清肝活血方及其拆方对脂多糖介导库普弗细胞活化ERK表达调控的研究

目的:探讨清肝活血方及其拆方对脂多糖介导库普弗细胞(Kupffercell,KC)活化表达细胞外受体活性激酶(ERK)的影响。方法:原代分离KC,筛选LPS(脂多糖)合适剂量和作用时间;制备清肝活血方及其拆方含药血清,以ELISA、Western方法检测该方及其拆方含药血清对KC表达肿瘤坏死因子α、细胞外受体活性激酶1/2(ERK1/2)和激活蛋白1(AP-1)的影响。结果:选择LPS合适剂量为100μg/L,合适时间为2小时。清肝方通过抑制磷酸化的ERK(P-ERK)来调节核因子AP-1的表达。活血方可以下调P-ERK,抑制效应产物TNF-α的表达。清肝活血方调控磷酸化P-ERK,抑制核因子AP-1的表达。结论:清肝活血方及其拆方含药血清通过影响ERK信号通路,抑制TNF-α的产生,从而起到保护肝细胞的作用。

Effects of Cumulus Cells on in vitro Fertilization of Bovine

[Objective] The aim was to explore the effect of cumulus cells on the in vitro fertilization of in vitro matured bovine oocytes. [Method] The in vitro matured oocytes were divided into three groups of cumulus cells removal, partial removal and no removal. [Result] In the co-culture with cumulus cells, the oocytes of the removal group had higher cleavage rate and blastocyst rate （74.4%±4.1, 53.7%±5.1） than those of the no removal group （72.7%±5.1, 52.4%±3.5）, but the difference was not significant （P〉0.05）, while both groups had better performances than the re- moval group （39.6%±4.5, 18.8%±4.6） with the difference reaching the significant level （P〈0.05）. All the three groups showed significant difference with the control. The combination of cumulus cells and melatonin achieved the best effects as the cleavage rate and blastocyst rate of the partial removal group （79.8%±3.7, 56.5%±5.1） were better than those of the no removal group （78.2%±2.6, 55.8%±4.6）, and the difference was not significant, while both group had better performances than the removal group （48.3%±5.5, 22.7%±4.3） and the control group with the differences reaching the significant level （P〈0.05）. [Conclusion] The study provided technical support for the production of dairy cows and beef cattle.

N-methyl-D-aspartate receptors mediate diphosphorylation of extracellular signal-regulated kinases through Src family tyrosine kinases and Ca^2＋/calmodulin-dependent protein kinase Ⅱ in rat hippocampus after cerebral ischemia

Objective： Extracellular signal-regulated kinases （ERKs） can be activated by calcium signals. In this study, we investigated whether calcium-dependent kinases were involved in ERKs cascade activation after global cerebral ischemia. Methods Cerebral ischemia was induced by four-vessel occlusion, and the calcium-dependent proteins were detected by immunoblot. Results Lethal-simulated ischemia significantly resulted in ERKs activation in N-methyl-D-aspartate （NMDA） receptor-dependent manner, accompanying with differential upregulation of Src kinase and Ca^2＋/calmodulin-dependent protein kinase Ⅱ （CaMKⅡ） activities. With the inhibition of Src family tyrosine kinases or CaMKⅡ by administration of PP2 or KN62, the phosphorylation of ERKs was impaired dramatically during post-ischemia recovery. However, ischemic challenge also repressed ERKs activity when Src kinase was excessively activated. Conclusions Src family tyrosine kinases and CaMKⅡ might be involved in the activation of ERKs mediated by NMDA receptor in response to acute ischemic stimuli in vivo, but the intense activation of Src kinase resulted from ischemia may play a reverse role in the ERKs cascade.

