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OBJECTIVE：To study t he effects of bro mophenylcurcumin（GL63）on the apoptosis ，migration and invasion of human cholangiocarcinoma RBE cells ，and to investigate its mechanism based on JAK/STAT signaling pathway. METHODS ：MTT assay was used to detect the effects of different concentrations of GL 63 [0（blank control ，similarly hereinafter ），1.25，2.5，5，10， 20，40 μ mol/L] on the proliferation of RBE cells after 48 h treatment ；the IC 50 was calculated. The effects of different concentrations of GL 63（0，5，10，20 μmol/L）on colony formation were detected by crystal violet staining after 48 h treatment. Flow cytometry ，Hoechst 33342 staining，cell scratch test and Transwell chamber invasion test were used to detect the effects of different concentrations of GL 63（0，5，10，20 μmol/L）on cell cycle distribution ，apoptosis，migration and invasion ability after 24 h treatment. Western blotting assay was adopted to detect the effects of different concentration of GL 63（0，5，10，20 μmol/L） on the expression of JAK 2/STAT3 signal pathway associated proteins. RESULTS ：The proliferation inhibition rates of RBE cells in different concentrations of GL 63 groups（1.25-40 μmol/L）were significantly increase d，compared with blank control group （P＜ 0.01），and showed a dose-dependent trend ，with IC 50 of （8.46±1.30）μmol/L. Compared with blank control group， 85917439。E-mail：zhaoji-an-88@163.com inhibition rates of RBE cell colony formation were significantly decreased in different concentrations （5，10，20 μmol/L）of GL 63 groups（P＜0.01）. The percentage of RBE cells at G 0/G1 phase increased significantly ，while that at S phase decreased significantly （P＜0.01）. The apoptotic rate increased significantly（P＜0.01），and the nucleus showed dense pyknosis and apoptotic bodies. The rate of cell migration and healing was significantly decreased （P＜0.01），and the number of invasive cells through basement membrane was significantly decreased （P＜ 0.01）. The protein expression of p-JAK 2， p-STAT3， Bcl-2， MMP-2， MMP-9， Pro-caspase-9 and P ro-caspase-3 were down-regulated significantly while the expression of Bax ，Cyt-c，Cleaved-caspase-9 and Cleaved-caspase- 3 were up-regulated significantly（P＜0.01）. CONCLUSIONS ：GL63 may inhibit the proliferation ，migration and invasion of RBE cells and promote its apoptosis by inhibiting JAK 2/STAT3 signal pathway.
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OBJECTIVE：To compare the an ti-hepatocarcinoma effects of curcumin （CUR）and its derivative hydrazincurcumin （HZC），and to explore the mechanism. METHODS ：MTT assay was used to detect the effects of CUR or HZC （2.5，5，10，20， 40，80 μmol/L）on the proliferation of HepG 2 cells. Flow cytometry was used to detect the effects of CUR or HZC （10，20，40 μmol/L）on cell cycle distribution and apoptosis of HepG 2 cells. Western blotting assay was used to detect the effects of CUR or HZC（10，20，40 μmol/L）on the expression of apoptosis-related protein in HepG 2 cells. The male SD rats were randomly divided into normal control group （n＝10），CUR control group （n＝10），HZC control group （n＝10），model group （n＝30），CUR protection group （n＝30）and HZC protection group （n＝30）. CUR control group and HZC control group were given CUR 85917439。E-mail：zhaoji-an-88@163.com or HZC （80 mg/kg） intraperitoneally. Model group ，CUR protection group and HZC protection group were given diethylnitrosamine （50 mg/kg）intraperitoneally to establish hepatocarcinoma model ；at the same time ，2 protection groups were given CUR or HZC （80 mg/kg）intraperitoneally，twice a day，for consecutive 12 weeks. During medication ，the change of body weight and death of rats were recorded. Twenty four weeks later，liver index of rats was calculated and appearance was observed ；the number of cancer nodules was counted ；HE staining was used to observe the pathological changes of liver tissue and calculate the nuclear division index of hepatocarcinoma ；the proliferating cell nuclear antigen （PCNA）index was detected by immunohistochemistry. RESULTS ：CUR and HZC could increase the inhibitory rate of HepG 2 cells（P＜0.05），and increased the percentage at G 0/G1 phase and apoptotic rate of HepG 2 cells（P＜ 0.05）. CUR and HZC could significantly decrease the protein expression of p-JAK 2，p-STAT3，Bcl-2 and Bcl-xl ，while increased the protein expression of Bax ，Cyt-c，Caspase-9，Caspase-3 and PAPR （P＜0.05）. Above effects of HZC were significantly better than those of CUR （P＜0.05）. The results of animal experiment showed that there was no death ，no liver canceration and no pathological changes in liver appearance and tissue section of the three control groups ；there was no statistical significance in body weight and its increased weight ，liver index ，nuclear division index of carcinoma or PCNA index （P＞0.05）. Compared with model group， survival rate of rats were increased significantly in CUR protection group and HZC protection group ， while hepatocarcinoma incidence and the number of cancer nodules were decreased significantly （P＜0.05）；body weight and its increased weight were increased significantly ，while liver index ，nuclear division index of carcinoma and PCNA index were decreased significantly （P＜0.05）. There were some pathological changes in liver appearance and tissue section ；cancerous lesions with focal necrosis or cancerous lesions with patchy necrosis were observed. There was no statistical significance in the improvement of above indexes in 2 protection groups （P＞0.05）. CONCLUSIONS ：HZC could inhibit the proliferation and induce apoptosis of HepG 2 cells by inhibiting JAK 2/STAT3 signaling pathway and regulating the activation of mitochondrial endogenous pathway，which shows stronger anti-hepatocarcinoma effect in vitro than CUR. On the other hand ，there was no significant difference in the improvement of liver caner indexes in hepatic cancer model rats between HZC and CUR.
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Objective:To study the anti-inflammatory effect of the ethanol extract of Coptis chinensis Franch.in vitro.Methods: An inflammatory cell model was established by stimulating the mouse peritoneal macrophages in vitro with lipopolysaccharide(LPS).LPS stimulated of RAW264.7 cells for a long time after administration of intervention.Effect of ethanol extract of Coptis chinensis Franch.on RAW264.7 cell growth activity was analyzed by MTT assay.The production of tumor necrosis factor-α(TNF-α),IL-1β,IL-6,NO,prostaglandin E2(PGE2) was determined by ELISA.The expression of mRNA of TNF-α,induced nitric oxide synthase(iNos) and HO-1 was detected by real-time fluorescent quantitative PCR(RT-PCR).Results: The ethanol extract of Coptis chinensis had no inhibition effect on the scope of RAW 264.7 cells the scope of 5-80 mg/L.Each treatment group concentration of interleukin-6 (IL-6),interleukin-1β (IL-1β),TNF-α,NO,prostaglandin E2 (PGE2) content of LPS stimulation model group were significantly (P<0.01),and content was not related to concentration.Real-time quantitative (RT-PCR) showed,the concentration of each treatment group were significantly lower iNos,HO-1 and TNF-α mRNA expression (P<0.05,P<0.05,P<0.01),and content was not related to concentration.Conclusion: Coptis chinensis Franch.ethanol extract has anti-inflammatory effects in vitro,the mechanism may be related to inhibition of TNF-α,NO and other inflammatory factors and the impact of the activation of arachidonic acid (AA) metabolism.
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Objective:To investigate the anti-fatigue phenomenon induced by forced swimming in the mice,and to explore the anti-fatigue effect of argininyl fructoyl glucose (AFG) from red ginseng in the mice and its mechanism.Methods:The AFG was extracted from red ginseng.The ICR mice were divided into blank control group,low dose of AFG group (100 mg · kg-1),middle dose of AFG group (200 mg · kg-1) and high dose of AFG group (400 mg · kg-1) (n=20).The mice mere given a forced swimming test after continuous gavage for 28 d.The weights,organ indexes,time of forced swimming,contents of lactic acid (LD),blood urea nitrogen (BUN),hepatic glycogen (Gly) and expressing levels of PGC-1α in gastrocnemius of the mice in various groups were detected.Results:Compared with blank control group,the weights and organ indexes of the mice in low,middle and high doses of AFG groups had no significant differences (P＞ 0.05).Compared with blank control group,the time of forced swimming,contents of Gly and expressing levels of PGC-1α of the mice in low,middle and high doses of AFG groups were significantly increased (P＜0.05 or P＜0.01) in a dose-dependent manner.Compared with blank control group,The contents of LD and BUN in serum of the mice in low,middle and high doses of AFG groups were significantly decreased (P＜0.01).Conclusion:AFG has anti-fatigue effect in mice,and its mechanism may be related to energy metabolism.
