Animals ; Arginine ; pharmacology ; Blood Pressure ; drug effects ; Hypertension ; etiology ; metabolism ; physiopathology ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley

AIM: To determine the evolutionary pattern of parenchymal cell apoptosis in multiple organs early after intestinal ischemia-reperfusion(I/R) and its induction mechanisms and the role of apoptosis in triggering SIRS/MODS. METHODS: An I/R model was reproduced by clipping and releasing the superior mesenteric artery in rats and mice. Flow cytometry, electron microscope, DNA agarose gel electrophoresis, TUNEL method, fluorescent and Gomori's silver-HE staining were used to detect apoptosis. Distribution features of apoptotic parenchymal cells in multiple organs were observed. Immunohistochemical staining of HSP 70 and Bcl-2 were performd to study the induction mechanisms of apoptosis.RESULTS and CONCLUSION: 1. Damage of the liver, lung, gut and kidney was appeared in early phase of I/R. The percentages of apoptosis in parenchyma organs increased progressively. The percentages of cell necrosis increased with the prolonged I/R duration. 2. Percentages of apoptosis were much higher near the central veins of liver lobules, in the outer medulla of the kidney, and the antimescenteric border of intestinal mucosal epithelium because of ischemia. 3. The expression of HSP 70 increased and Bcl-2 reduced in the areas mentioned above because of hypoperfusion. 4. Apoptosis of I/R hepatocytes, splenocytes and thymocytes was obviously increased after Kupffer cell blockage with GdCl3, showing the functional state of Kupffer cells may play an important role in SIRS/MODS.

Objective To prepare 5-aminolevulinic acid (ALA) and hematoporphyrin monomethyl ether (HMME) hydrogel suppositories and to evaluate their photosensitizer transfer efficiencies in rectal tumor tissue.Methods The BALB/c mice implanted SW837 rectal cancer cells subcutaneously were randomly divided into four groups:intrarectal suppository administration group,cutaneous administration group,intratumoral injection group and intravenous injection group.Fluorescence spectrophotometry was used to measure the concentration of protoporphyrin Ⅸ (PpⅨ) and HMME in rectal wall,skin and tumor tissue.The distribution of photosensitizer was determined by a fluorescence spectroscopy system.Results The concentration of PpⅨ in the ALA suppository administration group was 9.76 times (1 h) and 5.80 times (3 h) higher than that in the cutaneous administration group,and the differences were statistically significant (all P＜0.05).The maximal penetration depth of ALA in tumor tissue was about 3-6 mm at 2 h after the cutaneous administration.After the HMME suppository administration,the concentration of HMME in the rectal wall was very low.The maximal penetration depth of HMME in tumor tissue was less than 2 mm after the cutaneous administration.Conclusions ALA is more likely to penetrate mucosal barrier compared to skin tissue.The hydrogel suppository based rectal administration is expected to be a new administration method for the rectal cancer photodynamic therapy using ALA.

