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OBJECTIVE To investigate the inhibitory effect and the possible mechanism of hydrogen sulfide （H2S） on the proliferation of cardiac fibroblasts. METHODS The heart of neonatal SD rats was collected， and cardiac fibroblasts were separated with differential centrifugation. Using sodium hydrosulfide as the donor of H2S， the effects of H2S on the proliferation of cardiac fibroblasts induced by angiotensin Ⅱ （Ang Ⅱ）， hydroxyproline content and the expression of sirtuin 3 （SIRT3） protein were detected. After SIRT3 knockdown with siRNA technology， the effects of H2S on the proliferation of cardiac fibroblasts induced by Ang Ⅱ， hydroxyproline content， the expressions of collagen Ⅰ （Col Ⅰ）， collagen Ⅲ （Col Ⅲ ） and optic atrophy protein 1 （OPA1） were detected. RESULTS H2S could inhibit the proliferation of Ang Ⅱ-induced cardiac fibroblasts， reduce the content of hydroxyproline and increase the expression of SIRT3 （P＜0.05）. After down-regulating the expression of SIRT3 with siRNA technology， the inhibition of H2S on the proliferation of Ang Ⅱ-induced cardiac fibroblasts and the reduction of hydroxyproline content were both inhibited， and the effect of H2S on reducing the expression of Col Ⅰ and Col Ⅲ and enhancing the expression of OPA1 was also significantly weakened. CONCLUSIONS H2S inhibits the proliferation of Ang Ⅱ -induced cardiac fibroblasts through increasing the expression of SIRT3.
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BACKGROUND@#Fibroblast activation protein (FAP) is one of the surface markers of cancer-associated fibroblasts (CAFs) and is closely related to the malignant characterization of CAFs. SP13786 is a specific micromolecule inhibitor of FAP and this study is to investigate the effects and mechanism of SP13786 on the migration and invasion of A549 cells through regulating exosomes of CAFs.@*METHODS@#CAFs and paracancerous fibroblasts (PTFs) were isolated and subcultured from freshly resected lung adenocarcinoma tissues and paracancerous normal tissues separately. MTT assay was used to detect the proliferation of CAFs incubated by different concentrations of SP13786; PTFs-exo, CAFs-exo and CAFs+SP13786-exo were extracted by polymer precipitation method. The A549 cells were divided into Ctrl group, PTFs group, CAFs group and SP13786 group and each group was incubated with DMEM, PTFs-exo, CAFs-exo and CAFs+SP13786-exo separately. Laser confocal microscope was used to observe the endocytoses of exosomes by A549 cells. The expression of alpha-smooth muscle actin (α-SMA) and FAP in PTFs and CAFs and the expression of E-cadherin, N-cadherin, Slug, Stat3 and P-Stat3 in A549 cells were detected by immunofluorescence, immunohistochemistry and Western blot. The migration and invasion ability of A549 cells were detected by cell scratch and transwell methods.@*RESULTS@#α-SMA and FAP were expressed much higher in CAFs than that in PTFs which indicate that CAFs and PTFs were successfully obtained from lung adenocarcinoma and paracancerous tissues (P0.05). Finally, WP1066 (a specific inhibitor of Stat3) was used to comfirm whether SP13786 could influence EMT of A549 cells by inhibiting Stat3 phosphorylation via CAFs-Exo. The results showed that when the phosphorylation of Stat3 in CAFs group was inhibited by WP1066, SP13786 could not influence the P-Stat3 expression and EMT of A549 cells anymore (P>0.05).@*CONCLUSIONS@#As a specific micromolecule inhibitor of FAP, SP13786 indirectly inhibits the migration and invasion of A549 cells by affecting exosomes of CAFs. The possible mechanism is to inhibit the phosphorylation of Stat3 and thus affect the EMT of A549 cells.
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@#To perform the quality control of deferasirox and establish its quality criterion, three related substances which were determined by analyzing the synthetic route were synthesized, and their structures were confirmed by 1H NMR and MS. These substances were 2-(2-hydroxyphenyl)-4H-benzo［1, 3-e］oxazin-4-one(A), 2-hydroxy-N-(2-hydroxyben-zoyl)-benzamide(B), and methyl-4-［3, 5-bis(2-hydroxyphenyl)- 1H- ［1, 2, 4］triazol-1-yl］-benzoate(C).
