Abnormalities, Multiple ; Colon ; abnormalities ; Humans ; Intestinal Pseudo-Obstruction ; Urinary Bladder ; abnormalities

BACKGROUND:Adipose-derived mesenchymal stem cells are gaining widespread interest in the Achil es tendon tissue engineering and regeneration, and an enabling environment (oxygen concentration) for cellinduction and differentiation is particularly necessary.
OBJECTIVE:To co-culture adipose-derived mesenchymal stem cells with primary tenocytes in standard culture condition (20%O 2 tension) and in an atmosphere of reduced oxygen (2%O 2 tension) in order to determine whether the two conditions differ in their effect on tenogenic differentiation.
METHODS:Tenocytes were isolated via serial expansion in culture from several Sprague-Dawley rats’ Achil es tendons. Adipose-derived mesenchymal stem cells were purchased. After one passage, adipose-derived mesenchymal stem cells were indirectly co-cultured with tenocytes in standard culture condition (20%O 2 tension) and in an atmosphere of reduced oxygen (2%O 2 tension). Col agen 1, col agen 3, Tenomodulin, Thrombospondin-4, Scleraxis levels were compared for each culture condition at 7, 14 and 21 days fol owing co-culturing. Immunofluorescence staining was performed to evaluate production of col agen 1 and Thrombospondin-4.
RESULTS AND CONCLUSION:Fol owing indirect co-culturing, hypoxic differentiated adipose-derived mesenchymal stem cells expressed higher levels of tendon-related genes and proteins than normoxic controls, which suggest that oxygen levels can significantly affect tenogenic differentiation, and hypoxia is advantageous for efficient differentiation of adipose-derived mesenchymal stem cells in vitro for tendon tissue engineering.

Objective To explore the relationship between K-ras gene and cholangiocarcinoma by detecting the K-ras gene mutation of bile duct tissues.Method We studied all the patients who presented to our hospital from June 2010 to June 2012 with stenosis of the bile duct.There were 17 cases of cholangiocarcinoma and 17 cases of benign stenosis.From the DNA extracted from the paraffin tissues,we used the HRM assay for K-ras gene mutation.Result We found that the HRM method and the DNA typing had exactly the same result for the DNA content which confirmed the effectiveness of the HRM assay.Of note,the K-ras mutation rate was found to be significantly higher in the cholangiocarcinoma cases (11/17) when compared with the benign cases (3/17).Conclusion The mutation of the K-ras gene was closely related to cholangiocarcinoma.Our results suggest a new way to diagnosis cholangiocarcinoma.

Objective To investigate the value of timely duodenal papilla fenestration for reducing the incidence of pancreatitis after endoscopic retrograde cholangiopancreatography (ERCP).Methods The clinical data of 181 patients with difficult biliary cannulatian during ERCP at the Xinhua Hospital of Shanghai Jiaotong University from July 2006 to December 2009 were retrospectively analyzed.Of the 181 patients,98 patients who received traditional incubation were in the control group,and the other 83 patients who received early duodenal papilla fenestration were in the test group.The success rate of selective incubation and incidence of pancreatitis were compared between the 2 groups.All data were analyzed using the t test,chi-square test or Wilcoxon rank sum test.Results The success rate of incubation,incidences of hyperamylasemia and pancreatitis were 85.7％ ( 84/98 ),7.1％ (7/98) and 10.2％ ( 10/98 ) in the control group,and 94.0％ ( 78/83 ),18.1％ ( 15/83 ) and 2.4％ (2/83) in the test group,respectively,and there were significant differences between the 2 groups (x2 =10.12,5.03,4.41,P＜0.05).The numbers of patients with mild,moderate and severe pancreatitis were 3,5 and 2 in the control group,and 1,1,0 in the test group,respectively,and there was a significant difference between the 2 groups ( Z =- 2.11,P ＜ 0.05 ).Conclusion Timely duodenal papilla fenestration is safe and effective in reducing the incidence of pancreatitis for patients with difficult biliary cannulation during ERCP.