BACKGROUND:It is so difficult to have aortic valve replacement with smal aortic annulus. Improper treatment may lead to patients with valvular mismatch phenomenon, and thus make left ventricular outflow tract obstruction, increase transvalvular pressures, cause cardiac hypertrophy secondary to increased left ventricular afterload and even congestive heart failure.
OBJECTIVE:To summarize the treatment strategy for preventing valvular mismatch phenomenon caused by smal aortic annulus after aortic valve replacement.
METHODS:Eighty-five patients with smal aortic annulus underwent aortic valve replacement surgery. 19 mm SJM Regent valve was applied to the patients with orifice diameter>17 ≤ 19 mm;to the adult patients with orifice diameter ≤ 17 mm, we performed bovine pericardial patch enlargement of the smal aortic annulus and valve replacement using 19 mm SJM Regent valve. For those with orifice diameter>19 ≤ 21 mm, we selected 21 mm Hancock II ultra biological valve for valve replacement. Effective orifice area index, left ventricular mass index, inter-ventricular septal thickness, left ventricular wal thickness, trans-valvular peak velocity, the pressure difference across the valve and trans-valvular mean pressure were measured through echocardiography. After discharge, patients were fol owed up in out-patient clinic and evaluated regularly by echocardiography.
RESULTS AND CONCLUSION:There were no early deaths after operation and al cases were cured and discharged. Fol ow-up time was between 6 months and 3 years. The main complications included low cardiac output syndrome in two cases, reoperation due to bleeding in one case, and ventilator dependence in two cases. No cases occurred in cerebral complications such as cerebral hemorrhage or cerebral thrombosis, and no valvular dysfunction or card flap appeared. There was no bovine pericardium tearing, thrombosis, calcification, tumor-like bulge, infection or immune reactions. A total of 81 cases were fol owed up and the fol ow-up rate was 95%(81/85). There were NYHA class grade I in 65 cases, and grade II in 16 cases. Peak velocity across the aortic valve and the mean pressure were significantly decreased, effective orifice area index increased significantly, left ventricular mass index, left ventricular wal thickness and the thickness of the inter-ventricular septum were significantly reduced compared with pre-operation, and no valvular mismatch phenomenon occurred. Compared 21 mm Hancock II ultra biological valve with 21 mm SJM Regent group, the former got a better peak velocity and mean trans-valvular pressure, and better left ventricular remodeling index. Body weight and body surface area were significantly increased in 19 mm Regent valve group after operation. The results suggest that individualized treatment strategies should be taken to prevent the occurrence of postoperative valvular mismatch phenomenon for patients with smal aortic annulus.

Cirrhosis with portal hypertension is a common disease which has a significant impact on the quality of patients' life. Esophagogastric devascularization (EGDV) has been demonstrated to be an effective method to treat portal hypertension, however certain complications are associated with it. The purpose of this study was to evaluate the effectiveness and clinical outcome of the selective EGDV (sEGDV) for the treatment of portal hypertension. The study was conducted prospectively from Jan. 1 2011 to Dec. 31, 2012, and 180 patients were randomized to the sEGDV group (n=90) or the non-sEGDV (n-sEGDV) group (n=90). Patients' demographics, preoperative lab test results and operative details were comparable between the two groups. Postoperative and short-term complications were analyzed in two groups. There was statistically significant difference (P<0.01) in the PVF reduction between the two groups. Post-operative complications showed no statistically significant difference between the two groups in the incidence of bleeding, ascites, acute portal vein thrombosis, fever and hepatic encephalopathy. Mortality between two groups was comparable. The incidence of splenic fossa effusion after the surgery was lower in sEGDV group than in n-sEGDV group. There were no significant differences in the short-term follow-up data such as esophageal varices and portal hypertensive gastropathy (P>0.05). It is suggested that sEGDV is a safe, simple and effective surgical procedure. It has both the advantages of the shunt and devascularization because it preserves body's voluntary diversion. With the advantage of low incidence of postoperative complications, it is an ideal surgical approach for the treatment of portal hypertension.

Objective To investigate the expression of macrophage migration inhibitory factor(MIF) in renal tissue of children with Henoch-Schonlein purpura nephritis(HSPN),and its correlation with clinical indexes and pathological changes,and to explore its effect on the pathogenesis of HSPN.Methods According to the clinical manifestation,60 children with HPSN were divided into only purpura group,mixed group and HSPN group.MIF concentration of Henoch-Schonlein purpura(HSP) groups and healthy control group were detected with enzyme linked immunosorbent assay(ELISA).MIF protein expression and the marker of human macrophage(CD68) in renal tissues of HSPN and normal control group were detected with immunohistochemistry method.The total urine protein for 24 hours and urinary N-acetyl-beta-D-glucosaminidase (NAG) level were detected with laboratory routine method.Results MIF concentration in mixed group and HSPN group were significantly higher than that in only purpura group and healthy control group(Pa