Chronological and Morphological Progression of Nucleus during Mouse Oocyte Maturation and Fertilization in vitro

The chronological and morphological changes of the nucleus during mouse oocyte maturation and fertilization were systematically studied. Although most oocytes went through GVBD 2-4 hrs after culture, 13.6% remained at GV stage 8 hrs after culture.TEM observation revealed that nucleoli of oocytes which failed to go through GVBD were composed of fibrillar-granular component,small vacuoles and fibrillar centers or showed small vacuoles on nuclear surface. During GVBD, the nucleoli became smaller and smaller and finally disappeared with the nuclear-associated chromatin dislocated to the periphery. Nuclear membrane with attached chromatin became folded and electronic dense cores appeared in the center of chromatin clumps at the same time.The last event of GVBD was the disruption of nuclear membrane.At the end of the 5th hr after culture, meiosis progressed to prometaphase I.Chromosomes,distributed in the original GV area free of organelles,were surrounded by large quantity of mitochondria and small SER vesicles. At the end of the 12th hr after culture,48. 1% of the oocytes emitted PB1.Decondensing sperm head and early male pronuclcus(mPN)with condensed nucleoli were found 1-2 hrs after insemination.The formation and enlargement of female PN(fPN) occurred a little earlier than that of mPN. 33.3% finished syngamy at 8-9 hrs after insemination.The process of nucleolus formation was reverse to that in GVBD. The oolemma modification caused by cortical reaction could effectively inhibit polyspermy.in contrast,there were sperm binding to the oolemma where CGs failed to be released. In addition, PB2 was emitted 2-5 hrs after insemination. The difference between PB1 and PB2 as well as the abstriction of polar body were also discussed.

In Vitro Fertilization in Maize and Tobacco: The Fusion Patterns During Sexual Cell Fusion

Large scale of sexual cell fusion was carried out in order to observe the fusion pattern during in vitro fertilization with special interest in the influence of cell volume on the membrane behavior. Three patterns could be recognized in sexual cell fusion, which was supported by fluorescent microscopy with the aid of video enhanced system and cooled CCD. It was found that the fusion pattern and cell membrane behavior were related to the cell volume ratio of two fusing cells. The results reported here might be useful for interpreting and evaluating data from in vitro fertilization experiments and for explaining male cytoplasm exclusion during fertilization.

Detachment of esophageal carcinoma cells from extracellular matrix causes relocalization of death receptor 5 and apoptosis

AIM： To investigate the effect of detachment of esophageal cancer cells from extracellular matrix on the localization of death receptor 5 （DR5） and apoptosis. METHODS： Anchorage-dependent EC9706 cells of esophageal squamous cell carcinoma were pretreated or not treated with brefeldin A. Detached cells were harvested by ethylenediaminetetraacetic acid digestion. Expression and localization of DR5 in these cells were determined by immunocytochemical and immunofluorescence assays, as well as flow cytometry analysis. Apoptosis of EC9706 cells was detected by flow cytometry after stained with fluorescein isothiocyanate-labeled annexin V/propidium iodide. Activation of caspase 8 was detected by Western blot analysis. RESULTS： Immunocytochemical assay indicated that DR5 was predominantly perinuclear in adherent cells but was mainly localized in cell membrane in detached cells. In addition, immunofluorescence assay also confirmed the above-mentioned results, and further demonstrated that DR5 was present in the form of coarse granules in detached cells, but in the form of fine granules in adherent cells. Cytometry analysis revealed higher levels of DR5 expression on the surfaces of brefeldin-A-untreated cells than on the surfaces of brefeldin-A-treated cells, but brefeldin A treatment did not affect the total DR5 expression levels. Moreover, nocodazole did not influence the extracelluar DR5 expression levels in EC9706 cells. Apoptosis assay revealed that detached cells were more sensitive to DR5 antibody-induced apoptosis than adherent ceils. Western blotting showed that caspase 8 was activated in temporarily detached cells 4 h earlier than in adherent cells. CONCLUSION： Progress from adhesion to detachment of EC9706 cells causes DR5 relocalization, and promotes cytoplasmic translocation of DR5 to cell surfaces via a Golgi-dependent pathway. Moreover, it might also result in DR5 aggregation to render apoptosis of detached cells.

