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Arrhythmogenic cardiomyopathy (ACM), a fatal heart disease characterized by fibroadipocytic replacement of cardiac myocytes, accounts for 20% of sudden cardiac death and lacks effective treatment. It is often caused by mutations in desmosome proteins, with Desmoglein-2 (DSG2) mutations as a common etiology. However, the mechanism underlying the accumulation of fibrofatty in ACM remains unknown, which impedes the development of curative treatment. Here we investigated the fat accumulation and the underlying mechanism in a mouse model of ACM induced by cardiac-specific knockout of Dsg2 (CS-Dsg2 -/-). Heart failure and cardiac lipid accumulation were observed in CS-Dsg2 -/- mice. We demonstrated that these phenotypes were caused by decline of fatty acid (FA) β-oxidation resulted from impaired mammalian target of rapamycin (mTOR) signaling. Rapamycin worsened while overexpression of mTOR and 4EBP1 rescued the FA β-oxidation pathway in CS-Dsg2 -/- mice. Reactivation of PPARα by fenofibrate or AAV9-Pparα significantly alleviated the lipid accumulation and restored cardiac function. Our results suggest that impaired mTOR-4EBP1-PPARα-dependent FA β-oxidation contributes to myocardial lipid accumulation in ACM and PPARα may be a potential target for curative treatment of ACM.
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Objective:To explore the effect of miR-21 on cell proliferation, apoptosis, invasion and radiosensitivity of cervical cancer HeLa cells and unravel the underlying mechanism.Methods:RT-qPCR assay was used to detect the expression levels of miR-21 in cervical cancer tissues and adjacent non-tumor tissues, normal cervical epithelial cells (H8) and cervical cancer cell lines (HeLa, SiHa, ME180). HeLa cell line with inhibition of miR-21 or knockdown of RECK were constructed. CCK-8, Caspase3/7 live cell apoptosis detection, wound healing test, Transwell invasion, clone formation assay, Western blot and immunofluorescence were performed to detect cell viability, apoptosis, migration, invasion, radiosensitivity and related proteins. The dual luciferase assay verified whether miR-21 targeted RECK.Results:MiR-21 level in the cervical cancer tissues was significantly higher than that in its corresponding adjacent non-tumor tissues ( P<0.05). The expression levels of miR-21 in cervical cancer cell lines HeLa, SiHa and ME180 were significantly up-regulated compared with those in normal cervical epithelial cells H8(all P<0.05). MiR-21 knockdown significantly inhibited HeLa cell viability, promoted cell apoptosis, reduced radiation tolerance, down-regulated the expression of Cyclin D 1,Bcl-2, MMP-2 and MMP-9, and up-regulated the expression P21 and Bax proteins (all P<0.05). miR-21 targeted the 3’-UTR of RECK mRNA and negatively regulated the expression of RECK. Silencing RECK reversed the effects of miR-21 knockdown on HeLa cell apoptosis, migration, invasion and radiosensitivity. Conclusions:Inhibiting the expression of miR-21 significantly decreases cell viability, induces cell apoptosis, weakens cell migration and invasion capabilities, and enhances the radiosensitivity of HeLa cells. The potential mechanism is closely related to the targeted up-regulation of RECK.
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The clinical data of a child with systemic lupus erythematosus (SLE) and hypophosphatasia (HPP) admitted to the Department of Pediatrics, the Second Hospital of Jilin University in December 2015 were retrospectively analyzed.The patient was a 10-year-old boy who was hospitalized because of fever and facial rashes in the past 4 days.He had a history of HPP for 7 years.His clinical manifestations included skeletal and dental dysplasia, oral ulcers, buccal erythema and renal lesions.Laboratory examination showed a low level of serum alkaline phosphatase, whole blood cell count decreased, antinuclear antibody(ANA) 1∶1 000, anti-double strand DNA antibodies positive, anticardiolipin antibodies positive, complement 3 (C 3) and C 4 decreased.Therefore, he was diagnosed with SLE.After glucocorticoid, immunosuppressant and symptomatic treatment, the child′s condition improved and he discharged from the hospital.He was followed up regularly, and died 2 years after the diagnosis of SLE.SLE complicated with HPP is extremely rare in clinical practice, and the symptoms may overlap.Hence these two diseases should be differentiated.