Objective To investigate the combined effects of somatostatin (SST) and cisplatin (DDP) on proliferation and apoptosis in gallbladder cancer cells,and to investigate the mechanism of the combined effects.Methods We performed immunohistochemistry to detect the PTEN expression in gallbladder cancer.We then investigated the combined effects of SST and DDP on cell proliferation in vitro with CCK-8 assay and analyzed the interaction between these two drugs using isobologram analysis.We also investigated the combined effects on cell proliferation in vivo using a xenograft nude mouse model.FITC-Annexin V/PI assay and TUNEL staining assay were performed to detect the proportion of apoptosis after combined treatment in vitro and in vivo.Reactive oxygen species and mitochondrial membrane potential were detected with DCFH-DA assay and JC-1 staining assay after the combined treatment.We finally detected the PTEN and p-AKT associated proteins using western blotting after the combined treatment.Results PTEN was abnormally decreased in gallbladder cancer tissues.PTEN expression was negatively correlated with cancer differentiation and was positively correlated with patients'survival time.DDP treatment decreased while combined treatment with SST induced PTEN expression and inhibited AKT activation by reversing resistance to DDP.Isolated SST or DDP treatment inhibited gallbladder cancer GBC-SD and SGC996 cell proliferation which was dose-dependence.These two drugs synergistically inhibited gallbladder cancer cell growth in vivo and in vitro.Isolated SST or DDP treatment induced cell apoptosis and combined treatment induced cell apoptosis the most.SST inhibited AKT activation but did not induce ROS.DDP induced ROS resulting in increased cell apoptosis.Either SST or DDP alone increased the levels of cytoplasmic cytochrome C protein and activated caspase-3.Conclusions SST enhanced growth inhibition by cisplatin in gallbladder cancer cells through inducing PTEN expression.This study provides the theoretical basis for further combined clinical chemotherapeutic applications.

Objective To investigate the predictive value of preoperative serum tumor markers test for lymph node metastasis of intrahepatic cholangiocarcinoma (ICC).Methods The retrospecgtive cohort study was conducted.The clinicopathological data of 69 patients with ICC who were admitted to the Xinhua Hospital of Shanghai Jiaotong University between May 2006 and May 2016 were collected.Among 69 patients with pathological diagnosis,24 with lymph node metastasis were allocated into the lymph node metastasis group and 45 without lymph node metastasis were allocated into the non-lymph node metastasis group.Tumor markers of the 2 groups were preoperatively detected,including alpha-fetoprotein (AFP),carcinoembryonic antigen (CEA),prostate specific antigen (PSA),CA19-9,CA125,CA242,CA153,CA724,CA211,neuron-specific enolase (NSE) and squamous cell carcinoma antigen (SCC).Receiver operating characteristic (ROC) curve was built,and critical value,sensitivity and specificity were calculated based on ROC curve.Coincident rate between significant indicators and results of pathological examination was calculated.Observation indicators:(1) overall positive rates of tumor markers;(2) comparison of tmmor markers levels in the 2 groups;(3) tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Measurement data with skewed distribution were described as M (Q25,Q75) and comparison between groups was analyzed using the Wilcoxon rank sum test.Comparison of count data was analyzed by the chi-square test and Fisher exact probability.The statistically significant indicators were analyzed by the ROC curve.Results (1) Overall positive rates of tumor markers:positive rates of AFP,CEA,PSA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC in 69 patients were 27.5％ (19/69),29.0％ (20/69),4.3％ (3/69),69.6％ (48/69),36.2％ (25/69),50.7％ (35/69),26.1％ (18/69),21.7％ (15/69),62.3％ (43/69),31.9％(22/69) and 21.7％(15/69),respectively.Positive rates of AFP,CEA,CA19-9,CA125,CA242,CA153,CA724,CA211,NSE and SCC were more than 20％,which became comparison indicators of 2 groups.(2) Comparison of tumor markers levels in the 2 groups:levels of CA19-9,CA125,CA242 and CA211 were 284.9 U/mL (42.5 U/mL,730.3 U/mL),63.6 U/mL (23.4 U/mL,172.1 U/mL),71.7 U/mL (25.6 U/mL,138.9 U/mL),6.7 μg/L (3.9 μg/L,17.5 μg/L) in the lymph node metastasis group and 58.0 U/mL (25.9 U/mL,405.9 U/mL),18.2 U/mL (11.7 U/mL,33.8 U/mL),11.0 U/mL (3.7 U/mL,41.7 U/mL),3.7 μg/L (2.7 μg/L,6.9 μg/L) in the non-lymph node metastasis group,respectively,with statistically significant differences between the 2 groups (Z=2.016,3.213,3.143,2.482,P＜0.05).(3) Tumor markers predicted ROC curve of lymph node metastasis and coincident rate between ROC curve and results of postoperative pathological examination:area under the ROC curve of CA19-9,CA125,CA242 and CA211 were respectively 0.648 [95％ confidence interval (C I):0.515-0.781,P＜0.05],0.736 (95％ CI:0.608-0.864,P＜0.05),0.731 (95％ CI:0.603-0.859,P＜0.05),0.714 (95％ CI:0.581-0.847,P＜0.05).The positive critical value,sensitivity and specificity of CA19-9,CA125,CA242 and CA21 were 150.6 U/mL,35.7 U/mL,43.4 U/mL,6.0 μg/L and 62.5％,66.7％,70.8％,62.5％ and 71.1％,82.2％,77.8％,75.6％,respectively.The coincident rate between ROC curve and results of postoperative pathological examination of CA 19-9,CA 125,CA242 and CA211 were 68.1％ (47/69),76.8％(53/69),75.4％(52/69),71.0％(49/69),respectively.Conclusion The levels of serum CA19-9,CA125,CA242 and CA211 can effectively predict lymph node metastasis of patients with ICC.

