Effect of Acupuncture on Endometrial Blood Flow in Women Undergoing in vitro Fertilization Embryo Transfer: A Single Blind, Randomized Controlled Trial.

OBJECTIVE: To evaluate the effect of manual acupuncture on endometrial blood flow parameters by three-dimensional (3D) power Doppler ultrasound in women undergoing in vitro fertilization embryo transfer (IVF-ET). METHODS: Seventy patients undergoing IVF-ET were equally randomized into traditional or sham acupuncture treatment group for totally 4 days (from the day of oocyte aspiration to the day of embryo transfer) of treatment by random envelope method at the Reproductive Medicine Center and Outpatient Department of Integrated Traditional Chinese and Western Medicine, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology from January 2013 to December 2015. Patients in the traditional acupuncture group accepted traditional acupuncture methods with manual acupuncture, and Zhongji (CV3), Qihai (CV 6), Sanyinjiao (SP6), Taichong (LR 3), Tianshu (ST 25), Guilai (ST 29) and Zusanli (ST 36) were chosen. Patients at the sham acupuncture group accepted shallow acupuncture methods at 4 non-meridian points at each shoulder and upper arm. Outcome measures included endometrial ultrasonic indices such as vascularization index (VI), flow index (FI) and vascularization flow index (VFI), endometrial thickness and volume, subendometrial VI (sVI), subendometrial FI (sFI), subendometrial VFI (sVFI), implantation rate, clinical pregnancy rate, abortion rate, live birth rate and number of live births. RESULTS: Finally, 34 patients in the traditional acupuncture group and 35 in the sham acupuncture group completed this trial. VI, FI and VFI of the traditional acupuncture group were significantly higher than those in the sham acupuncture group (P<0.05). No significant differences were found in endometrial thickness, endometrial volume, sVI, sFI, sVFI, implantation rate, clinical pregnancy rate, abortion rate, live birth rate and number of live births (P>0.05). CONCLUSIONS: Manual acupuncture performed after oocyte aspiration and before transplantation improved the endometrial blood flow parameters VI, RI and VFI in women who underwent IVF-ET, instead of sVI, sFI and sVFI. Therefore, acupuncture might be beneficial in women undergoing IVF-ET by increasing endometrial blood flow and endometrial receptivity. (Registration No. ChiCTR2100053354).

[Clinical studies on acupuncture assisting in vitro fertilization-embryo transfer: a review and thoughts].

Acupuncture has a long history of application in the field of reproduction, and in the past 20 years, numerous clinical studies have been conducted to explore the value of acupuncture in assisted reproductive technology. However, due to inconsistent research findings, there are still controversies over the clinical safety and efficacy of acupuncture in assisting in vitro fertilization-embryo transfer (IVF-ET), which may be closely associated with lacking of standardized treatment regimen in clinical trials of acupuncture. For that reason, this article reviews 16 representative clinical trials of acupuncture assisting IVF-ET published worldwide in 1999-2020, summarizes the grouping design, acupuncture method (acupuncture timing, acupuncture frequency, acupoints, and acupuncture types), and pregnancy outcome (pregnancy rate, clinical pregnancy rate, ongoing pregnancy rate, and live birth rate), and discusses several key links in clinical protocol design, so as to provide references for future clinical research.

Efficacy of Electro-acupuncture in Treatment of Functional Constipation: A Randomized Controlled Trial.

Functional constipation (FC) is a chronic disease that significantly affects the life quality of patients. Acupuncture has been used for the treatment of FC for many years, but its effectiveness has not been scientifically assessed. The present study aimed to evaluate the efficacy of electro-acupuncture (EA) in relieving the symptoms, mental states and quality of life (QOL) of FC patients. A total of 96 FC patients were randomly allocated into EA, mosapride & sham EA group (MS) and mosapride control group (MC). In the EA group, patients were treated with 16 sessions of needling at Quchi (LI11) and Shangjuxu (ST37) bilaterally, 5 times a week in the first 2 weeks, and 3 times a week in the last 2 weeks. In the MC group, patients were treated with 5 mg mosapride citrate three times a day for 4 weeks. In the MS group, patients underwent sham EA and the same mosapride citrate treatment as in the MC group. The primary outcome was the number of weekly spontaneous bowel movements (SBMs). The secondary outcomes included stool consistency, intensity of defecating difficulty, 36-Item Short-Form Health Survey (SF-36), Self-rating Anxiety Scale (SAS), Self-rating Depression Scale (SDS), and the validated Patient Assessment of Constipation-Quality of Life (PAC-QOL). The results showed that as compared with the baseline, EA significantly improved the weekly SBMs, stool consistency and intensity of defecating difficulty (P<0.05). It also partly ameliorated the PAC-QOL, SF-36, SDS and SAS scores when compared with MC or MS group (P<0.05). However, no significant difference was observed between MS and MC groups in bowel function outcomes and QOL scores. It was concluded that EA could effectively improve bowel function, mental states and QOL of FC patients.

Effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation implantation dysfunction.

The effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation (COH) implantation dysfunction were studied. The COH implantation dysfunction model in mice was established by intraperitoneal injection of 7.5 IU pregnant mare's serum gonadotrophin (PMSG), followed by 7.5 IU human chorionic gonadotrophin (HCG) 48 h later. Then the female mice were mated with male at a ratio of 2:1 in the same cage at 6:00 p.m. The female mice from normal group were injected intraperitoneally with normal saline and mated at the corresponding time. Day 1 of pregnancy was recorded by examining its vaginal smears at 8:00 a.m. of the next day. Fifty successfully pregnant mice were equally randomly divided into 5 groups: normal control pregnant group (NC), COH implantation dysfunction model group (COH), low dosage of Bu-Shen-An-Tai recipe group (LOW), middle dosage of Bu-Shen-An-Tai recipe group (MID) and high dosage of Bu-Shen-An-Tai recipe group (HIGH). Then from day 1, the mice in different groups were respectively intragastrically given corresponding treatments at 9:00 a.m. for 5 consecutive days. The concentrations of 17ß-estradiol (E2) and progesterone (P4) were determined by radioimmunoassay (RIA). The ultrastructural changes of ovarian tissues were observed by transmission electron microscope (TEM). The histopathological changes of ovarian tissues were observed by HE staining. The number of atretic follicles and pregnant corpus luteum were also recorded. TUNEL was applied to measure apoptotic cells of ovarian tissues. Western blotting was used to detect the protein expression of apoptosis- related factors like Bax, Bcl-2 and cleaved-caspase-3 in ovarian tissue of mice. The results showed that ovarian weight, the concentrations of E2 and P4, the number of atretic follicles and pregnant corpus luteum, as well as the apoptosis of granulosa cells were significantly increased in the COH group. The ultrastructures of ovarian tissues in the COH group showed that chromatin in granulosa cells was increased, agglutinated, aggregated or crescent-shaped. The focal cavitation and the typical apoptotic bodies could be seen in granulosa cells in the late stage of apoptosis. After the treatment with different doses of Bu-Shen-An-Tai recipe, the ultrastructural changes of ovarian granulosa cells apoptosis were dramatically improved and even disappeared under TEM. Visible mitochondria and mitochondrial cristae were increased and vacuoles were significantly reduced. The lipid dropltes were shown in a circluar or oval shape. The protein expression levels of Bax and cleaved-caspase-3 were decreased, and the expression of Bcl-2 protein was increased after treatment. It was concluded that Bu-Shen-An-Tai recipe can inhibit the apoptosis of ovarian granulosa cells, probably by up-regulating the protein expression of Bcl-2 and down-regulating Bax and cleaved-caspase-3, which contributes to the formation and maintenance of ovarian corpus luteum. It's helpful to promote the embryonic implantation, to reduce embryo loss and ultimately to improve the success rate of pregnancy.

Effect and mechanism of Bushen Huoxue recipe on ovarian reserve in mice with premature ovarian failure.

