OBJECTIVE: To observe the effect of "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique on the reproductive function of the mice with poor ovarian response (POR) and explore the molecular mechanism of acupuncture on early embryos after in vitro fertilization-embryo transfer (IVF-ET) in POR mice. METHODS: Of 70 female C57BL/6 mice, 60 mice with regular estrous cycle were screened and 30 of them were randomly divided into a blank group, a model group, and an acupuncture group, 10 mice in each one. Mice in the model and acupuncture groups were administered with tripterygium glycosides suspension (50 mg·kg^-1·d^-1) via gavage for 2 weeks to prepare POR models; while in the blank group, the mice received an equal volume of 0.9% sodium chloride solution via gavage. After successful modeling, mice in the acupuncture group underwent "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique, once daily, for 20 min each time and lasting 2 consecutive weeks. After intervention completion, subsequently, all the three groups underwent ovulation induction, orbital blood collection, and ovary extraction and fresh denuded oocyte collection. Denuded oocytes, after incubated, together with the sperms from 15 male C57BL/6 mice, were transferred into the oviducts of 30 donor pseudopregnant C57BL/6 female mice via IVF-ET; and the embryonic tissue was collected on day 7 of implantation. After successful modeling and intervention completion, the general conditions of mice in each group were observed, and the estrous cycle disorder rate was compared among the groups. After intervention completion, the numbers of oocyte, ovarian wet weight and ovarian index were recorded in each group; the levels of anti-Müllerian hormone (AMH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E2) in the serum were measured using ELISA; and ovarian morphology and mitochondrial ultrastructure were examined using HE staining and transmission electron microscopy. The transcriptome sequencing technology was employed to identify differentially expressed genes (DEGs) in early embryos of each group, followed by bioinformatics analysis; and the reversed DEGs with significant difference were verified using quantitative real-time PCR (qRT-PCR). RESULTS: After intervention, compared with the blank group, the mice in the model group presented poor spirits and declined water and food intake, reduced activity, the higher rate of estrous cycle disorder (P< 0.01); the decrease of the numbers of oocyte, ovarian wet weight, and ovarian index (P<0.01), reduced AMH and E2 in the serum (P<0.01), elevated FSH and LH in the serum (P<0.01); more atresia follicles in ovarian tissue, disorganized granulosa cells, reduced the numbers of mitochondria, vacuoles appeared and cristae breakage. When compared with the model group, in the acupuncture group, the spirits of mice, food intake and activity were improved, the rate of estrous cycle disorder was reduced (P<0.01); the numbers of oocyte, ovarian wet weight, and ovarian index were higher (P<0.01, P<0.05), the levels of AMH and E2 in the serum increased (P<0.01), the levels of FSH and LH in the serum decreased (P<0.01); the atresia follicles were reduced, the numbers of follicle increased at all levels, the numbers of mitochondria got higher, vacuoles and cristae breakage were declined. In comparison between the blank group and the model group, and between the model group and the acupuncture group, a total of 258 intersecting DEGs were identified, and 194 DEGs of them exhibited a trend of reversion before and after intervention. The top 5 reversed DEGs with significant difference included dexamethasone-induced Ras-related protein 1 (Rasd1), gene regulated by estrogen in breast cancer 1 (Greb1), leucine-rich repeat-containing G protein-coupled receptor 6 (Lgr6), Fraser syndrome 1 (Fras1), and apolipoprotein D (Apod). Compared with the blank group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues decreased in the model group (P<0.01, P<0.05); and when compared with the model group, the mRNA expression of Rasd1, Greb1, Lgr6, Fras1 and Apod in embryonic tissues increased in the acupuncture group (P<0.05, P<0.01). These findings were consistent with the DEGs screening results. Gene ontology (GO) analysis revealed that the reversed genes of co-expression were primarily participated in the biological processes such as myoblast differentiation, endocardial cushion development, and cardiac morphogenesis. Kyoto encyclopedia of genes and genomes (KEGG) analysis indicated that the reversed genes of co-expression are enriched in the Wnt signaling pathway, and they were associated with various types of cardiac diseases. CONCLUSION The "Zhibian" (BL54)-to-"Shuidao" (ST28) needling technique may ameliorate the decline of ovarian response in mice with POR, promote normal follicle development and ovulation, thereby improve embryo quality and reduce the risk of developmental defects and deformity in the organs such as the embryonic heart. The underlying mechanism may be related to the regulation of the expression pattern of embryonic key genes through the Wnt signaling pathway.
Animals ; Female ; Mice ; Mice, Inbred C57BL ; Acupuncture Therapy ; Ovary/physiopathology* ; Humans ; Fertilization in Vitro ; Transcriptome ; Embryo Transfer ; Infertility, Female/physiopathology*

