Objective] To observe the influence of YIGU oral liquid on the pain threshold and bone mineral density in ovariectomized rats. [Methods] Eighty-four Female SD rats were randomly divided into 4groups:YIGU oral liquid group, calcitonin group,YIGU oral liquid combined with calcitonin group, saline control group.The ovariectomized rats were confirmed the successful model of osteoporosis,rats in the YIGU oral liquid group were administered with YIGU oral liquid, rats in the calcitonin group were administered with calcitonin, rats in the YIGU oral liquid combined with calcitonin group were administered with YIGU oral liquid group and calcitonin,rats in the saline control group were administered with saline.To detect the pain threshold and bone mineral density at the 10th day, 30th day, 60th day and 90th day respectively. [Results]At the 10th, 30th, 60th and 90th day after treatment, the pain threshold of the YIGU oral liquid group, calcitonin group,YIGU oral liquid combined with calcitonin group compare to saline control group, there was significant statistical difference( P<0.05).At the 10th, 30th day after treatment,the bone mineral density of the YIGU oral liquid group, cal-citonin group,YIGU oral liquid combined with calcitonin group compared with saline control group, there was no significant statistical difference( P>0.05). At the 60th,90th day after treatment, there was significant statistical difference( P<0.05).[Conclusion] YIGU oral liquid can effectively enhance pain thresh-old and bone mineral density of the ovariectomized rats to inhibit the osteoporosis pain.

To determine the (tttta)n repeat polymorphisms at the promoter region of CYP11alpha gene, and study its linkage to hyperandrogenism of polycystic ovary syndrome (PCOS) in Chinese women, a case-control study was conducted in the Reproductive Medical Center of the Second Affiliated Hospital of Zhengzhou University (Zhengzhou, China). 96 PCOS patients and 78 healthy control women were included. CYP11alpha (tttta)n repeat-polymorphism genotyping analysis was performed by using polymerase chain reaction (PCR). Serum pituitary hormone and total testosterone levels were measured by ELISA. 4 different CYP11alpha (tttta)n allelles were identified, corresponding to 4-, 6-, 8-, and 9-repeat-unit alleles. The frequency and distribution of these alleles are 0.16, 0.33, 0.38, and 0.13 respectively in PCOS patients, as compared with 0.20, 0.34, 0.35, and 0.11 respectively in healthy controls. There were no significant differences between these two groups. Moreover, no correlation between the polymorphism of CYP11alpha gene and serum testosterone level of patients with PCOS and controls was observed. It is concluded that microsatellite polymorphism (tttta)n of gene CYP11alpha exists in Chinese women and the polymorphism of CYP11alpha gene does not play an important role in the pathogenesis of Chinese patients with PCOS, especially in patients with hyperandrogenism.
Cholesterol Side-Chain Cleavage Enzyme/*genetics ; Hyperandrogenism/complications ; Hyperandrogenism/*genetics ; Microsatellite Repeats ; Polycystic Ovary Syndrome/complications ; Polycystic Ovary Syndrome/*genetics ; *Polymorphism, Genetic/genetics

Taking the mouse as a model, the experimental method of observing the morphology of meiotic spindles and chromosomes in mature oocytes were investigated in order to evaluate the effects of various interventions on the quality of oocytes accurately and rapidly. Laser scanning confocal microscope (LSCM) was used to examine the meiotic spindles and chromosomes by the technologies of optical section and three-dimensional (3D) image reconstruction. The results showed that the configurations of meiotic spindles and chromosomes could be observed clearly by LSCM. The normal rate of meiotic spindles and chromosomes was 82% and 86% respectively. It was concluded that the LSCM was a valid instrument to observe the meiotic spindles and chromosomes of mature oocytes and could be used as a valid method to evaluate the quality of M II oocytes.

