Establishment and verification of a mouse model of gene H435Y mutation / 南方医科大学学报

<p><b>OBJECTIVE</b>To establish a mouse model of H435Y mutation of gene using CRISPR/Cas9- mediated gene targeting.</p><p><b>METHODS</b>The single-stranded guide RNA (sgRNA) specific to the H435Y loci of gene was designed based on the sequence of gene. After activity assessment, the active sgRNA and Cas9 were transcribed into RNA and microinjected along with the donor DNA fragment with point mutations into fertilized mouse eggs. The microinjected eggs were transferred into pseudopregnant mice to obtain the F0 generation mice with the target gene mutation confirmed by PCR and gene sequencing. gene mutations in the offsprings of the F0 generation mice were analyzed.</p><p><b>RESULTS</b>Gene sequencing confirmed the successful establishment of mouse models carrying H435Y mutation of gene in 4 of the F0 generation mice. The positive F0 generation mice were crossed with wild-type C57BL/6J mice to obtain the F1 generation mice, and PCR confirmed the presence of H435Y mutations of gene in 6 of the F1 mice. Then F2 generation mice were obtained by F1 generation matting with each other. PCR showed that H435Y mutation of gene in F2 mice was found, indicating the mousemodel of gene mutation in H435Y was established and propagated successfully.</p><p><b>CONCLUSIONS</b>We successfully established gene H435Y mutant mouse models using CRISPR/Cas9 technique.</p>

Factors related to death of thoracic trauma emergency in children / 中华全科医师杂志

Objective To investigate the factors related to death of thoracic trauma emergency in children.Methods Total 528 children and infants aged 0-14 years with thoracic injuries , including 317 boys and 211 girls, admitted in emergency department from January 2010 to January 2014 were included in the study.The factors related to emergency death were investigated by using conditional logistic analysis.Results Among 528 cases, emergency death occurred in 34 cases with an emergency mortality rate of 6.44%.Single-factor analysis showed that emergency death was correlated with complication with other organ trauma, the time of first medical intervention , the time of arriving at first contact hospital , the rank of first contact hospital , hypoxemia and causes of injuries ( all P0.05).Logistic analysis showed that the time of first medical interventio n>30 min ( B=1.467,95%CI:0.412-0.975), complication with other organ trauma (B=2.342,95%CI:0.415-0.943), hypoxemia (B=2.915,95%CI:0.749-0.819), and first visiting to tertiary hospital (B=-1.861,95%CI:1.023-1.742) were influencing factors of emergency death.Conclusion The results indicate that to improve the success rate of emergency treatment of thoracic trauma in children , it is necessary to reduce the time of first medical intervention and to correct the hypoxemia promptly.

The investigation of methods for prevention of anastomotic stenosis after resection of esophageal cancer / 中国癌症杂志

Background and purpose:Esophageal cancer is one of the common malignant tumors in our country. Anastomotic stenosis is a common complication after resection of esophageal cancer, seriously affecting the quality of life of patients after operation. By changing anastomosis, this study explored the methods for prevention of anastomotic stenosis after esophageal cancer surgery.Methods:Patients were randomly divided into groups. Patients admitted on odd dates were placed in the control group whereas patients admitted on even dates were placed in the experimental group. Patients in the control group were treated with gastroesophageal anastomosis using anastomat for gastroesophageal anastomosis. Anastomotic stomach was contracted by purse string suture at first, and then treated with stapler gastroesophageal anastomosis, before the gastroesophageal anastomosis was carried out on patients in the experimental group. After 6 months’ follow-up, the incidences of anastomotic stenosis between the two groups were compared.Results:The postoperative anastomotic stenosis rate in the control group was 19.2%, while that in the exper-imental group was 0%. There were statistically signiifcant differences between them (χ2=22.8,P<0.005). The incidence of anastomotic stenosis in the control group was signiifcantly higher than that in the experimental group.Conclusion:Anastomotic stomach contracted by purse string suture before stapler gastroesophageal anastomosis can effectively reduce the occurrence of anastomotic stenosis after esophageal cancer surgery.