Thyroid hormone regulation of apoptotic tissue remodeling during anuran metamorphosis

Anuran metamorphosis involves systematic transformations of individual organs in a thyroid hormone (TH)-dependent manner. Morphological and cellular studies have shown that the removal of larval or- gans/tissues such the tail and the tadpole intestinal epithelium is through programmed cell death or apop- tosis. Recent molecular investigations suggest that TH regulates metamorphosis by regulating target gene expression through thyroid hormone receptors (TRs), which are DNA-binding transcription factors. Cloning and characterization of TH response genes show that diverse groups of early response genes are induced by TH. The products of these TH response genes are believed to directly or indirectly affect the expression and/or functions of cell death genes, which are conserved at both sequence and function levels in different animal species. A major challenge for future research lies at determining the signaling pathways leading to the activation of apoptotic processes and whether different death genes are involved in the regulation of apoptosis in different tissues/organs to effect tissue-specific transformations.

The matrix metalloproteinase stromelysin-3 cleaves laminin receptor at two distinct sites between the transmembrane domain and laminin binding sequence within the extracellular domain

The matrix metalloproteinase (MMP) stromelysin-3 (ST3) has long been implicated to play an important role in extracellular matrix (ECM) remodeling and cell fate determination during normal and pathological processes. However like other MMPs, the molecular basis of ST3 function in vivo remains unclear due to the lack of information on its physiological substrates. Furthermore, ST3 has only weak activities toward all tested ECM proteins. Using thyroid hormone-dependent Xenopus laevis metamorphosis as a model, we demonstrated previously that ST3 is important for apoptosis and tissue morphogenesis during intestinal remodeling. Here, we used yeast two-hybrid screen with mRNAs from metamorphosing tadpoles to identify potential substrate of ST3 during development. We thus isolated the 37 kd laminin receptor precursor (LR). We showed that LR binds to ST3 in vitro and can be cleaved by ST3 at two sites distinct from where other MMPs cleave. Through peptide sequencing, we determined that the two cleavage sites are in the extracellular domain between the transmembrane domain and laminin binding sequence. Furthermore, we demon strated that these cleavage sites are conserved in human LR. These results together with high levels of human LR and ST3 expression in carcinomas suggest that LR is a likely in vivo substrate of ST3 and that its cleavage by ST3 may alter cell-extracellular matrix interaction, thus, playing a role in mediating the effects of ST3 on cell fate and behavior ob- served during development and pathogenesis.

Dynamic Changes of β Tubulin during the Resumption of Meiosis of Mouse Oocyte

Morphological changes of tubulin during the resumption of meiosis in both mouse oocyte and fertilized egg were revealed by indirect immunofluorescent marking with monoclonal antibody against β tubulin. During germinal vesicle period (GV), tubulin was found to be distributed around the GV menibrane. With the disruption of GV membrane, microtubule complexes (MTCs) appeared in cytoplasm, first around GV membrane later to spread to other portions as well. Quantitative difference was noted among different oocytes. MTCs coexisted with spindlesformed by prometaphase tubulin, while metaphase tubulin polymerized into spindles and anaphase and telophase tubulin was concentrated in the two poles of the meiotic apparatus and the midbody. In egg arrested in the 2nd metaphase, whether maturing in vitro or vivo, all the tubulin went to form spindles with no MTCs left in the cytoplasm. After fertilization in vitro,MTCs reappeared in the egg cytoplasm activated by sperms while no MTCs could be revealed in cytoplasm after formation of pronucleus. As demonstrated by this experiment,cytoplasmic tubulin in eggs are polymerized chiefly into two forms: the star-shaped MTCs and the spindle.Cytoplasmic MTCs are the structure newly formed when the 1st and 2nd oocytes resumed meiosis. With colchicine disrupting the polymerization of tubulin, the maternal chromosomes,instead of orderly arrangement and orderly separation, either formed disordered mass or were divided into multiple chromatin masses. However, the penetration of sperm into egg, and decondensation and formation of pronuclei were not affected.

