Background Electroacupuncture pretreatment plays a protective role in myocardial ischemia/reperfusion (I/R) injury and microRNAs (miRNAs) could act on various facets of cardiac function. However, the role of miRNAs in the cardioprotection by electroacupuncture pre-treatment on myocardial I/R injury remains unknown. The purpose of the study was to examine whether miR-214 was involved in cardio-protection by electroacupuncture. Methods Using rat myocardial I/R model, we examined the role of electroacupuncture pretreatment in myocardial I/R injury and analyzed the changes in the expression of miR-214. In addition, I/R was simulated in vitro by performing oxy-gen-glucose deprivation (OGD) on H9c2 cell cultures, and the effect of electroacupuncture pretreatment on I/R injury as well as expressional level of miR-214 were examined in vitro. Furthermore, the miR-214 mimic was transfected into OGD-treated H9c2 cells, we analyzed the cell apoptosis, lactate dehydrogenase (LDH) and creatine kinase (CK) activities, intracellular free Ca2+concentration ([Ca2+]i) as well as the relative protein levels of sodium/calcium exchanger 1(NCX1), BCL2-like 11 (BIM), calmodulin-dependent protein kinase IIδ(CaMKIIδ) and Cyclophilin D (CypD). Results The in vivo results revealed that compared with the I/R group, the electroacupuncture pretreatment group showed significant decreased myocardial infarct size, as well as the increased indices of the cardiac function, including heart rate, mean arterial pressure, left ventricular systolic pressure and maximal rate for left ventricular pressure rising and declining (±dp/dt max). In addition, electroacupuncture pretreatment could inhibit the elevation of LDH and CK activities induced by I/R injury. The quantitative PCR (qPCR) results demonstrated electroacupuncture pretreatment could provide cardioprotection against myocardial I/R injury in rats with miR-214 up-regulation. In the meanwhile, in vitro, electroacupuncture pretreatment protected H9c2 cells from OGD-induced injury. Trans-fection of miR-214 mimic showed protective effects on OGD-induced injury to H9c2 cells by reducing apoptosis, decreasing LDH and CK activities, rescuing the OGD-induced Ca2+and down-regulating elevated protein levels of NCX1, BIM, CaMKIIδand CypD. Conclusions Our findings firstly demonstrated that electroacupuncture pretreatment promotes the expression of miR-214 in myocardial I/R injury and miR-214 contributes to the protective effect of electroacupuncture on myocardial I/R injury.

Objective:To investigate the role of long-chain non-coding RNA MEG3 and DNA demethylase TET2 in the invasive growth of pituitary growth hormone (GH) adenomas, and provide research directions for the subsequent exploration of molecular biological mechanisms.Methods:All of 60 patients with GH pituitary adenomas diagnosed in Dalian Central Hospital from January 2016 to November 2019 were collected, and the normal anterior pituitary tissue samples of 10 patients with traumatic brain injury without neurological or endocrine system diseases were collected as control group. Real-time reverse transcription-polymerase chain reaction (qPCR) was used to detect the expression of lnc RNA MEG3 and TET2 in normal pituitary tissue as well as invasive and non-invasive pituitary growth hormone adenomas, and the differences between each groups were analyzed. At the same time, age and gender were included in the research, and the effects of age and gender on the invasive growth of pituitary GH adenomas were analyzed.Results:The aggressive growth of MEG3 and TET2 was independent of patients′ age and gender. The expression of lncRNA MEG3 in invasive pituitary GH adenoma and non-invasive pituitary GH adenoma was significantly lower than that in normal control brain tissue. Analysis of the differences between the groups found that the expression level of lncRNA MEG3 in normal trauma control pituitary tissues, non-invasive GH adenomas, and invasive GH adenomas were sequentially decreased, and MEG3 expression level was related to the aggressive growth behavior of pituitary GH adenomas ( P<0.05). The expression levels of DNA demethylase TET2 in invasive pituitary GH adenomas and non-invasive pituitary GH adenomas were significantly lower than those in the normal control brain tissue, and the expression levels in the three groups of samples gradually decreased ( P<0.05). It suggested that the expression of lncRNA MEG3 was positively correlated with the expression of DNA demethylase TET2. Conclusions:The low expression of lncRNA MEG3 and DNA demethylase TET2 is closely related to the aggressiveness of pituitary growth hormone adenoma.

