95％ CI:1.02-1.06,P=0.001),neonatal head circumference (OR=1.20,95％ CI:1.05-1.39,P =0.010),mediolateral episiotomy (OR =4.96,95 ％ CI:3.05-8.07,P =0.0005 ),spontaneous vaginal delivery (OR=5.22,95％ CI:2.53-10.76,P=0.000) and forceps delivery (OR=9.20,95％ CI:4.07-20.79,P=0.000).UUI was related to maternal weight before delivery (OR=1.51,95％ CI:1.12-2.05,P=0.008).MUI was related to maternal weight before delivery (OR=1.06,95％ CI:1.00-1.11,P=0.049),duration of second stage of labor (OR=1.01,95％ CI:1.00-1.03,P =0.010),mediolateral episiotomy (OR =7.76,95％ CI:1.42-42.52,P=0.017) and forceps delivery (OR=15.21,95％ CI:1.61-143.44,P=0.018).(3) The prevalence of SUI was higher at 4 days and 42 days after delivery (7.95％ and 9.10％).Conclusions (1) F1 and UI prevalence is lower in this study than in other reports.(2) Vaginal delivery is a risk factor for women's FI and UI,especially forceps delivery and mediolateral episiotomy.(3) Maternal age,pre-delivery weight,newborn head circumference,spontaneous vaginal delivery,forceps delivery and mediolateral episiotomy might increase the risk of UI.

Objective In order to improve blood donors to understand the health education knowledge,this study designed and evaluated a new project,that is the health education pathway for primary apheresis donors.Methods A total of 2900 primary apheresis donors participated in the current study,who were randomly divided into the experimental group and the control group.The experimental group was performed the health education pathway for primary apheresis donors,while the control group was conducted in the traditional health educational ways.We compared the basic information,the awareness rate of apheresis donation knowledge,the number of regnlar/repeated donors,and the frequency of donations.Results Two groups were matched with no group differences in basic information (P＞0.05).After performed the health education pathway for primary apheresis donors,the awareness rate of apheresis donation knowledge was significantly improved from 23.6％ to 84.3％ (P＜0.01).Moreover,the percentage of regular donors (40.2％) in the experimental group higher than the percentage (26.7％) in the control group(P＜0.01).The average donation times of experimental group (3.8) was also higher than the control group.There were 79.2％ donors changed to regular/repeated donors higher than the percentage (66.4％) in the control group,and the average frequency of apheresis of those regular/repeated apheresis donors (7.4) in the experimental group higher than the control group (6.4) (P＜0.01).Conclusion As showed in our results,the health education pathway for primary apheresis donors could effectively help donors to understand the knowledge of blood donation and health care,and promote team construction of regular donors.We hope,in the future,the health education pathway for primary apheresis donors could be widely spread.

Objective:To generate sex hormone binding globulin(SHBG) conditional knockout mice model.In order to investigate the physiological function of SHBG in vivo and to provide experimental means for the study of the relationship between SHBG and gestational diabetes mellitus.Methods:The mouse genomic DNA sequence of SHBG was verified through bioinformatic analysis.According to the SHBG genomic DNA sequence,the gene targeting and knockout vector were constructed.Transfection of the vectors to ES cells by electroporation was performed according to common protocol.Positive ES cells were screened and identified by PCR.Therefore,the dual selected ES cells were microinjected into blastula,then blastula transplantations into the host mice.The chimeric mice were mated with C57BL/6J mice,and the Flox mice were obtained after screening.The Flox mice were hybridized with EIIA-Cre transgenic mice,and the progeny of the SHBG gene knockout (SHBG-/-) mice were obtained by autocopuation for several times.Results:Several Flox homozygous mice and SHBG gene knockout mice were successfully obtained.Compared with control mice,homozygous mice of SHBG gene knockout were well developed and had reproductive ability.The growth and development of SHBG knockout mice were not significantly different from that of wild type mice.Conclusion:Homozygous mice model of SHBG gene knockout was successfully established,which laid the foundation for further study of the role of SHBG in the gestational diabetes.The SHBG gene knockout mouse model was successfully established and the preliminary phenotypic analysis was performed,which laid the foundation for further study on the role of SHBG in gestational diabetes mellitus.SHBG gene knockout mice were normal in appearance.Due to the limited number of samples and many unknown biological characteristics of gene knockout mice,it needs further study.