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Objective To detect the expression of costimulatory molecules B7-H4 and programmed death ligand 1 (PD-L1) in bladder cancer,and to explore the correlation between them and clinicopathological features of bladder cancer.Methods Immunohistochemical staining was used to detect the expression of B7-H4 and PD-L1 in 98 cases of bladder urothelial carcinoma,which were confirmed by pathology from August 2014 to December 2015 in our hospital.There were 23 females,aged 45-82 years,with an average age of 67.8 years.Among them,42 cases of adjacent normal tissues were used as controls.The clinical stage,histological grade and recurrence of bladder cancer were collected,and the correlation between them was analyzed.Results The positive rates of B7-H4 and PD-L1 in bladder urothelial carcinoma were 54.1％ (53/98) and 59.2％ (58/98),respectively,and there was no expression in normal bladder tissues (P ＜ 0.05).The positive expression rate of B7-H4 in muscle invasive bladder cancer (MIBC) patients was higher than that in non-muscle invasive bladder cancer (NMIBC) [73.5％ (25/34) vs.43.8％ (28/64),P =0.005].The positive expression rate of B7-H4 in high-grade patients was higher than that of low-grade [70.0％ (21/30) vs.47.1％ (32/68),P =0.036].The expression rate of B7-H4 in high-risk group was higher than that of low-intermediate risk group [57.1％ (20/35) vs.27.6％ (8/29),P =0.018].The positive expression rate of PD-L1 in patients with MIBC was higher than that in NMIBC [79.4％ (27/34) vs.48.4％ (31/64),P =0.003].The PD-L1 expression rate of histological high-level group was higher than that of low-level group [73.3％ (22/30) vs.52.9％ (36/68)],but the difference was not statistically significant (P =0.058).The PD-L1 expression rate in high-risk group was 68.6％ (24/35),and also higher than low-middle group 24.1％ (7/29) (P ＜ 0.05).There was a positive correlation between the expression of B7-H4 and PD-L1 in bladder urothelial carcinoma (r =0.318,P=0.002).The combined recurrence rate of the two groups was significantly higher than that of the negative expression of the two groups [66.7％ (14/21) vs.30.8％ (8/26),P=0.014].Conclusions The expression of B7-H4 and PD-L1 is up-regulated in bladder urothelial carcinoma,which is closely related to the clinical stage,histological grade,risk classification and recurrence of NMIBC.
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Objective To study the protective effects of human umbilical cord-derived mesenchymal stem cells (ucMSCs)against severe acute pancreatitis (SAP) in rats.Methods A total of 135 Sprague-Dawley male rats were randomly divided into Sham group,SAP group and SAP+ucMSCs group (45 each).SAP+ucMSCs group:Severe acute pancreatitis was induced by injecting 5％ sodium taurocholate (0.1ml/100g) into the common bilio-pancreatic duct and then CM-DiI-labeled ucMSCs at 1 × 107cells/kg were injected via the tail vein.All the rats were sacrificed 12,24 and 72 hours after SAP.The 72h death rate was counted.Pathological changes in the pancrease were detected by HE staining and pathological score was graded,ucMSCs colonization was observed by fluorescence microscopy.The serum levels of amylase,lipase,TNF-α,IL-1β,IL-4 and IL-10 were determined by ELISA.Results ucMSCs colonize the injured area of pancreatic tissue,the 72h death rate was reduced,and the serum amylase and lipase were also reduced significantly.Moreover,ucMSCs significantly reduced the pathological score of the pancrea and the level of proinflammatory cytokines (TNF-α and IL-1β),but the levels of anti-inflammatory cytokines were increased (IL-4 and IL-10).Conclusion Transplantation of ucMSCs can reduce the severity of pancreatic injury and inflammation in SAP rats.
