[Objective]To study the effects of verapamil on the immediate-early gene(IEGs)expression of bone marrow mesenchymal stimulated by mechanical strain,and deduce the effects of calcium ion channel on the early responses of children bone marrow mesenchymal stem cells(MSCs)to mechanical strain in vitro.[Method]MSCs were isolated and cultured.The 3-6th generational MSCs were stimulated in different time by mechanical and(or)verapamil(20?mol/l).Flow cytometry was applied to measure intracellular free Ca2+ at once.Early-response genes /proteins(c-fos,c-jun and c-myc)were examined by RT-PCR and immunocytochemical stain.[Result]The application of mechanical strain increased intracellular c-fos,c-jun and c-myc mRNA /protein levels.The resulting effect is partially inhibited before MSCs were preincubated with verapamil(P

Objective To evaluate the efficacy of laparoscopic first-stage Fowler-Stephens for high cryptorchidism. Methods Seven children with high cryptorchidism (10 testes) were enrolled in this study. After detecting the testicles under a laparoscope, we dissected the spermatic cord vessels. Then, the spermatic duct was separated, and the gubernaculum testis was retained. Afterwards, the first-stage orchidopexy was performed laparoscopically. Results Nine of the 10 testes were successfully placed into the scrotum by first-stage Fowler-Stephens. In one patient who had bilateral cryptorchidism, the left testis was placed into the scrotum at the second stage operation. The seven patients were followed up for 6-24 months (mean, 14 months). None of them had retraction or atrophy of the testis. Conclusions Laparoscopic first-stage Fowler-Stephens is effective for high cryptorchidism. The procedure is worth being widely used.

In order to investigate the relationship between the expression of heme oxygenase-2 (HO-2) mRNA and the pathogenesis of Hirschsprung's disease (HD), total ribonucleic acid (RNA) was extracted in the aganglionic and ganglionic segments of colon respectively from 15 cases of HD. The single-stranded cDNA of HO-2 was synthesized and further amplified by reverse transcription-polymerase chain reaction (RT-PCR). The expression of HO-2 mRNA was normal in ganglionic segments, but absent in aganglionic segments. It is concluded that the absence of HO-2 mRNA expression may be an important mechanism responsible for HD.
Colon/enzymology ; Heme Oxygenase (Decyclizing)/biosynthesis ; Heme Oxygenase (Decyclizing)/*genetics ; Hirschsprung Disease/*enzymology ; Hirschsprung Disease/etiology ; Hirschsprung Disease/genetics ; RNA, Messenger/biosynthesis ; RNA, Messenger/genetics

Objective To evaluate the long-term results of partial internal sphincterectomy for the treatment of internal anal sphincter achalasia in childhood. Methods The clinical,radiographic,manometrical and histochemical data of 6 cases were reviewed retrospectively. All patients had received partial internal sphincterectomy and were followed-up for 2 to 8 years. Results All patients presented with severe constipation with or without soiling. No stenosis zone of intestine could be noted in 3 patients by barium enema examination. The rectoanal inhibition reflex on rectal balloon inflation was absent in all patients. The normal acetylcholinesterase activity on rectal biopsies was demonstrated by histochemical staining. Ganglion cells within internal anal sphincter was noted in all cases. On follow-up,all patients regained regular bowel habits and are not on any laxatives. Conclusion The long term results of partial internal sphincterectomy for the treatment of internal anal sphincter achalasia in childhood are satisfactory.

A case with the diagnosis of the onset of inflammatory bowel disease (IBD) with chronic granulo-matous disease (CGD) in Children′s Hospital Affiliated to Zhengzhou University in June 2016 was chosen, and the patient′s clinical data and whole treatment process were analyzed.According to the relevant literature from Chinese and foreign databases, the clinical characteristics were analyzed and summarized, principles of diagnosis and treatment for children who had the onset of IBD with CGD.This patient was a child of 1 year and 9 months old, and the initial symptoms included repeated diarrhea and bloody stools.He was diagnosed as " ulcerative colitis" in the local hospital.After admission, the neutrophil respiratory burst test was positive.The genetic analysis result suggested that the CYBB gene was mutated, thus obtaining the diagnosis of CGD.Then, he was given prophylactic antibiotic therapy and symptomatic treatment.After the 3 months of follow-up after discharge, the patient still had intermittent diarrhea and bloody stools.CGD is a rare primary immunodeficiency disease, and current treatment methods of CGD include hematopoietic stem cell transplantation and anti-infection treatment.When IBD patients have complications other than gastrointestinal symptoms, the IBD treatment is not effective, or there are suspected parents who are married to close relatives, or with the family history of IBD, CGD should be considered.

Bone marrow mesenchymal stem cells (BMSCs) have been shown to be multipotent cells that possess high self-replicating capacity. The purpose of our study was to investigate the feasibility of using enteric neuron-like cells obtained by in vitro induction and differentiated from rat BMSCs for the treatment of Hirschsprung's disease (HD). Glial cell-derived neurotrophic factor (GDNF) and neurotrophin-3 (NT-3) are neurotrophic factors that play important roles in neuronal development, differentiation, survival and function. Meanwhile, GDNF mutations are a major cause of HD. In this study, BMSCs were transfected with eukaryotic expression plasmids co-expressing GDNF and NT-3, and the transfected cells displayed neuron-like changes after differentiation induced by fetal gut culture medium (FGCM). Immunofluorescence assay showed positive expression of the neuronal marker NSE and the enteric neuronal markers PGP9.5, VIP and nNOS. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the expression of GDNF and NT-3 in transfected BMSCs. The present study indicates that genetically modified BMSCs co-expressing GDNF and NT-3 are able to differentiate into enteric neuronal cells and express enteric nerve markers when induced by FGCM. This study provides an experimental basis for gene therapy to treat enteric nervous system-related disorders, such as HD.

