Objective To analyze the clinical effect of minimally invasive percutaneous nephrolithotomy titanium laser lithotripsy(MPCNL)on upper ureteral calculi.Methods According to digital table,68 patients with upper ureteral calculi were randomly divided into the study group and control group,cach group in 34 cases.The study group received the MPCNL treatment,the control group received extracorporeal shock wave lithotripsy(ESWL)therapy. The therapeutic effects were compared.Results The success rate of gravel,stone clearance rates of the study group were 91 .1 8% and 91 .1 8%,which were higher than 70.59%,67.65% of the control group,there were significant differences between the two groups(χ2 =4.660,5.757,P =0.031 ,0.0164).Conclusion Percutaneous nerve titanium laser lithotripsy for upper ureteral stones can achieve good therapeutic effect.

Objective To compare the difference in quercetin against oxidative stress response in mouse and in NIH-3T3 cells before and after H2O2 treatment,to explore the underlying mechanism for the quercetin antioxidant.Methods The cultured NIH-3T3 cells were randomly divided into 4 groups: quercetin(Q) pre-protective group(Qb) firstly treated with quercetin for 24 h followed by incubation with H2O2 for 30 min;post-protective group(Qa) treated with H2O2 for 30 min followed by incubation with quercetin for 24 h;H2O2 group(H2O2) after exposure to H2O2 for 30 min,incubated with DMEM medium and the control group(C) only cultured with DMEM medium.The survival rate and apoptotic rate were detected respectively with MTT and TUNEL in NIH-3T3 cell sus-pension samples.The expression of cyclin D1,PTEN,NF-?B,HSP-70,BCl-2,BAX and caspase-3 were examined with immunocytochemistry and immunoblotting.Besides,20 Wistar rats were divided into control group and experimental group,the latter was given with quercetin in the doze of 0.13 mmol/kg.The levels of T-AOC,SOD,GSH-Px,GSH,MDA,NOS and NO2-/NO3-were detected both in the cleaved NIH-3T3 cells and in the plasma from both experimental and control animals prior to and post-1 h,2 h and after 24 h.Results When the Qb group was compared with H2O2 or Qa group,the survival rate was higher and the apoptotic rate was lower.When the H2O2 group was compared with C group,the expression of cyclin D1、PTEN or BCl-2 was down-regulated;while that of BAX、HSP-70、NF-?B or caspase-3 was up-regulated;the level of T-AOC,SOD,GSH-Px or GSH was decreased;that of NOS、NO2-/NO3-or MDA enhanced in the cleft NIH-3T3 cells.When the plasma level of the anti-oxidative enzyme system prior to-compared with post-1h and 2h-treatment with Q,the level of T-AOC,SOD,GSH-Px and GSH,especially the former two,were higher;MDA,lower;NOS or NO2-/NO3-promoted.However,the above parameters basically became normal 24 h after treatment with Q.Conclusion Quercetin down-regulates the promoted expression of HSP70,NOS,NO2-/NO3-and NF-?B etc.in H2O2-treatment NIH-3T3 cells.Qb could reverse the H2O2 damage effects more markedly.Moreover,the quercetin exerts anti-oxidant protective effect through modulating the anti-oxidative enzyme system both in vivo and in vitro.However,based on the cell heterogeneity in none-or pre/post-H2O2-treatment state,a difference in quercetin antioxidant response is noted.

