PurposeTo determine the correlation between expression of MHC classⅠon ovarian tumor cells and T-cell infiltration in vivo on ovarian tumor micro-environment.MethodsThirty three samples of primary epithelial ovarian carcers were analysed with flow cytometry for MHC class Ⅰ expression. Immunohistochemical analysis was performed using monoclonal antibodies that recognize leukocyte differentiation antigens ( LCA+ , CD3 + ) on cryostat sections of primary epithelial ovarian carcinoma. The results were compared with the expression of MHC class Ⅰ expression. ResultsClose correlation were revealed bewteen the expression of MHC class Ⅰ molecules and the numbers of infiltrating LCA+ positive cells ( r = 0. 846, P = 0. 000 1) and CD3 + cells ( r = 0. 738, P = 0. 000 1 ).ConclusionsExpression of MHC class Ⅰ molecules may influence the status of human immunosurveillance in tumor micro-environment on ovarian carcinoma and the assay may be helpful in selection patients for adoptive immnotherapy.

Objective To investigate the expression of neive growth factor (NGF) in the ectopic endometrium in adenomyosis patients,and explore the relationship between NGF expression and innervation or pain scales.Methods From Mar.2009 to Oct.2009,45 adenomyosis patients undergoing hysterectomy in Obstetrics and Gynecology Hospital of Fudan University were enrolled in this study,which were classified into 33 cases in pain group and 12 cases in non-pain group based on symptom.The degree of dysmenoreal,chronic pelvic pain and dyspareunia was evaluated by visual analogue scale,including no pain,mild to moderate pain and severe pain group.In the mean time,26 patients with leiomyoma or cervical intraepithelial neoplasia Ⅲ (CIN Ⅲ)undergoing hysterectomy were defined as control group.Ectopic endometrium from experimental group and eutopic endometrium from control group were collected in the surgery.The expression of NGF was examined by immunohistochemistry.The density of protein gene product (PGP)9.5 positive nerve fibers was detected by immuno-fluorescence.Results The NGF level and the density of PGP 9.5 positive nerve fibers in adenomyosis pain group (0.25 ± 0.08,16 ± 8) were higher than adenomyosis painless (0.19 ± 0.05,P =0.007 ; 11 ± 5,P =0.018) and control group (0.18 ± 0.05,P =0.000;9 ± 4,P =0.000).The NGF level and the density of PGP9.5 positive nerve fibers in severe dysmenorrheal group(0.29 ± 0.07,19 ± 10) were higher than mild to moderate dysmenorrheal (0.22 ± 0.07,P =0.018 ; 13 ± 4,P =0.035) and painless group (0.18 ± 0.05,P =0.000 ; 11 ± 5,P =0.006) of adenomyosis patients.There was no difference of NGF level and the density of PGP 9.5 positive nerve fibers in chronic pelvic pain group and no chronic pelvic pain group of adenomyosis patients,so was dyspareunia group and no dyspareunia group.Conclusion The increased NGF level of adenomyosis nodules and improving innervation might be involved in the mechanism of adenomyosis related pain.

To investigate different doses of daidzein (DAI) in regulating bone formation of osteoblasts, and the regulating mechanisms of estrogen receptors (ERs) and peroxisome proliferator-activated receptor γ (PPARγ) in bone formation.

