Study on Efficient in Vitro Embryo Production Technology of Lambs

The aim was to compare the effects of hormone treatments on the ovari- an follicular development and in vitro embryo development between 5-week and 13- week old lambs. FSH and pregnant mare serum gonadotropin （PMSG） treatment were used to compare the number of 2-8 mm follicles in the ovary and the in vitro embryo development. The results showed that： the 5-week old lambs had larger fol- licular numbers and collected oocytes numbers （58.6±1.9, 47.4±4.2） than the 13- week old lambs （14.4±4.1, 11.6±1.7）, and the difference was significant （P〈0.05）. However, there was no significant difference in the cleavage rate （60.0±1.9%, 61.6± 5.2%） and blastocyst rate （27.4±2.1%, 28.2±3.7%） of the fertilized oocytes between the 5-week and 13-week old lambs.

Kidding after Transfer of in Vitro Produced Saanen Goat Embryos into Local Ukrainian Breed Recipients in Different Seasons

In recent years, the demand for goat products has been growing due to the fact that goat milk has a number of advantages over cow milk, for example, it is low in lactose, and is considered less allergenic and easier to digest. To increase production during both breeding and non-breeding seasons and reduce the price of dairy products, it is necessary to effectively use reproductive management and assisted reproductive technologies. In vitro embryo production makes it possible to obtain a large number of eggs from goats, which for some reason are unable to conceive, but have genetic value. Afterward in vitro produced embryos can be transferred into recipient goats of other less genetically valuable breeds, such as the Ukrainian local breed. Therefore, the aim of the present study was to investigate the effectiveness of transfers of in vitro produced embryos of Saanen goats into surrogate sires of the Ukrainian local breed in different seasons. All manipulations with animals were carried out following ethical standards (Strasbourg, 1986). Six Saanen goats were selected as the oocyte donors. After the hormonal stimulation oocytes were retrieved by laparoscopic ovum pick-up. In vitro produced embryos were transferred laparotomically into 24 recipients of Ukrainian local breed. Fifty days after embryo transfers, pregnancies were determined by ultrasound diagnostics. Although the embryo development rate in the breeding season was 20% higher than in the non-breeding season, there was no difference in pregnancy and kidding rates between seasons. In conclusion, the transfer of in vitro produced Saanen goat embryos to recipients of the Ukrainian local breed gives the opportunity to achieve pregnancy and kidding regardless of the breeding season, which will enable a faster and more efficient increase in the livestock of highly productive goats in Ukraine in the post-war period.

Effect of Mitochondrial Function of Ovarian Granulosa Cells on In Vitro Fertilization and Embryo Transfer Outcomes in Obese Polycystic Ovary Syndrome Patients

Objective:To investigate the effect of abnormal ovarian granulosa cell metabolism on in vitro fertilization and embryo transfer(IVF-ET)outcomes in obese polycystic ovary syndrome(PCOS)patients.Methods:Patients with PCOS who met the study criteria were screened according to the inclusion criteria.A total of 32 patients with obese PCOS were recruited into the study group,and 39 patients with non-obese PCOS were recruited into the control group.The general data(age,body mass index,and years of infertility),insulin resistance index(HOMA-IR),follicle-stimulating hormone(FSH),luteinizing hormone(LH),granulosa cell mitochondrial function,and IVF-ET outcome of patients in the study group and control group were retrospectively analyzed.Results:The differences in age and years of infertility between the study group and the control group were insignificant(P>0.05),and the body mass index(BMI)of the study group and control group was 30.5±1.24 kg/m2 and 22.3±1.12 kg/m2,respectively,in which the difference was statistically significant(P<0.05);the HOMA-IR of the study group was significantly higher than that of the control group(P<0.05);the reactive oxygen species(ROS)in the study group was significantly higher than that in the control group(P<0.05),and the ATP content in the study group was significantly lower than that in the control group(P<0.05);comparing the FSH and LH levels between the two groups,the difference was not statistically significant(P>0.05);the rate of IVF-ET failure was significantly higher in the study group than in the control group.Conclusion:PCOS is a complex endocrine disorder,and obesity is one of the independent risk factors for the development of PCOS.

