AIM:To investigate whether gonadotropin-releasing hormone analogue(GnRHa)affect the sensitivity of breast cancer cells to 5-FU and epirubicin in vitro.METHODS:Two breast cancer cell lines(MCF-7 and MDA-MB-231)were treated with different concentrations of GnRH analogue,triptorelin acetate,or with a GnRHa+5-FU or GnRHa+epirubicin.The cellular growth profiles were determined by CCK-8.The mRNA levels of GnRH receptor,PCNA and MDR1 were measured by RT-PCR.RESULTS:Both cell lines had positive GnRH receptor mRNA expression detected by RT-PCR.GnRHa did not suppress cell growth after GnRHa exposure.IC50 of 5-FU and epirubicin was not changed in the presence of GnRHa.Suppression of cell growth by the exposure to 5-FU and epirubicin was not changed in the presence of GnRHa.GnRHa treatment up-regulated PCNA mRNA expression in MDA-MB-231 cells but not in MCF-7 cells.The expression of MDR1 mRNA was down-regulated by GnRHa in MCF-7 cell lines.No MDR1 mRNA expression in MDA-MB-231 cells was observed.CONCLUSION:The present data suggest that GnRH analogue(triptorelin acetate)does not affect the sensitivity of breast cancer cell lines MCF-7 and MDA-MB-231 to 5-FU and epirubicin.GnRHa may decrease the drug resistance by down-regulating MDR1 mRNA expression.

AIM: To identify the key factors responsible for drug resistance in different ovarian cancer cell lines using protein microarray system. METHODS: Six ovarian cancer cell lines were employed. The sensitivity of ovarian cancer cell line to common chemotherapeutic drugs was determined by using MTT assays. The expression of 78 cytokines and other factors was examined by using cytokine antibody array technology. RESULTS: Different ovarian cancer cell line responded to chemotherapeutic agents differently. The drug resistance was correlated with certain cytokine expression. Cell line SKOV3 was less sensitive to first line chemotherapeutic drug (ADM, CBPDA) and accumulated high amounts of GRO and TIMP-2 compared with other 5 cell lines. OVCAR4 cells were more resistant to second line chemotherapeutic drug (TAXOL, VP16) and had higher levers of IL-6 and IL-8 than IGROV1, OVCAR3 and OVCAR5. CONCLUSIONS: Among the most common excretive cytokines, increasing of GRO, IL-6, IL-8 and TIMP-2 might be related to drug-resistance of ADM and CBPDA in ovarian cancer cell, while IL-6 and IL-8 might also be related with drug resistance of TAXOL and VP16. The different types of ovarian cancer cell might have roughly similar excretive cytokines-induced mechanism of drug resistance.

Objective To discuss the clinical feature, diagnosis and treatment options of adolescent endometriosis Methods The records of adolescent patients with endometriosis (11 20 years old) who were admitted to First, Second and Third Affiliated Hospital of Zhongshan University and Guangdong Provincial People's Hospital between 1990 and 2003 were retrospectively reviewed Results Forty three patients were diagnosed as endometriosis either by laparotomy or laparoscopy The chief symptoms leading to the diagnosis were palpable pelvic mass (18/43), dysmenorrhea (15/43), chronic pelvic mass (10/43) and acute pelvic pain (4/43) The majority of patients (53%) presented with the revised AFS r classification stage Ⅲ, 8 cases (19%) presented with stage Ⅰ, 3 cases (7%) with stage Ⅱand 9 cases (21%) with stage Ⅳ. Nine cases (21%) had complicated genital tract abnormalities Conservative operations, including salpingo oophorectomy ins cases, ovarian cystectomy in 31 cases and laparoscopic vaporization in 8 cases, were performed Surgeries were followed by hormonal suppression using oral contraceptives in a continuous or cyclic manner Conclusions Adolescent endometriosis may occur around 4 6 years after menarche The chief symptoms are palpable pelvic mass and dysmenorrhea The treatment of endometriosis in adolescence does not differ principally from that in adult women In the treatment of endometriosis and for the prevention of recurrence, it is recommended to give 3 6 months of oral contraceptives