Objective To investigate the effects of different doses of estradiol on apoptosis of T lymphocytes from spleens in ovariectomized mice and explore the possible mechanisms.Methods The mice splenic T lymphocytes were isolated and divided into ten groups: young group, sham- ovariectomized group, ovariectomized group, ovariectomized plus estradiol(10-11, 10-10, 10-8 and 106groups, ovariectomized plus estradiol (10-10mol/L) plus 1CI182 780 (10-7tool/L) group, ovariectomized plus estradiol(10-10mol/L) plus 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPP, 10-6mol/L) group, ovariectomized plus estradiol(10-10mol/L) plus pyrroline dithiocarbamate(PDTC, 10-6mol/L) group. The apoptosis rates were determined by flow eytometry using Annexin V-FITC/ PI and the protein levels of ERa, ERβ, Bax, Bcl-2 and P65 were detected by Western blot. Results The apoptosis rate of ovariectomized group was(19. 4±2.5)%, which was higher than that of young group [(14.6±2.4%) 3 and sham-ovariectomized group [p (14.5±2.3)%], and the levels of Bcl-2 and nuclear P65 were lower than the young group [(0. 25±0. 05, 0. 09±0. 01) vs. (0. 40± 0.07,0. 15±0. 02), P<0.01]. The ovariectomized plus estradiol (10-10tool/L) group had lower apoptosis rate and higher Bcl-2 and P65 levels compared to the ovariectornized group[(16.6±1.8)% vs.(19.4±2.5)%,P<0.05;0.36±0.03 vs. 0.25～0.05, 0.14±0.01 vs. 0.09±0.01, P< 0. 01)], while the ovariectomized plus estradiol(10-8mol/L, 10-6tool/L) groups had higher apoptosis rates than the ovariectomized group[(22. 55±2. 5)% vs. (19. 4±2. 5)% ,P<0. 05;(27.8±3.1)% vs. (19.4 4±2. 5)%,P<0. 01, respectively]. The 2protein levels of ERa and ERβ of ovariectomized group were 0. 23±k0.01 and 0. 22±0. 03, respectively, which were lower than those of young((0. 27±0. 02) and (0. 29±0.04)] and sham-ovariectomized group [(0. 28±0. 03) and (0. 29±0.02)]. The ovariectomized plus estradiol(10-1110-1010-8tool/L) groups had higher while ovariectomized plus estradiol(10-6mol/L) group had lower ERα and ERβ protein levels (0. 09±0. 01,0. 14±0.02) than the ovariectomized group(P<0. 01). There was no significant difference between ovariectomized plus estradiol(10-10mol/L) plus ICI182 780 group or ovariectomized plus estradiol(10-10tool/L) plus PDTC group and ovariectomized group [(19.4±1.6)% vs. (19.4±2. 5)%, (21.0±2. 9)% vs. (19.4d±2. 5)%, P>0. 05). There were also no significant difference between ovariectomized plus estradiol(10-10mol/L) plus MPP group and ovariectomized plus estradiol (10-10mol/L) grou p[(16.9±2.2)% vs. (16.6±1.8)%, P>0.05]. Conclusions The ovariectomy of mice leads to increased apoptosis rates of splenic T lymphocytes. The effects of estradiol on the apoptosis of T lymphocytes in ovariectomized mice are dependent on doses: physiological dose of estradiol inhibits while higher dose of estradiol exacerbats the apoptosis of T lymphocytes in ovariectomized mice.Physiological dose of estradiol may act on Rice T lymphocytes via ERβ and NFkB signaling.

Objective To investigate the relationship between insulin resistance and endogenous androgens at early and late phase of postmenopause.Methods A total of 105 women with early postmenopause (≤5 years since menopause) and 107 women with late postmenopause (≥ 10 years since menopause) were enrolled in this study.In the mean time,those women were classified into normal weight [body mass index (BMI),BMI ＜24 kg/m2] group and overweight (BMI≥24 kg/m2) group.Sex hormonebinding globulin (SHBG),testosterone (T),dehydroepiandrosterone-sulfate (DHEA-S),fasting blood glucose(FBG),fasting insulin (FINS)levels were measured and then calculated free androgen index(FAI) and homeostatic model assessment of insulin resistance (HOMA-IR).The relationship between sex hormones and insulin resistance was analyzed by partial correlation and multiple linear regression analyses.Results Compared to early postmenopausal women,late postmenopausal women had higher FINS [(7.9 ± 6.6) mU/L versus (6.6 ±4.0) mU/L] and HOMA-IR(2.1 ± 1.9 versus 1.7 ± 1.1),but they had lower DHEA-S [(0.9 ± 0.5) mg/L versus (1.1 ± 0.5) mg/L,all P ＜ 0.05)].Both in early postmenopausal and late postmenopausal groups,overweight women had higher HOMA-IR (early group,2.2 ± 1.0 versus 1.2 ±0.9 ; late group,2.8 ± 2.6 versus 1.6±1.1)and FINS early group[(6.9±2.9) mU/L versus (4.6±2.0) mU/L] ;late group [(10.2 ± 9.3) mU/L versus (6.4 ± 3.6) mU/L] than those at women with normal weight group(all P ＜ 0.05).In early postmenopausal group,overweight women had lower SHBG [(52 ±37) nmol/L versus (71 ±37) nmol/L] and higher FAI(2.5 ±2.1) versus (1.3 ± 1.1) than those at normal weight women group(all P ＜ 0.05).In late postmenopausal group,overweight women had higher DHEA-S (1.0 ± 0.5) mg/L versus (0.8 ± 0.4) mg/L (P ＜ 0.05).The analyses suggested that in early postmenopausal group,SHBG was correlated negatively with FINS and HOMA-IR (β =-0.386,P ＜ 0.05 ;β =-0.553,P ＜0.05),DHEA-S was correlated positively with FBG (β =0.348,P ＜ 0.05) in early postmenopausal group.FAI was correlated positively with FBG in late postmenopausal group (β =0.505,P ＜ 0.05).Conclusions The increased androgenic activities are associated with insulin resistance after of menopause.These correlations are different at different stages of postmenopause,which SHBG levels correlate with high risk of insulin resistance and DHEA-S levels correlates with high blood glucose levels at early postmenopause and FAI correlates with high blood glucose levels at late postmenopause.