Objective To explore the killing effect of dielectric barrier discharge (DBD) plasma on tumor cells and to analyze the DBD-induced apoptosis mechanism.Methods Thiazolyl blue tetrazolium bromide (MTT) assay method was used to detect the killing effect of low temperature plasma on the cytotoxicity of normal spleen leukocytes and acute promyelocytic leukemia cells (LT-12) at different doses.The changes of reactive oxygen species (ROS) level were measured after plasma treatment.The cell apoptosis rate was detected by Annexin V/PI double staining at different doses.The expression of apoptosis-related genes and proteins was detected by qRT-PCR and Western Blot.Results MTT results showed that the killing effect of plasma treatment was dose-dependent and time-dependent.The cell survival rate after 8 hours of treatment decreased from 98％ to 63％ with the dose increasing from 30 s to 240 s.The survival rate decreased from 78％ (2 h) to 39％ (24 h) after the treatment with a same dose (e.g.240 s).Annexin V/PI double staining results demonstrated that the plasma effect can induce apoptosis,and the apoptosis rate was not only positively correlated with the plasma dose,but also with the post-plasma time.The longer the post-plasma time,the higher was the apoptosis rate.The apoptotic rate of the 60 s dose treatment after 12 h was 48％ that increased to 55.3％ with the dose of 120 s.The production of reactive oxygen species (ROS) detected by flow cytometry also showed a time correlation of the plasma treatment.After the plasma treatment,the ROS level immediately increased to 1.24 times,and sharply increased to 5.39 times after 20 h post-plasma.The experimental results of qRT-PCR and Western blot showed that the expression of the genes and proteins of Caspase family and Bcl-2 family was very active at 8 to 12 h post-plasma treatment.Conclusions Low-temperature plasma can effectively kill tumor cells,and apoptosis is the main mechanism of death.The molecular mechanism of apoptosis of tumor cells induced by low temperature plasma was preliminary confirmed.

Objective:To present evidence for the pathogenetic role of allergic factor,histamine,in type I allergy for induction of liver damage.Methods:Three groups of rabbits were fed normally and injected (iv) daily with 0, 0.04 or 0.08 ?g/kg phosphohistamine, respectively, for days. The serum level of ALT and AST in each group rabbits was assayed dynamically during the treatment. After treatment for days, the tested rabbits were sacrificed for pathological examination of the liver tissues.Results:The serum level of both ALT and AST in rabbits treated with phosphohistamine increased significantly during the tested periods, compared to that of the control group. However, both ALT and AST levels showed no significant difference between 0.04 ?g/kg and 0.08 ?g/kg groups. Liver microscopic examination, pathological damage could be observed in the tested groups in a time-and dose-dependent manner under microscopic examination. No evident pathological change appeared in the control group.Conclusion:Liver damage could be induced by histamine dosage-and time-dependently. This pathological action of histamine, a type I allergic factor, presents further evidence for a direct role of type I allergy in the pathogenesis hepatic injury.