In order To evaluate whether the parameters of spiral artery blood flow, as measured by transvaginal color Doppler, may be used to assess endometrium receptivity prior to embryo transfer (ET), a retrospective study of 94 infertile women who had undergone ART treatments with different outcomes (pregnant or nonpregnant) was done. Subendometrial blood flow was evaluated. The resistance index (RI), systolic/diastolic ratio (S/D) and pulsatility index (PI) were significantly lower in those who achieved pregnancy as compared with those who did not: 0.62+/-0.04 vs 0.68+/-0.04 (P<0.001), 2.66+/-0.33 vs 3.19+/-0.39 (P<0.01) and 1.15+/-0.17 vs 1.34+/-0.22 (P<0. 05), respectively. Furthermore, when RI>0. 2, PI>1. , and S/D>3. , no pregnancy occurred. These data suggest that the parameters of spiral artery blood flow could be used as a new assay in predicting endometrial receptivity before ET.
Arteries/ultrasonography ; Embryo Implantation/*physiology ; Embryo Transfer ; Endometrium/*blood supply ; Endometrium/*physiology ; Endometrium/ultrasonography ; Fertilization in Vitro ; Infertility, Female/physiopathology ; Regional Blood Flow ; Sperm Injections, Intracytoplasmic ; Treatment Outcome ; Ultrasonography, Doppler, Color

In order to establish a simple and useful way for preimplantation genetic diagnosis (PGD) of chromosomal diseases in general IVF laboratory, the methods that are most commonly used in the embryo biopsy, fixation of blastomere and fluorescence in situ hybridization were compared. The three aspects of PGD were analyzed respectively. There was no significant difference in further development capacity of embryos between mechanical (79.7%) and chemical biopsy group (78.6%) (P>0.05). In this study, more cells were successfully fixed with the Tween/HCL method (93.8%) than with the methanol/acetic acid method (80.5%, P<0.05). There was no significant difference in cytoplasm remains between methanol/acetic acid method and Tween/HCL method (P>0.05). The hybridization efficiency of fluorescence in situ hybridization was 89.5% in successive denaturation method and 90.9% in codenaturation method with the difference being not significant (P>0.05). In conclusion, the mechanical or chemical method, Tween/HCL fixation method and codenaturation fluorescence in situ hybridization method can constitute a simple and useful way for PGD of chromosomal diseases.

ObjectiveTo study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro.MethodsESC was induced decasualization in vitro and matched with non-decidualized cells as controls.The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR.Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it.Results (1) Between decidualized and undecidualized stromal cells,there were 49 miRNAs significantly different expression by microarray chip,including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b,30c,143,101,181 b,29b,30d,507,23 a,222,221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P ＜0.05).(2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC,ESC were cultured by FBS medium for 24 hours,the rate of transfection was 70％.ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours,the percentage of ESC at Sphase of (6.2 ± 0.7 ) ％ were significantly lower than ( 10.9 ± 0.8 ) ％ in control group ( P ＜ 0.05 ) ; the percentage of ESC at G0/G1 phase increased at transfection group [ (77.5 ± 1.3 ) ％ vs.(73.0 ± 1.6) ％ at control group ],but there was no significant difference (P ＞ 0.05 ).Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h,the percentage of ESC at S-phase was ( 3.3 ± 0.6) ％ in transfection group,which were significantly lower than (7.8 ± 0.9 ) ％ in control group ( P ＜ 0.05 ).The percentage of ESC at G0/G1 phase was ( 80.7 ± 1.6 ) ％ in transfection group and ( 74.9 ± 1.1 ) ％.In control group,which did not reached statistical difference ( P ＞ 0.05).ConclusionmiRNA was involved in ESC decidual process in vitro by regulating cell cycle.

Objective To observe and classify endometrial wavelike activities in controlled ovarian hyperstimulation (COH) cycles,and to study their effect on assisted reproductive techniques(ART) outcomes in COH cycles.Methods Patterns of endometrium wavelike movements were described,then their percentages of the total endometrium movements were calculated and compared retrospectively in 57 infertile women who had undergone ART treatments with different outcomes.Results Five types of endometrial wavelike movements could be distinguished:No activity, random,positive,negative,and opposing endometrium movements.The frequencies and types of endometrial movements varied in COH cycles.Endometrium movements,especially negative movements more frequently appeared in non-conception group than in conception group.Conclusions Abnormal endometrium activity could be seemed as underlying mechanisms in some cases of unexplained infertility or ART failure.