Reproductive fluids,used for the in vitro production of pig embryos,result in healthy offspring and avoid aberrant placental expression of PEG3 and LUM

Background:In vitro embryo production(IVP)and embryo transfer(ET)are two very common assisted reproductive technologies(ART)in human and cattle.However,in pig,the combination of either procedures,or even their use separately,is still considered suboptimal due to the low efficiency of IVP plus the difficulty of performing ET in the long and contorted uterus of the sow.In addition,the potential impact of these two ART on the health of the offspring is unknown.We investigated here if the use of a modified IVP system,with natural reproductive fluids(RF)as supplements to the culture media,combined with a minimally invasive surgery to perform ET,affects the output of the own IVP system as well as the reproductive performance of the mother and placental molecular traits.Results:The blastocyst rates obtained by both in vitro systems,conventional(C-IVP)and modified(RF-IVP),were similar.Pregnancy and farrowing rates were also similar.However,when compared to in vivo control(artificial insemination,AI),litter sizes of both IVP groups were lower,while placental efficiency was higher in AI than in RF-IVP.Gene expression studies revealed aberrant expression levels for PEG3 and LUM in placental tissue for C-IVP group when compared to AI,but not for RF-IVP group.Conclusions:The use of reproductive fluids as additives for the culture media in pig IVP does not improve reproductive performance of recipient mothers but could mitigate the impact of artificial procedures in the offspring.

Production of early monozygotic twin bovine embryos in vitro by the blastomere separation and coculture technique

The objective of this study was to establish an efficient system of producing early monozygotic twin bovine embryos in vitro using the blastomere separation and coculture technique. In this study, early eight-cell embryos were chosen to optimize the separation method, and multi-coculture tactics were applied to improve the efficiency of this production system. Bovine embryo blastomeres（groups of at least 30 at the eight-cell stage） were separated into eight segments（to regard an eight-cell embryo as a tangerine, a blastomere as one segment） and one, two and four segments（blastomeres） were cultured respectively in microwells on the bottom of the four-well dish（Nunc, Denmark） with 400 μL of culture medium under paraffin oil. Four different types of coculture tactics（cocultured with nothing, intact embryos, bovine cumulus cells（b CCs）, intact embryos ＆ b CCs） were applied to the group of four segments（blastomeres）. Finally, diameter and inner cell mass（ICM）：trophectoderm（TE） cell ratio was measured as a criterion to assess the quality of the twin embryos which were derived from bovine separated blastomeres. Our results showed that rate of blastocyst formation of the four segments group was significantly greater than one or two group（P〈0.05）. In addition, rate of blastocyst formation was significantly increased when the four segments were cocultured with intact embryo ＆ b CCs（P〈0.05）. Although the ICM, TE and total cells of blastocysts derived from separated blastomeres was less than the control group from intact embryo（P〈0.05）, more important quality indicator of the blastocyst diameter and ICM：TE cell ratio was similar between our experimental group and the control group（P〉0.05）. Thus, these results suggest that combined with intact embryos ＆ b CCs coculture system, culturing four isolated segments（blastomeres） per microwell is an efficient system of producing early monozygotic twin bovine embryos. Furthermore, our results also indicate that the quality of blastocysts derived from separated blastomere may be similar to those derived from intact eight-cell embryos.

Immature Embryo Rescue and in Vitro Development Evaluation of Intraspecific Hybrids from Brazilian Seedless Grapevine“Superior×Thompson”Clones

The fruit production for export is an economically significant activity in the Valley of S?o Francisco River, especially in the irrigated lands of Petrolina-PE/Juazeiro-BA, Brazil. The development of new genetic material most suitable to the tropical climate and the demands of the consumer market have led to the selection of new seedless grapes cultivars. In this case, the use of the embryo rescue technique has produced satisfactory results for obtaining such materials, especially in the semiarid region. This study aimed to evaluate the in vitro development of intraspecific hybrids of grapevine (Vitis vinifera L.), derived from the rescue of immature embryos resultant from the crossing of “Superior Seedless” and “Thompson Seedless” Brazilian clones. To establish and develop the cultivation, the culture media was supplemented with 30 g/L sucrose, 0.1 g/L myo-inositol, 0.002 g/L glycine, 0.1 mg/L indoleacetic acid (IAA), 6.5 g/L of agar, adjusted pH to 5.7. The experiment was evaluated after 90 days. The variables measured were: number of nodes, number of leaves, plant height (cm), number of roots and length (cm) of the root system and internodes. The period of 60 days of in vitro culture of ovules resulted in the highest values of embryos (about 50%), as well as better characterized developmental stages with higher germination (47.3%). The three types of hybrid grapes evaluated in micropropagation showed very similar values of the measured parameters, even having originated from embryos of different developmental stages.

