Objective To investigate mRNA expression of matrix metalloproteinase (MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1) in ectopic endometriosis tissue and uterine endometrium from women with and without endometriosis. Methods Thirty-eight women with endometriosis (Revised American Fertility Society classification, RAFSⅠ-Ⅳ) were selected as study group. Thirty-eight specimens of ovarian endometrioma(ovarian chocolate cysts, OCC), 16 red peritoneal endometriotic lesions(RPL), and 35 matched eutopic endometrium(Eu) were collected from them simultaneously at the time of surgery. Twenty specimens of endometrium from reproductive women undergoing laparoscopic surgery without endometriosis were obtained as control group. The mRNA expressions of MMP-9 and TIMP-1 were detected by reverse transcription polymerase chain reaction (RT-PCR). Results Expression of TIMP-1 mRNA was detected in all samples. The level from endometriosis patients and control group was similar(2.31?1.21, 2.40?0.89). However, ectopic endometrium expressed significantly fewer TIMP-1 mRNA(OCC 1.67?0.79, RPL 1.45?0.68)compared with eutopic endometrium from both endometriosis and endometriosis-free patients(P

Objective To investigate the effect of vagina reconstruction using tissue-engineering biological material (acellular dermal matrix) in an animal model.Methods Vagina excision and vagina reconstruction with tissue-engineering biological material were performed in 12 Chinese experimental miniature pigs.The control group was matched with two of normal vagina specimens resected.At week 1,2,4,6,8,12 after surgery,the animals were sacrificed,respectively,and the neovaginas were prepared for immunohistochemical and Van Gieson (VG) staining to evaluate the status of various layer growth of vagina.Epithelial broad spectrum of monoclonal antibodies of AE1/AE3 and α-actin were used to test the existence of epithelial and smooth muscle tissue by immunohistochemical staining.The ultrastructure of neovagina was studied by transmission electron microscope at week 1 and 12 after surgery.Contractile function of isolated smooth muscle of neovagina was evaluated by chemical and electronic stimulation after 12 weeks' reconstruction.Results (1) Epithelization of 2/3 neovaginal mucosa was observed within 1 week.Only 1-2 layer epitheliums were observed under the light microscopy and epithelial cells with characteristics of loose and disarrangement were shown with the electron microscopy.Within 4-6 weeks,epithelization in mucosa of neovaginal canal was intensified to 4-5 layers.After 12 weeks,the differences between the neovagina and the native vagina were harldy noted either in the gross or microscopically.(2) After 4 weeks,a few smooth muscle cells were observed with VG and immunohistochemical staining,and homogeneous muscle bundle was formed.(3) After 12 weeks,similar contractile responses between neovagina and native vagina were observed when KCI and electrical stimulation with different frequency and voltage were given [(2.96±0.29)g vs.(3.14±0.30)g,(3.43±0.34)g vs.(4.65±0.73)g,(4.92±0.38)g vs.(4.89±0.44)g].Conclusion The tissue-engineering biological material might be an ideal graft used in the reconstruction of vagina.

Objective To investigate clinical features and management of complicated or rare vulvovaginal anomalies derived from urogenital sinus malformation.Methods Medical documents of 6 cases with vulvovaginal anomalies treated in Department of Obstetrics and Gynecology in Peking Union Medical College Hospital in the past 10 years were reviewed retrospectively.According to embryological and anatomical characteristics,the classification system were established.Results This kind of vulvovngihal anomalies was very rare and not enrolled into current diagnosis and treatment system. All patients had a common clinical features:menstruation bleeding and urine from the same outflow tract;and two orifices on perineum were found by bimannal examination,one orifice located under clitoris,the other orifice Was anus.No vagihal vestibule and vulvar were observed between those two orifices.According to clinical features,those 6 cases were divided into 3 types:type Ⅰ Was persistence of distant urogenital sinus with clinical manifestation of simple labial fusion;type Ⅱ was low communication between vagina and urogenital sinus (infrasphincterie),presenting with distal vaginai atresia and urethrovagihal fistula;type Ⅲ was high communication (suprasphincteric) with distal vagihal atresia and more proximal urethrovavihal fistula.Type Ⅲ was frequently associated with severe rectum and anus anomaly,and other malformation as well.All patients were treated by surgical correction.Basically.the principle of operation wag to reconstruct perineal appearance,release the obstruction,and rear the defect and ultimately recovering the normal function.The alternative surgical methods should be considered individually.Conclusions The vulvovnginal anomalies derived from urogenital sinus malformation were complicated clinically.To establish classification system will be benefit for clinicians to make diagnosis and choose appropriate treatment. The individualized surgery should be performed on those patients to restore the structure and function,in the mean time,the factors associated with esthetics.sex and psychics should be considered.

