Novel mechanisms underlying anti-polycystic ovary like syndrome effects of electroacupuncture in rats: suppressing SREBP1 to mitigate insulin resistance, mitochondrial dysfunction and oxidative stress

BACKGROUND: Acupuncture, a therapy of traditional Chinese medicine, is confirmed to exert the therapeutic action on polycystic ovary syndrome (PCOS). However, the detailed therapeutic mechanisms of acupuncture in PCOS remain ambiguous. In this study, we further investigated whether electroacupuncture (EA) alleviated PCOS-like symptoms in rats via regulating a metabolic regulator, sterol regulatory element binding protein-1 (SREBP1). Methods: The PCOS-like rat model was built by hypodermic injection with dehydroepiandrosterone (DHEA). The rats were subjected to EA intervention (ST29 and SP6 acupuncture points) for 5 weeks. Primary granulosa cells were isolated from control and PCOS-like rats for evaluating insulin resistance, mitochondrial dysfunction and oxidative stress in vitro. RESULTS: The expression of SREBP1 was increased in PCOS-like rats, which was suppressed by EA treatment. In addition, lentivirus-mediated overexpression of SREBP1 restrained EA treatment-induced improvement in pathological changes, serum hormone levels and insulin resistance in rats. In addition, overexpression of SREBP1 repressed insulin-stimulated phosphorylation of insulin receptor ß (IR) and AKT in primary granulosa cells. Moreover, upregulation of SREBP1 further exacerbated mitochondrial dysfunction and oxidative stress in granulosa cells isolated from PCOS-like rats. Mechanically, EA treatment suppressed SREBP1 expression through inducing the activation of AMP-activated protein kinase (AMPK) signaling pathway in PCOS-like rats. CONCLUSION: EA intervention alleviated PCOS-like symptoms in rats via improving IR, mitochondrial dysfunction and oxidative stress through regulating SREBP1, a lipid metabolism regulator. Our findings illuminate the novel protective mechanisms of EA in the treatment of PCOS.

Linc-OIP5 in the breast cancer cells regulates angiogenesis of human umbilical vein endothelial cells through YAP1/Notch/NRP1 signaling circuit at a tumor microenvironment

BACKGROUND: LincRNAs have been revealed to be tightly associated with various tumorigeneses and cancer development, but the roles of specific lincRNA on tumor-related angiogenesis was hardly studied. Here, we aimed to investigate whether linc-OIP5 in breast cancer cells affects the angiogenesis of HUVECs and whether the linc-OIP5 regulations are involved in angiogenesis-related Notch and Hippo signaling pathways. METHODS: A trans-well system co-cultured HUVECs with linc-OIP5 knockdown breast cancer cell MDA-MB-231 was utilized to study the proliferation, migration and tube formation abilities of HUVECs and alterations of related signaling indicators in breast cancer cells and their conditioned medium through a series of cell and molecular experiments. RESULTS: Overexpressed linc-OIP5, YAP1, and JAG1 were found in breast cancer cell lines MCF7 and MDA-MB-231 and the expression levels of YAP1 and JAG1 were proportional to the breast cancer tissue grades. MDA-MB-231 cells with linc-OIP5 knockdown led to weakened proliferation, migration, and tube formation capacity of co-cultured HUVECs. Besides, linc-OIP5 knockdown in co-cultured MDA-MB-231 cells showed downregulated YAP1 and JAG1 expression, combined with a reduced JAG1 level in conditioned medium. Furthermore, a disrupted DLL4/Notch/NRP1 signaling in co-cultured HUVECs were also discovered under this condition. CONCLUSION: Hence, linc-OIP5 in MDA-MB-231 breast cancer cells may act on the upstream of the YAP1/Notch/NRP1 signaling circuit to affect proliferation, migration, and tube formation of co-cultured HUVECs in a non-cellular direct contact way through JAG1 in conditioned medium. These findings at least partially provide a new angiogenic signaling circuit in breast cancers and suggest linc-OIP5 could be considered as a therapeutic target in angiogenesis of breast cancers.