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BACKGROUND: A few devices have been reported to be used for studies on trauma, but these devices are unavailable for establishing the animal models of trauma because of their limited application range. OBJECTIVE: To develop a multifunctional impact system and evaluate its application effect, thus paying ways for establishing the animal models of trauma and basic experiments.METHODS: The multifunctional impact system was designed based on the theory of energy storage device, simple multifunctional impact device and impact parameter measuring equipment, and its effectiveness and stability were detected. The rat chest and different visceral organs were subjected to the closed impact experiment using a 5 cm2 impact at the predetermined parameter of 200, 300, 400, 500 kPa, respectively. Afterwards, the rats were sacrificed for morphological observation.RESULTS AND CONCLUSION: The multifunctional impactor was successfully developed, of which the maximum impact stress could be adjusted from 0 to 200 kg and compressive and extrusion stress also could be continuously adjusted from 0 to 100 kg. The experimental results showed that the impactor made certain damage to the rat lung, liver and spleen suggesting its favorable effectiveness (P 0.05). These findings suggest that the impactor is easy to operate in various ways and holds good effectiveness and stability, and its impact parameters can be detected in real time. Therefore, the impactor is suitable for both establishing the animal model of trauma and basic experiments.
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<p><b>OBJECTIVE</b>To explore whether microRNA-200a (miR-200a) could be used as a novel biomarker of liver cancer using a rat model system.</p><p><b>METHODS</b>Diethylnitrosamine abdominal injection was applied to induce liver cancer in the F344 rat strain (n =40); ten unmodeled rats served as controls. In addition, human subjects with normal healthy liver (n =10), liver cirrhosis (n =10), and liver cancer (n =10) were enrolled in the study. Blood samples from both rats and patients and rats' livers were collected for analysis. Real-time quantitative PCR and enzyme-linked immunosorbent assay were used respectively to measure the expressions of serum miR-200a and alpha-fetoprotein (AFP) for all rat and human subjects. In situ hybridization was used to detect the miR-200a expression in the rats' livers.</p><p><b>RESULTS</b>Comparison of normal rats and the liver cancer modeled rats showed that the latter had significantly lower expression of miR-200a (P less than 0.05), with decreasing expression following the progression of liver injury to cancer (liver cirrhosis rats less than early liver cancer rats less than advanced liver cancer rats); in contrast, the AFP levels were significantly higher in the liver cancer modeled rats only at the early and advanced stages of the liver cancer (P less than 0.05). These</p><p><b>RESULTS</b>suggested that miR-200a expression decreases during the developmental process of liver cancer, while AFP expression increases distinctly at the stage of tumor formation. Analysis of the human subjects' clinical samples showed that miR-200a expression was decreased in both liver cirrhosis patients and liver cancer patients (vs. normal liver subjects, P less than 0.05), while AFP showed abnormal expression only in the patients with liver cancer. Comparison of the normal rats and modeled rats using in situ hybridization showed the positive rates for miR-200a expression were 1.00% +/- 0.01% in rats with normal liver, 0.37% +/- 0.03% in rats with fibrotic liver, 0.14% +/- 0.01% in rats with cirrhotic liver, 0.05% +/- 0.00% in rats with early stage liver cancer, and 0.01% +/- 0.00% in rats with advanced stage liver cancer.</p><p><b>CONCLUSION</b>MiR-200a may play an important role in liver cancer development and may have diagnostic value for indicating early liver cancer.</p>
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Adult , Aged , Animals , Female , Humans , Male , Middle Aged , Rats , Young Adult , Case-Control Studies , Liver , Metabolism , Liver Neoplasms , Blood , Metabolism , MicroRNAs , Blood , Metabolism , Rats, Inbred F344 , alpha-Fetoproteins , MetabolismABSTRACT
With the continuous development of the cancer stem cell theory,biomarkers for CSCs (Cancer stem cells) have gradually become the hot issues in tumor research.As the most commonly used CSCs biomarker,CD44 gains lots of attentions.This paper will review recent research about CD44 molecule and its powerful role in regulating tumor progenesis and progression.