In vitro Oocyte Maturation in the Zebrafish Brachydanio rerio and Fertilization and Development of the Mature Egg

Maturation process of zebrafish oocyte was investigated using in vitro incubation.In medium EM-199 containing 0.5 μg/ml of 17α-hydroxyprogesterone incubated under 80％ O_2 and at 25°C,germinal vesicles(GV)of oocytes in stage Ⅳ migrated from midway between the center and theperiphery ofoocytes to the periphery in 40 minutes and the oocytes went into stage V.Half an hourlater,the oocytes underwent germinel vesicle breakdown(GVBD)with a breakdown rate of 59％.Two more hours were needed for such oocytes to complete their final maturation.The mature eggscould not come off from the follicle layer surrounding them by themselves(ovulation).By removingthe follicle and adding active sperms for insemination,we could make the mature eggs fertilized.Thechorion was elevated and blastoderm formed on the animal pole.The cleavage and development ofthese fertilized eggs followed the same course as the naturally matured and fertilized eggs.Usingblastula formation as a marker of successful fertilization of the in vitro matured egg,the fertilizationrate was 78％.This is the first report on the successful in vitro incubation of mature oocytes inzebrafish.The establishment of this in vitro oocyte maturation technology has laid the foundationfor further investigation of the transfer of foreign genes in the germinal vesicles of oocytes.

Vigorous,but differential mononuclear cell response of cirrhotic patients to bacterial ligands

AIM： To study the role of gram-positive and gram-negative bacteria in the pathogenesis of liver injury, specifically the activation of inflammatory mediators. METHODS： Peripheral blood mononuclear cells of 20 out-patients were studied, 10 of them with cirrhosis,Peripheral blood mononuclear cells were isolated and exposed to lipopolysaccharide or lipoteichoic acid. CD14, Toll-like receptor 2 and 4 expression was determined by flow cytometry, and tumor necrosis factor （TNF） α, interleukin （1L）-1β, IL-6, IL-12 and IL-10 secretion in su- pernatants was determined by ELISA. RESULTS： Higher CD14, Toll-like receptor 2 and 4 expression was observed in peripheral blood mononudear cells from cirrhotic patients, （P 〈 0.01, P 〈 0.006, P 〈 0.111） respectively. Lipopolysaccharide and lipoteichoic acid induced a further increase in CD14 expression （P 〈 0.111 lipopolysaccharide, P 〈 0.013 lipoteichoic acid）, and a decrease in Toll-like receptor 2 （P 〈 0.008 lipopolysaccharide, P 〈 0.008 lipoteichoic acid） and Toll-like receptor 4 （P 〈 0.008 lipopolysaccharide, P 〈 0.028 li- poteichoic acid） expression. With the exception of TNFα, absolute cytokine secretion of peripheral blood mononuclear cells was lower in cirrhotic patients under nonexposure conditions （P 〈 0.070 IL-6, P 〈 0.009 IL-1]5, P 〈 0.022 IL-12）. Once exposed to lipopolysaccharide or lipoteichoic acid, absolute cytokine secretion of peripheral blood mononuclear cells was similar in cirrhotic and non-cirrhotic patients, determining a more vigorous response in the former （P 〈 0.005 TNFα, IL-1β, IL-6, IL-2 and IL-10 lipopolysaccharide; P 〈 0.037 TNFα; P 〈 0.006 IL-113; P 〈 0.005 IL-6; P 〈 0.007 IL-12; P 〈 0.014 IL-10 lipoteichoic acid）. Response of peripheral blood mononuclear cells was more intense after lipopolysaccharide than after lipoteichoic acid exposure. CONCLUSION： Peripheral blood mononuclear cells of cirrhotic patients are able to respond to a sudden bacterial ligand exposure, particularly lipopolysaccharide, suggesting that immune regulation mechanisms are still present.