Objective To evaluate the clinical characteristics and surgical risks for elderly patients with gastric cancer. Methods The clinical data from 607 patients, who received surgical therapy for gastric cancer between January 2006 and December 2009, were retrospectively analyzed.Among them, 267 patients were over 65 years of age, and the other 340 patients (≤65 years of age)were served as control. Complete surgical resection was done by doctors in same group. The preoperative complications such as cardiopulmonary disease and post-operative complications as well as pathologic patterns were compared between two groups. Results Coexisted diseases were found more in elderly patients than in controls (68.91% vs 31.63 %, P＜0.01). Whereas there was no significant difference between two groups with respect to post-operative complication [28. 19% (75/267)vs 25.59 % (87/340)]. However, the incidence of cardiopulmonary complications, especially pulmonary infection, was higher in elderly patients than in controls (17.23% vs 5.29% ,P＜0.01). The digestive tract complications were related to the pattern of radical operation. Post-operative complication occurred less in patients treated with D1 radical operation than those with D2 and D3 radical operation.Conclusions The incidence of post-operative complication in elderly patients with gastric cancer was closely related to the surgical method. The elderly patients may has less complications if they were operated with D1 radical method which can reduce risk of complication and improve the quality of life.

Objective:To investigate the influence of miR-7 knock down on the development of CD 4+SP cells in the thymus in mice,and preliminary explore its possible mechanism.Methods:The changes of volume ,weight and total cell counts of thymus in miR-7 knock down (miR-7KD) mice were observed compared with Wild-type(WT)mice;the pathological changes of thymus were observed by HE staining.FACS analysis was performed on the proportion ,as well as the expression level of CD44 and CD62L,of thymus CD4+single positive (SP) cells.Meanwhile,the proliferation percentage of CD4+SP cells was measured by Ki-67 staining.The apoptosis percentage of CD4+SP cells was analyzed by FACS.The changes on the transduction of ERK 1/2 pathways were determined by Western blot.Results:Compared with WT mice ,the size,weight and total cell number of thymus were marked reduced in miR-7KD mice( P<0.05 );moreover ,pathological change also was presented.The proportion and total cell number of thymus CD 4+SP cells were marked decreased ( P<0.05 ).Furthermore ,the expression level of CD 44 and proliferation percentage ,as well as apoptosis percentage ,of CD4+SP cells were obviously increased (P<0.05),however,the expression level of CD62L of CD4+SP cells were decreased (P<0.05). Finally,the level of total ERK1/2 and phosphor-ERK1/2 was decreased obviously ( P<0.05 ).Conclusion: miR-7 knock down can affect the development of CD 4+SP cells in the thymus , which might be closely related to the cell activation state and altered the transduction of ERK1/2 pathways.

Objective · To investigate the difference in myocardial strain of left ventricle between obstructive hypertrophy cardiomyopathy (HCM) and nonobstructive HCM. Methods · Cardiac magnetic resonance imaging (MRI) exam was performed on 48 sequential enrolled patients with HCM (18 with obstructive HCM, and 30 with nonobstructive HCM), whose left ventricular ejection fractions (LVEF) were over 50%. Twenty-five healthy volunteers were examined as normal controls. Global longitudinal strain (GLS), global radial strain (GRS), global circumferentialstrain (GCS), LVEF, left ventricular end diastolic volume (LVEDV), left ventricularmass (LVM), left ventricular end diastolic volume index (LVEDVI), and left ventricular mass index (LVMI) were collected and compared. Radial strain, circumferential strain and peak radial displacement were also measured in medial segment of left ventricle according to American Heart Association (AHA) 17-segment model. Results · ① LVEF of the patients with obstructive HCM was bigger than those of nonobstructive HCM patients and control group (P<0.05). LVM and LVMI of the HCM groups were bigger than those of control group (P<0.01). ② Left ventricle GLS, GRS, and GCS significantly decreased in the patients with nonobstructive HCM compared to those with obstructive HCM (P<0.05). The three parameters of two HCM groups were significantly lower than those of healthy volunteers (P<0.05). ③ Compared with obstructive HCM patients,the segmental parameters of left ventricule, the medial segment circumferential strain and radial strain of nonobstructive HCM patients significantly decreased (P<0.05), and the two parameters of both HCM groups were lower than those in healthy volunteers. Compared with obstructive HCM patients and healthy volunteers, peak radial displacement of left ventricule medial segment in nonobstructive HCM witnessed a significant decrease, while no significant difference was observed between obstructive HCM patients and healthy volunteers. Conclusion · In the LVEF preserved HCM patients, the myocardial strain of left ventricle in nonobstructive HCM patients decrease significantly than that in obstructive HCM patients, which may result in the different clinical outcomes intwo types of HCM patients. It is suggested that the myocardial strain is more sensitive than ejection fraction in the evaluation of myocardial performance of HCM patients.