Expression of lncRNA DGCR5 in esophageal squamous cell carcinoma tissues and its clinical significance / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To investigate the expression of long non-coding RNA (lncRNA) DiGeorge syndrome critical region gene 5 (DGCR5) in esophageal squamous cell carcinoma (ESCC) tissues, and to analyze its relationship with clinicopathological features and prognosis of ESCC patients. Methods: The expression of DGCR5 in ESCC data set from TCGA database was analyzed by bioinformatics method. Sixty pairs of ESCC tissues and para-cancerous tissues resected at the Fourth Hospital of Hebei Medical University from August 2016 to March 2017 were collected for this study. The expression of DGCR5 in ESCC tissues was detected by qPCR. The correlation between the expression of DGCR5 and the clinicopathological features and prognosis of ESCC patients was analyzed. Results: TCGAdatabase analysis showed that the expression of DGCR5 in ESCC tissues was significantly higher than that in normal esophageal tissues (P<0.01). The expression of DGCR5 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.01). The expression level of DGCR5 was significantly correlated with TNM staging and lymph node metastasis in ESCC patients (all P<0.05). Kaplan-Meier univariate analysis showed that the 2-year survival rate of ESCC patients with high DGCR5 expression was significantly lower than that of patients with low expression (P<0.05). Conclusion: DGCR5 is highly expressed in ESCC tissues and is closely related to TNM staging, lymph node metastasis and poor prognosis, which may serve as a molecular marker for early diagnosis and prognosis prediction of ESCC.

Role of JNK and ERK1/2 MAPK signaling pathway in testicular injury of rats induced by di-N-butyl-phthalate (DBP)

BACKGROUND: Di-N-butyl-phthalate (DBP) is an endocrine disrupting substance. We investigated the adverse effect of DBP on testis of male rat and reveal its potential mechanism of MAPK signaling pathway involved this effect in vivo and in vitro. Gonadal hormone, sperm quality, morphological change and the activation status of JNK, ERK1/2 and p38 was determined in vivo. Primary Sertoli cell was established and cultivated with JNK, ERK1/2 inhibitors, then determine the cell viability, apoptosis and the expression of p-JNK, p-ERK1/2. Data in this study were presented as mean ± SD and determined by one-way analysis of variance (ANOVA) followed by Bonferroni's test. Difference was considered statistically significant at P < 0.05. RESULTS: In vivo experiment, DBP impaired the normal structure of testicular tissue, reduced testosterone levels in blood serum, decreased sperm count and increased sperm abnormality, p-ERK1/2 and p-JNK in rat testicular tissue increased in a dose-dependent manner. In vitro studies, DBP could decrease the viability of Sertoli cells and increase p-ERK1/2 and p-JNK. Cell apoptosis in SP600125 + DBP group was significantly lower than in DBP group (P < 0.05). p-JNK was not significantly decreased in SP600125 + DBP group, while p-ERK1/2 was significantly decreased in U0126 + DBP group. CONCLUSIONS: These results suggest that DBP can lead to testicular damage and the activation of ERK1/2 and JNK pathways, the JNK signaling pathway may be primarily associated with its effect.

HIV/AIDS knowledge, attitude and awareness of HIV/AIDS policy and regulations among medical students of higher vocational colleges in Sichuan / 中国学校卫生

Objective@#To understand HIV/AIDS knowledge, attitude, and awareness of HIV/AIDS policies and regulations among vocational medical students in Sichuan, and to provide scientific basis for AIDS prevention and treatment in higher vocational medical colleges.@*Methods@#A total of 1 458 medical students in a vocational college in Sichuan Province were selected through stratified cluster random sampling and investigated with questionnaire on HIV/AIDS related knowledge, policies and regulations.@*Results@#The average score of AIDS knowledge was(6.63±1.31), and the awareness rate was 82.9%; The average score of attitude towards AIDS was(2.17±0.45), with the highest score being fear and avoidance(2.56±0.61), and the lowest score responsibility perception(1.66±0.58); The total score of HIV/AIDS related policies and regulations was(19.17±2.91). AIDS knowledge was positively correlated with attitude and understanding of policies and regulations(r=-0.24, 0.28, P<0.01) , the higher the knowledge score, the better the attitude and the higher the understanding of policies and regulations.@*Conclusion@#HIV/AIDS knowledge of higher vocational medical students has been improved, the fear and avoidance of AIDS is widespread. There are insufficient awareness in AIDS related policies and regulations. Higher vocational medical colleges should be more targeted AIDS health education.

