Erratum: Moxibustion improves ovarian function based on the regulation of the androgen balance.

[This corrects the article DOI: 10.3892/etm.2021.10664.].

Moxibustion improves ovarian function based on the regulation of the androgen balance.

The effect of androgens on follicular development and female reproduction has become an active research topic. Moxibustion is a Traditional Chinese Medicine therapy that has been reported to be able to prevent and treat numerous ovary-related problems. However, studies on the effect of moxibustion for diminished ovarian reserve (DOR) on androgen balance are still lacking. The present study aimed to assess the efficacy of moxibustion intervention prior to disease onset and at the early stage of disease in a rat model of DOR and explore the mechanisms of its effect on ovarian function. A total of 32 rats were randomly divided into four groups: Blank group, Model group (a drug-induced model of DOR), Moxibustion group 1 and Moxibustion group 2. Moxibustion was performed on the BL23 and RN4 acupoints of female rats daily for a total of 20 days (once a day, five times a week for a total of 4 weeks). The two moxibustion groups were established with different intervention times: One group was subjected to pre-disease intervention and the other group to early-disease intervention. The ovarian function was evaluated by detecting anti-Mullerian hormone (AMH), follicle-stimulating hormone (FSH), estradiol (E2), testosterone (T), dehydroepiandrosterone (DHEA), dihydrotestosterone (DHT) and androgen receptor (AR) levels in the serum or the ovary samples. To further investigate the downstream regulatory factors for AR after moxibustion treatment for pre-disease or early-disease intervention, FSH receptor (FSHR) and microRNA (miR)-125b expression in ovaries were also analyzed. The results indicated that AMH and DHT levels were reduced in the model group compared with those in the blank group, while FSH, T and DHEA levels were increased. AMH and DHT levels were increased in Moxibustion group 1 compared with those in the model group, while FSH, T and DHEA levels were reduced. There was no difference in E2 levels between Moxibustion group 1 and the model group. Compared with that in the model group, the AR content in the ovary was increased in Moxibustion group 1. There was no difference in FSHR mRNA in the ovaries between Moxibustion group 1 and the model group. miR-125b levels were significantly increased in Moxibustion group 1 as compared with those in the model group. Furthermore, AMH and DHT levels were increased in Moxibustion group 2 compared with those in the model group, while FSH, T and DHEA levels were reduced. E2 levels were significantly decreased in Moxibustion group 2 compared with those in the model group. The relative mRNA expression of AR, FSHR and miR-125b was decreased following establishment of the model. Compared with that in the model group, the AR content in the ovary was increased in Moxibustion group 2. In comparison with the blank and model groups, the FSHR content in the ovary of Moxibustion group 2 was significantly increased. miR-125b levels were not obviously altered in Moxibustion group 2 as compared with those in the model group. In addition, there was no significant difference in AMH, FSH, T and DHEA levels between the two moxibustion groups. E2 and DHT levels were higher in Moxibustion group 1 than in Moxibustion group 2. There was no difference in AR mRNA expression between the two moxibustion groups. FSHR mRNA levels were lower in Moxibustion group 1 than in Moxibustion group 2, while miR-125b mRNA levels were higher in Moxibustion group 1 than in Moxibustion group 2. In conclusion, the present study suggested that moxibustion intervention prior to disease onset and at the early disease stage was able to improve ovarian function via modulation of the AR-mediated stable equilibrium of androgens. However, the effects and mechanisms of moxibustion intervention for pre-disease and early-disease intervention of DOR appear to be different. The appropriate duration of treatment and the time-effect relationship require to be further studied.

Integrated miRNA-seq analysis reveals the molecular mechanism underlying the effect of acupuncture on endometrial receptivity in patients undergoing fertilization: embryo transplantation.