Objective To evaluate the effect of captopril and isoflurane preconditioning on cell apoptosis during myocardial ischemia/reperfusion (I/R) in rabbits.Methods Forty New Zealand white rabbits of both sexes,weighing 1.8-2.5 kg,were randomly divided into 5 groups (n =8 each) using a random number table:sham operation group (group S),group I/R,isoflurane preconditioning group (group I),captopril preconditioning group (group C) and captopril and isoflurane preconditioning group (group C + I).The animals were anesthetized with 3％ pentobarbital sodium 30 mg/kg.Myocardial ischemia was induced by occlusion of the left anterior descending branch of coronary artery for 30 min followed by 120 min of reperfusion.1.1％ isoflurane was inhaled for 30 min followed by 15 min washout before myocardial ischemia in group I.Captopril 25 mg/kg was given through a gastric tube into the stomach at 24 h before myocardial ischemia in group C.Captopril 25 mg/kg was given through a gastric tube into the stomach,24 h later 1.1％ isoflurane was inhaled for 30 min followed by 15 min washout,and then myocardial ischemia was performed in group C + I.The rabbits were sacrificed at the end of reperfusion and myocardial specimens were removed for microscopic examination and observation of ultrastructure,and for determination of the expression of Bcl-2 and Bax proteins (by Western blot).The apoptosis rate was detected by flow cytometry.Bcl-2/Bax ratio was calculated.Results Compared with group S,the apoptosis rate was significantly increased,the expression of Bcl-2 and Bax proteins was up-regulated,and Bel-2/Bax ratio was decreased in I/R,I,C and C + I groups (P ＜ 0.05).Compared with I/R,I and C groups,the apoptosis rate was significantly decreased,Bcl-2 protein expression was up-regulated,Bax protein expression was down-regulated,and Bcl-2/Bax ratio was increased (P ＜ 0.05),and the pathological changes were significantly attenuated in group C + I.Conclusion Regulation of Bcl-2/Bax ratio and inhibition of apoptosis in myocardial cells are involved in the mechanism by which isoflurane and captopril preconditioning reduces I/R injury in rabbits.

The roots of Glycyrrhiza uralensis are widely used in Chinese medicine for their action of clearing heat, detoxicating, relieving cough, dispelling sputum and tonifying spleen and stomach. The reason why Glycyrrhiza uralensis has potent and significant actions is that it contains various active secondary metabolites, especially glycyrrhizic acid. In the present study, we cloned the cDNA coding 3-hydroxy-3-methylglutary CoA reductase (HMGR) involved in glycyrrhizic acid biosynthesis in Glycyrrhiza uralensis. The corresponding cDNA was expressed in Escherichia coli as fusion proteins. Recombinant HMGR exhibited catalysis activity in reduction of HMG-CoA to mevalonic acid (MVA) just as HMGR isolated from other species. Because HMGR gene is very important in the biosynthesis of glycyrrhizic acid in Glycyrrhiza uralensis, this work is significant for further studies concerned with strengthening the efficacy of Glycyrrhiza uralensis by means of increasing glycyrrhizic acid content and exploring the biosynthesis of glycyrrhizic acid in vitro.

