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Objective To study the effect and mechanism of Momordica anti-HIV protein of 30 ku (MAP30) on the migration of bladder cancer.Methods The IC50 of human bladder cancer 5637 and T24 cells was calculated by methyl thiazolyl tetrazolium (MTT) method.The migration ability of these two cells was evaluated by scratch migration test and Transwell cell migration test.The expression of migrating proteins such as matrix metalloproteinases (MMPs) and adhesion molecule N-cadherin were compared by Western blot.Results Scratch migration test:there were significant differences in migration rates of 5637 cells at 8 h and 22 h (P ＜ 0.05).There were significant differences in migration rates of T24 cells at 22 h (P ＜ 0.05),but no significant differences in migration rates at 8 h (P ＞ 0.05).The expression of Vimentin,Fibronectin,MMP-2,MMP-9 and N-Cadherin in 5637 cells and T24 cells of human bladder cancer decreased significantly after adding MAP30.The E-Cadherin expression in human bladder cancer 5637 cells were decreased,but no target band was detected in human bladder cancer T24 cells.Conclusions The ribosome-inactivating protein MAP30 can effectively inhibit the migration of human bladder cancer 5637 and T24 cells by inhibiting the EMT pathway and inhibiting the expression of MMPs.
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Objective To study the effect of enhancer of rudimentary homolog (ERH) gene on migration and invasion in human bladder cancer T24 and 5637 cells.Methods After knocking out the ERH gene of human bladder cancer T24 and 5637 cells,Wound healing assay,Transwell cell migration assay and Transwell cell invasion assay were used to verify the migration and invasion function.Cell migration related protein was detected by Western blot.Nude mouse tail vein transfer assay was used to study the metastasis ability of bladder cancer cells in vivo.Results (1) The Wound healing assay showed that there were significant differences in the migration cell counts of human bladder cancer 5637 and T24 (P ＜ 0.05).(2) There were significant differences in migration and invasion cell counts of Transwell assay (P ＜0.05).(3) Western blot showed that the expression of E-Cadherin in human bladder cancer 5637 cells and T24 cells was significantly increased (P ＜ 0.05) after knocking out ERH gene,while the expression of Fibronectin,Twist,Vimentin and Snail2 protein were significantly decreased (P ＜ 0.05).(4) Nude mouse tail vein transfer assay showed that lung metastases were significantly reduced in the ERH knockout group compared with the normal ERH group.Conclusions Both in vitro and in vivo experiments suggest that ERH knockout affects the migration and invasion of human bladder cancer T24 and 5637 cells.
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Objective To investigate the effect of ERH gene knockdown on the proliferation and apoptosis of human bladder cancer T24 cells. Methods T24 cells infected by lentivirus with interference on ERH gene sequence were cloned to establish stable T24 cells clone in ERH gene suppression. The expression of ERH mRNA gene in bladder cancer was detected by using quantitative real time polymerase chain reaction (qPCR). The effects of ERH knockout on the cell proliferation and apoptosis were examined by using methylthiazolyl tetrazolium (MTT) assay, colony formation assay and flow cytometry. The effect of ERH knockout on the tumorigenic effect of T24 cells in vivo was verified by subcutaneous tumor formation in nude mice. Results After lentiviral transfection, qPCR results showed that the knockdown effect of ERH mRNA in ERH normal group (untreated T24 cells) was better than that in ERH gene knockdown group, and the difference was statistically significant [(1.006±0.126) vs. (0.079±0.007); t=12.72, P=0.0002]. After knocking out ERH gene, MTT assay showed that the proliferation ability of T24 cells in ERH gene knockdown group was weakened compared with ERH normal group, and the difference was statistically significant [A490 value: (0.13±0.00) vs. (0.66±0.01);t=104.61, P<0.0001]. Colony formation assay indicated that the ability of clone in ERH normal group was weakened compared with ERH gene knockdown group [(10.5 ±1.2) vs. (196.4 ±4.0); t= 73.63, P< 0.0001]. Flow cytometry showed that the cell apoptosis rate in ERH gene knockdown group was higher than that in ERH normal group [(11.0 ±0.5) ％ vs. (4.2 ±0.5) ％; t= 16.06, P<0.0001]. Imaging results of subcutaneous tumor formation in nude mice showed that the total fluorescence intensity of the tumor area in ERH gene knockdown group was (4.67 ±0.59) × 1010 μW/cm2, and the corresponding part in ERH normal group was (9.54±4.20) × 1010μW/cm2 (t=3.64, P=0.0051);tumor weight in ERH gene knockdown group was (0.80±0.62) g, and in ERH normal group was (1.79±0.71) g (t=3.33, P=0.0037). Conclusion ERH gene knockout can inhibit the proliferation of human bladder cancer T24 cells, and promote the cell apoptosis.
