Objective To investigate the protective effects and possible mechanism of hydrogen sulfide (H2S) against oxidative stress injury induced by hydrogen peroxide (H2O2) in retinal ganglion cell-5 (RGC-5).Methods RGC-5 cells were divided into four groups:RGC-5 group (normal control group),RGC-5 + H2O2 (RGC-5 were cultured in 500 μmol · L-1 H2O2 for 24 hours) group,RGC-5 + NaHS (RGC-5 were cultured in 50 μmol · L-1 NaHS for 30 minutes) + H2O2 (RGC-5 were cultured in 500 μmol · L-1 H2 O2 for 24 hours) group,and RGC-5 + NaHS (RGC-5 were cultured in 50 μmol · L-1 NaHS for 30 minutes) group.Western blots were applied to measure the expression of cytochrome c (Cyt.c) and optic atrophy 1 (OPA1).The fluorescent dye JC-1 assay was chosen to detect the mitochondrial membrane potential (△Ψm).Furthermore,transmission electron microscope was used to observe the morphology of mitochondria.Results Compared with RGC-5 group,the expression of Cyt.c in RGC-5 + H2O2 group decreased in mitochondria,and increased in cytoplasm (all P ＜ 0.05),but there was no statistical difference between RGC-5 group and RGC-5 + NarHS + H2O2 group (all P ＞0.05).Compared with RGC-5 group,the expression of Cyt.c in RGC-5 + NaHS group increased in mitochondria,and decreased in cytoplasm (all P ＜ 0.05).Compared with RGC-5 group,the expression of OPA1 in RGC-5 + H2O2 group decreased in mitochondria,and increased in cytoplasm (all P ＜ 0.05).In RGC-5 + NaHs + H2O2 group and RGC-5 + NaHS group,the expression of OPA1 within and outside the mitochondria had no significant difference compared with RGC-5 group (all P ＞ 0.05).Compared with other three groups,the mitochondrial membrane potential in RGC-5 + H2O2 group obviously decreased,but there was no statistical difference among other three groups (P ＞ 0.05).The mitochondria were globular swelled in RGC-5 group,but in other three groups,the mitochondria had slightly swelled.Conclusion H2S can protect the mitochondrial morphology and functions of RGC? 5 from H2O2-induced oxidative stress via preventing OPA1 release from mitochondria.