Objective To investigate the influence of gonadotropin releasing hormone agonist (GnRH-a) on the expression mRNA of nerve growth factor (NGF) and its receptors (TrkA and P75NTR) in normal and eutopic endometrial stromal cells (ESC).Methods From January to April 2009,3 patients with endometriosis undergoing surgery in Peking Union Medical College Hospital were obtained eutopic endometrium as study group matched with eutopic endometrium from 3 parents with teratoma as control group.ESC were incubated with different concentration of GnRH-a (0,5 × 10-11,5 × 10-10,5 × 10-9,5 ×10-8,5 × 10-7 g/ml).The expression of mRNA of NGF,TrkA and P75NTR were measured by real-time-PCR.Results At concentration of 0 g/ml,the levels of NGF,TrkA and P75NTR mRNA in ESC were 6.32,8.55,8.08 in study group,which were significantly higher than 0.94,0.67,1.08 in control group (P ＜0.05).Treated by the following concentration of GnRH-a (5 × 10-11,5 × 10-10,5 × 10-9,5 × 10-8,5 ×10-7 g/ml),the median expression of NGF,TrkA and P75NTR mRNA was 1.00,0.96,1.05; 1.09,0.82,1.27 ; 1.04,0.52,0.81 ; 1.00,0.55,0.64; 0.78,0.49,1.02 in study group.Compared with the expressions of those untreated by GnRH-a in study group, they showed significantly lower trends (P ＜0.05).In control group,the median expression of NGF,TrkA and P75NTR mRNA was 0.98,0.37,0.92; 0.70,0.45,1.15; 1.55,0.80,1.35; 1.09,0.41,1.35; 0.90,0.82,1.18.Compared with the expressions of those untreated by GnRH-a in control group,there were no statistically differences ( P ＞0.05).And treated by the same concentration of GnRH-a,the expressions of NGF,TrkA and P75NTR mRNA did not show statistically difference between the two groups ( P ＞ 0.05 ).Conclusion The expression of NGF.TrkA and P75NTR mRNA were suppressed by GnRH-a.

Objective To investigate the efficiency of biological function of AT rich interaction domain 1A (ARID1A) gene silenced by small interfering RNA (siRNA) on ovarian clear cell carcinoma ES2 cell line. Methods (1) The three pairs ARID1A gene siRNA interference fragments siN1 (ARID1A-705), siN2 (ARID1A-1513), siN3 (ARID1A-2282) and one pair negative control were respectively designed, and transfected into ES2 cells by RNA interference max reagent transiently. Reverse transcription (RT)-PCR and western blot methods were used to detect the expression of ARID1A mRNA and protein in ES2 cells transfected with interference fragments respectively. So as to select the best silencing effect of siRNA interference fragment(that was siN3),and then was used in the following experiment. (2) The following experiment were divided into three groups, namely siN3 transfection group, negative control group and blank control group. The proliferative activity of three groups of cells after transient transfection ( 6, 24, 48, 72, 96 hours) was assessed by cell counting kit-8 (CCK-8) assay and expressed as absorbance (A) value; the apoptosis rate of three groups of cells transfected transiently with interference fragment was measured by flow cytometry with annexin V/propidium iodide (PI) staining;the ability of cellular invasion of three groups of cells transfected transiently with interference fragment was tested by transwell experiment;the expression of nuclear factor-kappa B (NF-κB), membrane type-1 matrix metalloproteinase (MT1-MMP) and matrix metalloproteinase-2 (MMP2) protein in ES2 cells transfected transiently with interference fragment was measured by western blot. Results (1) The RT-PCR results showed that the ARID1A mRNA relative expression levels in ES2 cells after transfected transiently with siN1, siN2 and siN3 were 0.007 8±0.005 7, 0.006 8±0.000 3 and 0.002 8±0.000 3 respectively. They were all apparently lower than that in the negative control group (0.034 6 ± 0.001 3;all P<0.01). The western blot results showed that the expression levels of ARID1A protein were 0.439 4±0.000 7, 0.424 4±0.005 0 and 0.386 0±0.005 8 respectively. They were also lower than that in the negative control group (0.732 4 ± 0.030 3; all P<0.01). The siN3 with the highest transfection efficiency was selected to use in the following experiment. (2) The CCK-8 method showed that the proliferative activity of siN3 transfection group cells after transfected transiently at 6 hours was not statistically significant difference compared with those in negative control group and blank control group (0.506 ± 0.010, 0.491 ± 0.006, 0.498 ± 0.009, respectively; all P>0.05). However, the proliferative activity of siN3 transfection group cells after transfected transiently at 24, 48, 72, 96 hours were higher than those in negative control group and blank control group (all P<0.01). The flow cytometry results showed that the apoptosis rate of siN3 transfection group cells was (20.0±3.9)%, which was significantly lower than those in negative control group and blank control group [(31.5 ± 5.0)%, (34.0 ± 4.2)%, respectively;all P<0.05]. The transwell experiment showed that the penetrated cell counts of siN3 transfection group was 60.4±2.9, which was apparently higher than those in negative control group and blank control group (54.2 ± 3.5, 52.1 ± 3.8, respectively; all P<0.01). Western blot experiment showed that the relative expression levels of NF-κB, MT1-MMP and MMP2 protein in siN3 transfection group were respectively 1.85 ± 0.16, 0.37 ± 0.08, 1.38 ± 0.11, which were apparently higher than those in negative control group (0.93±0.11, 0.17±0.05, 0.86±0.06;all P<0.05) and blank control group (0.94 ± 0.04, 0.15 ± 0.08, 0.85 ± 0.10, respectively; all P<0.01). Conclusions It would be to promote the cell doubling time, reduce cell apoptosis and increase the invasive capability in ES2 cells that ARID1A expression was down-regulating by ARID1A mRNA interference. The invasion mechanism may be related to the activation of NF-κB signal transduction pathway, up-regulation of MT1-MMP expression and then promoting the invasion of tumor cells via the up-regulation of MMP2 expression.

