Objective To evaluate the diagnostic significance of esophageal minimal change in gastroesophageal reflux disease (GERD) and explore its clinical characteristics.Methods From May to September in 2013,patients with minimal esophageal mucosa changes including esophageal mucosa rough,white secretin adhesion,erythema,edema,increased brittleness,blurring of the Z line or zigzag looking and blurring of paliform blood vessel,or patients with Los Angeles classification (LA) which were identified by endoscopy were enrolled.The subjects received gastroesophageal reflux disease questionnaire (GcrdQ) investigation and the related history were collected.The total score of GerdQ over eight was set as the criteria for GERD diagnosis.The R × C chi-square test was performed for statistical analysis.Results A total of 417 valid questionnaires were completed.Of which,202 cases were in minimal change group,176 were in LA A group and 36 were in LA-B group.The diagnostic rate of GERD in minimal change group was 20.3％ (41/202),which was lower than that of LA-A group (74.4％,131/176) and LA-B group (83.3 ％,30/36),and the differences were statistically significant (x2 =129.144,P＜0.01).The incidences of heartburn in minimal change group,LA A group and LA-B group were 25.7％ (52/202),62.5％ (110/176) and 86.1％ (31/36),respectively.The incidences of reflux were 29.7％ (60/202),67.6％ (119/176) and 75.0％ (27/36),respectively.The incidences of non cardiac chest pain were 5.4％ (11/202),22.2％ (39/176) and 22.2％(8/36),respectively.The incidences of heartburn,reflux and non cardiac chcst pain of minimal change group were all lower than those of LA A group and LA-B group,and the differences were statistically significant (x2 =75.775,64.120,24.016;all P＜0.01).The leading cause of endoscopy examination in minimal change group was abdominal discomfort,which accounted for 49.0％(99/202).The leading causes of endoscopy examination in LA A group and LA-B group were esophageal symptoms,which accounted for 52.8％ (93/176) and 61.1％ (22/36).Conclusions The diagnostic rate of GERD in patients of minimal change group is low and the clinical symptoms are not typical,which is insufficient for diagnosis of GERD and needed further investigation.

Objective To explore the role and mechanism of protein disulfide isomerase A3 (PDIA3) in abnormal colonic mucosa immune response of irritable bowel syndrome (IBS) rats with visceral hypersensitivity.Methods A total of 48 SD rats were divided into blank control group (n=12),empty virus group (IBS-1) (n=12),PDIA3 silencing group (IBS-2) (n=12) and model control group(IBS-3) (n=12).The expression of CD103 in ileocecus colonic tissues was detected by immunohistochemistry.Dendritic cells (D C) of mesenteric lymph node were isolated by flow cytometry.CD4+/CD8+ T cells of spleen were separated by immune-magnetic beads sorting technique.DC and CD4+/CD8+ T cells were co-cultured.The proliferation ability of lymphocytes promoted by DC was measured by methyl thiazolyl tetrazolium (MTT).The secretion levels of interlcukin (IL) 4 and IL-9 of CD4+/CD8+ T cells stimulated by DC were determined by enzyme linked immunosorbent assay (ELISA) method.Independent sample t-test was performed for statistical analysis.Results The cell number of CD103 positive DC of rats colon of blank control group,IBS-3 group was 6.25±1.14 and 10.83± 1.03(t=10.07,P＜0.05);that of IBS-2 group was 7.42 ± 0.90,and compared with that of IBS-3 group,the difference was statistically significant (t=-9.25,P ＜ 0.05).The MTT value of CD4+ T cells proliferation stimulated by DC of blank control group and IBm3 group was 0.54±0.01 and 0.60±0.01 (t=3.373,P＜0.05);that of IBS-2 group was 0.53±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t =-3.139,P ＜ 0.05).The MTT value of CD8+ T cells proliferation stimulated by DC was 0.52±0.01 and 0.59±0.00(t=3.539,P＜0.01);that of IBS-2 group was 0.54±0.01,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.183,P＜0.05).The level of IL-4 secreted by CD4+T cells promoted by DC of blank control group and IBS-3 group was 10.24±0.09 and 16.61±1.00 (t=3.222,P＜0.05);that of IBS-2 group was 11.75±0.54,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.539,P＜0.01).The level of IL-9 secreted by CD4+T cells stimulated by DC of blank control group and IBS-3 group was 15.86±10.19 and 43.51±11.32 (t=4.529,P＜0.05);that of IBS-2 group was 29.05±2.09,and compared with that of IBS-3 group,the difference was statistically significant (t=-6.841,P＜0.01).The level of IL-4 secreted by CD8+T cells promoted by DC of blank control group and IBS-3 group was 7.35±0.12 and 13.91±0.57 (t=19.557,P＜0.01);that of IBS-2 group was 8.63± 0.24,and compared with that of IBS-3 group,the difference was statistically significant (t =-14.782,P＜0.01).The level of IL-9 secreted by CD8+T cells stimulated by DC of blank control group and IBS-3 group was 29.12±5.14 and 60.70±11.02 (t=4.122,P＜0.05);that of IBS-2 group was 37.17±2.65,and compared with that of IBS-3 group,the difference was statistically significant (t=-3.255,P＜ 0.05).Conclusions Protein disulfide isomerase A3 may mediate the process of DC activating T lymphocyte levels of related cytokines,cause abnormal immune response of colonic mucosa and promote the genesis of IBS visceral hypersensitivity.