Progress of immunologic mechanisms of Vδ2-γδT cell in cytomegalovirus infection / 国际儿科学杂志

Recently,an unconventional T cell population (collectively designated as Vδ2 -γδT cell ) has been characterized during the anti-cytomegalovirus immune response in all organ transplant recipients,neo-nates,and healthy individuals.These cytomegalovirus-induced Vδ2 -γδT cells undergo a dramatic and stable ex-pansion after cytomegalovirus infection,in a conventional adaptive way.Similarly,as cytomegalovirus-specific CD8 +αβT cells,they exhibit an effector/memory TEMRA phenotype and cytotoxic effect or functions.This paper reviews the researchs and reports about cytomegalovirus induced-Vδ2 -γδT cell,including its location,phe-notype,and activation,as well as its immunologic mechanism in cytomegalovirus infection,acute or chronic re-jection,and anti-cancer function.

Experimental studies on the role of GATA4 in the endocardial cushions development / 中华心血管病杂志

<p><b>OBJECTIVE</b>To investigate the role of GATA4 gene in the endocardial cushions development.</p><p><b>METHODS</b>Target gene eukaryote expression vectors were constructed by pcDNA3.1(-) vector plasmid, and were identified by DNA sequence analysis. Recombinant plasmids were transfected into Hela cells with lipofectamine 2000, meanwhile Hela cells transfected with empty vector or those without transfection served as transfection control group and blank control group, respectively. Real-time PCR and Western blot were performed to detect the relative expression of mRNA and protein of transcription factors GATA4, Sox9, Scleraxis and ECM proteins Aggrecan, Tenascin in each group.</p><p><b>RESULTS</b>The relative mRNA expression of GATA4 in experimental group was significantly higher than in transfection control group and blank control group. GATA4 mRNA expression in Hela(GATA4), Hela(H436Y), Hela(Null) and Hela group was 310.83 ± 2.39, 146.35 ± 1.74, 0.94 ± 0.32, 1.00 ± 0.28, respectively (F = 72.508, P < 0.05). Western blot results were consistent with the results obtained by qRT-PCR. The relative mRNA and protein expressions of Sox9, Scleraxis, Aggrecan and Tenascin in both experimental groups were significantly higher than that in transfection control group and blank control group (P < 0.05), and above gene expressions were significantly downregulated in GATA4(H436Y) group, while they were similar between transfection control group and blank control group (all P > 0.05).</p><p><b>CONCLUSIONS</b>GATA4 H436Y mutation reduces it's transcriptional activation, which might serve as a theoretical framework to demonstrate the roles of GATA4 gene in endocardial cushion development.</p>

Analysis of ERCC1 mRNA expression to predict prognosis in patients with non-small cell lung cancer who received adjuvant chemotherapy / 重庆医学

Objective To discover the excision repair cross-complementing 1 (ERCC1) expression in non-small cell lung cancer (NSCLC) patients and explore the prognostic value of ERCC1 .Methods The ERCC1 mRNA expressions in NSCLC was tested from 85 tumor tissues and 34 adjacent tissue samples from patients who were after the surgery were used by semi-quantitative RT-PCR .The data of clinical features and progression-free survival (PFS) and overall survival (OS) were linked to ERCC1 expression by retrospective analysis .Results In 85 patients ,the ERCC1 negative ones had a significantly longer survival than the ERCC1 posi-tive expression ones (PFS ,P=0 .001;OS ,P=0 .001) .During the multivariate analysis ,ERCC1was found to be a significant factor in PFS and OS (P=0 .018 and P=0 .027) .Conclusion NSCLC patients who were undertaken platinum-based adjuvant chemother-apy after surgery could use the detection of ERCC1 mRNA as a determinant factor for the prognosis predicting of individualized treatment .

Expression level and correlation of IL-23 and MMP-9 in esophageal squamous cell carcinomas / 实用医学杂志

Objective To investigate the relationship of the serum level of IL-23 and MMP-9 with the clinicopathologic features in patients with esophageal squamous cell carcinoma (ESCC). Methods 48 pathologically confirmed ESCC patients and 30 Endoscopic biopsy of benign were included in this study. The serum levels of IL-23 and MMP-9 were examined by enzyme-linked immunosorbent assay (ELISA). Results Serum IL-23 level in patients with ESCC was significantly higher than that in controls (t = 26.66, 16.89, P<0.05). Furthermore, Pearson′s correlation analysis revealed that serum IL-23 was positively correlated with the serum MMP-9 level in ESCC patients (r = 0.790, P < 0.05). Statistical analysis showed that enhanced serum IL-23 significantly correlated with the degree of differentiation and lymph node metastasis. Conclusion Overexpression of IL-23 may involve in the occurrence and development of ESCC. IL-23 may contribute to tumorinvasion and metastasis by stimulating the expression of MMP-9.