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Objective To investigate the combined detection of serum vascular endothelial growth factor (VEGF),squamous cell carcinoma antigen (SCC-Ag)and cytokeratin 19 fragment (CYFRA21-1)value in assessing the efficacy of chemotherapy in patients with advanced squamous cell carcinoma of the lung.Methods 60 cases of lung squamous cell carcinoma patients in January 2011 between June 2013 to be admitted to hospital were treated with gemcitabine plus cisplatin or carboplatin chemotherapy.Detected the levels of serum VEGF,SCC-Ag and CYFRA21-1 expression in 60 cases before and after 1 cycle of chemotherapy in patients using ELISA and immunoradiometric assay,and evaluated the efficacy evaluation with reference to RECIST criteria after 2 cycle of chemotherapy.Results Effective group serum VEGF and CYFRA21-1 levels were 413± 114 pg/ml and 14.7±9.6 ng/ml;serum VEGF and CYFRA21-1 levels after one cycle of chemotherapy were 272±131 pg/ml and 10.8±7.1 ng/ml,which decreased after chemotherapy (P0.05). Serum SCC-Ag levels before and after each treatment group difference was not statistically significant (P>0.05).Multiple regression analysis showed that serum VEGF,CYFRA2 1-1 levels and the efficacy of lung squamous were related changed (t=5.86,P<0.001;t=2.26,P=0.027),and there was no significant correlation between SCC-Ag levels changes and the effect of chemotherapy (t=1.52,P=0.133).Conclusion Serum VEGF and CYFRA21-1 expression level changes associat-ed with squamous cell carcinoma of the lung chemotherapy effect,which can advance understanding of the effect of chemo-therapy,combined with advanced squamous cell carcinoma detection of serum VEGF and chemotherapy assessment CY-FRA2 1-1 has a certain reference value.
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Objective To investigate the values of serum vascular endothelial growth factor (VEGF) ,carcinoembryonic antigen (CEA) ,carbohydrate antigen 19-9 (CA19-9) and ferritin (SF) determination in assessing the efficacy of chemotherapy in patients with advanced gastric cancer .Methods Wedetermined the concentrations of serum VEGF ,CEA ,CA19-9 and SF for 85 patients with advanced gastric cancer ,before and after 1 cycles of chemotherapy and evaluated the the effect of chemotherapy after 2 cycle of chemotherapy .Results In effective group ,serum concentrations of VEGF ,CEA ,CA19-9 and SF significantly decreased after chem-otherapy cycle(P 0 .05) ,compared with those before chemotherapy cycle .Multiple regression analysis showed that serum CEA ,VEGF ,CA19-9 concentration changes correlated with chemotherapy efficiency ,while serum SF didn′t(t=1 .58 ,P=0 .119) . Conclusion Serum CEA ,VEGF ,CA19-9 concentration changes associated with gastric cancer chemotherapy efficiency ,which help predict the efficiency of chemotherapy .
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Objective To explore the prognostic value of APACHE Ⅱscore in patients with Severe acute organophosphorus poi-soning .Methods 42 patients with Severe acute organophosphorus poisoning ,in which 34 cases survived ,8 cases dead ,were select-ed .The APACHE Ⅱscores of patients in first 24 h of admission were collected ,and receiver operating characteristic curves (ROC curve) were drawn .Results APACHE Ⅱ score of the 42 patients with Severe acute organophosphorus poisoning was 18 ~30 (20 .11 ± 6 .32) ,in which the survival group was(16 .10 ± 3 .12) ,the dead group was(28 .01 ± 4 .46) (P<0 .01) .With the increase of APACHE Ⅱ score ,the fatality rate gradually increased .The total area under the ROC curves of APACHE Ⅱ score for death judgment was 0 .922 ,APACHE Ⅱ score of 21 .2 was the best diagnostic point ,the sensitivity was 95% ,and specificity was 89% . Conclusion The APACHE Ⅱscore could predict severity of patients with Severe acute organophosphorus poisoning ,and APACHEⅡscore ≥21 .2 could be used as the prognosis for death of the patients .