OBJECTIVE To study the disinfectant resistance of multi-drug resistant Escherichia coli strains isolated clinically,and to find out the efficacy of disinfectants commonly used in killing multi-drug resistant E.coli strains.METHODS Minimal inhibitory concentration(MIC) and suspension quantitative germicidal test were used.Compared with standard strains,strains of multi-drug resistant E.coli isolated clinically were determined the resistance to four kinds of disinfectants including benzalkonium bromide etc.RESULTS A higher MIC of benzalkonium bromide compared with standard strains was observed in 61.9% of all 21 multi-drug resistant E.coli strains,and as for povidone iodine and NaClO,the ratio was 71.4% and 14.3%,respectively.All multi-drug resistant E.coli strains had the same MICs of peroxyacetic acid with standard strains.The above-mentioned 4 disinfectants commonly used at the routine concentrations killed 100% of the resistant strains of E.coli within 5 minutes.CONCLUSIONS The resistance to benzalkonium bromide and povidone iodine of multi-drug resistant E.coli isolated clinically is higher than standard strains;and 4 kinds of disinfectants commonly used are effective for multi-drug resistant E.coli strains isolated clinically.

Objective: Lung metastases are common in patients with differentiated thyroid carcinoma (DTC). Post-therapeutic 131I-whole-body scan (WBS) was conventionally administered after the radioactive iodine treatment (RAI) of DTC lung metastases. This study aimed to investigate the influencing factors of WBS imaging on the RAI of DTC lung metastases. Methods:DTC patients (n=60) with lung metastases treated with 131I were retrospectively included. Before treatment, the thyroid function was assessed. Neck and chest computed tomography (CT) was performed, and WBS was inspected. Patients with lung metastases were classified into negative and positive subgroups according to the imaging of 131I WBS, and the relative influencing factors were analyzed. Results:Univariate analy-sis showed that age and chest CT imaging, which revealed pulmonary fibrosis, calcification, and patchy shadows, were related to WBS imaging. Binary variable logistic regression analysis revealed that pulmonary fibrosis (OR=0.175, P<0.001) and calcification (OR=0.088, P<0.05) went against the development of WBS. Conclusion:WBS imaging on RAI of lung metastases was not obvious in the el-derly. The fibrosis, calcification, and patchy shadows of the lung were not conducive for WBS imaging. The fibrosis and calcification of the lung were the main factors that affect WBS imaging.

Objective To investigate the impact of hepatitis B virus (HBV) infection on semen parameters,sperm DNA integrity,acrosin activity and sperm-nucleoprotein transition.Methods Semen samples from 527 subjects including 273 hepatitis B surface antigen (HBsAg) positive and 254 HBsAg negative,who sought medical attention and received in-vitro feritilization in reproductive medicine center of First Hospital of Wenzhou Medical University from Jan 2011 to Oct 2012 were collected.Semen parameters,sperm DNA fragmentation index (DFI),sperm-nucleoprotein transition and acrosin activity of both HBsAg-positive and HBsAg-negative subjects were analyzed.Results Semen parameters of both groups were within the normal range,but sperm concentration and percentage of forward moving sperms of HBsAg positive group were significantly lower than those of HBsAg negative group (P=0.000),while percentage of static sperms of HBsAg positive group were significantly higher than that of HBsAg negative group (P =0.000).DFI in HBsAg positive and negative group were (17.85 ± 0.70) ％ and (11.85 ± 0.50) ％,respectively,which was significantly different (t=6.951,P=0.000).Percentage of sperms with normal morphology in both groups were within the normal range,but sperms with neck and tail deformity in the HBsAg positive group was significantly higer than those in HBsAg negative group (all P＜0.05).Acrosin activity of sperms in HBsAg positive group was significantly lower than that in HBsAg negative group (t=3.756,P=0.000).Linear regression analysis indicated that serum HBsAg level was reversely correlated with sperm concentration (r=-0.140,P =0.021),but positively correlated to DFI (r =0.151,P =0.014).Conclusions HBV infection not only affects the routine semen parameters and sperm morphology,but also compromises sperm function including impaired DFI and acrosin activity.However,the impact of anti-HBV agents on sperm quality and male fertility requires further research.