AIM:To determine the function of peritoneal mesothelial cells on the inflammatory microenvironment by administration of endometrial cells,and further define the pathogenesis of endometriosis.METHODS:Homogenous mouse endometrial epithelial and stromal cells were injected into the peritoneal cavities of Swiss Webster mice.After 4,24,and 72 h,a number of endpoints evaluated:protein concentrations of cytokine MCP-1,IL-1 ?,IL-6 in peritoneal lavage and gene expressions of MCP-1,IL-1 ?,IL-6 in peritoneal mesothelial cells and macrophages.RESULTS:The intraperitoneal administration of endometrial cells increased the protein expressions of cytokines in the peritoneal lavage of the recipient mice,which increased at 4-hour points and subsequently decreased with time.Gene expressions of cytokines in peritoneal mesothelial cells paralleled with the protein quantities in peritoneal lavage.The peak time of gene expression of cytokines in peritoneal macrophages was at the 24-hour point.The endometrial epithelial cells stimulated stronger inflammatory responses in the peritoneal cavity than the endometrial stromal cells.CONCLUSION:The recipient mice have a non-specific inflammatory response to the presence of endometrial cells in the peritoneal cavity.Mesothelial cells may be the targets of early inflammatory stress initiated in the presence of endometrial cells.

A total of 1 107 Chinese women were retrospectively investigated for the effects of overweight and underweight on ovarian stimulation, as well as the outcomes of in vitro fertilization and embryo transfer (IVF-ET)treatment. It showed that overweight women required more ampoules of gonadotrophin [ (36. 87 ±11. 18 vs 33.57±10.96)/ampoule,P<0.01]and had lower peak of estradiol concentration [ (1 846.6±1 390.3 vs 2 337.2± 1 490.6)pg/ml,P<0.01].increased cycle cancellation due to insufficient follicle development(6. 5% vs 2. 8% , P<0.05) ,and a higher miscarriage rate( 10.5% vs 5.4% ,P<0.05)compared with normal weight women. But no differences were found in clinical pregnancy and live birth rates. Compared with normal weight women, underweight women showed no differences in ovarian stimulation and IVF outcome.

AIM: To investigate the effects of leptin (LEP) on the alveolar type Ⅱ cells(AECⅡ) apoptosis induced by Na2S2O4 and explore the molecular mechanisms. METHODS: Primary AECⅡ culture was prepared according to a specific immunosorption procedure with slight modification and the cells were identified by transmission electron microscope and immunocytochemistry. AECⅡ damage was induced by 5 mmol/L Na2S2O4. LEP group cells were treated with LEP at concentrations from 100 ?g/L to 1 600 ?g/L. The cell survival rate was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Cell cycle and apoptosis were analyzed by flow cytometry and the level of caspase-3 was measured by Western blotting. RESULTS: Highly purified AECⅡ, obtained by the method of modified immunosorption, were identified with the positive expression of SP-A and intracellular lamellarbodies were found under electron micrography. The cells, exposed to 5 mmol/L Na2S2O4, showed characteristic changes of apoptosis and activation of caspase 3. These damages were relieved by the treatment of LEP (100-1 600 ?g/L), with survival increasing, apoptosis peak decreasing, cell morphology restoring and caspase 3 activation inhibiting.CONCLUSION: Leptin prevents AECⅡ from apoptosis induced by Na2S2O4 or hypoxia. The potential mechanism of its action may be related to promoting cell cycle from G1 phase to S phase and inhibiting the activating of caspase 3.