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The clinical data of a child with systemic lupus erythematosus (SLE) and hypophosphatasia (HPP) admitted to the Department of Pediatrics, the Second Hospital of Jilin University in December 2015 were retrospectively analyzed.The patient was a 10-year-old boy who was hospitalized because of fever and facial rashes in the past 4 days.He had a history of HPP for 7 years.His clinical manifestations included skeletal and dental dysplasia, oral ulcers, buccal erythema and renal lesions.Laboratory examination showed a low level of serum alkaline phosphatase, whole blood cell count decreased, antinuclear antibody(ANA) 1∶1 000, anti-double strand DNA antibodies positive, anticardiolipin antibodies positive, complement 3 (C 3) and C 4 decreased.Therefore, he was diagnosed with SLE.After glucocorticoid, immunosuppressant and symptomatic treatment, the child′s condition improved and he discharged from the hospital.He was followed up regularly, and died 2 years after the diagnosis of SLE.SLE complicated with HPP is extremely rare in clinical practice, and the symptoms may overlap.Hence these two diseases should be differentiated.
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Objective:To investigate the clinical characteristics of patients with hepatic metastasis after cervical cancer operation, and analyze the risk factors and prognostic factors of hepatic metastasis.Methods:A total of 1 312 patients with stage Ⅰa2-Ⅱb2 cervical cancer received radical surgery from January 2013 to January 2016 in the First Affiliated Hospital of Zhengzhou University were collected, of which 13 cases (0.99%, 13/1 312) had hepatic metastasis after operation. A retrospective analysis was conducted on clinical features of patients with hepatic metastasis after cervical cancer operation. T-test, chi-square test, rank sum test and logistic regression were used to analyze the risk factors of hepatic metastasis after surgery of cervical cancer operation. Kaplan-Meier method was used for survival analysis. Log-rank test was used for screening of prognostic factors in patients with postoperative hepatic metastasis.Results:(1) Clinical features: there were 3 cases of simple hepatic metastasis without obvious clinical symptoms, 2 patients with perihepatic lymph node metastasis showed only low back pain, 8 patients with multiple extrahepatic metastases, and their clinical symptoms were related to the site of metastasis. Five cases out of 9 (5/9) with liver metastasis had abnormal tumor marker results. The abnormal kinds of tumor markers were mainly carcinoembryonic antigen (CEA), CA 125, CA 199, and CA 72-4. The interval time of hepatic metastasis after operation was 2-22 months. (2) Analysis of risk factors for hepatic metastasis: univariate analysis showed that lymph node metastasis, histological type, infiltration depth, and lymph-vascular space invasionwere associated with hepatic metastasis after cervical cancer surgery ( P<0.05).Multivariate analysis showed that lymph node metastasis and small cell carcinoma were independent risk factors for postoperative hepatic metastasis ( P<0.05). (3) Prognostic factors in patients with hepatic metastasis:among 13 patients with postoperative hepatic metastases from cervical cancer, 9 died during the follow-up period and 4 survived. The median total survival time after hepatic metastases was 7 months (range 3-32 months). Univariate analysis showed that multiple extrahepatic metastases and treatment after hepatic metastasis had significant effects on the prognosis of patients with hepatic metastasis after cervical cancer operation ( P<0.05). Conclusions:The interval of hepatic metastasis after surgery for cervical cancer operation is within 2 years. Patients with lymph node metastasis and small cell carcinoma are more prone to postoperative hepatic metastasis. The prognosis of patients with extrahepatic multiple metastases is poor, and individualized treatment should be carried out after comprehensive analysis for patients with hepatic metastasis after cervical cancer operation.