Objective To compare the results and safety between video-assisted thoracoscopic surgery ( VATS ) and conventional radical operation in patients with stage Ⅰ , Ⅱ esophageal cancer. Methods Retrospectively reviewed 43 patients with stage Ⅰ , Ⅱ esophageal cancer,underwent either VATS radical operation (VATS group,16 cases) or conventional radical operation (control group,27 cases ) from September 2007 to September 2009. Patient's operative characteristics and postoperative courses were compared between two groups. Results In VATS group the operation time was ( 115.6 ± 48.0) min,the peri-operative blood loss was ( 131 ± 71 ) ml,the first postoperative day chest lead quantity was (331 ± 170)ml, the time of postoperative chest tube was (7.25 ± 2.35) d,the postoperative 36 h visual analogue scale (VAS) was (3.4 ± 1.2) scores,the postoperative drainage of chest was ( 1281 ± 534) ml,the 72 h postoperative locomotor activity of right upper extremity was (5.1 ± 1.5) cm. While in control group was ( 145.6 ± 20.6)min, (292 ± 111 ) ml, (494 ± 194) ml, ( 10.00 ± 2.79 )d, (7.3 ± 1.4) scores, ( 1780 ± 731 ) ml, ( 15.6 ± 3.1 )cm respectively (P ＜ 0.01 or ＜ 0.05 ). The lymph node dissection number,the total cost of hospital between were no statistically significant differences in two groups (P ＞0.05). Conclusion Comparing with conventional radical operation, VATS radical operation for patients with stage Ⅰ , Ⅱ esophageal cancer appears to be as effective but less morbid.

Objective To analyze the relationship between polymorphisms of EDNRB gene and Hubei provincial patients of Han ethnicity with sporadic Hirschsprung disease(sHD). Methods Peripheral blood samples from 104 patients with sHD and 84 parents of 42 patients, and 120 normal children(as controls) were collected. PCR-SSCP and direct DNA sequencing were used to detect mutations and polymorphisms of exon-4 in EDNRB gene. The differences of allele frequencies and genotype distribution in polymorphic sites were further analyzed between the three groups. Allele frequencies of SNPs in forty-two sHD trios were analyzed by transmission disequilibrium test(TDT), and the association between phenotype of HD and SNPs was analyzed. Results No mutant site was detected and one polymorphic site of c831 G→A(L277L) was observed in Hubei provincial patients of Han ethnicity with sHD. The allele frequency of A(68% vs 53%) and genotype frequency of AA(49% vs 30%) were significantly higher in sHD group than that in control group(P

In order to confirm the existence of indoleamine 2, 3-dioxygenase (IDO) gene in swine, and to clone the novel gene followed by the molecule structure properties and expression pattern analysis, the porcine mRNA sequences homologous to human IDO were obtained from GenBank database by bioinformatics method. By using RT-PCR, the IDO gene was cloned from porcine endothelial cell line and the accuracy of the nucleic acid sequence was confirmed, and the expression pattern of the gene was detected. The three-dimensional structure model of porcine IDO was built referring to the tertiary structure of human IDO using biological sequence analysis software and database. The results showed that the porcine IDO was identified by sequencing. The nucleotide sequences were confirmed as a novel gene after submitted to Genbank. Porcine IDO was expressed in the lung, thymus, epididymis and anterior chamber with a basic level, however in peripheral blood mononuclear cells (PBMCs) the IDO gene was highly expressed. The three-dimensional structure model of porcine IDO was similar to that of human IDO. It was suggested that identification of the structure information of porcine IDO is essential to further investigate the immunologic function of the gene. Study of IDO on NK cells-mediated xenograft rejection will be a novel therapeutic target for the development of xenotransplantation.

Background and purpose:High expression of excision repair cross-complementing 1 (ERCC1) is related to resistance in patients treated with platinum-containing regimens. The ERCC1 antibody 8F1 was usually used in past studies, but it was found to have no-speciifcity recently. This study aimed to investigate the predictive role of a new ERCC1 antibody 4F9 to platinum chemotherapy in non-small cell lung cancer (NSCLC) patients. Methods:Expression of ERCC1 was detected using antibody 4F9 by immunohistochemistry (IHC) in 72 NSCLC tissues. The relationship between the expression of ERCCl and the clinical pathological parameters, the efficacy of platinum chemotherapy and overall survival of patients were explored by statistical analysis. Results: The high expression of ERCCl protein was 55.5%in 72 cases. There was no signiifcant correlation between the ERCC1 expression with gender, age, pathological type, clinical stage and lymphatic metastasis (P>0.05). Patients with low expression of ERCC1 had signiifcantly higher response rates to platinum chemotherapy, longer median survival time and 2-years survival rate comparing with those with high expression of ERCC1 (62.5%vs 37.5%;22.9 vs 18.4 month;46.9%vs 37.5%), respectively (P<0.05). Conclusion:The expression analysis of ERCC1 using new ERCC1 antibody 4F9 by IHC method is helpful to assign chemotherapeutic regimen, and guide individual platinum chemotherapy for post-operation patients.