Objective To evaluate the effectiveness and safety of triptorelin in the treatment of uterine leiomyoma. Methods A multi-center, prospective, randomly controlled clinical trial was carried out from Dec. 2002 to Mar. 2004 in three university hospitals. A total of 125 qualified patients with uterine leiomyoma were randomly divided into either triptorelin group (63 cases) treated with 3.75 mg triptorelin injected intramuscularly or leuprorelin group (62 cases) treated with 3.75 mg leuprorelin injected subcutaneously. Both drugs were injected every 28 days for a total of 3 months. Results All 125 patients finished the trial. The uterine volumes were similar before treatment between the triptorelin group and the leuprorelin group and were decreased significantly after drug therapy (P0.05).) The volumes of the largest leiomyoma decreased significantly after drug therapy (P0.05). Patients with serum level of 17?-estradiol 0.05). Dysmenorrhea, noncyclic pelvic pain and pressure-like symptoms were relieved quickly and remarkably in both groups after treatment. The rates of adverse event occurred in 71% of patients in both groups. The main side effects included flare-up effects and hypoestrogenic symptoms. Nine patients in the triptorelin group and 6 in the leuprorelin group received add-back therapy with tibolone 1.25-2.50 mg/d because of remarkable climacteric-like symptoms. Conclusion Treatment of uterine leiomyoma with triptorelin for 3 months is both effective and safe in Chinese women.

AIM: To investigate the optimal materials and culture system of human primordial germ cells (PGCs) in order to establish human embryonic germ (EG) cell lines. METHODS: Human embryos of different gestational age were collected to isolate human PGCs. The isolated human PGCs were cultured in different medium and on different feeder layers, then their growth, proliferation and differentiation in different culture systems were observed. RESTILTS: The formation rate of primary colonies was higher when human PGCs were obtained with enzyme-mechanical method from 8-and 9-weeks gestational age human embryos than that from 7-weeks. Human PGCs grew better and maintained undifferentiating when mouse embryonic fibroblast or STO cells served as feeder layers and in conditional medium with hLIF, hbFGF, hSCF. CONCLUSION: 8-and 9-week gestational age human embryo are optimal material for isolating human PGCs. Enzyme-mechanical method is simple and available to isolate human PGCs. Feeder layer and growth factors are necessary for human PGCs culture in vitro.

[Objective]To evaluate the terminal hair growth of different body positions among in pregnant women ,analyze the contribution of each area to the diagnosis to hirsutism ,to improve the mFG scoring system.[Method]Pregnant women aged 20~41 years,with normal range of total testosterone levels and non-hirsute were recruited at their prenatal care in Sun Yat-sen Memorial Hospital,Sun Yat-sen University. They were followed up before pregnancy,at the 5th and the 9th week,the 10th and the 14th week, the 15th and the 20th week,the 21th and the 24th week of gestation. Then 72 more cases of pregnant women were recruited and followed up at 15~24th week. At each time of followed up,their total testosterone(TT)levels was examined by liquid chromatography tandem mass spectrometry(LC/MS-MS)and terminal hair growth were assessed by mFG score. Significant difference procedure least(LSD) analysis of variance was used to compare the levels of testosterone and mFG score in different gestational weeks. Receiver operating characteristics(ROC)analysis and logistics analysis were conducted to evaluate the contributory strength of hair growth in each body position for the diagnosis of hirsutism. The scores in the body area which made a significant contribution to the total were summed up and termed the simplified mFG score(sFG score). Following,the sFG scores were subjected to ROC analysis to determine the thresh-old that would maximize both the sensitivity and specificity of the measure to accurately distinguish hirsute from non-hirsute patients.[Results]Among the forty three pregnant women who were followed-up from before pregnancy to 24th week,the mean±SD for TT was (1.09 ± 0.59)nmol/L before pregnancy,and(1.13 ± 0.40),(1.28 ± 0.38),(1.83 ± 0.63),(1.82 ± 0.52)nmol/L for 5~ 9th,10~14th,15~20th week,and the mFG score was 1.65 ± 0.60,2.30 ± 0.45,3.60 ± 0.68,4.20 ± 0.41 and 4.40 ± 0.77,respectively. The order of the facial and body sites presented with new terminal hair growth was upper abdomen,lower abdomen,lower back,up-per lip,thighs,upper back,chest,upper arm,and chin,in sequence. After analyzing 115 cases(including the 72 cases recruited later),ROC analysis showed that the diagnostic value of different sites for hirsutism(mFG≥5):upper lip>lower back>thighs>lower abdomen>upper arm>upper back>chest/upper abdomen. Four sites among them ,namely upper lip ,lower back ,thighs and lower abdomen had the best diagnostic value,and the AUC for ROC were 0.779,0.728,0.675,and 0.626,respectively. Both ROC and logistic analysis indicated that he most significant body areas in defining hirsutism(defined as an mFG score≥5)were the upper lip, lower back,thighs,and lower abdomen. Using a cut-off value of 3,the combination of the four areas has the best sensitivity and specificity in distinguishing hirsute from non-hirsute women. [Conclusion]The study suggested that the mFG score increased as pregnancy progressed before the 24 weeks of gestation. The subset of upper lip,lower back,thighs and lower abdomen may be a reli-able simplification of mFG system for the evaluation of excess hair growth. The cut-off value was of≥3.

