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Objective To investigate the value of using gonadotropin-releasing hormone agonist (GnRH-a) pretreatment in adenomyosis patients before adenomyomectomy. Methods From May 2012 to September 2015, 87 patients with adenomyosis who were non-effective to conservative therapy in Renmin Hospital of Wuhan University were enrolled in this study. According to the principle of randomized control, 41 patients were in the treatment group who were treated with GnRH-a 2-3 cycles before adenomyomectomy, while 46 patients in the control group. The control group paients were operated without any pretreatments. The blood loss, the number of penetrating into uterine cavity, duration of operation, duation of peritoneal drainage and the amount of drainage fluid, the difference of hemoglobin value before and after operation, total white blood cell count, duration of hospitalization, the maximum diameter of uterus and other indicators between the two groups were compared. Results In the treatment group, before and after treatment with GnRH-a, the uterus size, blood hemoglobinand CA125 value were statistically different (all P<0.05);between the treatment group of GnRH-a treated for 2 cycles and for 3 cycles, there were statistical differences of blood hemoglobin value [(108 ± 20) versus (118 ± 24) g/L], CA125 value [(26 ± 11) versus(19 ± 4) kU/L; all P<0.05]. There were statistical differences of blood loss in operation [(113 ± 32) versus (194 ± 42) ml], ratio of penetrating into uterine cavity [12%(5/41) versus 12%(8/46)], duration of operation[(79±23) versus (91±25) minutes], duration of peritoneal drainage after operation [(2.1±0.9) versus (3.0±1.2) days] and the amount of drainage fluid [(152±43) versus (232±32) ml], the difference of hemoglobin value before and after surgery [(-15.6±2.9) versus (-23.7±3.5) g/L], white blood cell count after 2-3 days of operation [(11.4±4.2)×109/L versus (13.5 ± 3.2) × 109/L], ratio of peri-operative blood transfusion [5% (2/41) versus 20% (9/46)] and duration of hospitalization [(11.2±1.9) versus (13.6±3.1) days] between the treatment group and the control group (all P<0.05). Conclusion The pretreatment of using GnRH-a before adenomyomectomy in adenomyosis patients has benefits for implementation of surgery and reducing peri-operative and postoperative complications.
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BACKGROUND:In vitro culture of sufficient vaginal epithelial cels and smooth muscle cels is the key for vaginal tissue engineering. However, the culture, purification and passage of vaginal epithelial celsin vitro are difficult. Primary culture and passage of vaginal epithelial cels from large animals such as canines has not been reported. OBJECTIVE:To establish a stable method of culturing canine vaginal epithelial cels and smooth muscle cels. METHODS: Vaginal epithelial cels were isolated from the vaginal specimens by enzymatic digestion with Dispase and trypsin separately, and cultured in keratinocyte serum-free medium. Vaginal smooth muscle tissue were minced and digested with colagenase type II; the colected smooth muscle cels were cultured in DMEM culture medium containing 10% fetal bovine serum. The cultured cels were passaged regularly. Cel morphology and proliferation characteristics were observed and cel phenotypes were confirmed by morphology and immunohistochemistry staining. RESULTS AND CONCLUSION: Primary vaginal epithelial cels began to adhere after 24-36 hours, grew logarithmicaly after 4-5 days, and reached 70% confluence after 7-8 days; the epithelial cels showed a typical cobblestone, with no fibroblasts. Cultured epithelial cels passaged every 4-5 days and subcultured to 6-7 generations continuously. Immunohistochemical staining confirmed a positive staining for anti-pancytokeratin (AEl/AE3). Primary cultured smooth muscle cels adhered and grew after 24 hours. The smooth muscle cels were spindle-shaped and proliferated logarithmicaly. After 4 days, primary cultured smooth muscle cels were confluent and showed a typical shape of “peaks and valeys”, and then the cels could be passaged every 3-4 days and passaged 7-8 generations. Immunohistochemistry staining showed α-actin staining was positive. These findings indicate that canine vaginal epithelial cels and smooth muscle cels could have a long-term stable culture and proliferation, to provide adequate seed cels for vaginal tissue engineering.