The aim of the present study was to explore the effect and mechanism of Bushen Huoxue recipe (BHR) on ovarian reserve in mice with premature ovarian failure (POF). Mice were divided into 3 groups: normal group, model group and BHR group. Intraperitoneal injection of cyclophosphamide was performed to create the POF model. Primordial follicular (PDF) number, ovarian wet weight, ovarian index, and estrous cycle were analyzed to evaluate the effect of BHR on POF. Meanwhile, the mRNA and protein level of Mouse Vasa Homologue (MVH) in the bone marrow, peripheral blood and ovary were detected, to explore the underlying mechanism of the treatment efficacy of BHR on ovarian reserve. By the time of BHR treatment for 28 days, BHR increased the PDF number and shortened the estrous cycle of POF mice. BHR also decreased the mRNA level of MVH in the bone marrow, and increased mRNA and protein level of MVH in the ovary of POF mice. Our results demonstrated a treatment efficacy of BHR on POF mice, and revealed that BHR might repair the dysfunction of germline stem cells in the bone marrow, and thus to improve the ovarian reserve and enhance the ovarian function of POF mice through neo-oogenesis.

[Effect of Guishen Pill on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve].

OBJECTIVE: To study the effect of Guishen Pill (GSP) on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve (DOR). METHODS: Totally 40 female C57BL/6J mice were randomly divided into 4 groups, the normal control group, the model group, the GSP group, and the dehydroepiandrosterone (DHEA) group, 10 in each group. Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG), and prostaglandin F2α (PGF2α) were sequentially administrated to produce superovulation. The DOR model was established by exposing to ozone inhalation. Mice in the GSP group were intragastrically administered with GSP at 0.3 mL. Those in the DHEA group were intragastrically administered with DHEA at 0.3 mL. Equal volume of normal saline was intragastrically administered to mice in the normal control group and the model group. All mice wer treated for 21 days. Serum levels of estrogen (E2), progestogen (P), and anti-Müllerian hormone (AMH) were measured by ELISA. Changes of Oct-4, anti-AMH, and early growth response gene-1 (Egr-1) mRNA in ovaries were dtected by Real-time PCR. RESULTS: Compared with the model group, serum levels of E2, P, and AMH, as well as contents of estrogen receptor (ER), progestogen receptor (PR), MVH, and Oct-4 mRNA significantly increased in the GSP group and the DHEA group (P < 0.05). CONCLUSION: GSP could improve expression levels of Oct-4, MVH, and Egr-1 mRNA in DOR mice and their ovarian function.

Effects of Bu-Shen-An-Tai recipe and its two components on endometrial morphology during peri-implantation in superovulated mice.

The aim of this study was to investigate the effect of Bu-Shen-An-Tai recipe (BSATR) and its two components (Bushen recipe, and Huoxue recipe) on endometrial morphology during peri-implantation in superovulated mice. Mice were randomly divided into five groups, including the normal (N), model (M), Bushen (BS), Huoxue (HX) and Bu-Shen-An-Tai (BH) groups. The uteri were collected on day 4 of pregnancy, and the endometrium thickness, microvessel density (MVD) and number of pinopodes observed. Compared with the M group, the endometrial thickness in the BS, HX and BH groups was significantly increased and there was a significant difference in endometrial thickness between the BS and the BH groups. The mean MVD was significantly lower in the M group than in the N group, and there was a significant increase in MVD in the BS, HX and BH groups as compared with the M group. Compared with the M group, the pinopode scores in the endometrium were significantly increased in the HX and BH groups; and the BS group had significantly higher pinipode scores than the HX and BH groups. In conclusion, the results of the present study demonstrated that the recipes (Bushen, Huoxue and BSATR) could improve the endometrial environment by regulating the endometrial thickness, MVD and the number of pinopodes at the window of implantation. Moreover, the Huoxue recipe and the BSATR were more efficient than the Bushen recipe, with the BSATR tending to have the most beneficial effects.

The effect of transcutaneous electrical acupoint stimulation on pregnancy rates in women undergoing in vitro fertilization: a study protocol for a randomized controlled trial.

