Construction and validation of molecular subtype and signature of immune cell-related telomeric genes and prediction of prognosis and immunotherapy efficacy in ovarian cancer patients.

BACKGROUND: Ovarian cancer (OVC) has emerged as a fatal gynecological malignancy as a result of a lack of reliable methods for early detection, limited biomarkers and few treatment options. Immune cell-related telomeric genes (ICRTGs) show promise as potential biomarkers. METHODS: ICRTGs were discovered using weighted gene co-expression network analysis (WGCNA). ICRTGs were screened for significant prognosis using one-way Cox regression analysis. Subsequently, molecular subtypes of prognosis-relevant ICRTGs were constructed and validated for OVC, and the immune microenvironment's landscape across subtypes was compared. OVC prognostic models were built and validated using prognosis-relevant ICRTGs. Additionally, chemotherapy susceptibility drugs for OVC patients in the low- and high-risk groups of ICRTGs were screened using genomics of drug susceptibility to cancer (GDSC). Finally, the immunotherapy response in the low- and high-risk groups was detected using the data from GSE78220. We conducted an immune index correlation analysis of ICRTGs with significant prognoses. The MAP3K4 gene, for which the prognostic correlation coefficient is the highest, was validated using tissue microarrays for a prognostic-immune index correlation. RESULTS: WGCNA analysis constructed a gene set of ICRTGs and screened 22 genes with prognostic significance. Unsupervised clustering analysis revealed the best molecular typing for two subtypes. The Gene Set Variation Analysis algorithm was used to calculate telomere scores and validate the molecular subtyping. A prognostic model was constructed using 17 ICRTGs. In the The Cancer Genome Atlas-OVC training set and the Gene Expression Omnibus validation set (GSE30161), the risk score model's predicted risk groups and the actual prognosis were shown to be significantly correlated. GDSC screened Axitinib, Bexarotene, Embelin and the GSE78220 datasets and demonstrated that ICRTGs effectively distinguished the group that responds to immunotherapy from the non-responsive group. Additionally, tissue microarray validation results revealed that MAP3K4 significantly predicted patient prognosis. Furthermore, MAP3K4 exhibited a positive association with PD-L1 and a negative relationship with the M1 macrophage markers CD86 and INOS. CONCLUSIONS: ICRTGs may be reliable biomarkers for the molecular typing of patients with OVC, enabling the prediction of prognosis and immunotherapy efficacy.

Neuroprotective effects of TRPV1 by targeting GDF11 in the Mpp+/MPTP-induced Parkinson's disease model.

Protecting dopaminergic neurons is a key approach in the prevention of Parkinson's disease (PD). Transient receptor potential vanilloid 1 (TRPV1) is a nonselective cation channel that is widely distributed in the mammalian nervous system. In this study, we designed experiments to investigate the effect and mechanisms of TRPV1 against DA neurons damage of PD. Our results showed that trpv1-deficient mice showed a significant loss of TH + neurons than PD mice after MPTP intraperitoneal injection, in addition, a significant decline in motor function was observed in trpv1-deficient mice versus the MPTP model. In addition, our study indicated that GDF11 overexpression inhibited MPP + - induced oxidative stress, cell senescence, and apoptosis in neurons. Results also showed that TRPV1 prevented the down-regulation of GDF11 expression in PD model, gdf11 knockdown blocks the effects of TRPV1 on the antioxidant, antiaging, and antiapoptotic activities of dopaminergic neurons. Consequently, our findings indicate that TRPV1 protects dopaminergic neurons from injury by promoting GDF11 expression in PD model.

P2Y6 regulates cytoskeleton reorganization and cell migration of C2C12 myoblasts via ROCK pathway.

Migration of skeletal muscle precursor cells is required for limb muscle development and skeletal muscle repair. This study aimed to examine the role of P2Y6 receptor in C2C12 myoblasts migration. C2C12 myoblasts were treated with P2Y6 agonist UDP, P2Y6 antagonist MRS2578, Ca2+ channel blocker BTP2, or ROCK inhibitor GSK269962 or Y27632, and the migration ability of C2C12 cells was assessed by wound healing assay. The cellular Ca2+ content was analyzed with fluo-4 probe and the activation of ROCK (phosphorlyation of LIMK and cofilin) was assayed by western blot. The cytoskeleton was labeled with Actin-Tracker Green and Tubulin-Tracker-Red. Silencing P2Y6 expression in C2C12 myoblasts reduced intracellular Ca2+ content and cell motility. Whereas UDP increased cellular Ca2+ content, actin filaments, and cell migration, MRS2578 had the opposite effects. The effects of UDP were abrogated by BTP2 and GSK269962 (and Y27632). Disruption of P2Y6 signaling pathway caused C2C12 myoblasts to have an elongated morphology. These results demonstrated that P2Y6 signaled through Ca2+ influx and RhoA/ROCK to reorganize cytoskeleton and promote migration in myoblasts.