Objective To explore the pathogenesis of irritable bowel syndrome (IBS) by detecting serum levels and the colonic mucosa expression of inflammatory cytokines,peptide YY (PYY),and claudin-1,and to analyze their correlation.Methods From April 2013 to April 2015,50 outpatients with IBS and 20 healthy controls were selected.Serum levels of PYY,interleukin (IL)-10,tumor necrosis factor (TNF)-α and claudin-1 were detected by enzyme-linked immunosorbent assay (ELISA).The expression of IL-10,TNF-α,PYY and claudin-1 in colonic mucosa was determined by immunohistochemistry.Single factor analysis of variance,least significant difference (LSD) method,chi-square test,and Pearson correlation analysis were performed for statistical analysis.Results Among the 50 patients with IBS,27 cases were diarrhea-type irritable bowel syndrome (D-IBS),and 23 cases were constipated-type irritable bowel syndrome (C-IBS).The serum level and the positive expression rate of PYY in colonic mucosa of D-IBS group were significantly higher than those of healthy control group ((16.28± 2.75) ng/L vs (10.12± 1.55) ng/L;66.7 ％ (18/27) vs 30.0 ％ (6/20)),and the differences were statistically significant (LSD-t=10.19,x2 =6.182,both P＜0.05).The serum level and the positive expression rate of IL-10 in colonic mucosa of D-IBS group were both significantly lower than those of healthy control group ((2.95 ±0.24) ng/L vs (3.58±0.35) ng/L;22.2％(6/27) vs 50.0％ (10/20)),and the differences were statistically significant (LSD-t =4.52,x2=3.948,both P＜0.05).The serum level and the positive expression rate of TNF-α in colonic mucosa of D-IBS group were both significantly higher than those of healthy control group ((8.73±0.41) ng/L vs (7.73±0.51) ng/L;66.7％(18/27) vs 30.0％(6/20)),and the differences were statistically significant (LSD-t=8.43,x2 =6.182,both P＜0.05).There was no statistically significant difference between C-IBS group and healthy control group in the serum levels of PYY ((10.24±1.34) ng/L vs (10.12± 1.55) ng/L),IL-10 ((3.43 ± 0.71) ng/L vs (3.58 ± 0.35) ng/L),TNF-α ((7.81±0.26) ng/L vs (7.73 ±0.51) ng/L),and thus the positive expression rate in colonic mucosa (39.1％(9/23) vs 30.0％(6/20),56.5％(13/23) vs 50.0％(10/20),34.8％ (8/23) vs 30.0％(6/20);all P＞0.05).The serum level of claudin-1 and its positive expression rate of PYY,IL-10,TNF-α in colonic mucosa in D-IBS group were both lower than those of healthy control group ((94.44 ± 6.61) ng/Lvs (103.64 ± 5.47) ng/L;11.1％ (3/27) vs 40.0％ (8/20)),and the differences were statistically significant (LSD-t=5.76,x2 =5.349;both P＜0.05).However,the serum level of claudin-1 and its positive expression rate in colonic mucosa in C-IBS group were both higher than those of healthy control group ((115.54±3.42) ng/L vs (103.64±5.47) ng/L;73.9％ (17/23) vs 40.0％(8/20)),and the differences were statistically significant (LSD-t=5.56,x2 =5.055;both P＜0.05).The serum levels of IL-10 and PYY,TNF-α and claudin-1 were negatively correlated in the D-IBS group (r=-0.874 and -0.863,both P＜0.05).While the serum levels of TNF-α and PYY,IL-10 and claudin-1 were positively correlated (r =0.865 and 0.876,both P＜ 0.05).Conclusions There may be the imbalance of proinflammatory factors and anti-inflammatory factors in IBS patients.PYY may decrease the expression of claudin-1 by promoting IL-10 and inhibiting TNF-α,and thus ameliorate the inflammation reaction of IBS patients.

Antigens, CD34 ; metabolism ; Cytokines ; metabolism ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Mucin-1 ; metabolism ; Perineum ; pathology ; Phosphopyruvate Hydratase ; metabolism ; Sarcoma ; metabolism ; pathology ; surgery ; Vimentin ; metabolism

Antigens ; chemistry ; Fixatives ; chemistry ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; methods ; Microwaves ; Staining and Labeling ; methods

Aged ; Antigens, CD20 ; analysis ; Gene Rearrangement ; Humans ; Immunoblastic Lymphadenopathy ; genetics ; metabolism ; pathology ; Immunoglobulin Heavy Chains ; genetics ; Immunohistochemistry ; Lymph Nodes ; metabolism ; pathology ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Male ; Polymerase Chain Reaction ; RNA, Viral ; analysis ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; Reed-Sternberg Cells ; metabolism ; pathology

Objective To study the features of CT and MRI diagnosis of intracranial germ cell tumors ,and to improve the un-derstanding of its imaging findings .Methods Fifteen cases of intracranial germ cell tumors proved by pathology and follow-up were retrospectively analyzed .Results Among the 15 cases with intracranial germ cell tumors ,9 cases in pineal body , tumours showed like-round masses,the margin of the masses were well defined .CT was slightly high density,MRI revealed isointense or hypointense signals on T1 weighted and homogeneous hyperintense signals on T 2 weighted image without edema around tumors and homogeneous en-hancement.One cases in CSF spread.Four cases were identified in the saddle-up area, among the 2 cases was cystic,1 cases was sol-id-cystic and 1 cases was solid,CT was isointense or high density in solid,low density in cystic,without edema around tumors.MRI re-vealed slight-hypointense signals on T 1 weighted and hyperintense signals on T 2 weighted image clearly enhanced in solid , cystoid with-out enhance .In the other 2 cases of intracranial germ cell tumors in the basal ganglia region ,CT was inhomogeneous high density ,inho-mogeneous signals on T2 and T1 weighted image, inhomogeneous enhancement .Conclusions Intracranial germ cell tumors show char-acteristic manifestations on CT and MRI,according to location,shape,signal,age characteristic,which is helpful in diagnosis and differ-ential diagnosis and guide treatment plan .