Mesenchymal stem cells (MSCs) have good immune regulatory function,can inhibit many immune cell proliferation,direct effects on activation and proliferation of T cell,play a role in immune regulation by Treg cells or by the secretion of soluble factors regulating Thl/Th2 secretion and reaction equilibrium,inhibit the inflammation through the anti-inflammatory,regulation of cytokines expression at the same time,so reduce the expression levels of matrix metalloproteinase-2 and-9,which may promote lacrimal gland tissue damage,and thus play a role in immune regulation.MSCs can reduce the autoimmune dry eye clinical index,recovery secretion function of part lacrimal gland.This article reviews the research advances in inmmmne regulation of MSCs on autoimmune dry eye.

Objective To explore the indication,surgical procedure and the postoperative treatment of breast conserving modified radical mastectomy for early stage breast cancer .Methods The clinical data of 21 patients with early stage breast cancer underwent breast conserving modified radical mastectomy were analyzed retrospectively. Results All patient recovered uneventfully. All the external configuration of the breast were fine. There were no recurrence and no complications in this series. Conclusions The breast conserving modified radical mastectomy is recommended for the early stage breast cancer. The external configuration of breast is fine postoperatively, and patients have higher survival quality.But follow up is necessary in order to find and treat the recurrence of breast cancer.

Objective To evaluate Chinese tinnitus handicap questionnaire (T HQ ) reliability and validity through clinical use and analysis .Methods A total of 230 tinnitus patients who were seen at our ENT clinic with tinnitus lasting longer than 3 months were administered with the THQ ,THI ,tinnitus questionnaire ,pure tone au‐diogram and tinnitus matching .Assessed THQ’s reliability and validity ,the relationship between THQ and its re‐lated factors .Results In the 206 tinnitus patients ,the Cronbach’sαwas 0 .93 ,split -half reliability was 0 .91 ,test-retest reliability was 0 .96 .In the 171 tinnitus patients ,the THI correlation coefficient was 0 .745 .The THQ scores were highly correlated with the hearing loss level of the worse ear .The age ,sex ,side ,complications ,dura‐tion and tinnitus matching were not correlated with THQ scores .Conclusion THQ Chinese version has good relia‐bility and validity .It is worthy of clinical application .

