Enhanced glycolysis in the myometrium with ectopic endometrium of patients with adenomyosis: a preliminary study.

OBJECTIVES: The objective of this study was to investigate the glycolytic activity of adenomyosis, which is characterized by malignant biological behaviors including abnormal cell proliferation, migration, invasion, cell regulation, and epithelial-mesenchymal transition. METHODS: From January 2021 to August 2022, a total of 15 patients who underwent total hysterectomy for adenomyosis and 14 patients who had non-endometrial diseases, specifically with cervical squamous intraepithelial neoplasia and uterine myoma, were included in this study. Myometrium with ectopic endometrium from patients with adenomyosis while normal myometrium from patients in the control group were collected. All samples were confirmed by a histopathological examination. The samples were analyzed by liquid chromatography-mass spectrometry (LC-MS), real-time quantitative PCR, NAD+/NADH assay kit as well as the glucose and lactate assay kits. RESULTS: Endometrial stroma and glands could be observed within the myometrium of patients in the adenomyosis group. We found that the mRNA expressions of HK1, PFKFB3, glyceraldehyde-3-phospate dehydrogenase (GAPDH), PKM2, and PDHA as well as the protein expressions of PFKFB3 were elevated in ectopic endometrial tissues of the adenomyosis group as compared to normal myometrium of the control group. The level of fructose 1,6-diphosphate was increased while NAD + and NAD+/NADH ratio were decreased compared with the control group. Besides, increased glucose consumption and lactate production were observed in myometrium with ectopic endometrium. CONCLUSIONS: We concluded that altered glycolytic phenotype of the myometrium with ectopic endometrium in women with adenomyosis may contribute the development of adenomyosis.

Electroacupuncture ameliorates hepatic defects in a rat model of polycystic ovary syndrome induced by letrozole and a high-fat diet.

BACKGROUND: This study was designed to evaluate the effects of low-frequency electroacupuncture (EA) on glucose and lipid disturbances in a rat model of polycystic ovary syndrome (PCOS) characterized by insulin resistance (IR) and hepatic steatosis. METHODS: The PCOS rat model was induced by continuous administration of letrozole (LET) combined with a high-fat diet (HFD). Female Sprague-Dawley rats were divided into the following four groups: control, control + EA, LET + HFD and LET + HFD + EA. EA was administered five or six times a week with a maximum of 20 treatment sessions. Body weight, estrous cyclicity, hormonal status, glucose and insulin tolerance, lipid profiles, liver inflammation factors, liver morphology and changes in the phosphatidylinositol 3-kinase (PI3-K)/Akt (protein kinase B) pathway were evaluated. RESULTS: The rat model presented anovulatory cycles, increased body weight, elevated testosterone, abnormal glucose and lipid metabolism, IR, liver inflammation, hepatic steatosis and dysregulation of the insulin-mediated PI3-K/Akt signaling axis. EA reduced fasting blood glucose, fasting insulin, area under the curve for glucose, homeostasis model assessment of IR indices, triglycerides and free fatty acids, and alleviated hepatic steatosis. Furthermore, low-frequency EA downregulated mRNA expression of tumor necrosis factor (TNF)-α and interleukin (IL)-6, upregulated mRNA expression of peroxisome proliferator-activated receptor (PPAR)-α, increased protein expression of phosphorylated (p)-Akt (Ser473), p-glycogen synthase kinase (GSK) 3ß (Ser9) and glucose transporter 4 (GLUT4), increased the ratio of p-GSK3ß to GSK3ß and downregulated protein expression of GSK3ß. CONCLUSION: An obese PCOS rat model with IR and hepatic steatosis was successfully established by the combination of LET and HFD. EA improved dysfunctional glucose and lipid metabolism in this PCOS-IR rat model, and the molecular mechanism appeared to involve regulation of the expression of key molecules of the PI3-K/Akt insulin signaling pathway in the liver.

Efficacy and safety of acupuncture in postoperative ileus after gynecological surgery: A protocol for system review and meta-analysis of randomized controlled trials.