Objective To investigate the effect of anti-Mlllerian hormone (AMH) on hormone secretion and P450 aromatase mRNA expression from cultured human luteinized granulosa cells. Methods Human luteinized granulose cells were derived from 10 patients treated by in vitro fertilization-embryo transplantation (IVF-ET) in the Second Affiliated Hospital of Sun Yat-sen University from June to December 2006. Granulose cells were divided into group A, B, C, D, E depending on different concentration of AMH,testosterone group and blank group. 1×10-7moL/L testosterone and 1,5,10,20,50 μg/L AMH were added into the culture medium of group A,B,C,D and E. 1×10-7mol/L testosterone was added into the culture medium of testosterone group while no other ingredient was added into the medium of blank group. Estrogen levels in supernates were measured at 24,48,72 hours after cell incubation. RT-PCR was performed to detect the P450 aromatase mRNA expression in group B, C, D, E and testosterone group at 72 hours after cell incubation. Results (1) Estrogen levels in supernates of granulose cell culture at 24,48,72 hours were (8.529±0.381)×104, (10.977±0.436)×104, (13.309±0.506)×104 pmol/L in group A, (7.027±0.276)×104, (9.167±0.300)×104, (10.794±0.555)×104 pmol/L in group B, (6.039±0.226)×104,(7.585±0.548)×104, (8.797±0.518)×104 pmol/L in group C, (5.118±0.460)×104, (5.716±0.496)×104, (6.205±0.667)×104 pmol/L in group D, (4.932±0.148)×104, (5.323±0.184)×104,(5.629±0.212)×104 pmol/L in group E. When compared with blank group [(0.001±0.001)×104,(0.006±0.003)×104, (0.029±0.011)×104 pmol/L], the statistical differences were observed in group A,B,C,D,E(P<0.01) ; when compared with testosterone group [ (8.418±0.569)×104, (10.841±0.689)×104, (13.301±0.637)×104 pmol/L], the statistical differences were observed in group B,C,D and E(P<0.01) ; statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C(P<0.01). No significant difference was observed between group D and E (P>0.05). In group A, B, C, D, E and testosterone group, the estrogen levels at 24 hours after cell culture were significantly lower than those at 48 and 72 hours (P<0.01) ; statistical difference was observed between estrogen levels at 48 and 72 hours(P<0.01). No significant difference was observed among 24,48 and 72 hours in blank group (P>0.05). (2) Relative ratios of intensity of P450 aromatase/β-actin at72 hours of cell culture in group B,C,D and E were 0.6148±0.0046, 0.5156±0.0012, 0.4698±0.0027 and 0.4282±0.0017, respectively, which were statistically lower than that in testosterone group (0.8224±0.0021, P<0.01) ;statistical differences were also observed in group C, D and E when compared with group B, and also group D and E when compared with group C(P<0.01). No significant difference was observed between group D and E (P>0.05). Conclusion It is suggested that AMH might affect estrogen synthesis by inhibiting P450 aromatose activity so that lead to hyperandrogenism microenvironment in local ovary.