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Objective To evaluate the feasibility of bladder extracellular matrix (BACM) seeding with autologous cultured vaginal smooth muscle cells (VSMC) repaired rabbit vaginal defect.Methods This study included 24 female rabbits.BACM and vaginal acellular matrix (VACM) were obtained from 8 rabbits by decellularization process.The other 16 rabbits were randomly divided into experimental and control groups.Vaginal tissue biopsies ( ～ 1 cm2) were harvested from female New Zealand rabbits.VSMC were cultured and stained with a-smooth muscle actin antibodies. Cultured VSMC cells were seeded on BACM ( experimental group) or VACM ( control group) at a cell density of 1 × 107 cells/cm2.The cell-seeded matrixes were cultured for 5days.In experimental group of 8 rabbits,a 2 cm segment of vagina was resected and replaced with BACM seeding with VSMC.Then the regenerative segment was studied with histological technique by hematoxylin-eosin staining after 3,6 and 12 weeks postoperative aud vaginography was performed at 12 weeks postoperative.The 8 rabbits in control group underwent the exact same procedure as above but the vaginal defect was repaired with VACM seeding with VSMC,Results The prepared BACM and VACM were transparent,HE staining and scanning electron microscopy showed the two acellular matrix was both consisted of abundant network of fibers with the regular arrangement,without cellular debris.Primary culture of VSMC was successfully established and passaged,and was uniformly spindle-shaped in the confluent state and showed a characteristic hill and valley'formation.Immunohistochemical staining showed VSMC in culture stained positively with a-smooth muscle actin antibodies.VSMC began to adhere to the BACM 5 hour after implanting and the number of the adhered cells increased with time.Cells gradually expanded and showed the typical morphology of smooth muscle cells.Three weeks after implantation in vivo,the luminal surface of matrix was completely covered by vaginal epithelial tissue,a layer of smooth muscle cells was formed in the outer surface of the matrix.Multilayered vaginal epithelial and improved development of organized muscle bundles was observed after 6 weeks.The regenerative tissue was equivalent to the normal vaginal tissue at 12 weeks postoperatively.Vaginography demonstrated the maintenance of full patency and a wide vaginal caliber without fibrosis and graft rejection.There was no significant difference in all evaluated items between experimental and control groups.Conclusions BACM has the same regenerative process as VACM in the replacement of vaginal defect.Moreover,BACM has wider source and appears to be an suitable material for vaginal replacement.
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ObjectiveTo explore the changes of the frequency and cytokine production of CD19 + CD5 + B cells in Endometriosis before and after treatment.MethodsFlow cytometry and quantitative PCR were used to assess the frequency,cytokine expression of CD19 + CD5 + B cells in the peripheral blood,peritoneal fluids and endometrium biopsies of 35 patients with Endometriosis and 29 patients with myoma of uterus (control subjects).Changes in the CD19+ CD5+ B cell compartment in the peripheral blood were compared before and after treatment.ResultsAll 35 endometriosis patients had higher frequencies(all P ＜0.01 ) of CD19+ CD5 + B cells in the peripheral blood,peritoneal fluids and endometrium biopsies [ (13.1 ± 1.9)％,(12.1 ±2.0)％,(11.7 ± 1.7)％] than 29 control subjects[ (2.9 ±0.8)％,(2.6 ±0.9)％,(2.8 ±1.1 )％ ].CD19 + CD5+ B cells from endometriosis patients expressed higher IFN-γ[ (7.2 ± 1.0) × 103,(7.9±1.3) ×103,(7.4±1.1) ×103 copies] than control subjects [ (1.9±0.7)×103,(2.2±0.8)×103,(2.0 ±0.5) × 103 copies).The endometriosis patients also had higher IL-10 production of CD19 + CD5 + B cells [ (6.4 ±0.9) × 103,(6.8 ± 1.2) × 103,(6.1 ±0.8) × 103 copies] than control subjects [ ( 1.7 ±0.5) × 103,(2.1 ±0.9) × 103,( 1.6 ±0.4) × 103 copies].Compared with control subjects,the endometriosis patients had a clinical response to treatment demomtrated a significant decrease in CD19 + CD5 + B cells in the peripheral blood[ ( 13.1 ± 1.9)％ vs (7.3 ± 1.2)％ ],expressed lower IFN-γ [ (7.2 ± 1.0)×103 copies vs (3.3 ±0.6) × 103 copies],produced less IL-10 [(6.4 ±0.9) × 103 copies vs (3.2 ±0.7) × 103 copies].ConclusionsCD19+ CD5 + B cells might play an important role in the pathogenesis of endometriosis through higher IFN-γand IL-10 expression.
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Medical postgraduates are the future core force of clinical diagnosis and treatment,scientific research,education and sanitary administration.Therefore,to cultivate qualified professionals,the key of contemporary medical postgraduate education is to pay more attention to comprehensive quality training in order to make students adapt to the continuous development of society and medical model.The First Clinical College in Wuhan University has implemented the hospital administration internship in combination with social needs and explored the education model of contemporary medical students with comprehensive quality.