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Objective To predict the severity of acute pancreatitis by detecting the changes of microRNA in the serum,and whether paracentesis catheter drainage (PCD) should be applied to the patients.Methods The peripheral blood of 120 patients with acute pancreatitis who were admitted to the General Hospital of Chengdu Military Command from October 2013 to March 2014 were collected.Thirty-five patients with severe acute pancreatitis (SAP) or moderately severe acute pancreatitis (MSAP) were in the group A,and 85 patients with mild acute pancreatitis (MAP) were in the group B.The scores of the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ),RANSON and bedside index for severity in acute pancreatitis (BISAP) were assessed.The expressions of the miR-146a,miR-10b,miR-21 and miR-26a in the serum were detected by the real-time quantitative PCR.The differences in the expressions of the 4 kinds of microRNAs in the 2 groups were compared,and the correlation between the 3 exaluation system was analyzed.Factors influencing the timing of the PCD for the PCD patients were analyzed.The measurement data were analyzed using the t test,and the relationship between the variables was analyzed using the linear correlation analysis or the rank correlation analysis.Univariate or multivariate analysis was done by the Logistic regression analysis.Results (1) The scores of the APACHE Ⅱ,RANSON and BISAP were 8.28 ±0.61,3.42 ±0.54 and 1.71 ±0.32 in the group A,and 3.18 ±0.52,1.43 ±0.25 and 0.37 ±0.06 in the group B,with significant differences between the 2 groups (t =4.266,7.809,4.113,P ＜ 0.05).(2) The expressions of the miR-146a,miR-10b,miR-21 and miR-26a were 1.41 ± 0.21,2.94 ± 0.49,1.62 ± 0.25,1.21 ± 0.20 in the group A,and 6.29 ± 0.91,0.52 ± 0.09,2.82 ± 0.33 and 3.57 ± 0.64 in the group B.There were significant differences in the expression of the miR-146a and miR-10b between the 2 groups (t＝-2.156,2.110,P ＜0.05),while no significant difference in the miR-21,miR-26a was detected between the 2 groups (t =-1.114,-1.571,P ＞ 0.05).(3) There was correlation between the expressions of the miR-146a,miR-10b and the APACHE Ⅱ,RANSON,BISAP in the group A (r =-0.826,0.837,-0.874,0.866,-0.833,0.899,P ＜ 0.05),while no correlation was detected between the expressions of the miR-21,miR-26a and the 3 exaluation systems in the group A (r =0.642,0.321,0.701,0.750,0.716,0.716,P ＞ 0.05).There was no significant difference between the miR-146a,miR-10b,miR-21,miR-26a and the APACHE Ⅱ (r =0.067,0.347,0.133,0.111,P＞0.05),RANSON (r=0.178,0.078,0.092,0.142,P ＞0.05) and BISAP (r =0.103,0.260,0.216,0.285,P ＞ 0.05) in the group B.(4) The results of univariate analysis showed that miR-10b,RANSON and BISAP were the factors influencing the timing of PCD (OR =4.170,5.612,2.500,95 ％ confidence interval:1.092-15.932,1.232-21.622,1.190-5.254,P ＜ 0.05).The results of multivariate analysis showed that miR-10b was the factor influencing the timing of PCD (OR =2.374,95％ confidence interval:1.115-5.056,P ＜ 0.05).Conclusions miR-10b and miR-146a might be the predictors of severity of severe acute pancreatitis; miR-10b might be the indicator in judging whether PCD should be applied.
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ObjectiveTo review the management and prevention of stenosis after hepaticojejunostomy. MethodsWe retrospectively analyzed 133 patients who developed stenosis after hepaticojejunostomy carried out for iatrogenic bile duct injuries. ResultThrough our active reoperative protocol, we were able to achieve satisfactory results in 131 patients, although the results were still unsatisfactory in 2 patients. ConclusionsStenosis after hepaticojejunostomy is a severe complication which should actively be prevented. Once it happens, doctors should apply active, appropriate and effective measures to correct the stenosis.