Adenocarcinoma ; diagnosis ; pathology ; Adult ; Aged ; Aged, 80 and over ; Ascitic Fluid ; chemistry ; Cadherins ; analysis ; Calbindin 2 ; Carcinoembryonic Antigen ; analysis ; Diagnosis, Differential ; Epithelial Cells ; chemistry ; Female ; Humans ; Immunohistochemistry ; Keratin-5 ; analysis ; Male ; Middle Aged ; Pericardial Effusion ; chemistry ; diagnosis ; Pleural Effusion ; chemistry ; diagnosis ; Pleural Effusion, Malignant ; chemistry ; diagnosis ; S100 Calcium Binding Protein G ; analysis

BackgroundEndostatin (ES) is currently the strongest endogenous angiognesis inhibitor,and it can inhibit the occurrence of neovascularization.Various studies demonstrated that the poly RGD sequence can enhance the function of the ES gene.ObjectiveThis study was to evaluate the use of gene therapy of modified ES for alkaline burn-induced corneal neovascularization (CNV).MethodsOne hundred and two clean SD rats were randomly divided into the normal control group,the pCI empty vector group,the pCI-ES group,and the pCI-RGDRGDES group.Corneal neovascularization models were established by placing a piece of 3 mm filter paper with 1 mol/L NaOH at the central cornea for 40 seconds.3 μg of the pCI blank vector,ES-tranfected pCI blank vector,or RGDRGD-ES-transfected pCI vector was injected into the superior bulbar conjunctiva after the alkali burn twice at 1-week intervals.Area of CNV and edema of the cornea in the various groups of rats were examined daily under the slit lamp biomicroscope.1,4,7 and 14 days after operation,the rats were sacrificed by the excessive anesthesia method and corneal tissues were obtained to evaluate pathological changes.The expression of CD34 in vascular endothelial cells was detected by immunochemistry to calculate the corneal neovascular density.The expressions of VEGF mRNA and Flk-1 protein in the corneas were detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis.The use and maintenance of animals followed the Statement of ARVO.Results Seven to fourteen days after corneal alkali-burning,the corneal neovascular area was smaller in the pCI-ES group and pCI-RGDRGD-ES group compared with the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and nevascular area in the pCI-RGDRGD-ES group was smaller than that in the pCI-ES group (P＜0.05).The expression level of CD34 was significantly lower in the pCI-ES group and pCI-RGDRGD-ES group than that in the normal control group and pCI blank vector group (P＜0.05,P＜0.01 ),and the expression level of CD34 was further declined in the pCI-RGDRGD-ES group compared with the pCI-ES group (P＜0.05 ).Compared with the normal control group and pCI vector group,the expressions of the Flk-1 protein and VEGF mRNA were decreased in the pCI-ES group and pCI-RGDRGD-ES group on the fourth day after corneal alkali-burning (P＜0.01,P＜0.05 ),and those in the pCI-RGDRGD-ES group were less than the pCI-ES group (P＜ 0.05,P＜ 0.05 ).Conclusions Subconjunctival injection of both ES and modified RGDRGD-ES genes result in significant suppression of CNV in vivo,and modified RGDRGD-ES appears to be more effective than native ES.The main mechanism of ES in inhibiting neovascularization is to downregulate the expression of VEGF and Flk-1.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; therapeutic use ; Doxorubicin ; therapeutic use ; Epstein-Barr Virus Infections ; Female ; Herpesvirus 4, Human ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; virology ; Lymphoma, T-Cell, Peripheral ; drug therapy ; pathology ; Neoplasms, Second Primary ; drug therapy ; pathology ; Prednisone ; therapeutic use ; Vincristine ; therapeutic use

Objective According to principle of forming primer dimer,a new cloning gene method was established. Methods Two and more oligonucleotide with complementary partnership in the 3′ end were synthesized by artifical method,oligonucleotide,Taq DNA polymerase and dNTP were mixed in the proportion of reasonable,then expanded and extended by PCR,result was identifed by agarose gel electrophoresis.Results A clear special band was appeared in the agarose gel. Conclusion Short fragment target gene less than 500 bp may be synthesized by this method.