Stimulation of staphylococcal enterotoxin A combined with PML-RARα peptide on the specifical T-cells against NB4 cell line

Objective： To investigate the effects of staphyococcal enterotoxin A （SEA） on the cytotoxicity of T cells stimulated by PML-RARa peptide in vitro. Methods： Peripheral blood mononuclear cells （MNC） from healthy donor were obtained by density gradient centrifugation on Ficoll-Hypaque, MNC were cultured with PML-RARa peptide and SEA for 20 days. After induction, the cytotoxicity of T cells induced against NB4 and K562 cell lines were examined by Cell Counting Kit-8 （CCK-8）. The CD4 and CD8 surface markers on the harvested CD3^＋ T cells were detected by flow cytometry （FCM）. Results： The cytotoxicity of T cells induced by PML-RARa peptide with SEA was higher than that of T cells induced only by PML-RARa peptide against NB4 cells. The FCM assay showed that the ratio of CD4^＋/CD8^＋ T cells were gradually decreased in both groups of PML-RARα peptide whether with SEA or not at the intervals of day 5,10 and 20 day after induction, but the most significantly decreased by PML-RARe peptide with SEA. Conclusion： The specific cytotoxicity of T cells induced by PML-RARa peptide against NB4 cells could be enhanced with superantigen SEA.

Effect of electroacupuncture on oocyte quality and pregnancy of patients with PCOS undergoing in vitro fertilization and embryo transfer

Objective To observe the impacts of electroacupuncture （EA） on oocyte quality and pregnant outcome for the patients with polycystic ovary syndrome （PCOS） undergoing in vitro fertilization and embryo transfer （IVF-ET） and explore its potential mechanism. Methods Sixty-six patients with PCOS and undergoing IVF-ET were divided into two groups randomly, including an observation group （34 cases） and a control group （32 cases）. Ethinylestradiol and cyproterone acetate tablets and gonadotropin-releasing hormone agonist were administered for long-program superovulation in either group. In the observation group, the intervention of EA was applied to Guānyuán （关元 CV 4）, Zhōngjí （中极 CV 3）, Sānyīnjiāo （三阴交 SP 6）, Zǐgōng （子宫 EX-CA 1） and Tàixī （太溪 KI 3） additionally for 30 min, once daily, 1 menstrual cycle before controlled ovarian hyperstimulation （COH） and during COH. The pregnant outcome, evaluation of kidney deficiency syndrome, blood hormone level on the day of human chorionic gonadotropin injection and the concentrations of stem cell factor （SCF） in the serum and follicular fluid on the day of oocyte collection were compared between the two groups. Results The score of kidney deficiency symptoms was reduced remarkably after treatment in either group and the improvement in the observation group was superior to that in the control group （P0.01）. The fertilization rate [（76.25？±？20.33）% vs （66.34？±？15.44）%], cleavage rate [（98.66？±？3.70）% vs （94.47？±？9.45）%] and the rate of high-quality embryos [（60.20？±？22.20）% vs （50.55？±？16.15）%] in the observation group were all superior to those in the control group separately （all P0.05）. Clinical pregnancy rate （46.67%, 14/30） in the observation group was higher than that （37.93%, 11/29） in the control group, but without statistical difference （P0.05）. SCF concentrations in the serum and follicular fluid on the day of oocyte collection in the observation group were higher obviously than those in the control group （both P0.05）. Conclusion Electroacupuncture plays an active role in the pregnant outcomes of PCOS patients undergoing IVF-ET and it can relieve the symptoms of kidney deficiency in terms of TCM and improve clinical pregnant rate. The mechanism is relevant to the overall adjustment of organic endocrinal system and the local micro-environment of ovary and the improvement of oocyte quality through the up-regulation of SCF concentration.

Therapeutic potential of extracellular ATP and P2 receptors in nervous system diseases

Extracellular adenosine 5’-triphosphate(ATP) is a key signaling molecule present in the central nervous system(CNS),and now is receiving greater attention due to its role as a messenger in the CNS during different physiological and pathological events. ATP is released into the extracellular space through vesicular exocytosis or from damaged and dying cells. Once in the extracellular environment,ATP binds to the specific receptors termed P2,which mediate ATP effects and are present broadly in both neurons an...