Supplementation of L-ascorbic acid improves the in vitro development of buffalo(Bubalus bubalis)embryos and alters the expression of apoptosis-related genes

Objective:To study the effect of L-ascorbic acid supplementation on the in vitro development of buffalo embryos and evaluate the relative mRNA abundance of some pro-apoptotic,anti-apoptotic,and embryonic development-related genes.Methods:In experiment 1,we evaluated the effect of the addition of 0(control),50,and 100μM L-ascorbic acid to the in vitro maturation medium on the developmental competence in terms of blastocyst rate and relative mRNA abundance of some pro-apoptotic(BAX,BID),anti-apoptotic(BCL-XL,MCL1),and embryonic development(GDF9,BMP15)related genes.Based on the results,we chose 50μM as the suitable dose of L-ascorbic acid for the subsequent experiments.We further evaluated the blastocyst rates following the addition of 50μM L-ascorbic acid to the in vitro culture medium(experiment 2),and in vitro maturation and in vitro culture media(experiment 3).In all three experiments,the maturation and culture media devoid of L-ascorbic acid served as the control group.Results:The blastocyst rate after adding 50μM L-ascorbic acid to the in vitro maturation medium was significantly higher than the control group(P<0.05),whereas 100μM L-ascorbic acid exhibited a negative effect on the blastocyst rate.The blastocyst rates for embryos cultured in 50μM L-ascorbic acid in the in vitro culture medium alone and both in vitro maturation and in vitro culture media were significantly higher than their corresponding control groups(P<0.05).The relative mRNA abundance of BAX significantly decreased in blastocysts produced after the addition of 50μM L-ascorbic acid as compared with the control group(P<0.05),whereas,for MCL1,it significantly decreased in blastocysts produced after the addition of 100μM L-ascorbic acid(P<0.05).Conclusions:The supplementation of 50μM L-ascorbic acid to in vitro maturation and in vitro culture media supports in vitro embryonic development in buffaloes by improving developmental competence and altering the expression of apoptosis-related genes.

Post-salpingectomy interstitial heterotopic pregnancy after in vitro fertilization and embryo transfer:A case report

BACKGROUND Heterotopic pregnancy(HP)refers to the coexistence of ectopic pregnancy and intrauterine pregnancy.Salpingectomy is proposed as a pretreatment before in vitro fertilization and embryo transfer(IVF-ET)to reduce the risk of HP.HP after IVF-ET occurs in women who had already underwent bilateral salpingectomy,even though it is extremely rare.CASE SUMMARY A case of a 29-year-old woman with recurrent interstitial HP after IVF-ET following salpingectomy is presented.The main symptom was a sudden and worsening pelvic pain.Physical examinations revealed signs of peritoneal bleeding and irritation with stable vital signs.Transvaginal ultrasound showed a live intrauterine pregnancy and another live embryo with cardiac activity in the left cornu extending beyond the lateral edge of the uterus.Her hemoglobin concentration was 8.0 g/dL,and serum human chorionic gonadotropin value was 171116.9 mIU/mL.With the diagnosis of ruptured HP with internal bleeding,an emergency laparoscopic resection of left cornu was performed.The interstitial pregnancy was removed with caution to protect the intrauterine pregnancy.After the surgical treatment,the intrauterine pregnancy continued with no complications.A healthy baby was delivered by caesarean section at 39 wk.Outcomes of another three cases are further summarized.CONCLUSION Post-salpingectomy HP is a rare but challenging condition.Surgical treatment is preferred in the case with a viable intrauterine pregnancy.