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Objective To investigate the expression and significance of the human telomeruse gene (hTERC)in the cytologic specimens of cervix for early cervical lesion screening.Methods Cytologic samples of cervix including 120 normal persons,86 CIN patients and 14 SCC patients were analyzed for aberrations of 3q26 using a commercially available two-color FISH probe.Results Positive expression rates of hTERC gene in normal,CIN Ⅰ ,CIN Ⅱ,CIN Ⅲ and SCC samples were 1.7%(2/120),2.6%(1/38),55.6%(10/118),87.5%(14/16),100.0%(14/14),100.0%(14/14),respectively.Compared with CIN Ⅰ/CIN Ⅱ group,hTERC gene positive expressions in SCC/CIN Ⅲ group were higher significantly(P=0.01),and them were also difference between CIN Ⅰ-CIN Ⅲ group and normal controls(X~2=113.52,P ＜0.01).With progression from CIN to carcinoma,polyploidization resulting in aneuploidy and high TERC gene copy numbers in SCC group ascended significantly.Among the total abnormal cells,the number of the cells percentage which hTERC amplification signals was more than 3:3 in CIN Ⅰ ,CIN Ⅱ,CIN Ⅲ and SCC groups were 6.12%,7.98%,28.07% and 33.97%,respectively.Condusion The hTERC gene is increasingly gained with progression of CIN and may appear to be another screening testmarker for early cervical cancer screening and accessory diagnosis.
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Objective To explore the changes of frequency, BAFF expression and sensitivity to apoptosis of CD19 + CD5 + B cells in endometriosis patients and investigate the relationship between CD19 +CD5 + B cells and endometriosis. Methods Flow cytometry and quantitative PCR were used to assess the frequency, BAFF expression and sensitivity to apoptosis of CD19+ CD5 + B cells in the peripheral blood,peritoneal fluids and endometrium biopsies of 39 patients with endometriosis and 31 patients with myoma of uterus ( control subjects). Results: All 39 endometriosis patients had higher frequencies ( all P ＜ 0. 01 ) of CD19 + CD5 + B cells in the peripheral blood, peritoneal fluids and endometrium biopsies than that in 31 control subjects. CD19 + CD5 + B cells from endometriosis patients expressed higher BAFF, and were more resistant to CD95L-induced apoptosis than the cells from control subjects ( P ＜ 0. 01 ). Conclusion CD19 + CD5 + B cells might play an important role in the pathogenesis of endometriosis through higher BAFF expression and more resistance to CD95L-induced apoptosis.
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Clinical data of 899 patients,who received colposcopic examination,were retrospectively analyzed.Both Reid's colposcopic index(RCI)score and biopsy were performed.The study showed that the value of RCI score was positively related to the severity of cervical lesions(F=2.043,P=0.03).The specificity of RCI for cervicitis,cervical intraepithelial neoplasia(CIN)Ⅰ were 98.8%,95.7% respectively;while sensitivity and negative predictive value for CIN Ⅱ,CIN Ⅲ were 77.7%,70.0%and 96.8%.97.4%,respectively.So RCI scoring can increase the diagnostic value of colposcope.
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Objective To explore the effects of quercetin and quercetin in combination with cisplatin on adhesion, migration and invasion of HeLa cells. Methods Adhesion, migration and invasion of HeLa cells treated with quercetin and quercetin in combination with cisplatin were measured by adhesion assay, wound healing assay, and transwell chamber method respectively. Results The results showed that quercetin and quercetin in combination with cisplatin could inhibit adhesion, migration and invasion of HeLa cells. The adhesive ratio, migration rate and the invasiveness were negatively proportional to concentration of quercetin and quercetin in combination with cisplatin. With increasing concentration of quercetin from 20 to 80 μmol/L, the adhesive ratio decreased from ( 82. 2±1. 5 ) % to ( 48. 4±1. 1 ) % ; the migration rote decreased from (7.26±0. 20) μm/h to (3.78±0. 64) μm/h; the invasiveness decreased from ( 124. 3± 1. 5) piece to (90. 7±2. 1 ) piece(P <0. 05). In quercetin and quereetin in combination with 10 μmol/L cisplatin treatment group, with quercetin concentration increasing from 20 to 80 μmol/L, the adhesive ratio decreased from (42. 6±1. 2) % to ( 27. 5±1. 7 ) %, the migration rate decreased from ( 2. 20±0. 33) μm/h to (0.72±0. 19) μm/h and the invasiveness decreased from (78.7±2.5) piece to (44.0±4.0) piece (P < 0. 05 ). Compared with the quercetin groups, querectin in combination with cisplatin groups had significantly higher inhibitory effects ( P < 0. 05 ). Conclusions Quercetin and quercetin in combination with cisplatin can inhibit adhesion and migration and invasion of HeLa cells. Quercetin can enhance the inhibitory effect of cisplatin on HeLa cell adhesion, migration and invasion.
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Objective To detect the in vitro effect of the traditional Chinese medicine on the tachyzoites of Toxoplasma gondii. Methods Supernatant (1