Objective To study the therapeutic effect of combined antihuman IgG antibody with mitomycin(MMC) on human bladder cancer ceils and the primary mechanism. Methods In vitro an-tiproliferation effects of goat antihuman IgG antibody(Ab) with MMC, alone or together, on human bladder cancer cell line T24 were tested by MTT assay. Flow cytometer(FCM) was used to detect T24 cell apoptosis. Detections of activated caspase-3 and PARP[poly(ADP-ribose) polymerase] were carried out by Western blot analysis. In vivo antitumor effects of Ab of anti-human IgG with MMC were assessed using T24 xenograft in BALB/c nude mice model. Results The inhibitory rates of tumor growth of Ab, M MC and Ab+ MMC on T24 cell were (25.02±6.71)%, (32.31±6.46)% and (73.66±5.81)%, respectively. The rates of apoptotic cell of PBS, normal goat IgG, Ab, MMC, MMC±normal goat IgG, and MMC+Ab were 1.7%, 2.3%, 20.7%, 22.4%, 28.3% and 53.8%, respectively. Western blot shows Caspase-3 and PARP were cleaved in T24 cell during the course of apoptosis induced by Ab and MMC, and indicated that cell apoptosis was associated with caspase-3 and PARP activation. Under the treatment of normal goat IgG, Ab, MMC, and MMC+Ab, the in-hibitory rates of T24 xenografts in BALB/c nude mice were 2.31%, 12.73%, 36. 81%, and 50.51%, respectively. Histological examination demonstrated significant necrosis and apoptosis in the mice treated with alone MMC or Ab but no control goat IgG or PBS, in addition, HE displayed more extensive necrosis and apoptosis in the mice with MMC+Ab.Conclusion Antihuman IgG Ab with MMC has in vitro and in vivo inhibitory effects of human bladder cancer T24 , which are related to in-ducing cell apoptosis.

Objective Increasing evidence suggest that aberrant activation of PI3K/Akt is involved in many human cancers, and that inhibition of the PI3K/Akt pathway might be a promising strategy for cancer therapy. The study is to evaluate the effects of combined therapy of PI3K inhibitor (LY294002) and Ad-PTEN in athymic mice xenogeneic transplant model of human glioma and to reveal the possible mechanisms involved.Methods Twenty-four athymic mice were randomly divided into 4 groups (DMSO、Ad-vector plus DMSO、LY294002 alone and Ad-PTEN plus LY294002), and were treated, respectively. Athymic mice xenogeneic transplant model was established by inoculation (sc) with LN229 glioma cells. Body mass (BM) and diameter of tumor mass were measured. Furthermore, The protein expressions of PTEN、p-Akt、CyclinD1、Caspase-3、MMP-2、p-FAK in tumor tissues were analyzed with immunohistochemistry.Results The tumor-inhibiting rate of was significantly higher in Ad-PTEN plus LY294002 than in the LY294002 alone (92.46 vs 65.59%)( P <0.05).The protein expressions of PTEN and Caspase-3 were significantly higher, while PCNA、CyclinD1、bcl-2 and MMP-2、p-FAK was significantly lower in Ad-PTEN plus LY294002 group than in the other three groups ( P <0.05).Conclusions LY294002 plus Ad-PTEN achieve better outcome than either alone in treating glioma possibly through enhancement of the inhibitory action of PI3K/Akt pathway and Ad-PTEN pathway.

To discuss clinical diagnosis and treatment of cow’s milk protein allergy and cow’s milk protein-induced FPIES (food protein induced enterocolitis syndrome). Methods We retrospectively analyzed clinical data of one infant with milk protein allergy-induced FPIES. Results A 67 days old female on mixed breast and formula feeding developed recurrent diarrhea, abdominal distension, vomiting, mucousy and bloody stools, feeding dififculty, anemia, and failure to thrive since 2 weeks after birth. Laboratory studies showed anemia, increased CRP level and elevation of peripheral white blood count and eosinophil proportion. Milk-speciifc IgE was negative. She was previously hospitalized 4 times, all with admitting diagnosis of“necrotizing enterocolitis”. We treated her with milk protein elimination for 4 weeks and all symptoms were resolved. Milk protein re-challenge test was positive, consistent with clinical features of cow’s milk protein allergy-induced infant FPIES. Conclusions Cow’s milk protein allergy and cow’s milk protein-induced FPIES can present with non-speciifc and variable clinical symptoms and signs, and should be considered in the differential diagnosis.