AIM: To study the changes of iNOS mRNA ,protein and the production of nitric oxide(NO) as well as whether puerarin regulates the expression of iNOS mRNA during the formation of diabetic rat cataract. METHODS: One hundred and eight Sprague-Dawley(SD) rats were randomly divided into three groups, thirty-six rats were taken as control group, seventy two rats were injected peritoneally with streptozotocin(STZ,45mg/kg) to establish animal model of diabetic cataract, and then divided into STZ (36) and puerarin(36) treatment groups. Morphologic changes of lens were observered with slit lamp and light microscope; Samples were taken at 20th, 40th, 60th day and the changes in iNOS mRNA and protein expression of lens epithelium cells(LEC)as well as production of NO and NOS activity were determined with reverse transcription -polymerase chain reaction(RT-PCR), western blot, and biochemical method ,respectively. RESULTS: Morphologyic changes of LEC, up-regulation of iNOS mRNA and iNOS protein as well as increase in NO production and NOS activity in the LEC were observered during the formation of rat diabetic cataract. Compared with TZ group, puerarin treatment group showed distinctly down-regulation of iNOS mRNA and iNOS protein and decrease in NO production and NOS activity as well as attenuation of morphologic changes. CONCLUSIONS: There are morphologic changes of LEC and up-regulation of iNOS mRNA and as well as increase in NO production and NOS activity in the LEC during the formation of diabetic rat cataract , and treatment with puerarin can reverse the above changes.

Objective:To investigate the ability of chymase inhibitors on tryptase release from human colon mast cells.Methods:Human mast cells were dispersed from colon tissue with collagenase and hyaluronidase,and were challenged with stimulus for 15 min at 37℃.Tryptase assay performed following previous procedures.In brief,a 96-well microtitre plate was coated with antiserum to human tryptase.The tryptase levels in the samples were detected with a monoclonal antibody specific to tryptase and the reaction was visualized by addition of OPD.Results:At 15 min and 35 min following incubation,anti-IgE and calcium ionophore were able to provoke significant tryptase release from human colon mast cells.Chymase inhibitors ZIGPFM,TPCK and ?1-antitrypsin had no stimulatory effect on colon mast cells at both 15 min and 35 min incubation periods.All the chymase inhibitors were able to inhibit anti-IgE induced tryptase release in a concentration dependent manner with a maximum of 37%,40% and 36.6% inhibition being achieved with 1 ?mol/mL of ZIGPFM,80 ?mol/mL of TPCK,30 ?mol/mL of ?1-antitrypsin,respectively.Preincubation of inhibitors of ZIGPFM and TPCK with cells for 20 min at 37℃ before challenging with anti-IgE was able to slightly enhance their inhibitory actions.Amastatin,a specific inhibitor of aminopeptidase,had no effect on anti-IgE induced tryptase release.All the chymase inhibitors were able to inhibit calcium ionophore induced tryptase release,the maximum inhibition were 23%-35.3%.And the extent of inhibition by ZIGPFM was increased when colon mast cells were preincubated for 20 min before calcium ionophore being added.However,the same treament failed to improve the action of TPCK.Conclusion:We found for the first time that inhibitors of chymase were able to inhibit anti-IgE and calcium ionophore induced tryptase release from human colon mast cells,which may indicated a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cell related diseases.

Aim To investigate the ability of chymase inhibitors o n histamine release from human colon mast cells. Methods Human ma st cells were dispersed from colon tissue with collagenase and hyaluronidase, an d were challenged with stimulus for 15 min at 37℃.A glass fibre-based fluorome tric assay was used to measure histamine in the supernatants of dispersed mast c ells.Results chymase inhibitors ZIGPFM, TPCK and ? 1-antitry psin failed to induce significant histamine release from colon mast cells. All t he chymase inhibitors were able to inhibit anti-IgE induced histamine release i n a concentration dependent manner with a maximum of 37%, 26% and 36.8% inhibit ion being achieved with 1 mmol?L -1 of ZIGPFM, 80 mmol?L -1 of TPCK , 30 mmol?L -1 of ? 1-antitrypsin, respectively. Preincubation of inhib itors of ZIGPFM and TPCK with cells for 20 min at 37℃ before challenging with a nti-IgE was able to slightly enhance their inhibitory actions. All the chymase inhibitors were able to inhibit calcium ionophore induced histamine release, th e maximum inhibition was 23.6%～35%.And the extent of inhibition by TPCK was in creased when colon mast cells were preincubated for 20 min before calcium ionoph ore being added. However, the same treament failed to improve the action of ZIGP FM. Conclusion In the current study, we found that inhibitors o f chymase were able to inhibit anti-IgE and calcium ionophore induced histamine release from human colon mast cells, which may indicate a potential of a novel therapy for the treatment of inflammatory bowel disease or other mast cell relat ed diseases.