Endostatin inhibits fibrosis by modulating the PDGFR/ERK signal pathway:an in vitro study

Accumulating evidence indicates that endostatin inhibits fibrosis. However, the mechanism is yet to be clarified. The aim of this study is to evaluate the effect of endostatin on platelet-derived growth factor-BB （PDGF-BB）- or transforming growth factor β1 （TGF-β1）-induced fibrosis in cultured human skin fibroblasts, and to further examine the molecular mechanisms involved. Human dermal flbroblasts were cultured in Dulbecco＇s modified Eagle＇s medium （DMEM） and serum-starved for 48 h before treatment. Cells were grouped as follows： ＂PDGF-BB＂, ＂PDGF-BB＋ endostatin＂, ＂TGF-β1＂, ＂TGF-β1＋endostatin＂, ＂endostatin＂, and ＂blank control＂. The fibroblasts were stimulated with either TGF-β1 or PDGF-BB for 72 h in order to set up the fibrosis model in vitro. The cells were co-cultured with either TGF-β1 or PDGF-BB and endostatin and were used to check the inhibiting effect of endostatin. A blank control group and an endostatin group were used as negative control groups. The biomarkers of fibrosis, including the expression of collagen I, hydrroxyproline, and α-smooth muscle actin （a-SMA）, were evaluated using an enzyme-linked immune- sorbent assay （ELISA） and Western blot. The expression of phosphorylated PDGF receptor β （p-PDGFRβ）, PDGFRβ, phosphorylated extracellular signal-regulated kinase （p-ERK）, and ERK was detected using Western blot and im- munofiuorescent staining was used to explore the mechanisms. Both PDGF-BB and TGF-β1 significantly up-regulated the expression of collagen I, hydroxyproline, and a-SMA. Endostatin significantly attenuated both the PDGF-BB- and TGF-β1-induced over-expression of collagen I, hydroxyproline, and a-SMA. PDGF-BB and TGF-β1 both promoted the expression of PDGFR, ERK, and p-ERK. Endostatin inhibited the expression of PDGFR and p-ERK but did not affect the expression of total ERK. Endostatin inhibited hypertrophic scar by modulating the PDGFRI3/ERK pathway. En- dostatin could be a promising multi-target drug in future fibrosis therapy.

Effect of Yikun Neiyi Wan on the Expression of Aromatase P450,COX-2,and ER Related Receptor in Endometrial Cells in Vitro from Patients with Endometriosis

To investigate the effect of Yikun Neiyi Wan （益坤内异丸YKNYW） and gestrinone on the expression of aromatase P450 （P450arom）, cyclo-oxygenase-2 （COX-2） and estrogen receptor （ER） in isolated ectopic and normal endometriat stroma cells in vitro. Methods： Digestion and serial filtration were used to isolate and culture the ectopic and eutopic endometrial cells from patients with chocolate cyst in virto Transformation of the cell morphology was observed in a inverted microscope. The effect of YKNYW on the expression of aromatase P450, cyclo-oxygenase-2, estrogen receptor in cultured endometriosis cells were detected by immunohistochemical method. Results： The expression levels of P450arom, COX-2 in glandular epithelium cells in vitro were decreased significantly by YKNYW compared with gestrinone （P〈0.05）. ER expression in mesenchymal cells of endometriosis was increased by YKNYW in the large and medium dosage groups compared with gestrinone. Conclusion： The mechanism by which YKNYW alleviates endometriosis pain is possibly related to the decrease in ectopic endometrial P450 arom and COX-2 expression in glandular epithelium, contrary to gestrinone, and the increase in ER expression in mesenchymalis, consistent with gestrione in patients with endometriosis.

A novel synthetic small molecule YF-452 inhibits tumor growth through antiangiogenesis by suppressing VEGF receptor 2 signaling