Objective To establish the model of autoimmune dry eye and analyze the level of cytokines and the pathological changes of autoimmune dry eye rabbits treated with human umbilical cord mesenchymal stem cells (MSCs).Methods Twentyfour female New Zealand rabbits were randomly divided into three groups:Normal control group,dry eye group and MSCs treatment group,8 cases in each group.After treated with MSCs for six weeks,the pathological changes were observed,and the expression of Th1 and Th17 cell differentiation related cytokines mRNA in lacrimal gland tissue was detected by real-time quantitative PCR.The percentage of regulatory T cells (CD4 +Foxp3 + cells) in the lacrimal gland and spleen tissues of the dry eye model group and MSCs treatment group was measured by flow cytometry.Results After treatment with MSCs for 6 weeks,HE staining from each rabbit lacrimal gland showed that,the normal control group showed no or only a few lymphocytes;The lacrimal gland cells were atrophied in the dry eye group,the distribution of lymphocyte scattered around the gland and small blood vessels;Compared with dry eye group,the lacrimal gland lymphocyte infiltration reduced in MSCs treatment group,and the cell morphology were better.Compared with dry eye group,lymphocyte infiltration and aggregation in the conjunctival conjunctiva in MSCs treatment group were significantly reduced,the epithelial structure was intact,and the degeneration and atrophy cells were rare.The expression of inflammatory factors including interferon-γ(IFN-γ),Interleukin-17 (IL-17) and transcription factor T-bet mRNA were decreased in MSCs treatment group,there were significant difference compared with dry eye group (all P ＜ 0.05);Thl7 cell associated cytokines IL-17 expression levels were decreased,but no significant difference was found between two groups (P ＞ 0.05);The expression of transcription factor RORrt mRNA in dry eye group was significantly decreased,the difference was statistically significant (P ＜ 0.05).The expression of inflammatory factors TNF-α in the lacrimal gland tissue of MSCs treatment group was significantly lower than that in the dry eye model group,while the expression of the anti-inflammatory factor TGF-β was significantly higher than that in the dry eye model group,and the differences were statistically significant (all P＜0.05).The regulatory T cells (CD4 + Foxp3 + cells) in the lacrimal gland of dry eye model group accounted for 10％ of lymphocytes was,while the treatment group accounted for 27.8％,MSCs group was significantly higher than dry eye group,the difference was statistically significant (P ＜ 0.05).Conclusion MSCs can reduce the histopathological changes of the immune dry eye,and they may have immunoregulatory effects on autoimmune dry eye.The mechanisms of immunomodulatory effects may be related to the balance of inflammatory cytokines and anti-inflammatory cytokines.

Objective To investigate the distribution of eosinphililic neurons ( ENs),reactive astrocytes ( RAs),and infarction after transient cerebal ischemia,and the time profile of pathomorphological changes.Methods Unilateral forebrain ischemia was induced in Mongolian gerbils by two 10 minutes unilateral common carotid artery occlusions with a 5 hours interval.Laser Doppler flowmetry was used to detect intra-ischemic anterior cortex blood flow.Animals were sacrificed at 24 hours,4 days,2 weeks,4 weeks,16 weeks and the brain were prepared for pathomorphological assay.Results Intra-ischemic laser Doppler flowmetry show significant ischemia during carotid artery occlusion:22.1％ ± 9.5％,26.3％ ± 4.9％,37.5％ ± 3.5％,F =67.219,P ＜ 0.01 ; the decrease was significantly greater in the anterior cortex.ENs appeared in middle and deep layers at 24 hours postischemia,and ENs area extend to all layers of cortex by 4 days.Large areas of high EN density ( ≥80/mm2) evolved to infarcts between 4 days and 4 weeks.Posterior cortex evolved to low EN area ( ＜ 80/mm2) without transformation into infarcts.RAs were consistently distributed in areas with ENs,and RA areas with high EN density were largely transformed into infarcts between 4 days and 4 weeks. Delayed astrocytic death took place in the RA areas with high EN density.Conclusion Density of ENs is an important indicator of delayed astrocytic death and infarction in postischemic tissue.