BACKGROUND: Acupuncture is widely used in treatment of postoperative ileus (POI), but the safety and efficacy of acupuncture in POI after gynecological surgery still lack of evidence-based basis. METHODS: PubMed, CINAHL, EMBASE, Web of science, Google Scholar, Wangfang database, Chinese Biomedical Literature Database (SinoMed), Chinese Science and Technology Periodical Database, and China National Knowledge Infrastructure database will be searched until December 31, 2020. Two independent investigators will screen the relevant randomized controlled trials from Data one by one by using prespecified criteria. The relevant data from included studies will be extracted and analyzed by using RevMan V.5.3 software. Quality of the included studies will be estimated by using the Cochrane Collaboration risk of bias tool, and publication bias will be assessed by using Egger test and Begg test. In addition, quality of evidence will be evaluated by using Grading of Recommendations Assessment, Development, and Evaluation. RESULTS: We will analyze the effect of acupuncture on time to first flatus and time to bowel sound recovery as the primary outcomes of this review. Meanwhile, frequency of bowel sounds, time to defecation, time of hospital stay, biochemical indicators related to gastrointestinal motility, inflammation factors, responder rate, and adverse events for patients receiving gynecological surgery. CONCLUSION: Our findings will benefit researchers and provide reference for the treatment and prevention of POI for the patients undergoing gynecological surgery.

[Traditional Chinese therapy of tonifying the kidney and invigorating blood circulation reduces sperm DNA damage and improves natural pregnancy in infertile males].

OBJECTIVE: To study the effect of the traditional Chinese therapy of tonifying the kidney and invigorating blood circulation (TKIB) on male infertility. METHODS: Forty-two infertile males with abnormal DNA fragmentation index (DFI) were randomly allocated into a TKIB (n = 22) and a control group (n = 20), the former treated by TKIB with an oral Chinese medicinal prescription while the latter with oral tamoxifen tablets and vitamin E capsules, both for 3 months. Before and after treatment, we obtained the semen parameters and sperm DFI from the patients and followed them up for the outcomes of natural pregnancy. RESULTS: Compared with the baseline, the patients in both the TKIB and control groups showed significant increases after medication in sperm concentration (ï¼»36.82 ± 29.16ï¼½ and ï¼»34.56 ± 37.03ï¼½ vs ï¼»50.00 ± 39.16ï¼½ and ï¼»40.72 ± 47.37ï¼½ ×106/ml, P<0.05), the percentage of progressively motile sperm (PMS) (ï¼»20.62 ± 9.10ï¼½% and ï¼»21.25 ± 9.11ï¼½% vs ï¼»36.82 ± 13.45ï¼½% and ï¼»26.18 ± 10.60ï¼½%, P<0.05) and the percentage of morphologically normal sperm (MNS) (ï¼»1.28 ± 1.00ï¼½% and ï¼»1.48 ± 0.91ï¼½% vs ï¼»3.44 ± 1.33ï¼½% and ï¼»2.57 ± 1.32ï¼½%, P<0.05), but remarkably decreased sperm DFI (ï¼»29.07 ± 11.52ï¼½% and ï¼»24.43 ± 8.46ï¼½% vs ï¼»15.51 ± 11.31ï¼½% and ï¼»18.53 ± 10.44ï¼½%, P<0.05). The patients of the TKIB group exhibited an even higher total sperm motility and percentages of PMS and MNS than those of the control group (P<0.05) but no statistically significant difference from the latter in sperm concentration or DFI (P>0.05). Besides, the former achieved higher rates of natural pregnancy (18.2%) and live birth (18.2%) than the controls (15% and 10%) though neither with statistically significant difference (P>0.05). CONCLUSIONS: The traditional Chinese therapy of tonifying the kidney and invigorating blood circulation can reduce sperm DNA damage and improve the outcomes of natural pregnancy in infertile men.

Chemerin isoform analysis in human biofluids using an LC/MRM-MS-based targeted proteomics approach with stable isotope-labeled standard.