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Objective To investigate the effect of ulinastatin combined pantoprazole on inflammatory factors and gastrointestinal tract in patients undergoing cardiopulmonary bypass ( CBP) cardiac surgery. Methods A total of 200 patients who suffered rheumatic heart disease were scheduled for valve replacement surgery with CPB, were randomly divided into four groups:control group (CON),ulinastatin (UTI),pantoprazole groups (PTZ) and ulinastatin+pantoprazole groups(UTI+PTZ),50 cases in each group.Before CBP,group UTI was given ulinastatin 10 000 U·kg-1,group PTZ was given pantoprazole 40 mg,group UTI+PTZ was given ulinastatin 10 000 U·kg-1and pantoprazole 40 mg,group CON was given 0.9% sodium chloride soution.The gastric mucosa pHi and blood samples would be collected in all four groups at the preoperative (t1),CPB 30 min (t2),after CBP (t3),6 h after surgery (t4),24 h (t5) five time points.The IL-6 and TNF-α would be detected by enzyme linked immunosorbent (ELISA) method,and abdominal distension,abdominal pain,hematemesis,black and defecate occult blood test positive for digestive tract related complications would be collected after the surgery 1,2 days. Results The concentration of TNF-α and IL-6 at t2,t3, t4,t5were higher than those at t1in all four groups(P<0.05).Compared with CON group,the concentration of TNF-α and IL-6 at t2, t3,t4,t5in UTI,PTZ and UTI+PTZ group were significantly decreased (P<0.05).The concentration of TNF-α and IL-6 in UTI and UTI+PTZ group were better than in PTZ group.The pHi at t2,t3,t4was lower than that at t1in four groups(P<0.05),and pHi at t5 was obviously lower than that at t1in group CON (P<0.05).The pHi at t2,t3,t4in UTI,PTZ and UTI+PTZ group was higher than that in CON group ( P<0. 05), and pHi in UTI+PTZ group was better than that in UTI and PTZ group. The postoperative gastrointestinal complications in CON group were higher than those in UTI,PTZ and UTI+PTZ group (P<0.05). Conclusion Ulinastatin combined with pantoprazole for patients undergoing CPB heart surgery,can significantly reduce the release of TNF-α and IL-6、increase gastric pHi and reduce the incidence of gastrointestinal complications.
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Objective To compare the treatment effects of different dosage of tamsulosin for patients with type Ⅲ prostatitis.Methods 150 patients with type Ⅲ prostatitis were selected as study objects.They were divided into control group and research group according to the digital table,each group in 75cases.The research group was treated with tamsulosin 2 tablets (0.4mg),1 time a day.The control group was treated with tamsulosin 1 tablet (0.2mg) + starch tablet 1 tablet,1 time a day.After treatment for 12 weeks,the clinical effects,NIH-CPSI score,white blood cell count and pH value,and adverse reaction between the two groups were compared.Results The total effective rate of the research group was higher than that of the control group(93.33％ vs.81.33％),the difference was statistically significant (x2 =4.881,P ＜ 0.05).After treatment,the pain,voiding symptoms,quality of life and NIH-CPSI scores in the research group were (5.13 ± 4.02) points,(2.52 ± 1.07) points,(3.64 ± 3.25) points,(9.19 ± 0.63) points,respectively,which were lower than those in the control group [(8.25 ± 3.54) points,(3.28 ± 1.87) points,(5.57 ± 3.62) points,(16.47 ± 0.38) points],the differences were statistically significant (t =5.044,3.055,3.436,85.693,all P ＜0.05).The white blood cell and pH value of the research group were (12.65 ± 5.88) x 109/L,(6.29 ±0.20),respectively,which were lower than those in the control group [(16.58 ±6.24) x 109/L,(6.73 ± 0.16)],the differences were statistically significant (t =3.970,14.878,all P ＜ 0.05).The single occurrence rate and total occurrence rate between the two groups had no statistically significant differences (x2 =0.340,0.207,0.000,0.000,0.362,all P ＞ 0.05).Conclusion The effect of tamsulosin in dose of 0.4 mg is better than 0.2 mg,and had no significant impact on safety.
