AIM:To construct a eukaryotic expression plasmid for enhanced green fluorescent protein(EGFP)and ZNF580 fusion protein,and study its subcellular localization in the transfected MGC803 cells.METHODS:The primers were designed according to the cDNA encoding sequence of ZNF580 full-length open reading frame(1-172aa),ZNF580 amino terminus(1-93aa)and ZNF580 carboxyl terminus(94-172aa).The three cDNA segments of PCR were cloned into pGEM-T vector.Then they were subcloned respectively into plasmid pEGFP-C1(enhanced green fluorescent protein).The subcellular localization of the fusion protein in MGC803 cells transfected with the vector was monitored by autofluorescence microscopy.RESULTS:Restricted enzymes analysis and DNA sequencing showed that the sequences of the pEGFP-ZNF580(1-172),pEGFP-ZNF580(1-93)and pEGFP-ZNF580(94-172)transgenic plasmid were correct.The fusion proteins of EGFP-ZNF580(1-172)and pEGFP-ZNF580(94-172)were localized in the nuclei.CONCLUSION:The recombinant eukaryotic expression vector pEGFP-ZNF580 has been successfully constructed.The nuclear localization signal is within amino acid residues 94 and 172 of ZNF580 carboxyl terminus(C2H2 zinc finger domain).

Constructing a research oriented learning platform of pathophysiology based on network environment has great significance in cultivating creative medical talents in quality oriented education.This paper aims at the problems of the research oriented learning platform application in students and teachers,and offers some useful advices to be consulted.

By discussing the importance of emotional education, the paper points out the existing problems of emotional education in nursing teaching as followings:the lack of teachers′emotional education awareness, the im-balance of cognitive development of nursing students and the lack of professional ideological education. Aiming at the above problems, this paper puts forward these specific countermeasures: strengthening the teachers′ emotional accomplishment so as to enhance their emotional education awareness and building a relaxed and pleasant diversi-fied educational environment so as to strengthen the professional thinking.

AIM:To elucidate whether ZFP580 is involved in the cardioprotective effects of hypoxic precondi-tioning (HPC) against hypoxia-reoxygenation (H/R) injury in H9c2 myocardial cells.METHODS: Rat heart-derived H9c2 cells were cultured in DMEM .H/R was induced by incubation under ischemic hypoxia for 3 h and reoxygenation for 2 h.HPC was induced by exposing the H 9c2 cells to 10 min of hypoxia and 20 min of reoxygenation for 3 cycles before H/R treatment.MTT staining and LDH leakage detection were used to evaluate the effects of HPC .Western blotting was used to detect the protein levels of ZFP580, phosphorylated ERK1/2 and cleaved caspased-3.The effects of ZFP580 overexpre-ssion or knockdown on H/R induced apoptosis were determined .RESULTS:The results of MTT staining and LDH leakage detection showed evidence of HPC cytoprotection against H /R-induced cell death in H9c2 cells.ZFP580 protein level and ERK1/2 phosphorylation were significantly increased in the HPC group compared with control group and H /R group. PD98059, an inhibitor of ERK1/2 phosphorylation , significantly suppressed the HPC-induced up-regulation of ZFP580 pro-tein expression.ZFP580 overexpression significantly inhibited apoptosis and caspase-3 activation in H9c2 cells.CON-CLUSION:HPC exhibits cytoprotection against H/R and leads to high level of ZFP 580 protein in H9c2 cells.ZFP580 is regulated by ERK1/2 activation and mediates the anti-apoptotic effect of the ERK1/2 signaling pathway in HPC cytoprotec-tion.

We took measures to construct sound learning mechanism and a set of effective learning system,which were connected tightly with various learning ways according to the idea of ‘ school-based ’ training.Goals of learning mechanism and the set of effective learning system were to improve teacher's learning ability,enhance knowledge and competence of medical college teachers and to construct learning teaching faculty.

Objective To evaluate the effects of dexmedetomidine (DEX) on cell apoptosis induced by endoplasmic reticulum stress and c-Jun N-terminal kinase (JNK) pathway during one-lung ventilation (OLV) in rats.Methods Sixty male Sprague-Dawley rats were randomly allocated into 6 groups (n =10 each):sham operation group (Sham group),OLV group,OLV + atipamezole (α2 receptor antagonist) group (AD group),OLV + atipamezole + DEX group (DEX+AD group),OLV + low-dose DEX group (DEX-L group) and OLV + high-dose DEX group (DEX-H group).The animals were anesthetized with 10％ chloral hydrate 4.5 ml/kg,tracheostomized and mechanically ventilated.Bilateral lungs were ventilated for 2.5 h in Sham group.The right lung was ventilated for 2.0 h followed by 0.5 h two-lung ventilation in OLV group.In DEX-L and DEX-H groups,DEX was infused intravenously for 1 h at a rate of 2.5 μg · kg-1 · h-1 and 5.0 μg · kg-1 · h-1,respectively,starting from 1 h prior to OLV.Atipamezole 250 μg/kg was injected intravenously at 1 h prior to OLV in AD group.Atipamezole 250 μg/kg was injected intravenously at the onset of DEX infusion (5.0 μg · kg-1 · h-1) in DEX+AD group.The rats were sacrificed and left lungs were removed for determination of weight to dry lung weight ratio (W/D),cell apoptosis in lung tissues (by TUNEL),and expression of glucose-regulated protein 78 (GRP78) mRNA and protein,JNK mRNA and phosphorylated JNK (p-JNK) protein (by RT-PCR and Western blot).Pathological changes of lungs were examined and the injured alveolus rate (IAR) was counted under light microscope.The changes in ultrastructure of lung tissues were observed under transmission electron microscope.Apoptosis index (AI) was calculated.Results W/D,AI and IAR were significantly higher in OLV,AD and DEX+AD group than in Sham group,while lower in DEX-L and DEX-H groups than in OLV,AD and DEX+AD groups.The pathological changes of the structure of lung tissues were observed in OLV,AD and DEX+AD groups,while the pathological changes were significantly alleviated in DEX-L and DEX-H groups.In OLV,AD and DEX + AD groups,there was apoptosis in lots of pulmonary vascular endothelial cells and alveolar epithelial cells,while cell apoptosis was significantly reduced after administration of DEX.The expression of GRP78 mRNA and protein,JNK mRNA and p-JNK protein was significantly higher in OLV,AD and DEX+AD groups than in Sham group,and lower in DEX-L and DEX-H groups than in OLV,AD and DEX +AD groups.Conclusion DEX pretreatment can protect lungs during OLV,and inhibited JNK signaling pathway and reduced cell apoptosis induced by endoplasmic reticulum stress may be involved in the mechanism.