BACKGROUND: The latest meta-analysis demonstrated that acupuncture improves pregnancy rates among women undergoing in vitro fertilization-embryo transfer (IVF-ET), and surface acupoint stimulation, such as transcutaneous electrical acupoint stimulation (TEAS), may have the same or better potential. METHODS/DESIGN: To explore the effect of TEAS on the clinical pregnancy rate (CPR) and live birth rate (LBR) compared with real acupuncture and controls in women undergoing IVF, a multicenter, randomized controlled trial will be conducted. The inclusion criteria are the following: infertile women <40 years of age undergoing a fresh IVF or intracytoplasmic sperm injection cycle, and the study will be restricted to women with the potential for a lower success rate as defined by two or more previous unsuccessful ETs (fresh or frozen). Those who have severe illnesses possibly precluding IVF or pregnancy, have FSH levels greater than 20 IU/L, received donor eggs, had been previously randomized for this study or had undergone acupuncture (in any modality) as infertility treatment will be excluded. The subjects will be randomly assigned to the TEAS group (IVF + TEAS), the electro-acupuncture (EA) group (IVF + EA), or the control group (only IVF). A total sample size of 2,220 women is required to detect differences in CPR among the three groups. TEAS or EA treatments will start once every two or three days from day 3 of menstruation in the ovarian stimulation cycle until the day of ET. The parameters of TEAS or EA will be the following: a frequency of 2/100 Hz, a moderate electrical current of 3 to 5 mA for TEAS and 0.8 to 1.0 mA for EA. The primary outcome is CPR. Secondary outcomes are LBR, the number of oocytes aspirated and the total gonadotropin dose used in the stimulation cycle. DISCUSSION: This study will provide significant evidence for using a new method (TEAS) in IVF. TRIAL REGISTRATION: ClinicalTrials.govID: NCT01608048 (05/24/2012).

Effect of acupuncture on CXCL8 receptors in rats suffering from embryo implantation failure.

To observe the effect of acupuncture on CXCL8 receptors (CXCR1 and CXCR2) in rat endometrium experiencing embryo implantation failure, 72 pregnant rats were randomly divided into four groups: normal group (N), embryo implantation failure group (M), acupuncture treatment group (A), and progestin treatment group (W). Then the rats in each group were equally randomized into a day-6 (D6) group, a day-8 (D8) group, and a day-10 (D10) group. The rats in group M, group A, and group W were treated with mifepristone-sesame oil solution on day 1, while the rats in group N were injected with the same amount of sesame oil. Meanwhile, "Housanli" and "Sanyinjiao" were selected for acupuncture. From day 1 to the time of death, the rats in group A were fastened up and then acupuncture was administered while the rats in group N and group M were only fixed, and the rats in group W were given progestin. The number of implanted embryos was calculated. The expression of CXCR1 and CXCR2 in rat endometrium was detected by immunohistochemistry, Western blotting and real-time PCR. Compared to group N, the average number of implanted embryos, the protein and mRNA expression of CXCR1 (D6, D8 and D10), and the protein and mRNA expression of CXCR2 (D8 and D10) in rat endometrium were significantly decreased in group M. Compared to group M, there was significant elevation in the average number of implanted embryos, the protein expression (D6, D8 and D10) and mRNA expression (D8) of CXCR1 in rat endometrium of group A, and the protein expression (D8 and D10) and mRNA expression (D8) of CXCR2 in rat endometrium of group W. These findings indicated that acupuncture can increase the number of implanted embryos in rats of embryo implantation failure, which may be relevant with up-regulation the expression of CXCR1 and CXCR2 at maternal-fetal interface of rats with embryo implantation failure.

Electro-acupuncture with different current intensities to treat functional constipation: a study protocol for a randomized controlled trial.

BACKGROUND: Functional constipation (FC) is highly prevalent in the general population of the world and has a substantial negative impact on the health-related quality of life of individuals. Many clinical trials have indicated that acupuncture is effective in the treatment of FC. However, the sample sizes of these previous studies were too small. Furthermore, there are no reports investigating the relationship between the stimulation parameter and the therapeutic effect. We therefore designed a multicenter randomized controlled trial to address these problems and hopefully provide a more conclusive answer to these questions. METHODS: Participants will be included if they meet all of the following conditions: (1) diagnosed with functional constipation according to the Roman III standard; (2) aged between 18 and 65 years; (3) not taking any drugs that promote gastrointestinal movements at least during the 1 week prior to randomization; (3) willing to sign an informed consent form; (4) willing to return to the study site for their study visits. The participants will be randomly assigned to three groups in a 1:1:1 ratio: high current intensity group, low current intensity group, and mosapride citrate control group. The total study period is 9 weeks for each patient, 1 week for baseline, 4 weeks for treatment, and 4 weeks for follow-up. The primary outcome in this trial is the number of defecating events per week. The secondary outcomes will include the shape and properties of the stool, intensity of defecating difficulty, Patient Assessment of Constipation Quality of Life (PAC-QOL), MOS item Short Form health survey (SF-36), Self-Rating Anxiety Scale (SAS), and Self-Rating Depression Scale (SDS). DISCUSSION: This study will provide significant evidence for the application of acupuncture in FC and will identify a suitable stimulation parameter for treatment. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT01274793.