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Objective To evaluate the efficacy and safety of PECS block under ultrasound guidance in multimodal analgesia after modified radical mastectomy.Methods Sixty female patients aged 18-65 years, ASA grade Ⅰ or Ⅱ, undergoing elective unilateral modified radical mastectomy were enrolled.Patients were randomly divided into PECS group (group P, n=30) or control group (group C, n=30).Two groups of patients were given flurbiprofen axetil 1 mg/kg via intravenous injection before operation.After general anesthesia induction, patients in group P received ultrasound guided pectoral nerves block with 30 ml of 0.375% ropivacaine.Patients in group C didn`t receive nerve block.Anesthesia maintenance was performed by combined intravenous-inhalation Anesthesia.Postoperative VAS pain scores (at 0, 3, 6, 12, and 24 postoperative hours), does of intraoperative remifentanil, rescue analgesic requirements in the first 24 h after surgery, adverse reactions were recorded.Results VAS score in group P was lower than that in group C at 0, 3, 6 and 12 h after surgery (P<0.05), there was no difference at 24 h.The dose of remifentanil and the rescue analgesic requirements in group P were lower than those in group C (P<0.05).There was no significant difference in postoperative adverse reactions between the two groups.Conclusion As a supplementary mode of multimodal analgesia, PECS block is a safe and reliable technique that provide better analgesia effect for modified radical mastectomy.
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Objective To evaluate the effect of recombinant human erythropoietin (rHuEPO) on lung injury induced by hepatic ischemia-reperfusion (I/R) in rats.Methods Sixty healthy male SpragueDawley rats, aged 6-8 weeks, weighing 220-280 g, were randomly divided into 3 groups (n=20 each) using a random number table: sham operation group (group S) , hepatic I/R group (group I/R), and rHuEPO group (group E).I/R and E groups underwent I/R of 70 percent of the liver.The rHuEPO 4 000 U/kg was injected intraperitoneally at 24 h before I/R in group E, while the equal volume of normal saline was given in S and I/R groups.The rats were sacrificed at 3 h of reperfusion, and lungs were removed and cut into sections which were stained with haematoxylin and eosin and examined under light microscope.Wet to dry lung weight ratio (W/D ratio) was calculated.The expression of heme oxygenase-1 (HO-1) and inducible nitric oxide synthase (iNOS) in lung tissues was determined by immunohistochemistry.The content of malondialdehyde (MDA), and activities of superoxide dismutase (SOD) and myeloperoxidase (MPO) in lung tissues were detected.Results Compared with group S, the W/D ratio, MDA content, and MPO activity were significantly increased, the SOD activity was decreased, the expression of HO-1 and iNOS was up-regulated (P＜0.05) , and the pathological changes of lung tissues were obvious in E and I/R groups.Compared with group I/R, the W/D ratio, MDA content, and MPO activity were significantly decreased, the SOD activity was increased, the expression of HO-1 was up-regulated, the expression of iNOS was down-regulated (P＜0.05) , and the pathological changes of lung tissues were reduced in group E.Conclusion The rHuEPO can alleviate hepatic I/R-induced lung injury in rats, and the mechanism may be related to up-regulated expression of HO-1 and down-regulated expression of iNOS.
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Objective To investigate the dilating effect of dexmedetomidine (DEX)on isolated vascular smooth muscles and to explore the mechanism.Methods Tension of isolated mesenteric arterial rings of male Sprague-Dawley rats was recorded.The effects of DEX on the rings and the effects of DEX on vascular reaction induced by various drugs were recorded. Results DEX completely relaxed the contraction induced by phenylephrine (PE)and KCl in a concentration-dependent manner in endothelium intact mesenteric arterial rings in rats.The vasodilating effect of DEX was increased by sodium nitroprusside.In phenylephrine (10-5 mol/L)based on pre-vasoconstriction,adding acetylcholine could not suppress DEX’s vasodilating effect.Vasodilation was not related to the endothelial cells.In physiological saline solution without calcium,DEX significantly inhibited the contraction induced by addition of CaCl2 .Conclusion DEX can induce vasodilation in a concentration-dependent manner,which is not dependent on the endothelial cells.