Endometrial receptivity (ER) is the main factor affecting in vitro fertilization (IVF) and embryo transplantation. Previous studies have shown that acupuncture might be useful for increasing ER. However, the underlying microRNA (miRNA) molecular mechanisms deserve to be further elucidated. In this study, we performed small RNA sequencing of endometrial samples from infertile women who either underwent acupuncture therapy or did not. Differentially expressed microRNAs (DEmiRNAs) were identified and used to predict target genes. Then, the functional and pathway were analyzed for the target genes. Moreover, quantitative reverse transcription PCR (qRT-PCR) was performed to validate the RNA-seq results. Finally, the miRNA-gene network was conducted by Cytoscape. A total of 39 DEmiRNAs were identified between the acupuncture group and the control group. The functional enrichment analysis suggested that the target genes of the DEmiRNAs were significantly enriched in GO biological process (BP) terms associated with transcription, such as regulation of DNA-templated transcription and positive regulation of DNA-templated transcription. The pathway analysis showed that DEmiRNAs might be involved in acupuncture therapy via Endocytosis, Axon guidance, Oxytocin signaling, the Hippo pathway, and Estrogen signaling pathways. Significant downregulation of hsa-miR-449a and hsa-miR-449b-3p, and significant upregulation of hsa-miR-3135b and hsa-miR-345-3p in the RNA-seq results were validated by qRT-PCR. Besides, these four DEmiRNAs and their 34 target genes conducted a miRNA-gene network. Our results predict that hsa-miR-449a, hsa-miR-3135b and hsa-miR-345-3p may underly mechanisms by which acupuncture therapy help increase ER and promote endometrium receptivity in preparation for in vitro fertilization and embryo transplantation.

[Effect on sperm quality of asthenospermia and oligospermia treated with grain-moxibustion combined with medicine therapy].

OBJECTIVE: To observe the clinical effect of grain-moxibustion combined with medicine therapy for asthenospermia and oligospermia. METHODS: A tatal of 60 patients were randomized into an observation group (30 cases) and a control group (30 cases) according to 1︰1 ratio. In the control group, vitamin E capsules were taken orally one capsule each time, twice a day, and Wuzi Yanzong pills 6 g each time, three times a day for a total of 3 months. In the observation group, grain-moxibustion was applied at Guanyuan (CV 4)，Shenshu (BL 23) and Zusanli (ST 36) based on the control group, once a week for 3 months, with a total of 12 times. The sperm concentration and sperm progressive motility were measured by automatic sperm quality analysis system in the two groups, and the clinical effects were compared. Sperm DNA fragmentation index (DFI) in the observation group was measured by sperm nucleus chromosome structure assay (SCSA). RESULTS: ①The sperm concentrations and sperm progressive motilities after 1-month, 2-month and 3-month of treatment were increased compared with those before treatment in the two groups (P<0.01), and they were increased with time. In the two groups, 2-month and 1-month of treatment, 3-month and 2-month of treatment were compared, the sperm concentrations and sperm progressive motilities were significantly increased (P<0.01). The sperm concentrations after 1-month, 2-month and 3-month of treatment in the observation group were higher than those in the control group (P<0.01), the sperm progressive motility after 3-month of treatment in the observation group was higher than that in the control group (P<0.05). ②After 3-month of treatment，the DFI in the observation group was significantly reduced compared with that before treatment (P<0.01). ③The total effective rate in the observation group after 3-month of treatment was 86.7% (26/30), which was superior to 63.3% (19/30) in the control group (P<0.05). CONCLUSION: Grain-moxibustion combined with medicine therapy can improve sperm concentration and sperm progressive motility, enhance the integrity of sperm DNA.

Circular RNA sequencing reveals the molecular mechanism of the effects of acupuncture and moxibustion on endometrial receptivity in patients undergoing infertility treatment.

The present study aimed to determine the profile of differentially expressed circular RNAs (circRNAs) in infertile patients treated with acupuncture and moxibustion and verify the role of acupuncture and moxibustion in altering endometrial receptivity (ER). High­throughput RNA sequencing and bioinformatics analysis of samples from six pairs of patients treated with or without acupuncture and moxibustion were conducted. The reliability of high­throughput RNA sequencing was validated using reverse transcription­quantitative PCR. The most significant circRNA functions and pathways were selected by gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. A circRNA­miR­mRNA interaction network was constructed to determine the connection between circRNAs, microRNAs (miRs), and mRNAs. High­throughput RNA sequencing identified 2,653 circRNAs. A total of 86 circRNAs was differentially expressed, of which 57 were upregulated and 29 were downregulated, between the acupuncture and moxibustion group and the control group. In the GO analysis, the identified BP terms were chromatin modification, positive regulation of transcription from RNA polymerase II promoter involved in unfolded protein response, oxidative DNA demethylation, regulation of transcription from RNA polymerase II promoter in response to hypoxia, and regulation of smooth muscle cell differentiation. The identified CC terms were nucleoplasm, nucleolus, nucleus, histone acetyltransferase complex, and annulate lamellae. The identified MF terms were methylcytosine dioxygenase activity, chromatin binding, zinc ion binding, histone binding, and protein binding. In the KEGG pathway analysis, the identified pathways were protein processing in endoplasmic reticulum, degradation of aromatic compounds, shigellosis, mTOR signaling pathway, bacterial invasion of epithelial cells, and prostate cancer. Circ­SFMBT2, circ­BACH1, and circ­LPAR1 were significantly upregulated (P<0.05) and associated with numerous miRs and mRNAs. Acupuncture and moxibustion could impact ER by regulating the expression of circRNAs.