Aim To investigate the effect of aloe-emodin on lipopolysaccharide(LPS)-induced production of nitric oxide and expression of inducible nitric oxide synthase in RAW264.7 cells.Methods RAW264.7 macrophage line in mice was induced by LPS to set up the inflammatory model.Nitric oxide(NO)production was examined by Griess reaction;the expression of iNOS mRNA was detected by RT-PCR analysis;NO radical generation was tested by sodium nitroprusside method.Results Aloe-emodin at the dose of 0.69～2.5 mg·L~(-1) exhibited the inhibitory effect on LPS-induced NO production in a dose-dependent and time-dependent manner;aloe-emodin at the dose of 0.63～5.00 mg·L~(-1) suppressed LPS-induced iNOS mRNA expression in RAW 264.7 cells.However,aloe-emodin had no scavenging effect on sodium nitroprusside-triggered NO production,and didn't affect iNOS enzyme activity.Conclusion Aloe-emodin inhibited signifi-cantly LPS-induced NO production through suppressing inducible NO synthase(iNOS)expression at mRNA level in a dose-dependent and time-dependent manner,but failed to affect sodium nitroprusside-triggered NO production and iNOS enzyme activity.

Extracorporeal membrane oxygenation (ECMO) is a technique in which breathing and circulation are supported extracorporeally. Severe trauma may induce cardiopulmonary failure, for which ECMO can play an adjunctive role in the salvage treatment of circulatory and respiratory failure when conventional treatments are ineffective. Bypass with ECMO can rapidly improve the state such as circulatory failure and hypoxemia in critically ill patients in short term and can partially or fully replace their cardiopulmonary function in long term, winning valuable time for normal recovery of cardiopulmonary function. Because of the physical state of severe trauma patients and the ECMO equipment, there are still various complications clinically. Trauma patients show high risk of bleeding, vulnerability to wound infection and probability of combined organ injury and dysfunction, so more comprehensive measures for the prevention and treatment of complications during the use of ECMO therapy are required. The authors review the research progress in complications and corresponding prevention and treatment strategies during ECMO support for severe trauma, aiming to provide a reference to prevent and treat these complications.

Objective To construct an professional quality training management system for in-service nurses and to evaluate its effect in clinical practice.Methods From June 2014 to October 2016, a total of 1233 nurses in a classⅢ grade A hospital were selected. Through the establishment and improvement of the organizational structure of the professional quality management of the in-service nurses, the various training content combining theory and practice were adopted, including case study, role playing and simulation scenario training to implement the professional training.Results The average score of nursing professional quality was (91.78±3.82) before training and (94.53±4.33) after training. The difference between before and after training was statistically significant (t=-3.156,P=0.006). The patient satisfaction rate was 90.57％ before training and 97.42％ after training, and the difference was statistically significant (Z=-2.516,P=0.006). The doctor satisfaction rate was 90.57％ before training and 94.88％ after training, and the difference was statistically significant (Z=-2.233,P<0.01).Conclusions The construction of nurses' professional quality training strengthens the importance of nurses' professional quality, improves the evaluation of professional quality, and has guiding significance for the training of in-service nurses' professional quality.

Polycystic ovary syndrome(PCOS)is the most common endocrine and metabolic disease in women of reproduc-tive age.PCOS is characterized by ovulatory disruption,which can lead to infertility.Patients with PCOS are also more likely to have poor pregnancy outcomes.For obese women,lifestyle interventions are recommended first,which have general health benefits.For women who have difficulty changing their lifestyle,drugs for the treatment of obesity or bariatric surgery could be considered.Clomiphene citrate is the first-line medication after weight loss that has been utilized in the past.Letrozole is supplanting clomiphene as the best option for ovulation induction for now,particularly in patients with PCOS.Metformin can improve ovulation and pregnancy rates;however,it has minimal effects in terms of raising live birth rates.Second-line therapies include gonadotropins and laparoscopic ovary drilling.In vitro fertilization can be utilized as a third-line treatment for patients with PCOS who have failed ovulation induction therapy or have other infertility factors.In summary,to achieve fer-tility,patients with PCOS require standardized individualized therapy.