The role of acupuncture in assisted reproductive technology.

The aim of this paper was to provide reliable evidence by performing a systematic review and meta-analysis for evaluating the role of acupuncture in assisted reproductive technology. All randomized controlled trials that evaluated the effects of acupuncture, including manual, electrical, and laser acupuncture (LA) techniques, on the clinical pregnancy rate (CPR) and live birth rate (LBR) of in vitro fertilization (IVF) or artificial insemination were included. The controlled groups consisted of no acupuncture and sham acupuncture groups. The sham acupuncture included sham acupuncture at acupoints, sham acupuncture at non- or inappropriate points, sham LA, and adhesive tapes. Twenty-three trials (a total of 5598 participants) were included in this paper. The pooled CPR from all acupuncture groups was significantly higher than that from all controlled groups, whereas the LBR was not significantly different between the two groups. However, the results were quite distinct when the type of control and/or different acupuncture times were examined in a sensitivity analysis. The results mainly indicate that acupuncture, especially around the time of the controlled ovarian hyperstimulation, improves pregnancy outcomes in women undergoing IVF. More positive effects from acupuncture in IVF can be expected if a more individualized acupuncture programs are used.

Effects of acupuncture on pregnancy rates in women undergoing in vitro fertilization: a systematic review and meta-analysis.

OBJECTIVE: To evaluate the effect of acupuncture on in vitro fertilization (IVF) outcomes. DESIGN: Systematic review and meta-analysis. PATIENT(S): Women undergoing IVF in randomized controlled trials (RCTs) who were evaluated for the effects of acupuncture on IVF outcomes. SETTING: Not applicable. INTERVENTION(S): The intervention groups used manual, electrical, and laser acupuncture techniques. The control groups consisted of no, sham, and placebo acupuncture. MAIN OUTCOME MEASURE(S): The major outcomes were clinical pregnancy rate (CPR) and live birth rate (LBR). Heterogeneity of the therapeutic effect was evaluated with a forest plot analysis. Publication bias was assessed by a funnel plot analysis. RESULT(S): Twenty-four trials (a total of 5,807 participants) were included in this review. There were no significant publication biases for most of the comparisons among these studies. The pooled CPR (23 studies) from all of the acupuncture groups was significantly greater than that from all of the control groups, whereas the LBR (6 studies) was not significantly different between the two groups. The results were different when the type of control was examined in a sensitivity analysis. The CPR and LBR differences between the acupuncture and control groups were more obvious when the studies using the Streitberger control were ignored. Similarly, if the underlying effects of the Streitberger control were excluded, the LBR results tended to be significant when the acupuncture was performed around the time of oocyte aspiration or controlled ovarian hyperstimulation. CONCLUSION(S): Acupuncture improves CPR and LBR among women undergoing IVF based on the results of studies that do not include the Streitberger control. The Streitberger control may not be an inactive control. More positive effects from using acupuncture in IVF can be expected if an appropriate control and more reasonable acupuncture programs are used.

De-qi, not psychological factors, determines the therapeutic efficacy of acupuncture treatment for primary dysmenorrhea.