Tumor angiogenesis is characterized by abnormal vessel morphology, endowing tumor with highly hypoxia and unresponsive toward treatment. To date, mounting angiogenic factors have been discovered as therapeutic targets in antiangiogenic drug development. Among them, vascular endothelial growth factor receptor 2 （VEGFR2） inhibitors exerts potent antiangiogenic activity in tumor therapy. Therefore, it may provide a valid strategy for cancer treatment through targeting the tumor angiogenesis via VEGFR2 pathway. In this study, we established a high-profile compounds library and certificated a novel compound named N-（N-pyrrolidylacetyl）-9-（4-bromobenzyl）-l,3,4,9-tetrahydro-^-carboline （YF-452）, which remarkably inhibited the migration, invasion and tube-like structure formation of human umbilical vein endothelial cells （HUVECs） with little toxicity invitro. Rat thoracic aorta ring assay indicated that YF-452 significantly blocked the formation ofmicrovascular exvivo. In addition, YF-452 inhibited angiogenesis in chick chorioallantoic membrane （CAM） and mouse corneal micropocket assays. Moreover, YF-452 remarkably suppressed tumor growth in xenografts mice model. Furthermore, investigation of molecular mechanism revealed that YF-452 inhibited VEGF-induced phosphorylation of VEGFR2 kinase and the downstream protein kinases including extracellular signal regulated kinase （ERK）, focal adhesion kinase （FAK） and Src. These results indicate that YF-452 inhibits angiogenesis and may be a potential antiangiogenic drug candidate for cancer therapy.

Effects of acupuncture on ovarian blood supply and pregnancy outcomes in patients receiving assisted reproduction

To observe the effects of acupuncture at the follicular phase on ovarian blood supply and pregnancy outcomes in patients who received in vitro fertilization/intracytoplasmic sperm injection-embry transfer （IVF/ICSI-ET） of assisted reproductive technology （ART）. Methods： A total of 169 IVF/ICSI-ET female recipients from the Reproductive Center of Xiehe Hospital, Tongji Medical College, Huazhong University of Science ＆ Technology were randomized into an observation group （57 cases）, a placebo group （54 cases） and a control group （58 cases）. The observation group received acupuncture during the follicular phase, meanwhile the placebo group received placebo-acupuncture, and the control group did not receive acupuncture. The hemodynamic index, biochemical pregnancy rate and clinical pregnancy rate of each group were observed, respectively. Results： As to the ovarian arterial hemodynamic index, the pulsatility index （PI）, resistance index （RI）, and the systolic-to-diastolic peak velocity ratio （S/D） of the observation group were （0.819＋0.131）, （0.552＋0.055） and （2.306＋0.512）, respectively, obviously lower than those in the placebo group and the control group, and the differences were statistically significant （all P〈0.05）, but there were no statistically significant differences between the placebo group and the control group （all P〉0.05）. As to the biochemical pregnancy rate and clinical pregnancy rate, the biochemical pregnancy rate in the observation group was 64.9% and the clinical pregnancy rate was 52.6%, which were significantly higher than those in the placebo group and the control group, and the differences were statistically significant （all P〈0.05）, while there were no significant differences between the placebo group and the control group （both P〉0.05）. Conclusion： Acupuncture treatment during the follicular phase can improve ovarian blood supply and pregnancy rate in those receiving IVF/ICSI-ET.

Effect of soothing liver therapy on oocyte quality and growth differentiation factor-9 in patients undergoing in vitro fertilization and embryo transfer

OBJECTIVE:To investigate the effect of Soothing liver therapy on infertile women undergoing in vitro fertilization and embryo transfer(IVF-ET)and to explore its mechanism.METHODS:Fifty-eight women with tubal infertility were randomized into two groups:30 in an experimental group treated with Xiaoyao powder(Shugan)plus gonadotropin-releasing hormone analog(GnRHa)/follicle-stimulating hormone(FSH)/human chorionic gonadotropin(hCG)and 28 in the control group who were treated with GnRHa/FSH/hCG only.The total gonadotropin(Gn)doses required,endometrial thickness,oocyte numbers,high quality embryo production rate and pregnancy rate of the two groups were compared.The concentration of growth differentiation factor-9(GDF-9)in follicular fluid was detected by western blotting and the expression of GDF-9 mRNA in granulosa cells was measured using reverse tran-scription-polymerase chain reaction amplification.RESULTS:In the experimental group,the Gn dose was significantly lower than that in the control group;the endometrial thickness,high quality embryo production and pregnancy rates were significantly higher and the expression of GDF-9 mRNA was also significantly higher than in the control group(all P<0.05).CONCLUSION:Shugan treatment can improve the pregnancy rate of women with tubal infertility;its mechanism is possibly related to the increased expression of GDF-9 in granulosa cells.