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Endoplasmic reticulum (ER) stress is mediated by disturbance of Ca²⁺ homeostasis. The store-operated calcium (SOC) channel is the primary Ca²⁺ channel in non-excitable cells, but its participation in agent-induced ER stress is not clear. In this study, the effects of tunicamycin on Ca²⁺ influx in human umbilical vein endothelial cells (HUVECs) were observed with the fluorescent probe Fluo-4 AM. The effect of tunicamycin on the expression of the unfolded protein response (UPR)-related proteins BiP and CHOP was assayed by western blotting with or without inhibition of Orai1. Tunicamycin induced endothelial dysfunction by activating ER stress. Orai1 expression and the influx of extracellular Ca²⁺ in HUVECs were both upregulated during ER stress. The SOC channel inhibitor SKF96365 reversed tunicamycin-induced endothelial cell dysfunction by inhibiting ER stress. Regulation of tunicamycin-induced ER stress by Orai1 indicates that modification of Orai1 activity may have therapeutic value for conditions with ER stress-induced endothelial dysfunction.
Subject(s)
Blotting, Western , Calcium , Endoplasmic Reticulum , Endoplasmic Reticulum Stress , Endothelial Cells , Homeostasis , Human Umbilical Vein Endothelial Cells , Tunicamycin , Unfolded Protein ResponseABSTRACT
<p><b>OBJECTIVE</b>To investigate the role of miR-199a-3p in cardiac fibrosis and the potential target of miR-199a-3p.</p><p><b>METHODS</b>Cardiac fibroblasts were isolated from C57BL/6 mice and cultured. The miR-199a-3p mimic and Smad1 siRNA were transiently transfected into the cardiac fibroblasts via liposome. Dual luciferase reporter assay was performed to confirm the interaction between miR-199a-3p and the 3'-UTR of Smad1. The expressions of Smad1 and fibrosis-related genes at the mRNA and protein levels in the cells after miR-199a-3p mimic transfection were determined using RT-qPCR and Western blotting, respectively. The expressions of Smad1, Smad3 and fibrosis-related genes at the protein level in cells transfected with miR-199a-3p mimic and Smad1 siRNA were detected using Western blotting.</p><p><b>RESULTS</b>Over-expression of miR-199a-3p significantly increased the expression of cardiac fibrosis-related genes in cultured mouse cardiac fibroblasts. Dual luciferase reporter assay revealed the interaction of miR-199a-3p with the 3'-UTR of Smad1. The results of RT-qPCR and Western blotting confirmed that miR-199a-3p inhibited Smad1 expression at the post- transcriptional level. Transfection with miR-199a-3p mimic and siRNA-mediated Smad1 silencing consistently activated the Smad3 signaling pathway and enhanced the expressions of cardiac fibrosis-related genes in the cardiac fibroblasts.</p><p><b>CONCLUSIONS</b>As the target gene of miR-199a-3p, Smad1 mediates the pro-fibrotic effect of miR-199a-3p by activating the Smad3 signaling in cultured mouse cardiac fibroblasts.</p>
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This paper aims to discuss the clinical significance of laparotomy nerve sparing radical hysterectomy [NSRH] on rectal function of early cervical cancer patients, compared with conventional radical hysterectomy. 30 cases of early cervical carcinoma patients who had received surgery in the First Affiliated Hospital of Zhengzhou University from June 2010 to June 2014 were selected as subjects. Patients were divided into two groups, with 15 in each group, in which Group A had received NSRH, B received CRH, and all them were in stage IB-IIA1. In the surgery, 2 cases of patients in NSRH group failed in nerve sparing operation, and were grouped into CRH group. The postoperative condition of two groups were observed, recorded and compared as well, especially the comparison between the postoperative recovery condition of rectal function of two groups. The comparison were conducted between two groups on the operation time, bleeding volume, quantity of cleaned pelvic lymph node, resection length of parametrium, resection length of vagina, etc. There was no statistical significance [P>0.05]. The postoperative urinary catheter indwelling time in NSRH group was shorter than CRH group, with statistical significance [P<0.05]. The postoperative maximum urine flow, maximum cystometric capacity, maximum detrusor pressure and urinary complications in NSRH group were significantly better than the postoperative condition in CRH group, with statistical significance [P<0.05]. NSRH surgery was safe and reliable, which not only had obvious advantage in improving postoperative rectal function and bladder function, but also had a significantly effect on improving postoperative life quality as well. The results proved that patients had low disease morbidity and with great clinical significance