Clinical Effects of the Follicular Phase Long Regimen and Luteal Phase Long Regimen on Ovulation Induction in IVF-ET Treatment: A Meta-Analysis

Objective:To systematically evaluate the clinical effects of the follicular phase long regimen and the luteal phase long regimen on ovulation induction in IVF-ET treatment.Methods:Databases including PubMed,Embase,Cochrane Library,CNKI,Chinese Biomedical Literature(CBM),VIP,Wanfang,and others were searched up to January 2021.Clinical studies on ovulation induction using the follicular phase long regimen and luteal phase long regimen in IVF-ET treatment were identified.Literature screening,data extraction,and quality evaluation were conducted based on inclusion and exclusion criteria.Meta-analysis was performed using RevMan 5.3 software.Results:After screening,a total of 11 studies were included,comprising 21,544 patients:9,974 in the follicular phase long regimen group and 11,570 in the luteal phase long regimen group.The meta-analysis results were as follows:(1)The number of Gn days and the total amount of Gn in the follicular phase long regimen were higher than those in the luteal phase long regimen(P<0.05);(2)The number of eggs obtained in the follicular phase long regimen was higher than that in the luteal phase long regimen(P<0.05).There were no significant differences in the rate of embryo optimization and cycle cancellation between the two groups(P>0.05);(3)The embryo implantation rate and clinical pregnancy rate in the follicular phase long regimen were higher than those in the luteal phase long regimen(P<0.05),while the abortion rate in the follicular phase long regimen was lower than that in the luteal phase long regimen(P<0.05).Conclusion:Compared to the luteal phase long regimen,the follicular phase long regimen involves more Gn days and a higher total amount of Gn.The optimal embryo rate and cycle cancellation rate were similar between the regimens,but the follicular phase long regimen resulted in more eggs,significantly improved the implantation and clinical pregnancy rates,and reduced the abortion rate.However,these conclusions require further validation through more multicenter,large-sample RCT studies.

Double Pulmonary Embolism and Left Common Iliac Vein Thrombosis after in vitro fertilization and embryo transfer： a Case Report

This article discusses the mechanism, diagnosis, and treatment of a case of double pulmonary embolism and left common iliac vein thrombosis following in vitro fertilization （IVF） and embryo transfer （ET） to arouse vigilance for unexpected thrombosis after ovarian hyperstimulation and to serve as a clinical reference of this condition. This paper reports the case of a woman with double pulmonary embolism and left common iliac vein thrombosis after IVF-ET and the successful management of this condition. The woman had primary infertility, and underwent IVF-ET with 14 ooeytes recovered and two embryos transferred. The patient suffered breathing difficulty 17 days after the ET, and was hospitalized 6 days later as her condition had exacerbated. Computed tomographic （CT） angiography of pulmonary arteries showed double pulmonary embolism, left common iliae vein thrombosis, abnormal density shadow in uterine appendages, and ascites. A healthy baby boy and girl were safely delivered by Caesarean section after successful treatment. Thrombosis after IVF-ET is an uncommon but life-threatening complication and concurrent pulmonary embolism is even rarer. It is suggested that careful thrombosis risk assessment be taken before therapy and particular attention be paid to patients with special body constitutions to develop thrombosis. Whenever patients experience breathing difficulty after IVF-ET, it is strongly advised to examine the possibility of pulmonary embolism. The main treatments for thrombosis are anticoagulant therapy by low-molecular-weight heparin and thrombolysis by urokinase.

Reproductive Outcomes in Women with Prior Cesarean Section Undergoing In Vitro Fertilization:A Retrospective Case-control Study