Objective To compare clinical effect of gonadotropin releasing hormone agonist(GnRH-a) alone and GnRH-a combined with low-dose dydrogesteronea and estradiol valerate on sex hormone, hypoestrogenic symptoms, quality of life and bone mineral density (BMD)in treatment of endometriosis.Methods Seventy patients with moderate or severe endometriosis, who were diagnosed by laparotomy or laparoscopic surgery within two months, were randomly assigned into two groups.35 patients in GnRH-a group were treated by goserelin (3.6 mg)for three months, and 35 patients in add-back group were treated by goserelin (3.6 mg)combined with estradiol valerate 0.5 mg and dydrogesteronea 5 mg daily.Before and after the treatment, clinical parameters were recorded and analyzed, including visual analog scale (VAS), medical outcomes survey short form 36 (SF-36), Kupperman menopausal index(KMI), BMD, the serum level of follicle stimulating hormone (FSH), estradiol (E_2) and bone gla-protein (BGP) .The first menstruation and VAS were also followed up after treatment.Results Every 3 cases in two groups lost follow-up.(1)Reproductive hormone: the level of E_2 in add-back group [(94 ± 71) pmol/L]was significantly higher than (54±52) pmol/L in GnRH-a group(P <0.01).The level of FSH in add-back group [(3.0 ± 1.9) U/L]was significantly lower than (5.7 ± 2.9) U/L in GnRH-a group (P < 0.05).(2) VAS: after treatment, VAS in both group decreased significantly when compared with that before treatment(P < 0.05), and remained until menstruated.(3) KMI: KMI in add back-group (10 ± 8) was significantly lower than (14 ± 6) in GnRH-a group (P < 0.05).(4) BMD: compared with that before treatment, BMD decreased significantly after treatment in GnRH-a group (P < 0.05), no remarkable difference of BMD was observed before and after treatment in add-back group.Before treatment, serum BGP in both groups did not show statistical difference.After treatment, the level of BGP in GnRH-a group [(7932±5206) ng/L]was significantly higher than (5419±2917) ng/L in add-back group (P <0.05).Conclusions GnRH-a combined with estrogen-progesterone regimen could relieve pain from endometriosis as effectively as GnRH-a alone and reduce hypoestrogenic symptoms and bone loss.Therefore,it is a safe and effective treatment.