Objective:To investigate the change of expression of miR-7 in activated CD4+ T cells in vitro, and preliminary explore its possible significance. Methods: CD4+CD62L+T cells was purified from splenocytes of FVB mice by magnetic cell sorting system (MACS). After stimulation with anti-CD3/CD28 antibody,the relative expression of miR-7 was examined by Real-time PCR, and the expression level of CD69 molecular was analyzed by FACS. Furthermore,the relative expression of miR-7 in CD4+T cells was detected at different time points during stimulation. With the treatment of ERK inhibitor PD98059,change of miR-7 expression was de-termined by Real-time PCR. Meanwhile, the proliferation of CD4+T cells was examined by CCK-8 assay and the expression level of CD69 and CD62L molecular were analyzed by FACS. Finally,the expression of cytokines IL-6,IL-10,and IFN-γ were determined by Real-time PCR. Results:Compared with control group,the relative expression of miR-7 was increased significantly after stimulation with anti-CD3/CD28 antibody,as well as expression level of CD69 molecular was augmented(P<0. 05). In contrasted with 0 h and 24 h,the expression of miR-7 was significantly increased after 48 h and 72 h during stimulation(P<0. 05). Furthermore,the relative expression of miR-7 was significantly declined in CD4+T cells in ERK inhibitor PD98059 treatment group. Finally, the expression level of CD69 molecular,as well as cytokines IL-6, IL-10 and IFN-γ, were also decreased significantly ( P<0. 05 ) . Conclusion: The relative expression of miR-7 was significantly increased in activated CD4+T cells,closely related to ERK pathway,which provided an important foundation for successive research work on exploring the functional role of miR-7 in the CD4+T cells.

Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.

OBJECTIVE To explore the application value of the cortical bone packing in external auditory bony canal and attic reconstruction in tympanoplasty.METHODS A total of 30 patients admitted during the period of middle ear cholesteatoma patients in our hospital from March 2014 to March 2016,were randomly divided to observation group and control group,the radical mastoidectomy with canal wall down was adopted in the control group,the radical mastoidectomy with canal wall down combined with bone autograft in the external auditory meatus and bony powder mastoid obliteration in observation group,the postoperative out-come between two groups were compared.RESULTS There was no significant difference between the speech frequency average air bone conduction and air conduction threshold before the treatment in two groups.There was significant difference between the speech frequency average air bone conduction and air conduction threshold after the treatment(P＜0.05),the air conduction threshold significantly reduced,the patients in the observation group were significantly lower than the control group,there was no recurrence of cholesteatoma in two groups of patients before and after the treatment during follow-up period;the dry ear time in observation group was significantly lower than that of the control group,and dry ear the incidence was significantly higher than that of the control group,the difference is statistically significant(t=2.758,4.865,P＜0.05).CONCLUSION Radical mastoidectomy with canal wall down combined with the cortical bone packing in external auditory bony canal and attic reconstruction,can effectively presesre the patient hearing,it should be recommended.