The impact of prior cesarean section（CS） on the pregnancy and neonatal outcomes of in vitro fertilization and embryo transfer（IVF-ET） was investigated. A retrospective analysis was performed on 144 patients with prior CS between January 2013 and December 2015. The pregnancy, delivery, and neonatal outcomes of patients who had previous CS delivery and received IVF-ET were analyzed. The control group comprised 166 patients who had only previous vaginal delivery（VD） and received IVF-ET during the same period. The results showed that the basal follicle stimulating hormone level, estradiol level on human chorionic gonadotropin（h CG） day, gonadotrophin dosage, duration of stimulation, retrieved oocytes, fertilization rate, high-quality embryo rate, multiple birth rate, abortion rate and ectopic pregnancy rate had no significant difference between the two groups（P〉0.05）. The pregnancy rate（40.28% vs. 54.22%） and implantation rate（24.01% vs. 34.67%） were significantly lower（P〈0.05）, and the ratio of embryo difficulty transfer（9/144 vs. 0/166） was significantly higher in CS group than in VD group. The risk of pernicious placenta previa and postpartum hemorrhage in twin deliveries was significantly increased in CS group as compared with that in VD group（P〈0.05）, and gestational age and neonatal birth weight were significantly reduced in twin deliveries as compared with singleton deliveries in both groups（P〈0.05）. It was suggested that the existence of CS scar may impact embryo implantation and clinical pregnancy outcome, and increase the difficulty of ET. We should limit the number of transfer embryos to avoid multiple pregnancies and strengthen gestational supervision in patients with cesarean scar.

Nerve growth factor promotes in vitro proliferation of neural stem cells from tree shrews

Neural stem cells promote neuronal regeneration and repair of brain tissue after injury,but have limited resources and proliferative ability in vivo.We hypothesized that nerve growth factor would promote in vitro proliferation of neural stem cells derived from the tree shrews,a primate-like mammal that has been proposed as an alternative to primates in biomedical translational research.We cultured neural stem cells from the hippocampus of tree shrews at embryonic day 38,and added nerve growth factor（100 μg/L） to the culture medium.Neural stem cells from the hippocampus of tree shrews cultured without nerve growth factor were used as controls.After 3 days,fluorescence microscopy after DAPI and nestin staining revealed that the number of neurospheres and DAPI/nestin-positive cells was markedly greater in the nerve growth factor-treated cells than in control cells.These findings demonstrate that nerve growth factor promotes the proliferation of neural stem cells derived from tree shrews.

Embryo culture is an efficient way to conserve a medicinally important endangered forest tree species Strychnos potatorum

The present study reports a protocol for germination of Strychnos potatorum （ver. Tel. Chilla） using zygotic embryo culture as an embryo rescue method. A 100% germination rate was obtained by culturing the embryos on full-strength Murashige and Skoog＇s medium （MS） containing 20 g/L sucrose in comparison to McCown and Lloyd＇s Woody Plant Medium （WPM）. Germination rates decreased when the sucrose concentration was lower or higher than 20 g·L-1 . WPM/MS medium containing glucose at levels 30, 20, 15 g·L-1 showed a smaller percentage of germination and at quarter strength, WPM/MS medium with glucose did not respond. Multiple shoot formation was found at 1.0 2.0 mg/L BAP; 3.0 mg/L Kn; 2.0 mg/L TDZ on MS medium with 20 g·L-1 sucrose. Germination rates improved when the embryos were placed upright （vertically） in the medium. The in vitro germinated seedlings were acclimatized in a walk-in-chamber and maintained in the green house with the survival rate of 65% 75%. These plants were transferred to the field and were found to be phenotypically normal, healthy and similar to donor plants. This protocol will be useful to overcome seed dormancy and for rapid multiplication and conservation of S. potatorum using zygotic embryo culture.

Assessment of the Quality of Frozen-Thawed Semen or Epididymal Sperm in Three Native Vietnamese Pig Breeds