Objective To evaluate effects and safety of gonadotrophin-releasing hormone agonist (GnRH-a) combined with transdermal estradiol and medroxyprogesterone acetate in the treatment of endometriosis. Methods From January I st, 2007 to July 31 st, 2007, 28 endometriosis patients underwent laparnscopic or transabdominal surgery in Obstetrics and Gynecology Hospital affiliated to Fudan University were randomly divided into group A and group B. 14 patients in group A received 3.6 mg goserelin once every 4 weeks, 12 weeks in all 14 patients in group B received goserelin and added 1/2 piece of half-hydrate estradiol every week and 6 mg oral medroxyprogesterone acetate per day, 12 weeks in all. Serum estradiol (E2 ), follicle stimulating hormone(FSH), bone gla protein levels, visual analogue scale (VAS) of pain, bone mineral density of lumbar spine, vaginal exfoliate cell spurs and the form of Kupperman were compared in patients before and after treatment. Results (1 ) After treatment, the level of FSH and E2levels were (5.0 ± 2. 6 ) U/L and (29 ± 17 ) pmol/L in group A and (3.0 ± 1.5 ) U/L, and (87 ± 53 ) pmol/L in group B, which were significantly lower than those before treatment [FSH (17. 0 ± 12. 2) U/L, and E2 (184 ± 194) pmol/L in group A and FSH :(15.3±13.6)U/L and E2: (281±242) pmol/L in group B, P ＜ 0. 01]. On the seventh day after three-month GnRH-a treatment, it was observed that the level of E2 was higher and FSH was lower in group B than the level of E2 and FSH of group A (P ＜ 0. 01 ). (2 ) After treatment, the basal vaginal exfoliate cell proportion in group A [(66. 2 ± 29. 0) %] was significantly lower than that in group B [(11.8 ± 28. 0) %, P ＜ 0. 01] ; while patients in group A owned a lower proportion of the middle [(29. 1 ± 23.1 ) %], superficial layers [(4. 0 ± 5.5 ) %] and esinophilic cells [(2. 3 ± 2. 6)%]than patients group B [middle layer: (73. 0 ± 25.2)% ; superficial layer: (15. 2 ± 10. 9)% ; esinophilic cells: (10. 8 ± 7.9 ) % ; P ＜ 0. 01]. (3) Before the treatment, patients' VAS scores of total, pelvic pain, dysmenorrheal and dyspareunia were 7.43±3. 20,2. 35 ± 1.82, 4. 93 ± 1.98 and 0. 14±0. 53 in group A and were 7.71±2. 02, 2. 57 ± 1.60, 4. 86 ± 1.56 and 0. 29 ± 1.07 in group B; after treatment, the scores above were changed to 0. 14±0. 36,0. 07±0. 27,0. 07±0. 27and 0 in group A and 0. 36±0. 50, 0. 29±0. 47, 0. 07±0. 27 and 0 in group B, which were all significantly lower than those before treatment separately (P ＜0. 01 ). When menstruation recovered, the scores were 0. 21±0. 43, 0. 07±0. 27, 0. 14 ± 0. 36, and 0 in group A and 0. 50±0. 65, 0. 29±0. 47, 0. 21±0. 43 and 0 in group B, which were also significantly lower than those before treatment (P ＜ 0.01 ), however, no statistical difference was found between groups at any time spot(P ＞ 0. 05). (4) In group A, the bone density after treatment [(0. 96 ± 0. 06 ) g/cm2] was lower than that before treatment [(0. 99 ± 0. 06 ) g/cm2, P ＜ 0.01 )]. In group B, the index was (0. 98 ± 0. 09) g/cm2, which was lower than that before treatment [(0. 99 ± 0. 10 ) g/cm2, P = 0. 201]. No statistical difference was found between groups(P ＞ 0. 05 ). The bone loss rate were (- 2. 77 ± 1.97 ) % in group A and (- 0. 93 ± 2. 86 ) % in group B (P = 0. 058 ). Before treatment, the bone gla protein was (13±3) μg/L in group A and (13±6) μg/L in group B. After treatment, the bone gla protein levels was (17±6)μg/L in group A, which was higher than that before treatment (P ＜ 0. 01 ), the level was (16±6)μg/L in group B, which was higher than that before treatment, however showed no statistical difference(P =0. 053). No difference was found in bone gla protein before and after treatment between two groups (P＞0. 05). (5) The form of Kupperman after treatment were 15±7 in group A and 11±6 in group B, which did not show significant difference (P ＞ 0. 05 ). The incidence of flash and sweat were 93% (13/14)in group A, which was significantly higher than that 57% (8/14) in group B(P ＜0.01 ). Conclusion The add-back therapy that consists of an estradiol patch and oral medroxyprogesterone acetate is effective and safe treatment for endometriosis.