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Objective:To investigate the anti-fatigue phenomenon induced by forced swimming in the mice,and to explore the anti-fatigue effect of argininyl fructoyl glucose (AFG) from red ginseng in the mice and its mechanism.Methods:The AFG was extracted from red ginseng.The ICR mice were divided into blank control group,low dose of AFG group (100 mg · kg-1),middle dose of AFG group (200 mg · kg-1) and high dose of AFG group (400 mg · kg-1) (n=20).The mice mere given a forced swimming test after continuous gavage for 28 d.The weights,organ indexes,time of forced swimming,contents of lactic acid (LD),blood urea nitrogen (BUN),hepatic glycogen (Gly) and expressing levels of PGC-1α in gastrocnemius of the mice in various groups were detected.Results:Compared with blank control group,the weights and organ indexes of the mice in low,middle and high doses of AFG groups had no significant differences (P＞ 0.05).Compared with blank control group,the time of forced swimming,contents of Gly and expressing levels of PGC-1α of the mice in low,middle and high doses of AFG groups were significantly increased (P＜0.05 or P＜0.01) in a dose-dependent manner.Compared with blank control group,The contents of LD and BUN in serum of the mice in low,middle and high doses of AFG groups were significantly decreased (P＜0.01).Conclusion:AFG has anti-fatigue effect in mice,and its mechanism may be related to energy metabolism.
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Objective To investigate the improvement of postpartum pelvic floor by rehabilitation training assessed with three-dimensional transperineal ultrasound . Methods One hundred cases of healthy postpartum women were randomly divided into two groups :control group and training group .The control group received the customary education ,and the training group received pelvic floor rehabilitation training . At 6 and 12 weeks postpartum ,levator hiatus area ,thickness of the levator ani muscle ,bladder neck mobility ,and bladder posterior horn were measured with three-dimensional transperineal ultrasound in all the subjects . Meanwhile ,the muscle strength situations were tested . Results At 12 weeks postpartum ,the anal levator hiatal area ,bladder neck mobility and bladder posterior horn in the training group were lower than those of the control group[ ( 21 .6 ± 3 .2) cm 2 vs ( 25 .6 ± 2 .4 ) cm 2 ,( 27 .9 ± 5 .3) mm vs ( 31 .5 ± 5 .9) mm ,( 126 .3 ± 21 .2)° vs (135 .3 ± 11 .6)°] ( P < 0 .05) . Compared with control group ,the thickness of the levator ani muscle increased in training group [ ( 13 .6 ± 2 .3) mm vs ( 15 .3 ± 2 .5) mm ] ( P < 0 .05) . The incidence of stress urinary incontinence in the training group ( 5% ) was significantly lower than the control group ( 12 .5% ) at 12 weeks postpartum ( χ2 = 5 .487 , P = 0 .025) . The muscle strength had no significant difference at 6 weeks postpartum . At 12 weeks postpartum ,the pass rate of class Ⅰ muscle fiber was 78 .5% ,and that of class Ⅱ muscle fiber was 83 .3% in the training group ;the pass rate of class Ⅰ muscle fiber was 28 .5% ,and class Ⅱ muscle fiber was 37 .3% in the control group , the improvement was significant at 12 weeks postpartum . Conclusions The result of the transperineal real-time ultrasonographic evaluation of post-natal pelvic floor rehabilitation training has high consistency with the measurement of muscle strength . The ultrasound examination is simple and accurate ,and has highly applicable value in evaluating the effect of post-pelvic rehabilitation training .