Whole Transcriptome Sequencing Reveals How Acupuncture and Moxibustion Increase Pregnancy Rate in Patients Undergoing In Vitro Fertilization-Embryo Transplantation.

BACKGROUND: In vitro fertilization and embryo transfer (IVF-ET) technology has been widely used in the therapy of refractory infertility. Previous studies showed that acupuncture can effectively increase the clinical pregnancy rate of IVF-ET. However, the molecular mechanism is unknown. MATERIALS AND METHODS: In this study, we performed whole transcriptome sequencing for endometrial samples from infertile women who underwent acupuncture and moxibustion therapy or not. Differentially expressed noncoding RNAs (ncRNAs) and mRNAs were identified and their functions were predicted. Besides, a competitive endogenous RNA network was constructed to further interpret the molecular mechanism of acupuncture and moxibustion therapy on infecund patients. In addition, real-time PCR was applied to validate the RNA-seq results. RESULTS: We identified 317 differentially expressed mRNAs and 82 ncRNAs in acupuncture and moxibustion therapy group compared with control group. Functional enrichment analysis suggested that these genes were significantly enriched in GO-BP terms associated with cellular transport, such as ATP hydrolysis coupled proton transport, vacuolar acidification, transferrin transport, and proton transport and metabolic process, including small molecule metabolic process and metabolic process. Pathway enrichment analysis enriched 11 terms, including oxidative phosphorylation, synaptic vesicle cycle, mineral absorption, and metabolic pathways. Four of five selected differentially expressed genes were validated by real-time PCR. CONCLUSION: Our results suggested that acupuncture and moxibustion therapy might increase the pregnancy rate of patients undergoing IVF-ET by the regulation of ncRNAs.

The effects of acupuncture on pregnancy outcomes of in vitro fertilization: a systematic review and meta-analysis.

BACKGROUND: The effects of acupuncture on in vitro fertilization (IVF) outcomes remain controversial. And the variation in participant, interventions, outcomes studied, and trial design may relate to the efficacy of adjuvant acupuncture. METHODS: We searched digital databases for relevant studies, including Embase, PubMed, Cochrane Library and some Chinese databases up to December 2018, for randomized controlled trials (RCTs) evaluating the effects of acupuncture on women undergoing IVF. We included studies with intervention groups using needling, and control groups consisting of no acupuncture or sham (placebo) acupuncture. Primary outcomes were clinical pregnancy rate (CPR) and live birth rate (LBR). Meta-regression and subgroup analysis were conducted on the basis of eight pre-specified covariates to investigate the variances of the effects of adjuvant acupuncture on pregnancy rates and the sources of heterogeneity. RESULTS: Twenty-seven studies with 6116 participants were included. The pooled clinical pregnancy rate (CPR) from all of acupuncture groups was significantly greater than that of control groups (RR 1.21, 95% CI: 1.07-1.38), whereas the pooled live birth rate (LBR) was not. Meta-regression subgroup analysis showed a more significant benefit of acupuncture for repeated IVF cycle proportion (number of women with a history of prior unsuccessful IVF attempt divided by number of women included in each trial) ≥ 50% group (CPR: RR 1.60, 95% CI: 1.28-2.00; LBR: RR 1.42, 95% CI: 1.05-1.92), and this covariate explained most of the heterogeneity (CPR and LBR: adjusted R2 = 100 and 87.90%). Similar results were found between CPR and number of acupuncture treatments (CPR: p = 0.002, adjusted R2 = 51.90%), but not LBR. CONCLUSIONS: Our analysis finds a benefit of acupuncture for IVF outcomes in women with a history of unsuccessful IVF attempt, and number of acupuncture treatments is a potential influential factor. Given the poor reporting and methodological flaws of existing studies, studies with larger scales and better methodologies are needed to verify these findings.

[Discussion on the impact of acupuncture for the pregnancy of in vitro fertilization-embryo transfer].