Objective To study the relativity analysis of abnormal cervical pathology results in cytology and histology.Methods With retrospective analysis of 31,634 cases of fluid-based thin-layer method(ThinPrep Cytology Test,TCT)of PUMC Hospital from January,2001 to March,2003,which reported in the Bethesda System,we checked the abnormal results and advised different diagnose biopsy of vaginoscopy and/or conization,and got the relativity description of abnormal results in TCT and CINⅢ/CIS results validated by vaginoscopy,match analysis of the CINⅡ～Ⅲ and CINⅢ/CIS results validated by vaginoscopy and conization labeled by the age group,and relativity analysis of abnormal results in TCT and CINⅢ/CIS results validated by vaginoscopy.Results Among 31,634 cases of TCT test,948 cases had confirmed biopsy results validated by vaginoscopy,of which 70 cases were of CINⅡ～Ⅲ,56 cases were of CINⅢ/CIS.The risk ratio(RR)of different abnormal TCT results in predicting CINⅢ/CIS results validated by vaginoscopy is:ASCUS group,14.7(95% confidence interval 8.0～27.0,P=0.00);CINⅠ group,13.9(6.3～30.9,P=0.00);CINⅡ group,44.2(15.5～126.5,P=0.00);CINⅢ group,272.2(161.6～458.6,P=0.00);Cancer group,unmeasured.As noted,there is no significant difference between the RR of ASCUS group and CINⅠ group(P=0.951)in predicting CINⅢ/CIS results.Conclusions Vaginoscopy examination and biopsy could verify histology abnormity of CINⅡ～Ⅲ及CINⅢ/CIS from abnormal results of TCT,and has a good accordance along with biopsy results of conization.There are significantly greater risk of being CINⅢ/CIS validated by vaginoscopy in the abnormal TCT patients,among which ASCUS group and CINⅠ group have the coequal risk.

Objective To investigate characteristics of endometriotic lesions clone with various pathological subtypes by analyzing the polymorphism of X chromosome linked human androgen receptor allele (HUMARA). Methods Eight frozen tissues of endometriotic leisons were collected from 6 patients who received laparotomy or laparoscopy surgery in Peking Union Medical College Hospital from Nov. 2008 to Jun. 2009. Fifty specimens of epithelial cells from single endometrial glands were isolated and collected from endometriotic lesions by using laser capture microdissection. HUMARA was applied as the gene marker of clonal analysis. Nested palymerase chain reaction, double-enzyme digestion reaction with two methylationsensitive restriction endonuclease (Hha Ⅰ and Hpa Ⅱ), and the automated gene sequencing technique were utilized in this study to evaluate the characteristics of endometriotic lesions clone. Results Of 50 specimens of isolated glands, 34 were informative for clonal analysis, and all of which showed monoclonality. Of 3 ovarian endometriotic tissues, one tissue of HUMARA showed unuseful information, the other 2 ovarian endometriotic tissues respectively had 4 and 7 informative specimens of gland epithelial cells, and all of the glands from each tissue showed uniform clonal pattern. Two peritoneal endometriotic tissues had 1 and 5 informative specimens from individual glands, respectively; and the clonal patterns in 5 glands from the single lesion were divergent. Two abdominal wall endometriotic tissues had 7 and 6 informative specimens,respectively; and variable clonal patterns were seen in different glands from each lesion. One deep infiltrating endometriotic lesion had 4 informative specimens of isolated glands, and all of them showed unique clonal pattern. The disparate clonal patterns were found in endometriotic lesions with variable pathological subtypes, even arising from the same patient. Conclusions The epithelial cells from individual endometriotic gland showed monoclonality , and different glands from the same endometriotic lesion might show divergent patterns.