Background and purpose: AMP-activated protein kinase (AMPK) plays an important role in the regulation of cell metabolism and energy balance and is associated with cell proliferation, survival and multiple signaling pathways. Recent reports found that AMPK is involved in tumor suppression and drug resistance. The aim of this study was to explore the effect of AMPK on the anti-tumor effect of adriamycin and underlying mechanism in breast cancer MCF-7/adr cells. Methods:The anti-proliferative effects of adriamycin was detected by methyl thiazolyl tetrazolium (MTT) assay in MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKαcells. The cell morphology in each group was stained with the lfuorescent dye Hoechst 33528, and the effects on apoptosis induction were examined by lfow cytometry (FCM). The intracellular concentration of adriamycin was detected by lfuorescence assay. The resis-tance-and apoptosis-related proteins were analyzed by Western blot. Results:The growth of breast cancer MCF-7/adr cells was inhibited by adriamycin in a dose-and time-dependent manner. The IC50 values at 24 and 48 h were (36.8±2.1) and (28.8±1.3) μg/mL, respectively. AMPKαover-expression enhanced the cytotoxic effect of adriamycin in MCF-7/adr-AMPKαcells in a dose-and time-dependent manner. Its IC50 values at 24 and 48 h were (16.0±0.7) and (4.2±0.2) μg/mL, respectively. Fluorescent morphological assay showed that AMPKαoverexpression contributed to adriamycin induced apoptosis in MCF-7/adr-AMPKαcells. After treatment with 1.0 μg/mL adriamycin for 48 h, the apoptosis rates of MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKα cells were (12.0±1.4)%, (12.7±1.6)% and (32.0±4.2)%, respectively, indicating that overexpression of AMPKα enhanced the adriamycin-induced apoptosis in MCF-7/adr cells. Fluorescence microplate assay showed that over expression of AMPKαsigniifcantly increased the intracellular accumulation of adriamycin, in a concentration dependent manner. Western blot analysis showed that, compared with MCF-7/adr and MCF-7/adr-vector cells, the expressions of Bax, caspase-3 and cleaved PARP proteins were increased. Meanwhile, Bcl-2 and P-gp protein expressions were decreased in MCF-7/adr-AMPKαcells. Furthermore, the release of cytochrome c from mitochondria into the cytosol was also observed in MCF-7/adr-AMPKαcells. Conclusion:AMP-Kαoverexpression can enhance the chemosensitivity of breast cancer MCF-7/adr cells to adriamycin through inhibiting the drug effux transporter and regulating the expression of apoptosis-related proteins.

Taking the mouse as a model, the experimental method of observing the morphology of meiotic spindles and chromosomes in mature oocytes were investigated in order to evaluate the effects of various interventions on the quality of oocytes accurately and rapidly. Laser scanning confocal microscope (LSCM) was used to examine the meiotic spindles and chromosomes by the technologies of optical section and three-dimensional (3D) image reconstruction. The results showed that the configurations of meiotic spindles and chromosomes could be observed clearly by LSCM.The normal rate of meiotic spindles and chromosomes was 82% and 86% respectively. It was concluded that the LSCM was a valid instrument to observe the meiotic spindles and chromosomes of mature oocytes and could be used as a valid method to evaluate the quality of M Ⅱ oocytes.

Objective To investigate the effects of total flavonoids of litchi chinensis sonn (TFL) on cell proliferation and the molecular mechanism in rat hepatic stellate cells (HSC-T6) activated by growth factor-β1 (TGF-β1). Methods HSC-T6 cells were treated by 0.25%Trypsin-EDTA and then were digested into single cell suspension by DMEM (10%FBS included), which were mixed with TGF-β1 (5μg/L). (1) MTT method was used to detect the proliferation of HSC-T6 cells. Cells were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (80, 160, 320, 640 and 800 mg/L TFL). Each group has three wells. The absorbance (A) value was measured by enzyme standard meter at the 490 nm wavelength after 24 h, 48 h and 72 h treatment. The cell inhibitory rate was calculated. The subsequent experimental drug concentration and drug treatment time were determined according to half inhibitory concentration (IC50). (2) The expression levels of NF-κB andα-SMA mRNA were detected by PCR (for mRNA) and Western blot assay (for protein). Cells were cultured in the 10 cm culture dish and were divided into different TGF-β1 groups, including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (125, 250 and 500 mg/L TFL). After 48 h, related indicators were measured. Results At the same treatment time point, with the increased concentrations of TFL, A values were gradually decreased, and the cell inhibitory rates were gradually increased. There were no significant differences in the expressions of NF-κB andα-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-κB andα-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB andα-SMA mRNA and protein with the increased concentrations of TFL. Conclusion TFL can inhibit TGF-β1-induced HSC-T6 cell proliferation, which is involved in the inhibited expressions of NF-κB andα-SMA to anti-fibrotic effects in liver fibrosis.