AIM: To investigate the ability of Chinese cobra snake venom-metalloproteinase(MT) to induce the histamine release from human mast cells and its potential mechanisms.METHODS: MT was purified from the snake venom by using heparin agarose and Superdex75 chromatography.Mast cells were dispersed from human lung, colon and tonsil tissues after digestion with collagenase and hyaluronidase.The dispersed mast cells were then challenged with MT,stimulus and control in LP4 tubes for 15 min at 37 ℃.A glass fibre-based fluorometric assay was used to measure histamine in the supernatants of dispersed mast cells.RESULTS: MT induced a dose-dependent release of histamine from human colon,lung and tonsil mast cells.As low as 0.03(mg/L) of MT was able to stimulate significant histamine release from human colon mast cells,but a minimum of 0.3 or 30 mg/L of MT was required to stimulate a similar level of histamine release from lung or tonsil mast cells,respectively.The release of histamine from colon and lung mast cells in response to MT was maximized at 12 min following the addition of the stimulus.This was quite different from the picture of the peak histamine release from tonsil mast cells,in which histamine release was maximized at 8 min following the addition of MT.Pretreatment of cells with metabolic inhibitors and pertussis toxin reduced dramatically histamine release from human colon,lung and tonsil mast cells by MT.In exogenous Ca~(2+) and Mg~(2+) free experiments,the release of histamine induced by MT was significantly decreased.CONCLUSION: Cobra snake venom MT induces human mast cells to release histamine through a G-protein-related mechanism,which may contribute to the pathogenesis of venomous snake bite.

Background and objective Targeting the mutations and amplifications in the epidermal growth factor receptor (EGFR) gene has curative effects on cancers of the lung, oral cavity, and gastrointestinal system. However, a systemic immune inflammation is an adverse effect of this therapeutic strategy. In this study, we aimed to identify the possible changes in the tumor microenvironment that contribute to the anti-cancer activity of EGFR inhibition.Methods Squamous-cell cancers were induced by the syngeneic transplantation of either EGFR-null or wild-type mouse primary keratinocytes that had been transduced with an oncogenic H-ras retrovirus. The mice were treated with gefinitib. Then, flow cytometric was used to detect the ratio of T cells and the expression of programmed cell death receptor 1 (PD-1). RT-PCR was used to detect the expression of cytokines and chemokines.Results Tumors that formed from EGFR-null keratinocytes were smaller, had fewer infilltrat-ing FoxP3+ Treg cells, lower Foxp3 RNA, and lower percentage of PD-1 positive CD4 cells than those formed from wild-type keratinocytes. These results indicated that tumor cells can autonomously regulate the tumor microenvironment. Hosts with wild-type cancers and that were treated with gefitinib for 1 week tended to have smaller tumors. The treated mice in the short-term pharmacological model tended to have reduced FoxP3+ cells and FoxP3 RNA in the tumor microenvironment, as well as a substantially increased ratio of IL-1A/IL-1RA transcripts. These results suggested that the brief systemic inhibition of EGFR signaling alters the immune environment of the targeted cancer.Conclusion The autonomous (genetic) or systemic (pharmacologic) inhibition of EGFR signaling in tumor cells reduces tumor growth and Treg infilltration in the tumor micro-environment. An EGFR-dependent Treg function supports the growth of squamous cancers. Therefore, Treg is a target in the therapeutic strategy of EGFR inhibition.