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.

[Analysis on CHENG Dan-an's exploration of quantification acupuncture and moxibustion stimulation].

As a physical therapy, there is a certain relationship between the stimulating quantity and the efficacy of acumoxibustion therapy. However, because of the thinking mode of traditional Chinese medicine in the ancient times, qualitative descriptions about acu-moxibustion stimulation in the classical literature are predominating. Mr. CHENG Dan-an, a famous master and educator of acupuncturology, conducted a series of exploration in both theory and clinical practice. In the present paper, the authors summarize Mr. Cheng's viewpoints about the quantification of acupuncture intervention from 1) the number of the selected acupoints in a session of treatment, 2) gauges and materials of the filiform acupuncture needles, 3) stimulating strength of the needling, including the duration of acupuncture needle retaining, depth of needling and intensity of the needling manipulation, 4) frequency of the acupuncture intervention (interval between every two sessions of treatment). Regarding the moxibustion therapy, the stimulating quantity including the duration of the ignited moxa and the number of moxa cones applied at the acupoint area should be varied according to the patients' physical constitutions, ages and the state of the illness or clinical conditions. Mr. Cheng also put forward his opinions about the quantification standards of moxibustion intervention.

[Comparative study of bloodletting therapy between traditional Chinese medicine and Tibetan medicine].

The comparative study of bloodletting therapy between traditional Chinese medicine and Tibetan medicine in view of history development, theoretic basis, bloodletting location, bloodletting tool, operation method, bloodletting amount, indications, contraindications and the others are conducted in this paper. It is pointed out that the bloodletting therapy could be better carried forward and developed through the interaction and integration of bloodletting therapy between traditional Chinese medicine and Tibetan medicine in term of the theoretic, practical and development patterns under the guidance of these two different medical theoretical systems.

[Professor YANG Zhao-min's contribution to the education of modern acupuncture and moxibustion].

It was completely collected and summarized the teaching history and the relevant historical materials of Professor YANG Zhao-min via consulting the literatures, investigation, interview, etc. in this paper. The summaries indicated that from the beginning of running a school, Professor YANG Zhao-min adhered to the CHENG's moral, worked hard and successfully explored and practiced the training pattern of teaching tour for the personnel training of acupuncture and moxibustion. He established the specialty of acupuncture and moxibustion, formatted courses and inherited the feature of practice teaching. He was in charge of the compilation of textbook and engaged in scientific innovation. He inherited the past and forged ahead into the future and had made the decision to cultivate the personnel of acupuncture and moxibustion. Professor YANG Zhao-min had practiced and developed the modern educational thought from the CHENG Dan-an and had made the important contribution to the training of acupuncture and moxibustion professionals.

[Clinical observation on acupotomy for treatment of simple obesity].

OBJECTIVE: To compare the efficacy differences among acupotomy, electroacupuncture and acupuncture for treatment of simple obesity. METHODS: One hundred and five cases were randomly divided into an acupotomy group, an electroacupuncture group and an acupuncture group, 35 cases in each group, and Zhongwan (CV 12), Tianshu (ST 25), Shangjuxu (ST 37), Sanyinjiao (SP 6), etc, were selected in three groups and also with selection of acupoints according to symptoms. The acupotomy group was treated with acupotomy 40 mm in length and 0.6 mm in diameter, the electroacupuncture group with the Han's LH402A electroacupuncture stimulator and the acupuncture group with simple acupuncture treatment. The clinical therapeutic effects of three groups were compared. The obesity signs such as body weight, body mass index (BMI), obesity degree, etc., and blood lipid and fasting blood sugar (FBS) were observed. RESULTS: The markedly effective rate of 91.4% (32/35) in the acupotomy group was higher than that of 71.5% (25/35) in the electroacupuncture group and that of 42.9% (15/35) in the hand acupuncture group (both P<0.05). There were significant differences in the obesity signs and blood fat and FBS of the three groups before and after treatment (P<0.05, P<0.01). The differences rates of body weight, BMI, obesity degree, chest circumference, waistline, thighline, waist-hip ratio, total cholesterol before and after treatment in the acupotomy group were all better than those in the electroacupuncture group and in the acupuncture group (all P<0.05), and the differences rates of hipline, FBS in the acupotomy group were better than those in the acupuncture group (both P<0.001). CONCLUSION: Acupotomy treatment can obviously reduce fat, FBS and blood lipid and has obvious therapeutic effects on simple obesity.