The impact of acupuncture for the pregnancy of in vitro fertilization-embryo transfer (IVF-ET) is discussed in the paper. Nowadays there are various conclusions about the impact of acupuncture for IVF-ET, and it may result from the differences in research designs. The effect is closely related to the demographic and clinical characteristics of subjects, such as age, the diagnosis of barrenness, blood flow index of uterine spiral arteries, the cycle of IVF, etc. Besides, the efficacy is influenced by treatment based on syndrome differentiation or not, the frequency and course of acupuncture in both the treating group and the control group, etc. If more reasonable design is achieved in the further study based on them, more reliable evidence will be provided for the effect and mechanism of the pregnancy of IVF-EF by acupuncture.
Acupuncture Therapy ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; physiopathology ; therapy ; Male ; Pregnancy ; Pregnancy Rate ; Randomized Controlled Trials as Topic ; Treatment Outcome

OBJECTIVETo observe the effects of "menstrual cycle-based acupuncture therapy" on ovarian function and pregnancy results of in vitro fertilization-embryo transfer (IVF-ET) in patients with decline in ovarian reserve (DOR).

METHODSA total of 63 patients of DOR who received treatment of IVF/intracytoplasmic sperm injection (ICSI) were randomly divided into an observation group (30 cases) and a control group (33 cases). The patients in the observation group were treated with "menstrual cycle-based acupuncture therapy". The syndrome differentiation and treatment were given based on different phases of menstruation. Shiqizhui (EX-B 8) and Mingmen (GV 4) were selected during menstrual phase, Shenshu (BL 23), Geshu (BL 17), Sanyinjiao (SP 6) and Taixi (KI 3) were selected after menstruation, Qihai (CV 6), Guanyuan (CV 4), Zigong (EX-CA 1), Zusanli (ST 36) were selected during ovulatory period, Qihai (CV 6), Guanyuan (CV 4), Yanglingquan (GB 34), Taichong (LR 3) were selected before menstruation. The acupuncture was given twice a week until second menstrual cycle of oocyte retrieval. The total times of acupuncture was (15 ± 2). After acupuncture, patients were treated with IVF-ET. The patients in the control group were treated with IVF-ET but no acupuncture. The indices of ovarian reserve function, including basic follicle-stimulating hormone (FSH), estradiol (E2), antral follicle count (AFC), number of retrieved oocytes, number of fertilization and number of high quality embryo, were compared and analyzed before and after acupuncture in the observation group. The differences of outcomes of IVF-ET, including the cycle cancellation rate, implantation rate, the clinical pregnancy rate, were compared between the two groups.

RESULTSCompared before acupuncture, the E2, AFC, number of retrieved oocytes, number of high quality embrgo and number of fertilization were all increased after acupuncture in the observation group (all P< 0. 05). Compared with the control group, levels of the E2, the number of retrieved oocytes, number of fertilization and number of high quality embryo were all increased in the observation group (all P < 0.05). Also, the implantation rate, the clinical pregnancy rate were improved (both P < 0.01) and cycle cancellation rate was reduced (P< 0.01).