Clinical analysis of 28 cases of non benign neonatal arrhythmias / 临床儿科杂志

Objective To discuss the etiology, clinical features and prognosis of non benign neonatal arrhythmias. Method Clinical data of 27 cases of non benign neonatal arrhythmias diagnosed from January 2005 to January 2010 were retrospectively analyzed. Results Among 27 neonats, there were 15 male and 12 female. Fourteen cases were early neonatal and 13 were late neonatal. Gestational age was less than 32 weeks in 6 cases, and more than 32 weeks in 21 cases. In 19 cases with tachyarrhythmia, 14 cases were induced by respiratory infection. The causes of 8 newborns with bradycardia arrhythmia were congenital heart disease (3 cases), electrolyte disturbance (2 cases), severe asphyxia with sepsis (2 cases), and severe asphyxia (1 case). The onset age and gestational age were lower in cases with bradycardia arrhythma that those in cases with tachyarrhythmia (P<0.005). The cure rate and effective rate of tachyarrhythmia was 89.5%and 100%, of bradycardia arrhythmia was 0%and 12.5%, respectively, and the differences were signiifcant (P<0.005). Conclusion Clinical characteristics, pathogenesis and prognosis were different between tachyarrhythmia and bradycardia arrhythmia in neonates.

Expression and methylation of TCF21 gene in non-small cell lung cancer / 中华胸心血管外科杂志

Objective Background and objective Bioinformatics technology found the TCF21 gene has difference expression in non-small cell lung cancer (NSCLC) and benign lung tissue.To explore TCF21 mRNA and protein expression in nonsmall cell lung cancer and its methylation of the promoter region is the aim of this study.Methods Use RT-PCR,Western blot and Pyrosequencing detected TCF21 gene mRNA,protein and the methylation of the promoter region respectively in 97 cases of non-small cell lung cancer and 21 cases of benign lung tissue.Results TCF21 gene mRNA-positive expression were detected 23 cases (23.71％) and 14 cases (66.67％) in 97 cases of NSCLC cancer tissue and 21 cases of benign lung tissue,the average gray value of TCF21 protein expression levels in NSCLC cancer tissue is 0.49 ± 1.78,while it is 1.48 ± 1.58 in benign lung tissue,the TCF21 gene promoter region have varying degrees methylation in NSCLC cancer tissue and benign lung tissue,and the significant of methylation frequency was found statistically significant between NSCLC cancer tissue and benign lung tissue,also it has higher frequency of 49.04％ and 51.37％ respectively at point 1 and 5 in NSCLC cancer organizations.Conclusion TCF21 gene mRNA and protein expression were statistically significant in NSCLC cancer tissue and benign lung tissue,the TCF21 gene promoter region average methylation frequency was significantly higher than that in benign lung tissue cells.

A logistic retrospective analysis about the risk factors of cerebral hemorrhage after the liver transplantation / 中国实用护理杂志

Objective Analysis the risk factors which maybe caused the cerebral hemorrhage after the liver transplantation, and then make out the counter nursing measures. Methods Selected 12 patients with cerebral hemorrhage after the liver transplantation as the experimental group, and then matching 48 patients without cerebral hemorrhage after the liver transplantation as the control group. Analysis the factor which related the cerebral hemorrhage. Results The factors which related the cerebral hemorrhage after the liver transplantation included in the Intensive Care Unite, heme level, Child level, APTT, albumin, blooding volume during the operation, operation time and the postoperative blood platelet level. The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage. Conclusion The factors of Child level and the blooding volume during the operation were the independence risk factors of cerebral hemorrhage, we should carry out counter nursing measures to prevent them.

Animal experimental research of the endothelialization of home-made atrial septal defect occluder device / 介入放射学杂志

Objective To evaluate the endothelialization of Chinese nitinol atrial septal defect occlud-er device. Methods Atrial septal defect with controllable size was created by the Brockenborough needle and Rashkind balloon atrial septostomy, the occluder devices were implanted in six piglets (mean weight 7. 5 kg). Two pigs were killed each time after 1 month, 3 months and 6 months after the device implantation and then the explanted devices were examined by scanning electron microscope (SEM). Results The devices were found covering with collagen fibrosis together with diffuse endothelial cells spreading over the primer 1 month after implantation. The implants were covered mostly by ncointima 3 months after implantation and completely covered by confluent endothelial cells 6 months after the implantation. Endothelial cells were not found on the smooth marker band at 3 months, however, did exist by 6 months. Conclusions Home-made atriatseptal defect occluder devices were mostly endothelialised 3 months after the implantation and did completely at 6months.