OBJECTIVE: To study the impact of De-qi (, obtaining qi) and psychological factors on the efficacy of acupuncture treatment for primary dysmenorrhea, with an attempt to explore the relationship among De-qi, psychological factors, and clinical efficacy. METHODS: The patients with primary dysmenorrhea were randomly assigned to a group of acupuncture with manual manipulation (manipulation group, n=67) and an acupuncture group without manipulation (non-manipulation group, n=64). Pain intensity and pain duration were used as measures for evaluating the therapeutic efficacy of the acupuncture treatment. De-qi, the sensations a patient experienced during the acupuncture treatment, was scored on a 4-point scale by the subjects. In addition, the psychological factors, including belief in acupuncture, the level of nervousness, anxiety, and depression, were quantitatively assessed. The personality of the subject was assessed using the Eysenck personality questionnaire (EPQ) and 16 personality factor questionnaire (16PF). RESULTS: Complete data were obtained from 120 patients, 60 patients in each group. There were statistically significant differences in pain intensity (W=2410.0, P<0.01) and pain duration (W=3181.0, P<0.01) between the two groups. The number of De-qi acupoints (W=1150.5, P<0.01) and the average intensity of De-qi (W=1141.0, P<0.01) were significantly higher in the manipulation group as compared with their non-manipulation counterparts. The correlation coefficients between De-qi and therapeutic efficacy of acupuncture were greater than those between psychological factors and therapeutic efficacy. CONCLUSIONS: Compared with the psychological factors, De-qi contributed more to the pain-relieving effect of acupuncture in subjects with primary dysmenorrhea. Moreover, manual manipulation is a prerequisite for eliciting and enhancing the De-qi sensations, and De-qi is critical for achieving therapeutic effects.

[Effect of bushen antai recipe and its various compositions on endometrial sexual hormones and receptors in mice with blastocyst implantation dysfunction].

OBJECTIVE: To compare the effects and mechanisms of Bushen Antai Recipe (BAR) and its different compositions, Bushen Recipe (BSR, the Shen-nourishing part) and Huoxue Recipe (HXR, the blood-activating part) on the endometrial receptivity in mice with blastocyst implantation dysfunction (BID). METHODS: Model mice of BID induced by indomethacin were treated respectively with BAR, BSR and HXR from the first day of pregnancy, and killed on day 5 or 6. Samples of their serum and uterine were collected for detecting serum estradiol (E2), progesterone (P) contents using radioimmunoassay; and endometrial expression levels of their receptors, ER and PR, using immunohistochemistry. RESULTS: Serum levels of E2 and P, and endometrial expression of ER and PR increased significantly on day 6 in mice after treated by BAR, but in those treated by BSR and HXR, the improvements were significantly lesser. CONCLUSION: BAR, by combined application of both Shen-nourishing and blood-activating methods, could impact E2, P, ER and PR to improve the endometrial receptivity to a higher extent in BID mice so as to promote embryo implantation.

[Effects of gap junction blocking on the oxygen partial pressure in acupoints of the bladder meridian].

OBJECTIVE: To explore the relation between gap junction and meridian phenomenon. METHODS: The oxygen partial pressure in acupoints [see text for formula] and in their corresponding non-acupoints of the Bladder Meridian was observed with the needle-type tissue oxygen tension sensor in the gap junction blocking goats by 1-Heptanol injection and the Connexin 43 (Cx43) gene knockout mice. RESULTS: (1) The oxygen partial pressure in acupoints of Bladder Meridian on goats was higher than that in non-acupoints after 1-Heptanol injection with significant differences between them (both P < 0.01). (2) The oxygen partial pressure in acupoints of Bladder Meridian on goats increased significantly after injecting 1-Heptanol as compare with that either injecting normal saline or injecting nothing with significant differences between them (all P < 0.01). (3) The oxygen partial pressure in acupoints of the Bladder Meridian was significantly higher than that in the non-acupoint controls in Cx43 wild type (WT) mice (all P < 0.01). In Cx43 heterozygote (HT) mice, the oxygen partial pressure between acupoints and non-acupoint controls showed no significant differences (all P > 0.05). (4) In acupoints, the oxygen partial pressure in Cx43 WT mice was significantly higher than that in Cx43 HT mice (all P < 0.05), while in the corresponding non-acupoints, this difference had no statistically significant (all P > 0.05). CONCLUSION: Gap junction maybe the essential factor in signal transduction of acupuncture.

[Effects of acupuncture on the luteal function of rats with dysfunctional embryo implantation].

