Objective Salidroside is a major active component of integripetal rhodiola herbal medicine, which has a significant activity against hypoxia and ischemia.This study was to investigate the effects of side chain carbon losssalidroside analogues (N04) on the expressions of HIF-1α-related factors in the hypoxia-injured EAhy926 human vascular endothelial cells.Methods EAhy926 human umbilical vein endothelial cells in the logarithmic growth phase were randomly divided into a normal control, a hypoxia model control, a salidroside, a high-dose N04, a medium-dose N04, and a low-dose N04 group.The hypoxia model was established by depriving the culture medium of sugar and serum and culturing the EAhy926 cells in an environment of 95%N2+5%CO2 for 2 hours, followed by intervention with salidroside at 1×10-6 mol/L and N04 at 1×10-6, 1×10-7, and 1×10-8 mol/L, respectively.Then, the activity of the cells was detected by MTT assay, their LDH activity examined by spectrophotometry, the mRNA expressions of HIF-1α and VEGF measured by RT-PCR, the protein expressions of HIF-1α, VEGF and pVHL determined by Western blot, and the activity of eNOS measured by ELISA.Results Compared with the normal control group, the hypoxia model cells showed significantly reduced activity (0.51±0.05 vs 0.27±0.02, P<0.01), an elevated LDH level ([6.65±1.43] vs [78.82±2.33] U/L, P<0.01), and decreased eNOS activity ([1.56±0.23] vs [1.16±0.20] U/100 mL, P<0.01).In comparison with the hypoxia model group, the cells treated with high-, medium-, and low-dose N04 exhibited remarkably increased activity (0.27±0.02 vs 0.0.42±0.05, 0.40±0.03 and 0.37±0.04, P<0.01), a reduced LDH level ([78.82±2.33] vs [53.05±3.90], [58.42±4.45] and [62.73±3.63] U/L, P<0.01), and increased eNOS activity ([1.16±0.20] vs [3.01±0.47], [2.60±0.26] and [2.32±0.29] U/100 mL, P<0.01).The activity of eNOS was also increased in the salidroside group ([2.32±0.29] U/100 mL, P<0.01).The cell activity in the high-and medium-dose N04 groups was markedly higher than that in the salidroside group (P<0.05), and so was the eNOS activity in the high-dose N04 group and the LDH level in the medium-and low-dose N04 groups (P<0.05).In comparison with the normal control group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were significantly up-regulated in the hypoxia model group (P<0.01) while that of the pVHL protein markedly down-regulated (P<0.01).Compared with the hypoxia model group, the expressions of HIF-1α mRNA, HIF-1α protein and VEGF protein were remarkably reduced (P<0.05), while that of the pVHL protein markedly elevated (P<0.05).Both the expressions of VEGF mRNA and HIF-1α protein were significantly lower in the medium-and low-dose N04 groups than in the salidroside group (P<0.05).Conclusion N04 can protect vascular endothelial cells against hypoxia-induced injury by regulating the expression of HIF-1α-related factors and eNOS.

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.