Targeted proteomics has advantages over earlier conventional technologies for protein detection. We developed and validated an LC/MRM-MS-based targeted proteomic method combined with immunoaffinity precipitation for the enrichment and detection of low abundance chemerin isoforms in human biofluids. After tryptic digestion, each chemerin isoform was characterized by isoform-specific peptides, and the absolute quantification was achieved by using stable isotope-labeled peptides as internal standards. In serum, follicular fluid and synovial fluid, a total of 6 chemerin isoforms were identified and quantified, among which a novel natural isoform 153Q was discovered for the first time. The relative content of the six chemerin isoforms in human serum was 157S ≫ 156F ≫ 158K > 154F ≥ 155A > 153Q in the ratio of 25:17:5:2.5:2.2:1, respectively. The absolute contents were in the range of 88-3.5 ng/mL. This distribution remained consistent among the 3 biofluids analyzed. Total chemerin were found to be increased in both polycystic ovary syndrome (serum and follicular fluid) and rheumatoid arthritis (serum) patients. However, chemerin isoform analysis revealed that only 156F & 157S were increased in the former, while 155A, 156F & 157S were increased in the latter. This demonstrates the potential of this method in detailed characterization of changes in chemerin isoforms that may be of clinical relevance.

Effects of Bushen Huoxue method for female with decreased ovarian reserve: A protocol for systematic review and meta-analysis.

BACKGROUND: Decreased ovarian reserve (DOR) is a common reproductive barrier in female. Bushen Huoxue (BSHX) method of TCM is widely applied to treat DOR clinically. The purpose of this study is to provide a systemic and comprehensive evaluation of BSHX in the treatment of DOR. METHODS: We have registered this protocol with OSF registry and the DOI is 10.17605/OSF.IO/QNUE2. We will search 4 English databases (PubMed, EMBASE, MEDLINE, Cochrane Library) and four Chinese databases (China national knowledge infrastructure database, Wanfang database, VIP and Superstar database) from their inception to August 10, 2020. Two authors will search and extract independently all related studies. RevMan 5.3 software will be applied to synthesize data. RESULTS: The results of this study will be published in a scientific journal after peer-review. CONCLUSION: This systematic review will provide reliable evidences for clinicians, and help them make decisions in DOR management.

A systematic review of the efficacy comparison of acupuncture and traditional Chinese medicine in the treatment of primary dysmenorrhea.

BACKGROUND: Dysmenorrhea is one of the most common symptoms in gynecology. It refers to people who experience lower abdominal pain, swelling, backache, or another discomfort before and after menstruation or during menstruation, which seriously affects the quality of life and work. Clinically, there are many methods to treat primary dysmenorrhea, among which acupuncture and traditional Chinese medicine (TCM) are beneficial. This work aimed to test the efficacy of acupuncture and TCM in the treatment of primary dysmenorrhea and hope to supply more reliable evidence for clinical treatment. METHODS: We searched articles from the Wanfang database and China National Knowledge Infrastructure and collected the clinical, randomized, or quasi-randomized controlled trials of acupuncture compared with TCM for primary dysmenorrhea. We tested the quality and data of the included studies according to the Cochrane criteria and compiled detailed systematic reviews and meta-analysis. RESULTS: Seven articles (including 492 patients) on acupuncture and TCM for primary dysmenorrhea were included in the systematic review. Seven studies were included in the final analysis, and there was no heterogeneity among the studies (P=0.98, I2=0%). The results showed that the clinical effectiveness of acupuncture was better than TCM (OR: 4.86, 95% CI: 2.84-8.33, Z =5.75, P<0.00001). CONCLUSIONS: The efficacy of acupuncture is superior to TCM in the treatment of primary dysmenorrhea.

MiRNAs expression profiling of rat ovaries displaying PCOS with insulin resistance.