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This paper aims to investigate the clinical curative effect and adverse reactions of cryotherapy combined with interferon in the treatment of chronic cervicitis complicated with HPV infection. 100 cases diagnosed with chronic cervicitis complicated with HPV infection from August 2014 to August 2015 in our hospital were selected and randomly divided into observation group [50 cases] and control group [50 cases]. The preoperative and postoperative HPV-DNA changes were observed, and the vaginal discharge, time of decrustation and hemostasis, HPV negative conversion ratio and clinical efficacy were compared, to record the adverse reactions during treatment. After treatment, the level of RLU/CO value of the observation group was significantly lower than that of the control group [P<0.05]; the vaginal discharge and time of decrustation and hemostasis of the observation group were shorter than that of the control group [P<0.05]; 3 months after treatment, the HPV negative conversion ratio in the observation group was significantly higher than that in the control group; the total efficiency of the observation group was higher than that of the control group, with statistical significance [P<0.05]. There were no serious adverse reactions in the two groups during the treatment. The efficiency of cervicitis complicated with HPV infection in the treatment of cryotherapy combined with interferon was more significantly. It can effectively reduce the load of HPV, promote the recovery of patients with pathological changes. Therefore, it is worth promoting
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AIM:To investigate the effect of microRNA-214 ( miR-214) on cardiomyocyte hypertrophy and the expression of the potential target genes .METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular cardiomyocytes ( NMVCs) .Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Western blot , respectively .RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-induced hypertrophic cardiomyocytes .Dual lu-ciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly increased in the hypertro-phic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hypertrophy-re-lated genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .
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AIM:To investigate the effect of miR-214 on cardiomyocyte hypertrophy and the expression of the potential target genes . METHODS:A cell model of hypertrophy was established based on angiotensin-Ⅱ( Ang-Ⅱ)-induced neonatal mouse ventricular car-diomyocytes (NMVCs).Dual luciferase reporter assay was performed to verify the interaction between miR-214 and the 3’ UTR of MEF2C.The expression of MEF2C and hypertrophy-related genes at mRNA and protein levels was determined by RT-qPCR and Wes-tern blotting, respectively.RESULTS:The expression of ANP, ACTA1,β-MHC and miR-214 was markedly increased in Ang-Ⅱ-in-duced hypertrophic cardiomyocytes .Dual luciferase reporter assay revealed that miR-214 interacted with the 3’ UTR of MEF2C, and miR-214 was verified to inhibit MEF2C expression at the transcriptional level .The protein expression of MEF2C was markedly in-creased in the hypertrophic cardiomyocytes .Moreover, miR-214 mimic, in parallel to MEF2C siRNA, inhibited the expression of hy-pertrophy-related genes in Ang-Ⅱ-induced NMVCs.CONCLUSION:MEF2C is a target gene of miR-214, which mediates the effect of miR-214 on attenuating cardiomyocyte hypertrophy .
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AIM:MicroRNAs ( miRNAs) were recognized to play significant roles in cardiac hypertrophy .But, it remains unknown whether cyclin/Rb pathway is modulated by miRNAs during cardiac hypertrophy .This study investigates the potential roles of microRNA-1 (miR-1) and microRNA-16 (miR-16) in modulating cyclin/Rb pathway during cardiomyocyte hypertrophy .METHODS:An animal model of hypertrophy was established in a rat with abdominal aortic constriction (AAC).In addition, a cell model of hypertrophy was also achieved based on PE-promoted neonatal rat ventricular cardiomyocyte .RESULTS:miR-1 and-16 expression were markedly de-creased in hypertrophic myocardium and hypertrophic cardiomyocytes in rats .Overexpression of miR-1 and -16 suppressed rat cardiac hypertrophy and hypertrophic phenotype of cultured cardiomyocytes .Expression of cyclins D1, D2 and E1, CDK6 and phosphorylated pRb was increased in hypertrophic myocardium and hypertrophic cardiomyocytes , but could be reversed by enforced expression of miR-1 and -16.CDK6 was validated to be modulated post-transcriptionally by miR-1, and cyclins D1, D2 and E1 were further validated to be modulated post-transcriptionally by miR-16.CONCLUSION: Attenuations of miR-1 and -16 provoke cardiomyocyte hypertrophy via derepressing the cyclins D1, D2, E1 and CDK6, and activating cyclin/Rb pathway.