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Objective To investigate the roles of a epoprostenol(PGI2) analog (Iloprost) in regulating the differentiation of CD4+ T cells to Treg cells and the possible mechanism.Methods Naǐve CD4+ T cells were isolated from human peripheral blood samples by using the magnetic-activated cell sorting (MACS) and then cultured under Treg-polarizing condition.The percentages of Treg cells and the expression of Foxp3 at mRNA level were respectively measured by using flow cytometry and RT-PCR for evaluation the effects of Iloprost on the differentiation of CD4+ T cells to Treg cells.The cAMP accumulation assay was used to detect the level of intracellular cAMP.Flow cytometry analysis was performed to detect the phosphorylation of signal transducer and activator of transcription 5 (STAT5).Results lloprost decreased the percentage of Treg cells and inhibited the expression of Foxp3 at mRNA level in a dose dependent manner (P＜0.05).However,the inhibitory effects of Iloprost were weakened when IP receptors were blocked by IP antagonist (CAY10449).A six-fold increase in the levels of intracellular cAMP in Treg cells was induced by Iloprost (P＜0.05) and a similar effect could be achieved by using a cAMP agonist,db-cAMP (P＞0.05).H-89,a protein kinase A inhibitor,inhibited the Iloprost-induced expression of cAMP in Treg cells.Moreover,Iloprost inhibited the IL-2 mediated phosphorylation of STAT5 (P＜0.05) and a similar effect could be achieved by using db-cAMP (P＞0.05).The Iloprost-mediated down-regulation of pSTAT5 was blocked by using H-89.Conclusion PGI2 could activate the cAMP-PKA signaling pathway by binding to the IP receptor,resulting in inhibited phosphorylation of STAT5 and suppressed differentiation of naǐve CD4+ T cells to Treg cells.
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Objective To summary the experience of laparoscopic cystectomy ileal conduit (Bricker) and orthotopic ileal neo‐bladder (Hautmann) and compare the short‐term efficacy of the two types of urinary diversion for invasive bladder cancer . Methods Retorspective analysis of the patients in our hospital who accepted laparoscopic radical cystectomy from 2010 to 2014 was performed ,74 of them accepted ileal conduit ,and 30 of them accepted orthotopic ileal neobladder .The general clinical data ,postop‐erative recovery ,postoperative complications and Oncology feature were analyzed and compared between the two groups .Results There was no demonstrable difference was found in operation time ,blood loss ,intraoperative blood transfusion rate ,the number of removed lymph node ,average hospital stay ,specimens positive margin rate and postoperative pathology results between the two groups (P>0 .05) .But there were significant difference in postoperative intestinal function recovery time[(4 .2 ± 1 .4)d ,(5 .3 ± 2 .2)d] ,(P=0 .002) ,and the complication rates 31 .9% (23 cases)vs .53 .3% (16 case) ,P=0 .043 .After 6 months ,the daytime and nighttime urinary control were 76 .9% ,57 .7% ,after 12 months ,the daytime and nighttime urinary control increased to 90 .9% , 81 .8% .2 cases(7 .7% ) were diagnosed with recurrence or metastasis during follow‐up in Hautmann group ,while 9 cases(14 .1% ) were diagnosed with recurrence or metastasis in Bricker group .Conclusion Two kinds of surgical procedures both have the similar therapeutic effect ,but the postoperative quality of life is better for Hautmann orthotopic neobladder patients .
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Objective To evaluate the effects between video endoscopic inguinal lymphadenectomy (VEIL ) and open inguinal lymphadenectomy(OIL) to provide the evidence-based basis for the selection of the clinical therapy schemes .Methods The related clinical controlled trial literature on the effective comparison of VEIL and OIL were retrieved from the databases of PubMed ,Co-chrane library ,Elsevier ,CNKI and Wanfang database .The screening was independently performed by 2 reviewers according to the including and excluding criteria .The related data were extracted and performed the meta analysis by the RevMan 5 .2 software .Re-sults A total of 4 trials were included .There were 146 cases of inguinal lymphadenectomies ,in which 61 cases were VEIL and 85 cases were OIL .The meta-analysis results showed that there were no statistical differences between the two operation modes in terms of the operative time(WMD=32 .33 ,95% CI -25 .70-90 .36 ,P=0 .27) ,intraoperative blood loss(WMD=9 .10 ,95% CI -76 .03-94 .23 ,P=0 .83) ,number of removed lymph nodes(WMD=0 .77 ,95% CI -1 .66-3 .20 ,P=0 .53) ,number of positive re-moved lymph nodes(WMD=0 .08 ,95% CI -0 .23-0 .40 ,P=0 .61) ,postoperative drainage time(WMD= -1 .30 ,95% CI -6 .40 -3 .80 ,P=0 .62) ,postoperative hospital stay (WMD= -4 .02 ,95% CI -10 .19-2 .15 ,P=0 .20) ,but the difference between VEIL and OIL in term of surgical complications had statistical significance (OR=0 .08 ,95% CI 0 .03-0 .26 ,P<0 .01) .Conclusion VEIL has equivalent efficacy to OIL ,but has less surgical complications .