OBJECTIVE: To observe the influence of acupuncture on the luteal function of rats with dysfunctional embryo implantation, and investigate its mechanism. METHODS: The early pregnant female rats were randomly divided into a normal control group, a model group, an acupoint group and a non-acupoint group. The model rats with dysfunctional embryo implantation were established with Mifepristone. For acupuncture treatment, bilateral "Housanli" (ST 36) and "Sanyinjiao" (SP 6) were selected in the acupoint group, while the points around the acupoint were used in the non-acupoint group. In this experiment, the levels of luteinizing hormone (LH), estrodiol (Es) and progesterone (P) in serum were tested with radioimmunity method, the expression of vascular endothelial growth factor (VEGF) in ovary was examined with immunohistochemical assay and Western-blot, and also, the mRNA expression of VEGF and luteinizing hormone receptor (LHR) in ovary were detected with RT-PCR. RESULTS: The levels of LH and P in serum, as well as the expression of VEGF, VEGF mRNA and LHR mRNA in ovary in the acupoint group were significantly higher than those in the model group and the non-acupoint group (P < 0.05), but there was no obvious difference from that of the normal control group. CONCLUSION: Acupuncturing at "Housanli" (ST 36) and "Sanyinjiao" (SP 6) can increase the levels of LH and P in the serum, and up-regulate the expression of VEGF, VEGF mRNA and LHR mRNA in ovary, which may enhance the luteal function of rats with dysfunctional embryo implantation and improve its embryo implantation.

[Effect of connexin 43 knockout on acupuncture-induced down-regulation of c-fos expression in spinal dorsal horn in visceral pain mice].

OBJECTIVE: To explore the effect of connexin 43 gene knockout on acupuncture analgesia. METHODS: Seventy-two wide type (WT) and connexin 43 gene knockout mice were separately and randomly divided into: WT control group, WT model group, WT acupuncture group, heterozygous (HT) control group, HT model group and HT acupuncture group, with 12 cases in each. Visceral pain model was established by intraperitoneal administration of acetic acid. "Zhongwan" (CV 12) and bilateral "Zusanli" (ST 36) were punctured with a filiform needle for 30 min and stimulated by manipulating the needle 30 s every 5 min. The expression of c-fos in the spinal dorsal horn was assayed by using RT-PCR and western blot techniques. RESULTS: There was no significant difference between HT and WT control mice in relative grey value of spinal c-fos mRNA expression (P>0.05), in which few c-fos mRNA and protein expressed. The expression of c-fos mRNA and protein was increased significantly following intraperitoneal acetic acid injection compared with control groups in both HT and WT mice (P<0.01). And no significant difference was found between HT and WT model groups in c-fos mRNA expression (P>0.05). Compared with WT model group, the expression of both c-fos mRNA and c-fos protein in WT acupuncture group was down-regulated significantly (P<0.01). In comparison with HT model group, the expression of both c-fos mRNA and protein in HT acupuncture group was down-regulated but without statistically significant difference (P>0.05). And the expression of c-fos mRNA and protein in HT acupuncture group was significantly higher than that in WT acupuncture group (P<0.05, 0.01). CONCLUSION: Acupuncture has a marked antinociceptive effect in visceral pain mice, and simultaneously suppresses the expression of c-fos mRNA and protein evoked by noxious stimulation in the spinal dorsal horn. Connexin 43 gene knockout may weaken acupuncture analgesia and reduce EA-induced down-regulation of c-fos expression, suggesting an involvement of connexin 43 in the analgesic effect of acupuncture.

[Effect of Bushen Antai Recipe on prostaglandin I2 expression and its nuclear receptor at implantation site in mice with blastocyst implantation dysfunction].

OBJECTIVE: To observe the effect of Bushen Antai Recipe (BAR) on expression of prostaglandin I2 (PGI2) and its nuclear receptor peroxisome proliferators-activated receptor delta (PPARdelta) at implantation site in mice with blastocyst implantation dysfunction. METHODS: Pregnant mice were divided into three groups randomly, the normal group, the model group and the BAR group. The pregnant uterus of all mice was cut off on the 4th (D4), 5th (D5), 6th (D6) and 8th (D8) day of pregnancy for determining the PGI2 expression with radio immunoassay; and the mRNA and protein expression of PPARdelta with RT-PCR and immunohistochemistry at implantation site. RESULTS: PGI2 expression in the model group was obviously lower than that in the normal group (P < 0.01), and also lower than that in the BAR group (P < 0.01), while the index was insignificantly different between the normal and the BAR group. Compared with the normal group, the expression of PPARdelta in the model group was delayed temporally and spatially (P < 0.05), while that in the BAR group was not significantly different. CONCLUSION: BAR can improve the implantation in mice with blastocyst implantation dysfunction through promoting the PGI2 expression and its nuclear receptor PPARdelta at implantation site.