Electroacupuncture attenuates ac4C modification of P16 mRNA in the ovarian granulosa cells of a mouse model premature ovarian failure.

BACKGROUND: Premature ovarian failure (POF) is a type of pathological aging, which seriously interferes with the fertility of affected women. Electroacupuncture (EA) may have a beneficial effect; however, its mechanism of action is unknown. The purpose of this study was to determine the effect of EA on ovarian function in ovarian granulosa cells (OGCs) in a cyclophosphamide (CTX)-induced mouse model of POF. METHODS: Mice were divided into three groups: wild type (WT) group, CTX group and CTX + EA group. EA was administered under isoflurane anesthesia at CV4, ST36 and SP6 for 30 min every 2 days, 2-3 times per week for a total of 4 weeks. Effects of EA on ovarian weight and level of estrogen were examined. The mRNA and protein expression levels of cell cycle-associated proteins were detected and mRNA modifications were analyzed. RESULTS: EA significantly increased ovarian weight and reduced the proportion of atretic follicles in mice with CTX-induced POF (p < 0.05). EA increased the level of estrogen in the peripheral blood of mice and inhibited the modification of total mRNA N4-acetylcytidine (ac4C). A significant increase in the expression of P16 and N-acetyltransferase 10 (NAT10) and a significant decrease in the expression of Cyclin D (CCND1) and cyclin-dependent kinase 6 (CDK6) were observed in the OGCs of POF mice (p<0.05). After EA, P16 and NAT10 expression was decreased, and CCND1 and CDK6 expression was increased. Finally, EA reduced the ac4C modification of P16 mRNA-specific sites in the OGCs of POF mice. CONCLUSION: This study demonstrated that EA promoted the repair of the ovarian microenvironment by inhibiting the ac4C modification of P16 mRNA to decrease its stability and expression intensity, and by altering the activity of the P16/CDK6/CCND1 axis in OGCs.

Moxibustion attenuates inflammation in intestinal mucosal by regulating RAGE-mediated TLR4-NF-κBp65 signaling pathway in vivo and in vitro.

This study was performed to investigate the effect of moxibustion on the RAGE/TLR4-NF-κBp65 pathways and mucosal damage in rat model of 5-fluorouracil (5-Fu)-induced intestinal mucositis (IM) and the underlying mechanisms. 5-Fu treatment significantly increased the expression of the receptor for advanced glycation end products (RAGE) and its ligand, thehigh-mobility group box 1 protein (HMGB1), in the rat intestinal tissue. The inhibition of RAGE could induce the repair of intestinal mucosal damage and downregulate the expression of Toll-like receptor (TLR)-4 and nuclear factor kappa-B (NF-κB) p65 in intestinal tissues of 5-Fu-treated rats. Moxibustion treatment significantly improved the physical symptoms and repaired the intestinal mucosal damage of IM rats and increased the expression of tight junction proteins in these rats. The expression of RAGE, HMGB1, TLR4, NF-κBp65, and related downstream inflammatory factors, namely, tumor necrosis factor (TNF)-α, interleukin (IL)-6 and IL-1ß, were significantly decreased after moxibustion treatment. A moxibustion dose of 15 min/day exerted a better therapeutic effect than a dose of 30 min/day. The phosphorylation of NF-κBp65 and IκBa is involved in reducing inflammation by regulating the RAGE signaling pathway. Moxibustion can reduce intestinal mucosal damage and inflammation in 5-Fu-induced IM rats via modulation of the RAGE/TLR4-NF-κBp65 signaling pathways.

Human Amniotic Fluid Mesenchymal Stem Cell-Derived Exosomes Inhibit Apoptosis in Ovarian Granulosa Cell via miR-369-3p/YAF2/PDCD5/p53 Pathway.