CONCLUSIONThe "menstrual cycle-based acupuncture therapy" can effectively improve the ovarian reserve function in DOR patients, leading to an improved clinical pregnancy rate of IVF-ET.

Acupuncture Therapy ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; physiopathology ; therapy ; Menstrual Cycle ; Middle Aged ; Ovarian Reserve ; Ovary ; physiopathology ; Pregnancy ; Young Adult

OBJECTIVESTo investigate the clinical effect of sequential therapeutic intervention Yupei Qisun [compensating for weakness by invigorating Kidney (Shen) and Spleen (Pi) in advance] in Chinese medicine (CM) and hysteroscopic endometrial mechanical stimulation on the treatment of infertile patients with repeated implantation failure (RIF); and to study the differences in patients' endometrial thickness and type on the day of embryo transfer, serum hormone levels on embryo transfer day and clinical pregnancy outcomes.

METHODSIn the clinical study, 168 frozen-thawed embryo transfer (FET) cycles for couples with RIF conforming to the research protocol were randomly divided into three groups: a CM group with 56 cycles (CM combined with FET), a hysteroscopy group with 55 cycles (hysteroscopic endometrial mechanical stimulation), and a control group with 57 cycles (conventional FET). Differences in endometrial thickness on the embryo transfer day, levels of serum estradiol (E2) and progesterone (P) on the embryo transfer day, the E2/P ratio on the embryo transfer day, biochemical and clinical pregnancy rates, implantation rate, abnormal pregnancy rate and other indices were compared among the three groups.

RESULTSEndometrial thickness, E2 and P levels, and the E2/P ratio on embryo transfer day and other factors had no significant differences among groups. The biochemical pregnancy, clinical pregnancy, and implantation rates of the CM and hysteroscopy groups were significantly higher than the control group (P<0.05), and there were no significant differences between these two groups. The abnormal pregnancy rate had no significant difference among the three groups.

CONCLUSIONSSequential therapy of Yupei Qisun could significantly improve the clinical outcomes of rif-fet cycles, being equivalent to hysteroscopic endometrial mechanical stimulation, and provided a reliable method to treat such infertile couples.

Abortion, Habitual ; therapy ; Adult ; Embryo Implantation ; Embryo Loss ; therapy ; Embryo Transfer ; Endometrium ; pathology ; physiopathology ; Female ; Humans ; Hysteroscopy ; Infertility, Female ; pathology ; therapy ; Medicine, Chinese Traditional ; methods ; Physical Stimulation ; methods ; Pregnancy ; Retreatment ; statistics & numerical data

Dingkun Dan is a representative of the classic gynecological medicine. With Tonifying the liver and kidney, supplementing Qi and nourishing blood, regulating menstruation Shuyu, promoting the role of pain. Used in treatment of liver and kidney deficiency, deficiency of both qi and blood, Qi stagnation and blood stasis caused by irregular menstruation, menstrual pain, uterine bleeding, leukorrhea with reddish discharge, bruise blood removal, infertility, and various postpartum deficiency and bone steaming hot flashes of gynecological common disease. In recent years, Dingkun Dan's new uses have been reported, the clinical application value is worth further digging. In this paper, clinical application and research progress of Dingkun Dan since the founding of new China were briefly discussed, and summarize for the randomized controlled trials.
China ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Infertility, Female ; drug therapy ; physiopathology ; Kidney ; drug effects ; physiopathology ; Liver ; drug effects ; physiopathology ; Menstruation ; drug effects ; Randomized Controlled Trials as Topic

OBJECTIVEThe multi-center, randomized, double-blind, double-simulated and positive-control trial was used to verify the contribution degree of Bushen Huoxue for the treatment of ovulatory dysfunction caused infertility, which provided scientific basis for clinical treatment.

METHODAccording to diagnostic, inclusion and exclusion criteria, we observed 349 patients which were divided into the treated group (n = 177, treated with Bushen Huoxue ricipe) and control group (n = 172, treated with clomiphene). Ovulation rate, pregnancy rate, clinical effective rate of traditional Chinese medicine, endometrium and diameter of dominant follicle were observed. Serum reproductive endocrine hormones were assayed before and after treatment.