Bloodstream Infections Caused by Enterococcus spp: A 10-year Retrospective Analysis at a Tertiary Hospital in China / 华中科技大学学报（医学）（英德文版）

In order to discover the risk factors for 30-day mortality in bloodstream infections (BSI) caused by Enterococcus spp.strains,we explored the clinical and therapeutic profile of patients with Enterococcus spp.BSI and the characteristics of this condition.A total of 64 patients with BSI caused by Enterococcus spp.who were treated in our hospital between 2006 and 2015 were included in the study.The clinical features of patients,microbiology,and 30-day mortality were collected from the electronic medical records database and analyzed.The results showed that there were 38 patients infected by Enterococcus faecalis (E.faecalis),24 by Enterococcus faecium (E.faecium),1 by Enterococcus casseliflavus (E.casseliflavus),and 1 by Enterococcus gallinarum (E.gallinarum).A Charlson comorbidity score ≥5,corticosteroid treatment,placement of catheters or other prosthetic devices and history of antibiotic use were found more frequently in E.faecium BSI patients than in E.faecalis patients (P=0.017,P=0.027,P=0.008 and P=0.027,respectively).Furthermore,the univariate and multivariate analysis showed that corticosteroid treatment (OR=17.385,P=0.008),hospital acquisition (OR=16.328,P=0.038),and vascular catheter infection (OR=14.788,P=0.025) were all independently associated with 30-day mortality.Our results indicate that E.faecalis and E.faecium are two different pathogens with unique microbiologic characteristics,which cause different clinical features in BSI,and the empiric antimicrobial treatments are paramount for patients with enterococcal BSI.

Silencing MTA1 by RNAi Reverses Adhesion, Migration and Invasiveness of Cervical Cancer Cells (SiHa) via Altered Expression of p53, and E-cadherin/β-catenin Complex / 华中科技大学学报（医学）（英德文版）

It has been reported that metastasis-associated gene 1 (Mtal) is overexpressed in many malignant tumors with high metastatic potential.In addition,some studies indicated that MTA1 participated in invasion,metastasis,and survival of cancer cells by regulating cell migration,adhesion and proliferation.But the role of MTA 1 is unclear in vitro in the development of cervical cancer cells.This study investigated whether and how MTA1 mediated cell proliferation,migration,invasion and adhesion in cervical cancer.MTA1 expression level was detected by Western blot in two cervical cancer cell lines of different invasion potentials.The effects of MTA1 expression on SiHa cell apoptosis,cycle,proliferation,migration,invasion and adhesion were tested by flow cytometry,MTT,wound-healing assay,Transwell assay and adhesion assay,respectively.The expression levels of p53,E-cadherin,and β-catenin activity were evaluated in untreated and treated cells.The results showed that MTA1 protein expression was significantly higher in SiHa than in HeLa,which was correlated well with the potential of migration and invasion in both cell lines.Furthermore,the cell invasion,migration and adhesion capabilities were decreased after inhibition of MTA1 expression mediated by Mtal-siRNA transfection in SiHa.However,no significant differences were found in cell apoptosis,cycle,and proliferation.In addition,E-cadherin and p53 protein levels were significantly up-regulated,while β-catenin was significantly down-regulated in SiHa transfected with the siRNA.These results demonstrated that MTA1 played an important role in the migration and invasion of cervical cancer cells.It was speculated that the decreased migration and invasion capability by inhibiting the MTA1 expression in the SiHa cell line may be mediated through the altered expression of p53,and E-cadherin/β-catenin complex.MTA1 could serve as a potential therapeutic target in cervical cancer.