Human amniotic fluid stem cell-derived exosome (HuAFSC-exosome) transplantation is considered a promising treatment for premature ovarian failure (POF). However, its mechanism remains unclear. In this study, exosomes were isolated and enriched from HuAFSC subsets of CD44+/CD105+, and the exosomes were transplanted into a POF model in vitro and in vivo. Our results confirmed that the exosomes produced by CD44+/CD105+ HuAFSCs could achieve therapeutic effects in a mouse POF model. Our research also showed that CD44+/CD105+ HuAFSC-exosomes carrying miR-369-3p could specifically downregulate the expression of YAF2, inhibit the stability of PDCD5/p53, and reduce the apoptosis of ovarian granulosa cells (OGCs), thereby exerting therapeutic effects on POF. Knowledge of these mechanisms demonstrates that miRNAs carried by CD44+/CD105+ HuAFSC-exosomes are critical to the therapy of POF. This will be useful for the clinical application of stem cells.

Electroacupuncture May Inhibit Oxidative Stress of Premature Ovarian Failure Mice by Regulating Intestinal Microbiota.

Premature ovarian failure (POF) is the leading cause of female infertility, and there is no optimal treatment or medication available currently. For POF, electroacupuncture (EA) has been considered a promising therapeutic approach, but the mechanism for this is not clear. In this study, we explored the effects of EA (CV4, ST36, and SP6) on oxidative stress and intestinal microbiota of high-fat and high-sugar- (HFHS-) induced POF mice. The development of mice follicles was observed by hematoxylin and eosin (HE) staining. The serum levels of estrone (E1), estrogen (E2), estriol (E3), and 21-deoxycortisol (21D) were measured by the HPLC-MS/MS method. The concentrations of Fe2+, superoxide dismutase (SOD), hydroxyl radical (·OH), glutathione (GSH), superoxide anion, and malondialdehyde (MDA) were measured by spectrophotometry. The 16S-rDNA sequencing was used to measure many parameters related to the host gut bacteriome and mycobiome composition, relative abundance, and diversity. mRNA expression levels of ferroptosis-related genes were determined by RT-qPCR. After 4 weeks of EA intervention in POF mice, mature follicles were increased and the levels of the sex hormone were improved. SOD activities, antisuperoxide activities, and GSH increased while MDA, ·OH, and Fe2+ decreased. In addition, EA also altered the intestinal microbiota. These results reveal that EA can effectively inhibit ovarian oxidative stress and the accumulation of Fe2+ in POF mice. It may be that the alteration in the intestinal microbiota is one of the potential mechanisms of EA treatment. These findings suggest that EA has clinical potential as a safe treatment for POF.

Efficacy of Acupuncture Combined with Rehabilitation Training for Intensive Care Unit-Acquired Muscle Weakness: A Protocol for a Randomized, Sham-Procedure-Controlled Clinical Trial.

To evaluate the efficacy of acupuncture combined with rehabilitation training in patients with intensive care unit (ICU)-acquired muscle weakness (ICUAW), a single-blinded, randomized, sham-controlled clinical trial is designed for execution. In total, 56 participants with ICUAW will be randomly assigned to the treatment and control groups with 28 participants in each group. The participants will be treated with acupunctures or sham procedures at LI15, LI11, ST36, GB34, and ST31, 5 times per week for a total of 20 sessions in 4 weeks while they will receive rehabilitation training. Patients will be followed up every month for 3 months after treatment. The primary outcomes include changes in quadriceps femoris muscle area, thickness, vastus intermediate muscle thickness, subcutaneous tissue thickness, and ultrasonic intensities of the rectus femoris. The secondary outcomes consist of the modified Barthel index score and the Medical Research Council total score. Participants' mechanical ventilation, the rate of detachment at the second week, the 28-day survival rate, and the occurrence of adverse reactions will be measured, and any side effects will be reported and recorded. Patient outcomes between the treatment and control groups will be compared and statistically tested. We anticipate that the therapeutic regimen of acupuncture combined with rehabilitation training would be more effective than the rehabilitation training alone for the treatment of the ICUAW. The findings of this study could help develop a better strategy for the treatment of the ICUAW disease and explore a clinical application of an acupuncture technique. Trial registration: Chinese Clinical Trial Register ChiCTR2000038779. Registered 30 September, 2020, https://www.chictr.org.cn/showproj.aspx?proj=62284.

Efficacy of acupuncture at three nasal acupoints plus acupoint application for perennial allergic rhinitis: A multicenter, randomized controlled trial protocol.