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Objective To evaluate the effect of propofol on liver injury in mice with acute liver failure (ALF).Methods Eighty adult male ICR mice,aged 1 months,weighing 20-25 g,were randomly divided into 3 groups (n =20 each) using a random number table:control group (group Ⅰ),ALF group (group Ⅱ),and ALF + propofol group (group Ⅲ).ALF model was established with intra-peritoneal D-galactosamine (D-GaIN) and lipopolysaccharide (LPS).Propofol 5 mg/kg was injected via the tail vein every 1 h within 6 h after injection of DGaIN and LPS in group Ⅲ,while the equal volume of normal saline was given instead in the other groups.Venous blood samples were taken from the tail vein at 1,3 and 6 h after injection of D-GaIN and LPS (T1-3) to detect the activities of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and serum tumor necrosis factor-alpha (TNF-α),interleukin-1β (IL-1β) and IL-10 concentrations (by ILISA).The survival within 12 h after injection of D-GaIN and LPS was observed and the survival rates were calculated.The mice were sacrificed and livers were removed for microscopic examination of pathologic changes.Results Compared with group Ⅰ,the activities of AST and ALT were significantly increased at each time point in Ⅱ and Ⅲ groups and the serum TNF-α concentrations at T1,2 and IL-1β and IL-10 concentrations at each time point were significantly increased in group Ⅱ,and the serum TNF-α concentrations at T1,and IL-1β and IL-10 concentrations at T2,3 were significantly increased in group Ⅲ (P ＜ 0.05).Compared with group Ⅱ,the activities of AST and ALT at each time point,serum TNF-α concentrations at T1,2 and IL-1β and IL-10 concentrations at T2,3 were significantly decreased and the survival rate within 12 h after injection of D-GaIN and LPS was increased in group Ⅲll (P ＜ 0.05).The pathologic changes of liver tissues were gradually attenuated in Ⅱ and Ⅲ groups.Conclusion Propofol can reduce the liver injury in mice with ALF through inhibiting inflammatory responses.
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Objective To investigate the effects of ulinastatin pretreatment on oxidative stress response and hepatocyte growth factor (HGF) after hepatectomy in rats.Methods One hundred and twelve pathogen-free male Sprague-Dawley rats,aged 3 months,weighing 230-280 g,were randomly divided into 2 groups (n=56 each): group hepatectomy (group H) and group ulinastatin pretreatment (group U).Left and median lobe resection was performed and then liver ischemia was induced by blood flow occlusion of right and caudate lobes for 30 min,followed by perfusion in both groups.Ulinastatin 50 000 U/kg was injected intravenously at 5 min before occlusion in group U.Eight rats in each group were chosen and the blood samples were taken from the inferior vena cava for measurement of serum ALT and AST activities and HGF concentration before ischemia and at 1,6,12,24and 48 h of reperfusion.Then the right lobe were removed for determination of apoptosis,SOD and myeloperoxidase (MPO) activities,MDA content,expression of proliferating cell nuclear antigen (PCNA) and liver regeneration.Apoptotic index was calculated.Another 8 rats in each group were chosen and the 7 day survival rate was recorded.Results Compared with group H,the levels of ALT,AST,MPO aud MDA and apoptotic index were significantly decreased,and the levels of HGF and SOD,PCNA expression and liver regeneration were significantly increased at different time points in group U (P ＜ 0.05).There was no significant difference in 7 day survival rate between group H and group U (P＞ 0.05).Conclusion Ulinastatin pretreatment can strengthen liver regeneration after hepatectomy in rats,the underlying mechanism may be related to inhibition of oxidative stress response and increase in HGF production.
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<p><b>OBJECTIVE</b>To investigate the effect of pretreatment with ulinastatin on liver regeneration and TNF-α/IL-6/STAT-3 signal pathway in rats after 70% hepatectomy combined with ischemia-reperfusion injury.</p><p><b>METHODS</b>A total of 120 normal male SD rats weighing 230-280 g were randomized into 3 groups (n=40), namely simple partial hepatectomy (PH) group, partial hepatectomy with ischemia-reperfusion (PHIR) group, and ulinastatin group. All the rats received resection of the left and middle liver lobes. In PHIR group, the remnant right lobes were subjected to blood flow occlusion for 30 min; in UTI group, the rats were given 50 000 U/kg UTI intravenously prior to the occlusion, and in PH group, the blood flow was not occluded. At 1, 6, 12, 24, and 48 after the reperfusion, the remnant liver tissues were examined for regenerated liver weight, PCNA staining, TNF-α and IL-6, STAT-3, cyclin D1, and Cdk4 expressions.</p><p><b>RESULTS</b>The regenerated liver weight and PCNA positivity rates were significantly higher in ulinastatin group than in PHIR group at 24 h and 48 h after the reperfusion (P<0.05). In ulinastatin group, the levels of TNF-α and IL-6 were significantly lower, and IL-6 level and the expressions of STAT-3, cyclin D1, and Cdk4 mRNA and cyclin D1 and Cdk4 proteins were significantly higher in ulinastatin group than in PHIR group at 24 h and 48 h (P<0.05).</p><p><b>CONCLUSION</b>Ulinastatin can promote liver regeneration after major hepatectomy and ischemia-reperfusion injury, and the effect is possibly related with activation of IL-6/STAT-3 signal pathway, which promotes the synthesis of cyclin Dl-Cdk4 complex and hepatocyte proliferation.</p>