BACKGROUND:Occluder implantation in patients with congenital heart disease can increase in vivo platelet adhesion and aggregation, resulting in thrombosis on the occluder surface. OBJECTIVE:To investigate the effect of the occluder on platelet function in patients with congenital heart disease undergoing transcatheter closure. METHODS: Clinical data from 124 patients with congenital heart disease undergoing transcatheter closure were retrospectively analyzed. These patients were divided into groups of atrial septal defect in 46 cases, patent ductus arteriosus in 43 cases and ventricular septal defect in 35 cases according to the types of congenital heart disease. The positive rates for peripheral blood CD62p, CD63 RESULTS AND CONCLUSION:There was no difference in the positive rates of peripheral blood CD and thrombin sensitive protein were compared before and 6 hours, 24 hours, 12 months after occluder implantation. 62p, CD63 and thrombin sensitive protein among three groups prior to occluder implantation. Up to 6 hours after occluder implantation, the expression levels of peripheral blood CD62p, CD63 and thrombin sensitive protein reached peak in the three groups, especialy in the patients with atrial septal defect and ventricular septal defect, then gradualy decreased. After 12 months, the expression levels of CD62p and CD63 recovered in the patients with patent ductus arteriosus and ventricular septal defect, but stil maintained a higher level in those with atrial septal defect (P < 0.05). The expression of thrombin sensitive protein showed no difference among the three groups at different time. These findings indicate that after occluder implantation, the platelet activation is more remarkable and lasts longer in the patients with atrial septal defect and ventricular septal defect, especialy in those with ventricular septal defect.

Objective To construct a new recombinant immunotoxin expression vector by using human VEGF165 and a truncated pseudomonas exotoxin A ramification (PE38) gene, and explore the expression of the VEGF165-PE38 fusion protein in HEK293 cells. Methods VEGF165 was cloned by polymerase chain reaction (PCR). PE38 gene was gained from an vector plasmid pRB391 by restriction endonuclease digestion, and then inserted to the eukaryotic expression vector pIRES2-EGFP. After the eukaryotic recombinant vector pIRES2-VEGF165-PE38-EGFP was identified by restriction endonuclease digestion and sequence analysis, the vector was transfected into HEK293 cells by liposome protocol. RT-PCR and ELISA method was used to confirm the expression of the fusion gene in the HEK293 cells. Results Restriction endonuclease digestion and sequence analysis revealed the VEGF165-PE38 fusion gene was cloned into the eukaryotic expression plasmid vector pIRES2-EGFP successfully. The pIRES2-VEGF165-PE38-EGFP fusion gene could express in the HEK293 cells. Conclusion The result provide the basis for search of the targeted cytotoxic activity to tumor vascular endothelial cells and may have some potential value in clinical application.

Objective To investigate the feasibility of enhancing therapeutic effects on goats' liver in vivo treated by high intensity focused ultrasound(HIFU)combined with microbubble contrast agent SonoVue.Methods Fifteen Nanjiang goats were adopted,own control was adopted in this study.One group received purely HIFU therapy(control group),another one was radiated with HIFU combined with SonoVue contrast agent(SonoVue group).The HIFU irradiating mode was dotted with 30 mm focal length and 150 W,250 W,350 W acoustic power respectively,and the exposure time was 15 s.Animals were sacrificed and dissected 24h later,and the sizes of necrotic region were measured.Specimens were collected for histological inspection.Results Under the same parameter of exposure,the maximum length,width,depth,and the mean volume of the tissue coagulated by HIFU in the SonoVue group were all significantly greater than those in control group(P＜0.05),the change of volume coagulated tissue was much more significant with the increase of the acoustic power.The energy effciency factor (EEF)of SonoVue group was significantly smaller than that of control group(P＜0.05).The demarcation line between the necrosis and normal region was clear and the necrosis was complete,no living cell.The boundary exist amount of bubbles.Conclusions The therapy of HIFU combined with microbubble eontrast agent can substantially enhance the biological effects of the treatment and improve the efficiency of HIFU therapy on goats liver.