BACKGROUND: Many studies have shown the potential therapeutic effect of acupuncture on allergic rhinitis. Most of these studies were limited by low-quality evidence. Preliminary experiments showed that the use of acupuncture at three nasal acupoints plus acupoint application (AAP) achieves a more persistent effect in the treatment of perennial allergic rhinitis than acupuncture alone. In this study, a multicenter, single-blind, randomized controlled trial will be performed, in which acupuncture at nonmeridian acupoints and sham AAP will be used as the control group to evaluate the effect of AAP through long-term observation. METHODS: The trial is designed on the basis of the Consolidated Standards of Reporting Trials 2010 guidelines and Standards for Reporting Interventions in Controlled Trials of Acupuncture. A total of 120 participants with perennial allergic rhinitis will be randomly assigned to a treatment or control group. A specially appointed investigator will be in charge of randomization. The participants in the treatment group will be treated with acupuncture at EX-HN3, LI20, and EX-HN8 thrice per week for a total of 12 sessions. In addition, they will undergo AAP at DU14, BL13, EX-BI, and RN22. The participants in the control group will be treated with sham AAP. The primary outcome will be the change in the Total Nasal Symptom Score from baseline to the completion of 4-week treatment. Secondary outcomes include changes in visual analog scale and total non-nasal symptom scores from baseline to the second and fourth weeks of treatment, as well as 1, 3, and 6 months after the completion of treatment. Peripheral blood IL-4, IL-5, IL-6, IL-8, and IL-10 levels will be measured, and any side effects related to treatment will be observed and recorded. DISCUSSION: It is expected that this randomized clinical trial will provide evidence to determine the effects of AAP compared with acupuncture at nonmeridian acupoints and sham AAP, particularly the long-term effect. These findings will help improve the clinical application of this technique. TRIAL REGISTRATION: Acupuncture-Moxibustion Clinical Trial Registry AMCTR-ICR-18000179. Registered on 12 April 2018.

[Effects on polycystic ovary syndrome of abdominal obesity treated with the dialing needling method at the abdomen].

OBJECTIVE: To observe the clinical effects on polycystic ovary syndrome (PCOS) of abdominal obesity treated with the dialing needling technique at the abdomen. METHODS: A total of 33 patients of PCOS of abdominal obesity were treated with the dialing needling method. The reinforcing technique was applied to Zhongwan (CV 12), Qihai (CV 6), Guanyuan (CV 4) and Zhongji (CV 3) and the reducing technique was applied to Huaroumen (ST 24), Tianshu (ST 25), Shuifen (CV9), Luanchaoxue (Extra), Huangshu (KI 16) and Daimai (GB 26). The electric stimulation was added at Huaroumen (ST 24), Tianshu (ST 25) and bilateral Daimai (GB 26). The treatment was given once every two days, three times a week. The treatment was discontinued during the menstruation, 10 treatments made one course. Totally, 6 courses of treatment were provided. The body mass index (BMI) and the ovarian volume were observed before and after treatment. The levels of serum fasting insulin (FINS), fasting blood glucose (FBG), serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T) and prolactin (PRL) were determined before and after treatment. RESULTS: As compared with those before treatment, BMI was reduced obviously (P<0.01), the ovarian volume was shrunk obviously (P<0.01) and the levels of serum FINS, LH and T were reduced obviously (P<0.05, P<0.01) after treatment. The differences in serum PRL and FSH were not significant before and after treatment (both P>0.05). CONCLUSION: The dialing needling method reduces the body mass and ovarian volume in the patients of PCOS of abdominal obesity type. The effect mechanism is relevant with the reducing of FNS, LH and T.

[A case-control study on the treatment of carpal tunnel syndrome with needle Dao].