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Objective To construct a sequence tag library of HepG2 cells by serial analysis of gene expression (SAGE). Methods Total RNA of HepG2 cells was extracted using a Trizol agent. The double strand cDNA were reverse-translated from mRNA which was isolated from total RNA. According to the protocol of SAGE, a sequence tag library of HepG2 cells was constructed. The transcript, which was represented by each tag, was analyzed by consulting the GenBank and UniGene, and the abundance of the tags was analyzed by SAGE analysis software. Results A total of 14 181 SAGE tags out of 15 586 tags were steadily and effectively amplified and the lengths of the tags were between 200-3000 bp. After sequence analysis of the tags, 2023 specific genes were revealed. Conclusions The use of improved reverse transcription agent kit may guarantee obtaining the full length cDNA, which provides materials for the construction of SAGE library. SAGE is a powerful method to investigate the gene expression profile, and it establishes a foundation for gene researches.
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Objective To study the proliferation and differentiation patterns of hepatic stem cells from mice cultured in vitro isolated identify their characteristics.Methods Mice were divided into 5 groups according to the pregnant (embryo days,ED13,ED15 and ED18) or born age (day0 and 3).Hepatic stem cells were isolated and cultured in vitro.The amount of the stem cells as well as their growing situation and differentiation pattern were observed and compared among different groups and markers of stem cells (CD117,CD90.1,CD49f,c-Met),hepatic cells(AFP) were used to identify the cultures.Resulls The cells with best situation of growing as colonies were obtained from ED15 group.Their expression of specific markers suggested that they were hepatic stem cells.The stem cells isolated from ED15 mice in subculturing proliferated in line pattern and differentiated in reverse line pattern.The expression of AFP varied as normal distribution as cell differentiation development.Conclusion Most cells have characteristics of hepatic stem cells isolated from fetal liver and the number of these cells decreases gradually as embryo duration prolongs.The hepatic stem cells proliferate in line pattern and differentiate into hepatic cells after in vitro culture.
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Objective:By analyzing the arrangement of the military medical curriculum for cadets,some advantages and disadvantages about the current course arrangements are discussed.Methods: To ensure the regain rate,188 cadets,who are undergraduates in our university,answered questionnaires in class.Then the results were analyzed by statistics.Later cadet representatives had an informal discussion with investigators.Results:According to the results,the arrangements of the military medical curriculum for cadets in the military medical universities are qualified on the whole,can meet cadets' cultivation needs.But to accommodate the requirement of military medical talented men in the 21-century,some suggestions about courses information,such as curriculum category,curriculum sequence,curriculum contents,curriculum duration and etc,are given to the related teaching departments.Meanwhile,the results of this study can provide some information for the cultivation of national defense students in the military medical university.
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Objective To investigate the effect of ipratropi um bromide and atropine on airway mucin hypersecretion in the chronic bronchiti s model of rats. Methods The model was established by intratrac heal instillation of small dose of lipopolysaccaride (200 μg) ipratropium bromi de and atropine were administrated 3-21 d after the model was established. Patho logical changes, mucin in bronchoalveolar lavage fluid (BALF) and tracheal ring culture medium were examined before and after the adnsinistration of iprat ropium bromide and atropine. Results Characteristic pathologica l manifestions of chronic brochitis were found after instillation of LPS. Sig nificant decrease in the number of tracheal epithelia goblet cells, secretion of mucin were observed in ipratropium bromide and atropine treated rats. The eff ect of inhibiting the secretion of mucin of ipratropium bromide was higher than atropine. Conclusion It suggests that muscarinic acetylcholine receptor plays an important role in airway mucin hypersecretion in chronic bro nchitis model of rat. Ipratropium bromide exhibit a stronger inhibition effect on mucin hypersecretion than atropine, moreover no inhibition effect on mucocil iary clearance which was observed in atropine.