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Objective To investigate the mechanism of anti-breast cancer cell proliferation induced by doxorubicin (DOX).Methods AMP-Activated Protein Kinase (AMPK) activator AICAR,AMPK siRNA,AMPK inhibitor compound C(AMPKi) and doxorubicin treated MCF-7 cells at different time points; AMPK,acetyl CoA carboxylase (ACC),p38 activation were detected by Western blot.MTT was used as cell viability assay. Results Doxorubicin-induced activation of AMPK, AMPK agonist (AICAR)or in combination with doxorubicin activated AMPK and increased MCF-7 cell proliferation rate [the difference of cell viability between group AICAR+DOX(17.7±1.6 ) ％ and group DOX(71.4±1.8 ) ％ was significant(P＜0.001)].After AMPKi or AMPK siRNA and doxorubicin combined administration, P-AMPK and P-ACC expression was significantly decreased,the level of p38 was not affected,and MCF-7 cell proliferation inhibition rate decreased [the cell viability of group AMPKi+DOX(72.7±1.8 ) ％ vs group DOX(96.3±1.7 ) ％,P＜0.001 ;group AMPK siRNA +DOX( 76.9±2.2 ) ％ vs group scramble siRNA+DOX(95.9±1.8) ％,P＜0.001].Conclusion AMPK is involved in doxorubicin-induced anti-breast cancer cell proliferation.
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Objective To investigate the effect of hexosamine biosynthesis pathway on the development of insulin resistance induced by high fat diet.Methods Normal male SD rats were randomly divided into three groups:control(fed with normal chow),high fat(fed with high fat diet for 13 weeks),and rosiglitazone (intragastric administration with rosiglitazone for 5 weeks)groups.After 13 weeks,all the rats were sacrificed,serum and muscle triglycerides(TG),serum total cholesterol(TC),and serum and muscle free fatty acids(FFA) were measured.Insulin sensitivity wss evaluated by insulin sensitivity index(ISI)and glucose infused rat(GIR) with the hyperinsulinemic englycemic clamp technique.The flux of HBP in skeletal muscle was detected with the expression level of glutamine-fructose-6-phosphate transaminase(GFAT)mRNA(RT-PCR),the content of UDPGlcNAc(HPLC)and the level of O-GlcNAc glycosylation in skeletal muscle proteins(Western blot). Results Compared with control group,senlm TG,TC,FFA and muscle TG,FFA levels of high fat group increased(aII P<0.01).both ISI and GIR decreased(both P<0.01),and the leveIs of GFAT mRNA(0.51±0.05 vs 0.18±0.02),UDP-GlcNAc[(6.18±0.86 vs 2.42±0.36)nmol/g],and O-GIcNAc glycosylation of skeletal muscle proteins in high fat group were raised(all P<0.01).In rosiglitazone group,serum and muscle TG.FFA welc deceased(all P<0.01).insulin sensitivity was increased(P<0.05)and the flux of HBP[GFAT mRNA 0.27±0.03,UDP-GIcNAc(2.62±0.32)nmol/g]was reduced(all P<0.05)as compared with high fat group. Conclusions High fat diet-induced insulin resistance in rats is correlated with the increased flux of HBP in skeletaI muscle.which is decreased by rosiglitazone.
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OBJECTIVE To investigate the changes in serum HBV-DNA level in HBsAg positive patients before and after operation and their infectious risk in hospital.METHODS HBV markers(HBV-M) in serum was detected in 58 HBsAg positive patients by time-resolved fluoroimmunometric assay before operation.HBV-DNA level in serum of them before operation and at 3rd,and 7th day after operation was detected by real time fluorescent quantitative polymerase chain reaction.We also detected HBV-DNA in gastric drainage juice and abdominal drainage after operation.RESULTS HBV-DNA was detected in 27 of 58 HBsAg positive patients' serum,the positive rate was 46.1%.After operation,serum HBV-DNA was increased remarkably at 3rd and 7th day compared with before operation in these patients respectively(P
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Objective To investigate the correlation between bacteria and upper urinary tract calculi (UUTC). Methods 530 cases of UUTC were enrolled in the study.Mid-stream urine culture was obtained before operation.Intraoperative pelvic urine culture,cotton swab culture of stone-incarcerated mucosal surface,pre- and post-sterilized stone culture were also carried out. Results Among 530 cases of UUTC,the stone culture showed positive (infectious stone) in 120 cases (22.6%),more prevalent in females than in males. Staphylococcus epidermidis and Escherichia coli were the predominant bacteria, accounting for 60.1% and 32.5%,respectively.The infection rate with which a stone size was ≥1.0 cm with coarse surface,loose density or radiolucent was much higher than that of size