Objective This study was designed to investigate the correlationship between plasma metastin and pathogenesis of adolescent women with polycystic ovary syndrome(PCOS).MethodsFrom Jan 2006 to Jun.2006.42 PCOS patients including 19 adolescent women and 23 adults with syndrome were treated in Second Affiliated Hospital of Sun Yat-Sen University.According to the range of age,those patients were divided into 19 cases in adolascent group(≤19 years)and 23 cases in aduh group(＞19 years).Meanwhile,20 adolescent women were matched as controls.Blood samples were collected between day 1 and day 5 of a spontaneous bleeding episode in the PCOS groups and a menstrual cycle of the controls.The Jevels of luteinizing hormone(LH)，follicle-stimulating hormone(FSH)，testosterone(T)，free T(FT)，dehydroepiandrosterone sulfate(DHEAS)，sex hormone-binding globulin(SHBG)，insulin，glucose，and metastin were detected from day 1 to day 5 of spontaneous bleeding or withdrawal bleeding by progesterone.On the next day,oral glucose tolerance test(75 g)and insulin release test were performed on those above patients and controls.The area under carve(AUC),the ratio of fasting blood glucose to insulin(GIR)and homeostasis model assessment insulin resistance jndex ( HOMA-IR)were calculated.Results(1)The level of hormone:the level of LH was in(12±7)U/L in adult group and(12±8)U/L in adolescent PCOS group，which were significantly higher than(6±4)U/L in controls(P＜0.05).The level of FT was(2.3±1.2)pmol/L in adult group，which was significantly higher than(1.3±0.8)pmol/L in adolescent group and(1.1±0.5)pmol/L in control roup(P＜0.05).It was observed that the level of(3.1±2.7)μmol/L in adolescent group was significantly lower than(6.3±2.7)μmol/L in control group(P＜0.05).Meanwhile,the level of FAI of 5.6±4.1 in adult group was significantly higher than 3.0±1.3 in control group(P＜0.05).No significant difference in FSH，T and SHBG levels among three groups were observed (P＞0.05).(2)Metastin and metabolism:Both the levels of fasting blood insulin,2-hour insulin and AUC of insulin were(13±7)mU/L，(88±59)mU/L and(133±80)mU·L-1·min-1 in adolescent group，which were significantly higher than(7±3)mU/L，(57±29)mU/L and(82±34)mU·L-1·min-1 in control group.The fasting blood insulin of(13±7)mU/L in adolescent group was significantly higher than (9±5)mU/L in adult group.The level of fasting blood glucose and 2-hour glucose were(5.01±0.44)mmol/L and(6.48±1.16)mmol/L in adult group，which were significantly higher than(4.68±0.29)mmol/L and(5.44±0.83)mmol/L in control group and(4.67±0.30)mmol/L and(5.93±1.44)mmol/L in adolescent group.The glucose AUC of(9.99±1.85)mmol·L-1·min-1 in adult group was significantly higher than(8.42±1.53)mmol·L-1·min-1 in control group(P＜0.05).HOMA-IR of 2.6±2.0 in adolescent group was significantly higher than 1.4±0.7 in control group.GIR of 10±8 in adolescent group was significantly lower than 16±10 in control group(P＜0.05).The metastin level of (0.25±0.19)pmol/L in adolescent group and(0.29±0.29)pmol/L in adult group were all significantly higher than(0.18±0.23)pmol/L in control group(PPh glucose were observed(r=0.256，0.286 and 0.267.P=0.044.0.025 and 0.043).Conclusions The expression of metastin in adolescent PCOS women was significantly higher that of normal adolescent women The increased level of metastin might be associated with pathogenesis of adolescent women with PCOS.

Objective To compare the difference of serum adiponectin levels between polycystic ovary syndrome (PCOS) patients and age, boby mass index (BMI) and insulin-resistance index matched controls, and explore its influence factors.Methods Case-control study, involving 97 women with PCOS and 116 age, BMI, fasting plasma glucose and insulin levels, homeostasis model assessment-insulin resistance index (HOMA-IR) matched controls.Hormone profiles, and serum adiponectin levels were measured and compared.Hormone profiles and serum adiponectin levels were compared among the four PCOS phenotypes.Multiple regression analysis was used to evaluate the factors affecting serum adiponectin levels.Results (1) Serum adiponectin level was significantly lower in PCOS group [(21 ± 16) mg/L] than controls [(25± 13) mg/L, P=0.038], and the same result in stratified analysis on weight height ratio (WHR, ≥0.8 and ＜0.8).(2) There was statistical differences in testosterone among different four PCOS phenotypes (P=0.001), there were no statistical differences in FSH, LH, WHR and serum adiponectin levels among four PCOS phenotypes (P＞0.05).(3) WHR and PCOS status were independent determinants of serum adiponectin levels (P＜0.05).Conclusions Low serum adiponectin levels in the women with PCOS is correlated with PCOS per se, independent of insulin resistance and obese.This fact supports the further study of the effect of adiponection in the pathophysiology of PCOS and its log-term impact.