PURPOSE: The present study established microRNA (miRNA) expression profiles for rat ovaries displaying polycystic ovary syndrome (PCOS) with insulin resistance and explored the underlying biological functions of differentially expressed miRNAs. METHODS: A PCOS with insulin resistance rat model was created by administering letrozole and a high-fat diet. Total RNA was extracted from the ovaries of PCOS with insulin resistance rats and normal rats. Three ovaries from each group were used to identify differentially expressed miRNAs by deep sequencing. A hierarchical clustering heatmap and volcano plot were used to display the pattern of differentially expressed miRNAs. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were conducted to explore the potential target genes of the differentially expressed miRNAs and identify their putative biological function. Nine of the differentially expressed miRNAs were selected for validation by Real-time Quantitative PCR (qRT-PCR). RESULTS: A total of 58 differentially expressed miRNAs were identified in the rat ovaries exhibiting PCOS with insulin resistance compared with control ovaries, including 23 miRNAs that were upregulated and 35 miRNAs that were downregulated. GO and KEGG pathway analyses revealed that the predicted target genes were related to metabolic processes, cellular processes, and metabolic pathways. Furthermore, qRT-PCR confirmed that miR-3585-5p and miR-30-5p were significantly upregulated and miR-146-5p was downregulated in the ovaries of PCOS with insulin resistance rats compared with the controls. CONCLUSION: These results indicate that differentially expressed miRNAs in rat ovaries may be involved in the pathophysiology of insulin resistance in PCOS. Our study may be beneficial in establishing miRNAs as novel diagnostic and therapeutic biomarkers for insulin resistance in PCOS.

Reagentless and sensitive determination of carcinoembryonic antigen based on a stable Prussian blue modified electrode.

Reagentless and sensitive detection of tumor biomarkers using label-free electrochemical immunosensors is highly desirable for early and effective cancer diagnosis. Herein, we present a label-free electrochemical immunoassay platform based on surface-confined Prussian blue (PB) redox probes for sensitive and reagentless determination of carcinoembryonic antigen (CEA). To facilitate the electron transfer of probes and improve sensitivity, Au nanoparticles and PB (Au-PB) are electrochemically co-deposited on a carbon nanotube (CNT) modified glassy carbon electrode (GCE). A polydopamine (pDA) layer is coated on the Au-PB nanocomposite layer in situ as a bifunctional linker. In addition to improving the stability of PB, pDA also provides reducibility for the preparation of gold nanoparticles, which offers an interface for anti-CEA antibody immobilization. The fabricated immunosensor has good stability and is able to reagentlessly detect CEA over a wide range (0.005-50 ng mL-1) with high reproducibility. Furthermore, the immunosensor was used for determination of CEA in human serum samples.

Effects of Jiawei Shaoyao-Gancao Decoction and Its Drug-Containing Serum on Proliferation, Apoptosis, and Ultrastructure of Human Adenomyosis Foci Cells.

OBJECTIVE: The present study aimed to investigate the effects of Jiawei Shaoyao-Gancao Decoction (JSGD) and its drug-containing serum (CDS) on the proliferation, apoptosis, and ultrastructure of human adenomyosis foci cells. METHODS: Primary cultures of human adenomyosis foci cells were prepared from hard uterine lesions of adenomyosis patients. The cells were treated with JSGD (10 and 20 mg/ml), CDS, and mifepristone (MIF) for 24 or 48 h. Cell proliferation was detected using CCK-8 assay, cell apoptosis was measured by flow cytometry, and the cell ultrastructure was observed by transmission electron microscopy (TEM). RESULTS: JSGD and CDS significantly induced cell apoptosis and inhibited cell proliferation for 24 h or 48 h, in which the effects of JSGD were in a dose-dependent manner. The effect of CDS for 24 h was higher than that of CDS for 48 h. Moreover, JSGD and CDS treatments induced marked apoptosis in adenomyosis foci cells, characterized by nucleus chromatin, condensation, fragmentation, mitochondria and endoplasmic swelling, and autophagy-lysosome. CONCLUSIONS: JSGD and CDS can suppress proliferation and induce apoptosis in adenomyosis foci cells, through altering their ultrastructure. The results provided support for JSGD and CDS in the treatment of adenomyosis and gained further insight into the effect of this prescription.

[Role of estrogen, estrogen receptors, and aromatase in the pathogenesis of uterine adenomyosis].