[Influence of connexin 43 gene knockout on the analgesic effect of acupuncture in visceral pain mice].

OBJECTIVE: To explore the possible central and peripheral mechanisms of connexin 43 gene knockout in acupuncture analgesia. METHODS: Fifty-four wide type (WT) mice and 54 connexin 43 gene knockout (heterozygote, HT) mice were randomly divided into WT control group, WT model group, WT acupuncture group, HT control group, HT model group and HT acupuncture group (n = 18/group). Visceral pain model was established by intraperitoneal injection of 0.6% acetic acid (0. 1 mL/10 g). "Zhongwan" (CV 12) and bilateral "Zusanli" (ST 36) were punctured with filiform needles and stimulated for 30 min by manipulating the needle for 30 s every 5 min. The latency and the number of body-writhing response were observed and the contents of beta-endorphin (beta-EP) in hypothalamus and serum prostaglandin E2 (PGE2) were detected with radioimmunoassay (RIA). RESULTS: There was no significant difference in the latency and the number of body-writhing response, and contents of hypothalamic beta-EP and serum PGE2 between HT and WT control groups (P > 0.05). Compared with the corresponding control groups, the latency of body-writhing in WT model and HT model groups shortened significantly and the number of body- writhing increased considerably in two model groups (P < 0.01). While in comparison with WT model group, the latency of body-writhing prolonged significantly and the number of body-writhing decreased apparently in WT acupuncture group (P < 0.01); but no marked differences were found between HT model and HT acupuncture groups in these two indexes (P > 0.05). Compared with the corresponding control groups, the contents of both beta-EP and PGE2 increased obviously in WT model and HT model groups (P < 0.05). In comparison with WT model group, beta-EP levels in WT acupuncture group increased further significantly (P < 0.05), and serum PGE2 in WT acupuncture group decreased obviously (P < 0.05); but no significant changes were found between HT model and HT acupuncture groups in beta-EP and PGE2 levels (P > 0.05). CONCLUSION: Acupuncture has a marked antinociceptive effect in WT mice with visceral pain, which may be related to its effects in increasing hypothalamic beta-EP and decreasing serum PGE2; while in connexin 43 gene knockout mice, all the above-mentioned effects of acupuncture are eliminated, indicating an important role of connexin 43 in the analgesic effect of acupuncture.

[Effect of Ca2+ complexation on oxygen partial pressure in the Bladder Meridian in goats].

OBJECTIVE: To explore the relationship between energy metabolism and meridian phenomenon. Methods Ten Boer hybridism goats mitigated under chlorpromazine (0.85 mg/kg, intramuscular injection) were subjected to the present study. Changes of oxygen partial pressure (PO2) in "Ganshu" (BL 18), "Dachangshu" (BL 25) and "Guanyuanshu" (BL 26) of Bladder Meridian and in 3 non-acupoints (3 cm apart from BL18, BL25 and BL26 separately on the unilateral side) were recorded by using a needle-type tissue oxygen tension sensor before and following local injection of EDTA-Na2 (200 microL, 0.05 kg/mol, for complexing Ca2+) and 0.9% saline. RESULTS: (1) Compared with the corresponding non-acupoints, Ca2+ potentials in BL18, BL25 and BL26 were all significantly higher (P < 0.05, 0.01). (2) In comparison with the corresponding non-acupoints, the values of tissue PO2 of BL18 and BL26 were significantly higher (P < 0.01). (3) After injection of EDTA-Na2, PO2 values in both BL18, BL26, and their respective non-acupoints increased significantly in comparison with those of either injection of saline or their respective basal values (P < 0.01, 0.05). Compared with those of their corresponding non-acupoints PO2 values in both BL18 and BL26 increased markedly (P < 0.01). CONCLUSION: Ca2+ potential of the Bladder Meridian was significantly higher than that of non-acupoints, and after complexing Ca2+, PO2 in acupoints and non-acupoints increased markedly, suggesting that Ca2+ may play an important role in meridian signaling and production of acupuncture effect.