Our aim was to evaluate the quality of ejaculated and epididymal frozen-thawed pig sperm of endangered Vietnam native pig breeds. Ejaculated sperm was collected from live boars and epididymal sperm was collected from slaughtered boars of the MuongTe, Kieng Sat and Co BinhThuan breeds and frozen in 0.25 ml straws using a protocol established earlier for modern pig breeds. We evaluated the sperm quality after thawing in terms of motility and rates of viable and abnormal spermatozoa. Our results revealed that the sperm motility and rates of viable and abnormal frozen-thawed sperm were >30%, >44%, and <14%, respectively. The origin of sperm had an effect on the production of pig embryos in vitro. In the Co BinhThuan breed, ejaculated sperm generated higher cleavage, blastocyst and hatching rates than did the epididymal sperm (60.11% vs 56.02%, 17.23% vs 14.31%, 3.78% vs 2.34%, respectively, P < 0.05). Although no difference in cleavage rate, blastocyst formation rate and the average number of cells/blastocysts, the hatching blastocyst rate was different between the breeds (P > 0.05). In the Co BinhThuan breed, the rate of pregnancy of ejaculated groups was similar to that of the epididymal group. In conclusion, the ejaculated and epididymal sperm of native Vietnamese pigs were successfully frozen. We succeeded in creating embryos in vitro and pregnant pigs after artificial insemination from frozen-thawed semen in three native Vietnamese pig breeds for the first time. The use of the ejaculated sperm improved the production of native pig embryos in vitro efficiency.

Cross Breeding of Populus and Its Hybrids for Cold Resistance

Populus tomentosa was crossed with P.tremuloidis, P.grandidentata, P.alba×P.grandidentata and P.alba×Ulmuspumila in order to maintain its rapid growth and high wood quality and improve its resistance to cold. Two methods were used to increase the germination rate from 1.5% to 41.1% and the remaining rate from 1.7% to 44.2%. Forty crossing combinations were conducted and 2744 hybrid seedlings were obtained. MX4×P.grandidentata （G-1-58）, MX3×P.tremuloidis （T-44-60）, MX2×P.tremuloidis （1-13-87-37） and MX2×（P.alba×P.grandidentata） were regarded as superior combinations after analysis and selection. Thirty seedlings of these combinations and 11 triploid seedlings identified by counting their chromosomes were selected as super plants.

Effect of Progestin-primed Ovarian Stimulation Protocol on Outcomes of Aged Infertile Women Who Failed to Get Pregnant in the First IVF/ ICSI Cycle： A Self-controlled Study

This study aimed to explore the outcomes of progestin-primed ovarian stimulation protocol （PPOS） in aged infertile women who failed to get pregnant in the first IVF/ICSI-ET cycles with GnRH-a long protocol. A self-controlled study was conducted to retrospectively investigate the clinical outcomes of 104 aged infertile patients who didn＇t get pregnant in the first IVF/ICSI-ET treatment by stimulating with GnRH-a long protocol （non-PPOS group）, and underwent PPOS protocol （PPOS group） in the second cycle between January 2016 and December 2016 in the Center for Reproductive Medicine, Renmin Hospital of Wuhan University. The primary outcomes included clinical pregnancy rate of frozen-thawed embryos transfer （FET） in PPOS group, and good-quality embryo rate in both groups. The secondary outcomes were fertilization rate, egg utilization rate and cycle cancellation rate. The results showed that there were no significant differences in basal follicle stimulating hormone （bFSH）, antral follicle count （AFC）, duration and total dosage of gonadotropin （Gn）, number of oocytes retrieved, intracytoplasmic sperm injection （ICSI） rate, fertilization rate, and cycle cancellation rate between the two groups （P〉0.05）. However, the oocyte utilization rate and good-quality embryo rate in PPOS group were significantly higher than those in non-PPOS group （P〈0.05）. By the end of April 2017, 62 FET cycles were conducted in PPOS group. The clinical pregnancy rate and embryo implantation rate were 22.58% and 12.70%, respectively. In conclusion, PPOS protocol may provide better clinical outcomes by improving the oocyte utilization rate and good-quality embryo rate for aged infertile patients who failed to get pregnant in the first IVF/ ICSI-ET cycles.

Comparison between a GnRH Agonist and a GnRH Antagonist Protocol for the Same Patient Undergoing IVF

In order to compare GnRH agonist with antagonist protocol for the same patient during controlled ovarian stimulation cycles, the in vitro fertilization and embryo transfer （IVF-ET） outcome was retrospectively studied in 81 patients undergoing 105 agonist protocols and 88 antagonist protocols. The results showed that there was no statistically significant difference in duration of ovarian stimulation, number of ampoules, oocytes retrieved, serum estradiol （E2） and progesterone （P） levels, thickness of endometrium, the zygote- and blastocyst-development rate between GnRH agonist and antagonist protocols （P〉0.05）. High quality embryo rate was higher in antagonist protocols, but there was no significant difference between two protocols. Implantation rate and clinical pregnant rate were significantly higher in antagonist protocol （15.82% and 30.26%, respectively） than in agonist protocol （5.26% and 10.64% respectively （P〈0.05）. It was concluded GnRH antagonist protocol probably improved the outcome of pregnancy of older patients with a history of multiple failure of IVF-ET in a GnRH protocol.