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Objective: To explore the effects of letrozole [LE] in combination with low-dose intramuscular injection of human menopausal gonadotropin [HMG] on the ovulation induction and pregnancy of patients with polycystic ovary syndrome [PCOS]
Methods: A total of 156 patients with PCOS infertility were randomly divided into an LE group, a clomiphene citrate [CC] group and an LE + HMG group [n= 52]. LE and CC were orally taken according to the prescribed dosage on the 3rd-5th days of menstruation respectively, and 75 IU HMG was given through intramuscular injection. The ovulation induction parameters and pregnancy outcomes were observed
Results: The number of ovulation cycle of LE + HMG group was significantly higher than that of LE group [chi[2]=8.451, P<0.001]. After injection of human chorionic gonadotropin, both endometrial thickness and number of mature follicles of LE + HMG group were significantly higher than those of other two groups [P<0.001], and the daily estradiol [E2] level was also higher [q=4.531, P<0.05]. The pregnancy rate of LE + HMG group was 55.7%, which exceeded those of other two groups [compared to LE group, chi[2]=4.012, P<0.05]. In LE + HMG group, the average medication cycle of clinically pregnant patients was [2.9 +/- 0.3] weeks, which was significantly shorter than those of CC and LE groups [F=17.241, P<0.001]
Conclusion: The regimen using LE in combination with low-dose intramuscular injection of HMG has satisfactory therapeutic effects on ovulation induction, short medication cycle and high clinical pregnancy rate, which is promising for treating patients with PCOS infertility
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Objective To explore the correlation of the TIAM2 functional gene mutations with the bleeding events in patients with coronary heart disease who undergone PCI and postoperative taken anti-platelet drug clopi-dogrel. Methods One hundred and twenty Chinese Han patients who had undergone PCI and postoperative taken anti-platelet drug clopidogrel were orderly enrolled from Guangdong General Hospital. Followed for 6 months after PCI, PCR sequencing was applied to test TIAM2 promoter region genotyping assay. Results After follow-up six months , 113 cases were remained lost of 7 cases , , including 19 cases with bleeding and 94 cases without bleed-ing; Bleeding risk in patients with diabetes mellitus (OR=3.115) or taking statins (OR=11.539), may be high, but there was no significant difference (P > 0.05); TIAM2 promoter region had three variants (c.3168+3116C>T, c.3168+3261A>G,c.3168+3596A>C), including wild-type, heterozygous, and homozygous. The probability of pa-tients with bleeding were 36.84%, 52.63%and 10.53%, and there is a certain chain of state , but the genotype was not significantly correlated with bleeding events of clopidogrel antiplatelet therapy (P>0.05). Conclusion Clopi-dogrel antiplatelet therapy bleeding events were not significantly correlated with TIAM2 functional gene mutation.
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Objective To study telomerase activity in cervical cancer and it′s precursor lesion Methods Thirty six cervical cancer and 16 cervical intraepithelial neoplasia (CIN) specimens were measured for telomerase activity using TRAP ELISA, and 11 normal cervix, 6 chronic cervicitis and 8 adjacent normal tissue specimens as controls Results Mean telomerase activity in CIN, cervical cancer, and controls were 0 398?0 293, 1 580?0 819, 0 050?0 012 There was statistically significant difference among three groups ( P