OBJECTIVE: To study clinical effects of needle Dao for the treatment of carpal tunnel syndrome. METHODS: From July 2014 to December 2016, 60 patients with carpal tunnel syndrome were divided into two groups: needle Dao treatment group(treatment group), and brace and medicine treatment group(control group), 30 cases in each group. There were 8 males and 22 females in the treatment group, with an average age of (49.38±7.43) years old. There were 7 males and 23 females in the control group, with an average age of (50.23±8.71) years old. There were no statistically significant differences between two groups before treatment in age, sex, course of disease, sensory nerve conduction velocity (SCV), sensory nerve action potential (SNAP), and distal motor latency(DML). The patients in the treatment group were treated with needle Dao for 1 to 2 times, while the patients in the control group were given celecoxib, vitamin B1 and vitamin B12, with the brace fixed. After treatment, the changes of SCV, SNAP and DML in two groups were observed. RESULTS: After treatment, the SCV were (48.63±7.33) m/s and (41.24±6.15) m/s in treatment group and control group separately, the SNAP were (9.89±3.71) m/s and (8.22±2.19) m/s, and the DML were(5.11±2.28) m/s and (6.13±2.23) m/s. The SCV, SNAP and DML after treatment were better than those before treatment(P<0.05). And the results of SCV, SNAP and DML of patients in the treatment group were better than those of patients in the control group(P<0.001). CONCLUSIONS: Needle Dao can effectively improve the overall treatment efficiency of patients with carpal tunnel syndrome and reduce the symptoms of patients, which is feasible and suitable to use in clinical.

[Clinical application of non-collinear spinous process in the diagnosis of cervical vertebra "Gu Cuo Feng and Jin Chu Cao"].

OBJECTIVE: To analyze value of X-ray in diagnosis of cervical vertebral rotation. METHODS: From May 2010 to June 2011,129 patients with cervical spondylosis were collected and examined by X-ray. Among them, 119 cases were collinear spinous, other 10 cases were excluded with collinear. Of involved patients, 28 cases were male,91 cases were females with an average age of (48.53 +/- 14.32) (ranged, 24 to 65) years. The segments and numbers of vertebral body spinous process which were not centered were recorded, and then CT scan was performed to examine segments of vertebral body and spinous process which were not centered to observe rotation of cervical vertebra and spinous process deviation. The relationship between numbers of spinous not centered and vertebral rotation was statistical analyzed. RESULTS: When the number of spinous not centered ranged from 1 to 6, vertebral rotation rate were 45.45%, 46.67%, 56.86%, 62.07%, 77.14%, 85.19% respectively. CONCLUSION: The more numbers of vertebral spinous process not centered, the more chance of corresponding vertebral rotation.

[Study on the correlation between "Gucuofeng and Jinchucao" and cervical spondylosis].

OBJECTIVE: To explore the relation between "Gucuofeng and Jinchucao" and cervical spondylosis. METHODS: From March 2006 to October 2011,333 patients with cervical spondylosis were collected in the study as cervical spondylosis group; 30 subjects of control group come from student of Shuguang Hospital and other health volunteer. There were 119 males and 214 females with a mean age of (48.11 +/- 12.21) years in cervical spondylosis group and there were 6 males and 24 females with a mean age of (45.27 +/- 10.12) years in control group. In aspect of the symptom and sign, dynamic palpation, X-ray examination wer performed to find the incidence rate of "Gucuofeng and Jinchucao" in two groups. RESULTS: There was significant difference in symptom and sign, dynamic palpation, X-ray examination between two groups (P<0.01). In cervical spondylosis group, "Gucuofeng and Jinchucao" occurred in 293 cases (87.99%),there was significant difference compared with control group (P<0.01). CONCLUSION: Cervical "Gucuofeng and Jinchucao" is one of pathological point of cervical spondylosis,and it can provide a guidance for the diagnosis and treatment of cervical spondylosis.

[Clinical significance of vertebral artery MRA to vertebral artery type of cervical spondylosis' diagnosis and treatment].

OBJECTIVE: To study clinical significance of vertebral artery magnetic resonance angiography (MRA) to vertebral artery type of cervical spondylosis' diagnosis and treatment. METHODS: There were two groups in the study, which were observation group and control group. There were 53 patients in observation group,including 19 males and 34 females,with a mean age of (52.51 +/- 11.29) years. There were 30 subjects in control group,including 10 males and 20 females,with a mean age of (48.11 +/- 12.21) years. Based on the vertebral artery MRA picture,the course and caliber of vertebral artery were compared between two groups. RESULTS: The abnormal incidence of course and caliber of vertebral artery in observation group was higher than that of control group, which had statistic difference (P=0.000). Furthermore, the patterns of abnormal course and caliber of vertebral artery in observation group were complicated and diverse, but the regional circuity and stegnosis was the most common pattern,about 47.18%(25/53). The cause of circuity and stegnosis was vertebra Gu-Cuofeng,about 43.41% (23/53). CONCLUSION: The vertebral artery MRA provides a guidance for the diagnosis of abnormal course and caliber of vertebral artery in vertebral artery cervical spondylosis.