Professor Qi Cong’ s therapeutic strategy of traditional Chinese medicine combined with different stages of assisted reproduction technology

The success rate of in vitro fertilization and embryo transfer is only about 28.5%.The combination of traditional Chinese medicine and assisted reproduction technology can effectively improve the pregnancy rate,which has good application prospect.However,there is no relevant record in traditional Chinese medicine.Unified diagnosis and treatment system has yet been established.According to many years of clinical experience,Professor Qi Cong summarized the treatment strategy of traditional Chinese medicine.Treatment of invigorating the spleen to nourish blood,reinforcing the spleen and kidney,tonifying the kidney and activating blood circulation,enriching the kidney to prevent miscarriage,cooling blood to stop bleeding are given based on the characteristics of each stage and patients'symptoms during the transplantation.Disease differentiation matters during intermittent stage.Besides,she advocated treating husband and wife together.All of the above are characterized by Chinese medicine concept of pre cultivating the loss,treating the same disease with different methods,and three categories of etiologic factors.

Curative effect of assisted reproduction technology by Traditional Chinese Medicine multi-channel interventional therapy on 95 cases of in vitro fertilization and embryo transfer failure

OBJECTIVE: To evaluate the curative effect of Traditional Chinese Medicine(TCM) multi-channel interventional therapy on women with Assisted Reproductive Technology(ART) failure, to compare the curative effect of the dual therapy and triple therapy on women with ART Failure, and to choose the best TCM interventional therapeutic plan.METHODS: The 95 cases with ART Failure from West China second University Hospital of Sichuan University meeting the inclusion criteria were randomly divided into three groups:dual therapy group(31 cases), triple therapy group(33 cases)and control group(31 cases). According to the intervene treatment of in vitro Fertilization and Embryo Transfer(IVF-ET) long cycle scheme, the control group wait naturally for 3 months before IVFET. The dual therapy group take TCM prescriptionⅡ of cultivating emotion and assisting reproduction and auricular acupoint therapy for 3 months before IVF-ET, then Western Medicine treatment progestin supporting as well as auricular application and Antai Recipe after IVF-ET transplantation.The triple therapy group take TCM prescription Ⅱof cultivated emotion and assisted reproduction,auricular acupoint therapy and retention enema of TCM, and combination treatment the same as dual therapy group after transplantation. The natural pregnancy number, the period condition of secondary IVF-ET and the improvement of the kidney deficiency, liver depression and blood stasis syndrome among those three groups were compared.RESULTS: It was showed from analysis in 95 cases with ART failure that the number of natural preg-nancy was as followings: 3 patients from the dual therapy group, 10 patients from the triple therapy group, and no patient from the control group. The comparison among three groups have statistical significance(P < 0.05). The treatment group is superior to the control group, while the triple therapy is superior to the dual therapy. The comparison of the condition of the fertility rate, clinical pregnancy rate, biochemical pregnancy rate and early abortion rate during the period of secondary IVF-ET between pre-therapy and post-treatment of both the dual therapy group and the triple therapy group have statistical significance(P < 0.05). The comparison of the improvement of the kidney deficiency,liver depression and blood stasis syndrome between pre-therapy and post-treatment of both the dual therapy group and the triple therapy group have statistical significance(P < 0.05). The comparison between three groups after treatment have statistical significance(P < 0.05).CONCLUSION: TCM multi-channel interventional therapy can increase the natural pregnancy rate of patients with ART Failure(the triple therapy is superior to the dual therapy); it can increase the fertility rate, clinical pregnancy rate, and decrease the early abortion rate during the period of secondary IVF-ET; it can improve syndromes of kidney deficiency, liver depression and blood stasis.

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.