Background and objective Tumor-associated fibroblasts (TAF) is an important part of TME, which inhibits the function of immune cells. CD8+ T cells play a significant role in tumor immunity. T-cell membrane possesses a distinct type of molecule with a negative regulatory function. Upon interaction with its corresponding ligand [programmed death factor ligand 1 (PD-L1)], programmed death factor 1 (PD-1) is activated and thus inhibits the kinase activity of T cells. This study aims to explore the possible effects of TAF on PD-L1 expression in lung cancer cells. Methods Lung cancer cell lines H1975 and H520 were co-cultured with (experiment) or without TAF (control) via Transwell assay for through 48 hours under the same culture condition. H1975 and H520 cells were counted using a microscope. The protein and mRNA expression levels of PD-L1 were detected by FCM assay and PCR analysis, respectively. Results The numbers of lung cancer cells in 100μm2 for H1975 and H520 cells are (46±21) and (38±10) in the experiment group, respectively, and (16±5) and (12±5) in the control group, respectively (P<0.05). The expression levels of the PD-L1 protein in H1975 and H520 cells are (20.93%±3.54%) and (19.26%±3.04%) in the experiment group, respectively, and (12.58%±2.52%) and (11.60%±2.65%) in the control group, respectively (P<0.05). The mRNA expression levels in H1975 and H520 cells are (16.45±1.25) and (15.38±2.02) pg/mL in the experiment group, respectively, and (7.78±1.27) and (7.20±1.58) pg/mL (P<0.05) in the control group, respectively (P<0.05). Conclusion TAF promotes the growth and increases the expression of PD-L1 in H1975 and H520 cells.

OBJECTIVE: To determine the effect of human corticotrophin-releasing hormone (CRH) on the expression of connexin-43 phosphate (P-Cx43) in human myometrial smooth muscle cells (SMCs) and the function of cell gap junction intercellular communication in SMCs. METHODS: Human non-conceive myometial SMCs were cultured with different concentrations of CRH (0, 5.85, 58.5, 585 and 5850 pmol/L). Western blot was used to test P-Cx43 and Cx43 non-phosphate (NP-Cx43) of protein expression. Cell scratch was used to test cell gap junction intercellular communication opening status in human myometrial SMCs. RESULTS: Compared with the control group, the expression of P-Cx43 was higher in the CRH groups (P<0.01), and was concentration-dependent. There was no significant difference in NPCx43 between the control group and the CRH groups (P>0.05). The transmission of cell layers in the CRH groups was higher than that in the control group (P<0.01), and as the concentration of CRH increased, the time was concentration-dependent (P<0.01). CONCLUSION CRH can enhance the expression of P-Cx43 and the function of gap junction intercellular communication in the primary cultured myometrial SMCs.
Cell Communication ; drug effects ; Cells, Cultured ; Connexin 43 ; metabolism ; Corticotropin-Releasing Hormone ; pharmacology ; Female ; Gap Junctions ; drug effects ; Humans ; Myocytes, Smooth Muscle ; metabolism ; Myometrium ; cytology ; Phosphorylation ; drug effects

Objective To evaluate the protective effect of Bletilla striata polysaccharide ( BSPS) on immunological and chemical liver injury in mice. Methods Thirty Kunming male mice were randomly divided into five groups, including the normal control group,model control group,and low-,middle-,and high-dose BSPS groups (n=6 each).Tail vein injection of ConA was carried out to establish the ConA-induced liver injury model.After different treatments,all the animals were sacrificed,and the plasma levels of ALT and AST were tested.Additionally,sixty Kunming male mice were randomly divided into six groups,including the normal control group,model control group,silymarin group,and low-,middle-,and high-dose BSPS groups (n=10 each).Tail vein injection of CCl4 was performed to establish the CCl4-induced acute liver injury model.After different treatments,the plasma levels of ALT and GSH were tested.The effects of BSPS on the weights of the liver and spleen were examined. Results The levels of ALT and AST were reduced in BSPS-treated mice when compared with those experiencing only ConA-induced liver injury ( model control group) ,and significant difference was found between the middle-and high-dose BSPS groups and the model control group (P<0.01,P<0.05).The weights of the liver and spleen and the level of ALT were reduced in BSPS-treated mice as compared with those with only CCl4-induced acute liver injury (model control group),while the level of GSH was significantly increased in middle-and high-dose BSPS groups (P<0.05). Conclusion BSPS at low,middle,and high doses can prevent against the ConA-induced immunological liver injury and CCl4-induced acute liver injury in mice.