Objective To study the influence of residual methylene blue after plasma viral inactivation on the human immune cell function by using the peripheral blood mononuclear cell(PBMC) .Methods PBMC were isolated by adopting the Ficoll‐Hypaque density gradient centrifugation method and co‐cultured for 72 h in presence of specific T cell stimulating factors(Anti‐CD3/28 and Anti‐CD28) ,with or without different concentration of methylene blue .The culture supernatant was collected and detected the cyto‐kines secretion situation by ELISA .After 66 h culture ,CCK‐8 dye was added and continueously cultured for 4-6 h ,the prolifera‐tion was determined at A450 .Results The high‐concentration doses of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) had signifi‐cantly inhibiting effect on the proliferation of PBMC stimulated by Anti‐CD3/28(P< 0 .01) ,its OD value was decreased from 0 .897 ± 0 .385 to 0 .632 ± 0 .334 ,0 .524 ± 0 .254 and 0 .445 ± 0 .287 respectively ,showing certain dose dependent effect .The high concentrations of methylene blue (1 .25 ,2 .5 ,5 μmol/L groups) could down‐regulate interleukin(IL)‐17a ,IL‐10 and interferon (IFN)‐γ secreted by anti‐CD28 induced PBMC ,moreover showing a dose dependent effect .1 .25 ,2 .5 ,5 μmol/L methylene blue af‐fected the IL‐17a level secreted by PBMC from (406 ± 57)pg/mL descending to (276 ± 38) ,(192 ± 31) ,(134 ± 24)pg/mL respec‐tively ;affected PBMC to secrete IL‐10 ,its level was reduced from (184 ± 15) pg/mL to (132 ± 13) ,(110 ± 12) ,(42 ± 8)pg/mL ;af‐fected PBMC to secrete IFN‐γ,its level was deduced from (4 512 ± 187)pg/mL to (2 876 ± 143) ,(2 234 ± 153) ,(1 988 ± 112)pg/mL respectively .Conclusion High concentrations of methylene blue (≥1 .25 μmol/L ) has the significant inhibiting effect on the proliferation and cytokine secretion functions of PBMC .In other words ,the residual methylene blue concentration in viral inactiva‐tion plasma (≤0 .33 μmol/L) has no obvious effect on the immune function of PBMC ,but whether this concentration of methylene blue having the effect on human pure T cell immune function needs to be further evaluated and studied .

To investigate the roles of daidzein in the expressions of steroid receptor coactivator-1 (SRC-1) and nuclear receptor corepressor (NcoR) in MC3T3-E1 osteoblastic cells.

Objective To explore the effects of daidzein (DA) on the expressions of estrogen receptors (ER) and peroxisome proliferator-activated recepor γ (PPARγ) in osteoblasts and the influence of estrogen on these effects.Methods A mouse osteoblastic cell line MC3T3-E1 cultured in α-MEM containing 2％ FBS was treated by 0.1 and 10 μmol/L DA.ER antagonist ICI182780 and PPARγ antagonist GW9662 in 0.1 μmol/L was added as required,and an equivalent amount of phosphate buffer solution (PBS) was used as control.For the study on estrogen effect,the cells were treated by DA in the serum-free medium with or without 10 nmol/L 17β-estradiol (E2).The expressions of ERa,ERβ and PPARγ were determined by real-time RT-PCR and Western blot analysis,respectively.Results DA inhibited ER,expression but stimulated PPARγ expression in the cells at the concentration of 0.1 and 10 μmol/L.The down-regulation of ERα by DA could be blocked by ICI182780,whereas the up-regulation of PPARγ could be repressed by GW9662 in transcription levels.Furthermore,the inhibitory effect of DA on ERβ expression was markedly enhanced,while its stimulatory effect on PPARγ expression was almost lost in serum-free medium with 10 nmol/L 17βestradiol as determined by real-time RT-PCR.Conclusions Besides its direct roles in ERs and PPARγ mediated gene transcriptions,DA could exert indirect effect on cellular pharmacological responses by altering ER and PPARγ expressions.The predominant influence on receptors expression probably involved in the time-related biphasic effects of DA on osteogenesis,which was supposedly influenced by estrogen level.