OBJECTIVE: To study the role of estrogen (E2), estrogen receptor (ER) and aromatase (P450arom) in the pathogenesis of uterine adenomyosis. METHODS: Paraffin-embedded specimens of the uterine tissue from patients with uterine adenomyosis and patients with cervical lesions (CIN; control) were examined for expressions of E2, ER and P450arom by immunohistochemistry and ELISA. The cells isolated from the lesions of patients with adenomyosis were cultured in vitro, and the changes in cell growth in response to treatments with E2, ER inhibitor, ER inhibitor + E2, estrogen deprivation, and estrogen deprivation+ ICI182780 were assessed using CCK-8 method. RESULTS: The expression levels of E2, ER, and P450arom were significantly higher in adenomyosis ectopic lesions and eutopic endometrium than in the myometrium and endometrium in the control group (P<0.05); no significant difference in E2 and P450arom expressions was found between adenomyosis ectopic lesions and eutopic endometrium (P>0.05), while the expression levels of ER in ectopic lesions was significantly higher than that in eutopic endometrium. The cell inhibition rates were similar between ER inhibitor group and ER inhibitor + Estrogen activation group (P>0.05), and was significantly higher in estrogen deprivation+ ER inhibitor group than in estrogen deprivation group (P<0.05). CONCLUSION: The high expression levels of E2, ER, and P450arom in adenomyosis ectopic lesions and eutopic endometrium promote uterine adenomyosis cell proliferation, in which process E2 combines with ER to execute its biological effect; ER also promotes the occurrence and development of uterine adenomyosis through other pathways.

Potential mechanisms of an antiadenomyosis chinese herbal formula shaoyao-gancao decoction in primary cell culture model.

Background. Shaoyao-Gancao Decoction (SGD), a well-known traditional Chinese medicine prescription, has been widely used to treat adenomyosis, dysmenorrhea, abdominal pain, and inflammation in Asia. However, the mechanism underlying the effectiveness of SGD in the treatment of adenomyosis still remains elusive. The present study aimed to investigate the bioactivity of SGD and its underlying molecular mechanisms using cultured human adenomyosis-derived cells. Methods. Human adenomyosis-derived cells were treated with SGD and its major constituents (paeoniflorin and liquiritin) in vitro. Effects of SGD, paeoniflorin, and liquiritin on cell proliferation and apoptosis were examined by MTT assay and flow cytometry analyses. The effects of SGD, paeoniflorin, and liquiritin on the production of PGE2 and PGF2α were assayed using ELISA. ER-α and OTR mRNA expression levels were also evaluated by real-time qRT-PCR. Results. SGD, paeoniflorin, and liquiritin inhibited proliferation and induced apoptosis of human adenomyosis-derived cells in a dose-dependent manner. SGD and paeoniflorin significantly reduced the PGE2 and PGF2α production. Furthermore, they remarkably decreased the mRNA levels of ER-α and OTR. Conclusions. The results of this study provide possible mechanisms for the bioactivity of SGD for treating adenomyosis and contribute to the ethnopharmacological knowledge about this prescription.

Perimenopause Amelioration of a TCM Recipe Composed of Radix Astragali, Radix Angelicae Sinensis, and Folium Epimedii: An In Vivo Study on Natural Aging Rat Model.

Traditional Chinese medicine (TCM) has been extensively applied as preferable herbal remedy for menopausal symptoms. In the present work, the potential of a TCM recipe named RRF, composed of Radix Astragali, Radix Angelicae Sinensis, and Folium Epimedii, was investigated on a natural aging rat model. After administration of RRF (141, 282, and 564 mg/kg/d), the circulated estradiol (E2) level increased accompanied by a reduction of serum follicle stimulating hormone (FSH). But no significant impact on serum lutenizing hormone (LH) level was observed. As a result of the E2-FSH-LH adjustment, the histomorphology degenerations of ovary, uterus, and vagina of the 11.5-month female rats were alleviated. And lumbar vertebrae trabecular microstructure was also restored under RRF exposure by means of increasing the trabecular area and area rate. Moreover, levels of hypothalamic dopamine (DA) and norepinephrine (NE) rallied significantly after RRF treatment. Results from our studies suggest that RRF possesses a positive regulation on the estrogen imbalance and neurotransmitter disorder, thereby restoring reproductive organ degeneration and skeleton deterioration. The above-mentioned benefits of RRF on the menopause syndromes recommend RRF as a potential candidate for the treatment of perimenopausal syndrome.