RESULTThe treated group showed ovulation rate of 69.34%, with pregnancy rate of 41.35%. The clinical effective rate of treated group and control group were 91.73% and 80.77%. There was remarkable difference in endometrium (P < 0.05) and remarkbale difference in sex hormones PRL and E₂in treated group at prior-treatment and post-treatment (P < 0.05). No adverse effects were found in the experiment. Security indicators did not show abnormal change.

CONCLUSIONThe comparison between the two groups showed that the treated group was significantly different from control group in the pregnancy rate (P < 0.05), without notable difference in ovulation rate. There was significant difference in clinical effective rate between the treated group and control group. Both the two groups could contribute to the mature development and discharge of the follicles. The growth of endometrium and endometrial receptivity in the treated group were higher than control group. The treated group has regulatory effect on PRL and E₂.

Adult ; Double-Blind Method ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Fertility Agents, Female ; administration & dosage ; Humans ; Infertility, Female ; drug therapy ; physiopathology ; Ovary ; drug effects ; physiopathology ; Ovulation ; drug effects ; Pregnancy ; Treatment Outcome ; Young Adult

High quality of fertility and pregancy is a specific presentation of family planning in this new historic environment. Our country adopted two-child policy from November, 2013. This symbolized an adjustive stage of reproduction policy in 21st century. It is a major adjustment in the development of national population. As a contry of tremendous population, it is very important to have a high- quality fertility and pregancy for the future of the whole nation, which would improve population quality and restrict development of population. The morbidity of infertility has increased significantly, which brings a huge crisis for national population. Under the guidance of high-quality fertility and pregancy in Chinese medicine theory, improving fertile women's ability of high-quality fertility and pregancy, and follow-up studying is of vital importance for evaluating the treatment of abortion prevention by traditional Chinese medicine and human health of high-quality fertility and pregancy.
Adult ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Follow-Up Studies ; Humans ; Infertility, Female ; drug therapy ; physiopathology ; Kidney ; drug effects ; physiopathology ; Medicine, Chinese Traditional ; Pregnancy ; Pregnancy Outcome ; Reproduction

OBJECTIVETo explore the impact and effect mechanism of electroacupuncture (EA) on oocyte quali ty in the patients with infertility of kidney deficiency pattern.

METHODSSixty-six cases differentiated as kidney de ficiency and with in vitro fertilization-embryo transplantation (IVF-ET), aged fromnt 35 to 42 years were rando- mized into an observation group and a control group, 33 cases in each one. The IVF-ET therapy of the long proto- col with gonadotrophin releasing hormone agonist was adopted in the two groups. In the observation group. on the 5th day of menstruation in IVF cycle, EA was applied to Sanyinjiao (SP 6). Zigong (EX CA 1), Zhongji (CV 3) and Guanyuan (CV 4). In the control group, the sham-acupuncture was applied to the same acupoints. The treatment was given once every two days till the date of egg collection and the needles were retained for 30 min each time. The change in the score of kidney deficiency syndrome, the high-quality oocyte rate, the high-quality embryo rate and clinical pregnant rate were observed in the two groups. The levels of insulin-like growth factor-i (IGF-1) and IGF-2 in follicular fluid and the serum β-endorphin β-EP) on the date of egg collection and the correlation with oocyte quality were compared bIetween the two groups.

RESULTS1) In the observation group, the kidney deficiency syndrome score after treatment was reduced apparently as compared with that before treatment (P<0. 05), the score after treatment in the observation group was reduced much more apparently as compared with the control group (P<0.05). 2) The high-quality egg rate and the high-quality embryo rate in the observation group were both higher than those in the control group [81.3% (161/198) vs 57.6% (98/170), 59.8% (58/97) vs 37.7% (26/69), both P<0.05]. 3) Compared with the control group. the levels of IGF-1 and IGF-2 in follicular fluid and serum β-EP on the day of egg collection were all increased obviously in the observation group (all P<0. 05). 4) The levels of IGF-1 and IGF-2 in follicular fluid and serum β-EP presented the linear positive correlation with the high-quality egg rate.