Objective To investigate the expression of transient receptor potential vanilloid subtype 1 (TRPV1) in uterosacral ligament and its correlation with pain in endometriosis.Methods Total of 54 patients undergoing endometriotic lesions excision in uteroscaral ligament by laparoscopy due to pelvic pain were enrolled in this study.According to visual analogue scale(VAS) scores,27 patients with VAS 5 - 10 were in group A and 27 patients with VAS 0 - 4 were in group B.In the mean time,20 patients with dysmenorrhea without endometriosis (VAS:0 -4 ) were matched as group C.Specimens (including the sacro-ligaments of 20 women without endometriosis) were immunostained with specific antibodies of TRPV1.Western blot and real time PCR were performed to detect TRPV1 expression in endometriosis lesions and control group.Results( 1 ) Immunohistochemnistry:the positive area of TRPV1 was found in endometriotic lesions in uterosacral ligament in group A,B and tissue of uterosacral ligament group C.The semi-qualitification of TRPV1 expression were 3 in group A, 1 in group B and 1 in group C by immunohistochemistry staining.There was significantly different expression between group B and group A ( P =0.005 ) or group C ( P =0.027 ).(2) mRNA expression:the expression of TRPV 1 was 1.84 in group A,0.80 in group B,0.24 in group C,respectively.With higher VAS scores,the expression of TRPV1 exhibited increasing trends.The expression of TRPV1 mRNA was higher in group A than thai in group B ( P =0.022).There was statistically different expression between group B and group C ( P =0.031 ).( 3 ) Western blot:the expression of TRPV1 protein was 0.63 in group A,0.19 in group B,0.02 in group C.There was significant differences between group A and group B ( P =0.022 ),and between group B and group C (P ＜ 0.01 ).Conclusion The expression of TRPV1 was correlated with the degree of pain in patient with endometriosis.

Objective To investigate the pathogenesis of endometriosis by studying endometriosis coexisting with variable genital tract anomalies and analysis the association between obstructive or nonobstructive anomalies with endometriosis.Methods From January 2000 to April 2010,a total of 67 cases of endometriosis coexisting with genital tract anomalies undergoing treatment in Peking Union Medical College Hospital were studied retrospectively.Results According to subtypes of concurrent genital tract anomalies,67 cases were divided into 19 cases in obstructive group and 48 cases in non-obstructive group.(1) Age and symptoms:the mean age were (22 ± 8) years in obstructive group and (32 ± 7) years in non-obstructive group,which reached statistical difference (P ＜ 0.05).The major symptoms were dysmenorrheal or chronic pelvic pain (14/19) and amenorrhea (8/19) in obstructive group.However,in non-obstructive group,the major symptoms were dysmenorrheal or chronic pelvic pain (31％,15/48) and infertility (35％,17/48) and abortion (21％,10/48).(2) Degree and incidence of endometriosis:the moderate or severe endometriosis was 11/19 in obstructive group and 40％ (19/48) in non-obstructive group,which did not show statistical difference (P ＞ 0.05).The incidence of peritoneal endometriosis,ovarian endometriosis,adenomyosis did not show significant difference between two groups (all P ＞ 0.05).The rate of moderate or severe endometriosis coexisting with duplex uterus,uterus bicornis and uterus septus did not show significant difference in non-obstructive group (P ＞ 0.05).Conclusions The severity of endometriosis showed no association with obstructive anomalies.The results implied that there might be other factors involved in pathogenesis of endometriosis.

Objective　To evaluate the ability of a risk of malignancy index (RMI), based on a serum CA125 level, ultrasound findings and menopausal status,to discriminate a benign from a malignant pelvic mass. Methods　One hundred and forty women with a pelvic mass, 30 years or older,admitted between January 1998 and June 1999, were studied. The sensitivity,specificity and positive predictive value of serum CA125 level, ultrasound findings and the menopausal status separately and combined into the RMI,to diagnose ovarian cancer. Results　RMI was more accurate than any individual criterion in diagnosing cancer. Using a RMI cut-off level of 200 to indicate malignancy, the RMI derived from this data set gave a sensitivity of 87.3%, specificity of 84.4%, and positive predictive value of 82.1%. Conclusions　RMI is able to correctly discriminate between malignant and benign pelvic mass. It can be introduced easily into clinical practice to facilitate the selection of patients for primary surgery.