TLR9 Expression and Its Rol*e in Chemosensitivity to DDP in Human Cervical Cancer Cells in vitro / 华中科技大学学报（医学）（英德文版）

Inflammation and infection play an important role in the pathogenesis of many cancers.Toll-like receptors (TLRs) are a class of pattern recognition receptors that recognize conserved components of microbes and trigger the immune response against invading microorganisms.Toll-like receptor 9 (TLR9) recognizes non-methylated cytosine-phosphateguanosine (CpG) DNA sequences which are the surrogate for viral DNA.TLR9 may react to tumor development and progression during chronic inflammation that involves the tumor microenvironment.In order to study the role of TLR9 in cervical cancer,we analyzed the TLR9 expression in different types of HPV infection cervical cancer cells.Then we detected if CpG sequences influenced the TLR9 expression and the sensitivity to cisplatin (DDP) of these cervical cancer cells in vitro.The expression of TLR9 mRNA and protein in SiHa,Hela and C33A cells was detected by RT-PCR and Western blotting.Real-time PCR was used to examine the TLR9 expression changes induced by CpG.Chemosensitivity of the cervical cancer cells to cisplatin (DDP) was measured by MTT.It was observed that the expression of TLR9 mRNA and protein was increased gradually in SiHa (HPV16+),Hela (HPV18+) and C33A (HPV-) cells.Low doses of CpG increased the TLR9 expression only in C33A (HPV-) cells,but not in SiHa (HPV16+) and Hela (HPV18+) cells.Furthermore,low dose of CpG significantly increased the sensitivity ofC33A (HPV-) cells,but not that of SiHa (HPV16+) and Hela (HPV18+) cells.These results indicated that TLR9 may serve as a protective agent in HPV negative cervical cancer cells.It was concluded that TLR9 could improve the sensitivity to DDP in HPV negative cervical cancer cells and might represent a potential therapeutic option in clinical practice.

Relationship between the Expression of Thymidylate Synthase,Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase and Survival in Epithelial Ovarian Cancer / 华中科技大学学报（医学）（英德文版）

The mRNA and protein expression of thymidylate synthase(TS),thymidine phosphorylase(TP)and dihydropyrimidine dehydrogenase(DPD)and their relationship with prognosis were investigated.Real-time quantitative RT-PCR(Taqman)was used to detect the mRNA expression of TS,TP and DPD in formalin-fixed and paraffin-embedded 106 samples of epithelial ovarian cancer and 29 normal ovaries.A TATA box-binding protein(TBP)was used as an endogenous reference gene.A relationship between TS,TE DPD expression and clinicopathologic features was investigated.The protein location and expression of TS,TP and DPD was examined in the same patients by an avidin-biotin-peroxidase immunohistochemistry.TS and TP mRNA expression levels were significantly higher in tumor group than in normal controls,with the average value of TS and TP mRNA being 6.14±0.62 and 0.59±0.06 in tumor tissue,and 0.71±0.14 and 0.16±0.04 in normal tissue,respectively.DPD mRNA expression levels were significantly lower in tumor group(0.11±0.02)than in normal controls(0.38±0.05).There was statistically significant difference in TS and TP mRNA expression levels among different pathological grades and clinical stages(P<0.05),but histological subtype was not significantly associated with TS and TP mRNA expression.DPD gene expression was not significantly associated with any clinicopathological parameters.Immunohistochemistry revealed that TP protein was mainly distributed in nucleus,and TS and DPD mainly in cytoplasm.The protein expression intensity of TS,TP and DPD was coincided with the mRNA expression levels.It was concluded that TS,TP mRNA and protein expression levels were significantly higher in epithelial ovarian cancer,and DPD mRNA and protein expression levels were significantly lower.The expression levels of TS and DPD were related to the patients' prognosis and survival.Combined gene expression levels of TS,TP and DPD represent a new variable to predict the clinical outcome in ovarian cancer.The association of TS,TP and DPD expression levels with survival suggests an importance of these genes for tumor occurrence and progression.