CONCLUSIONEA effectively improves the expressions of IGF in follicular fluid and serum β-EP, increases the high-quality egg rate and high-quality embryo rate and relieves the symptoms of kidney deficiency.

Acupuncture Points ; Adult ; Electroacupuncture ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; metabolism ; physiopathology ; therapy ; Insulin-Like Growth Factor I ; metabolism ; Kidney ; physiopathology ; Oocytes ; Pregnancy ; beta-Endorphin ; metabolism

OBJECTIVETo observe the impacts on endometrial and pregnancy outcomes treated with acupuncture and moxibustion in the patients of in vitro fertilization-embryo transfer (IVF-ET) and explore the application value, of acupuncture and moxibustion in IVF-ET treatment.

METHODSOne hundred and fourteen patients of IVF-ET treated with standard long-term program at luteal phase were randomized into an observation group and a control group, 57 cases in each one. In the observation group, at the beginning of ovulatory induction, moxibustion was applied to Shenque (CV 8) and acupuncture was to Zhongji (CV 3), Guanyuan (CV 4), Qihai (CV 6), Zigong (EX-CA 1), Xuebai (SP 10), etc. till the transfer time for one session of treatment. Totally, 3 sessions were required. In the control group, no intervention of acupuncture and moxibustion was applied. The endometrial morphology, subendometrial blood flow index, the levels of serum estradiol (E2), progesterone (P) and luteinizing hormone (LH) on the day of injection of human chorionic gonadotropin (hCG), the dosage and time of gonadotropin (Gn), oocyte count, high-quality embryo number, embryo cultivation rate and clinical pregnant rate were observed in the two groups.

RESULTSThe A type endometrial proportion on hCG day and high-quality embryo rate in the observation group were higher than those in the control group, indicating the significant differences (both P< 0.05). The difference in endometrial thickness on hCG day was not significant between the two groups (P> 0.05). In the observation group, endometrial hemodynamic index (peak systolic blood velocity/end-diastolic blood velocity, S/D), resistive index (RI) and pulse index (PI) were lower than those in the control group (P<0. 01, P<0. 05). The levels of serum E2 and P on hCG day in the observation group were higher than those in the control group (both P<0. 05). The differences were not significant in Gn dose, Gn medication time, numbers of follicles >1. 6 cm on hCG day, oocyte count, embryo cultivation rate and clinical pregnancy rate and LH level on hCG day between the two groups (all P>0. 05).

CONCLUSIONIn IVF-ET treatment, acupuncture and moxibustion affect estrogen level on hCG day, improve high-quality embryo rate, endometrial blood flow state and morphology so that the endometrial receptivity is increased and the method is expected to be the assistant therapeutic approach for the improvement of IVF-ET outcome.

Acupuncture Therapy ; Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; physiopathology ; therapy ; Moxibustion ; Pregnancy ; Pregnancy Outcome ; Young Adult

With the ascending attack rate of anovulatory infertility year by year, people also began to pay attention to its treat methods. According to Doctor Ma Kun,who are engaged in clinical work about the treatment for anovulatory infertility, kidney deficiency is the basic pathogenesis and blood stasis is an important factor that has been through. Flexible use of tonifying the kidney and promoting blood circulation treatment of anovulatory infertility in clinic, has achieved remarkable curative effect. Director Ma adjusts menstruation by the different periods, and regulates both patients' negative emotions and sleep quality. Through years of clinical experience accumulation, Director Ma gradually formes special treatment of anovulatory infertility by flexibly using of tonifying the kidney and promoting blood circulation individually.
Adult ; Anovulation ; drug therapy ; physiopathology ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Infertility, Female ; drug therapy ; physiopathology ; Kidney ; drug effects ; physiopathology