ObjectiveTo study the effects and mechanism of survivin gene on invasion of prostate cancer cell.MethodsAfter PC-3 prostate cancer cell lines were transfected by survivin small interfering RNA (siRNA), the mRNA and protein of survivin and matrix metalloproteinase-2 and-9 were determined by real time RT-PCR and western blot assay, respectively. The anchorage-independent growth was examined by clon formation in soft agar, and invasion ability was evaluated by boyden chamber model.The invasion ability of cancer cells in vivo was determined by nude mice model. ResultsThe results from clon formation in soft agar showed that the colonies numbers of group of 3.125,6.250 and 12.500 nmol/L of siRNA were 17.8±1.6,13.6±1.5 and 8.8±1.4, and the control group was 22.6±1.8(P<0.05). The results from boyden chamber model exhibited that the cells numbers crossing filter membrane in group of 3.125,6.25 and 12.5 nmol/L of siRNA were 33.6±2.1,19.5±1.9,8.1±1.83, and the control group was 49.4±2.3(all P<0.05). The results in vivo showed that cancer cells of control groups invaded into striped muscle and blood vessel, and there were no these phenomenons in transferred group with survivin siRNA. Survivin siRNA could reduce expression level of MMP-2 and MMP-9 in prostate cancer cells(P<0. 01).Conclusions Survivin-directed RNA interference can inhibit invasion of human prostate cancer cell through down-regualting MMP-2,-9 genes.

Objective To study the influence of combined spinal-epidural anesthesia on perioperative coagulation function,RAAS activity and postoperative analgesia effect in parturients with cesarean section.Methods One hundred and eighteen parturients of cesarean section in our hospital from June 2013 to January 2016 were collected and divided into the observation group and control group according to the random number table method,59 cases in each group.The observation group received the combined spinal-epidural anesthesia and the control group received epidural anesthesia.The coagulation function indicators on preoperative 1 d (T0),at 10 min before operation end(T1) and postoperative 6 h (T2) were detected by adopting the automatic blood coagulation analyzer,the renin angiotensin aldosterone system (RAAS) function indices were detectd by radioimmunoassay.The pain indicators at postoperative 6 h(T2),12 h (T3) were detected by the pain threshold test instrument.Results The levels of prothrombin time(PT),activated partial thromboplastin time (APTT) and thrombin time (TT) at T1 and T2 in the observation group were significantly higher than those in the control group,while the PTA level was lower than that in the control group(P<0.05);serum RAAS indices such as (renin),angiotensin Ⅱ(ANG II) aldosterone(ALD) in the observation group were lower than those in the control group(P<0.05).The VAS score at T2,T3 in the observation group was lower than that in the control group,while the pain threshold and pain tolerance threshold levels in the observation group were higher than those in the control group(P<0.05).Conclusion Combined spinal-epidural anesthesia can reduce the blood coagulation and RAAS activation caused by cesarean section trauma,and the effect of postoperative analgesia is more significant.

AIM: To study the effects of Cripto gene on vascular endothelial growth factor(VEGF) of colon carcinoma cells.METHODS: Cripto siRNA was designed and constructed.Colon cancer LS-174T cells were divided into 4 groups: control group and different dose (3.125,6.25 and 12.5 nmol/L) of siRNA groups.After transfected for 24,48 and 72 h,colon cancer cells were harvested to carry on the next tests.Expression of Cripto mRNA was determined with real-time PCR,and immunofluorescence isothiocyanate(FITC) labeling assay and Northern blotting were performed to examine the expression of protein and mRNA of VEGF,respectively.The cells in control group and cells transfected with 12.5 nmol/L siRNA were inoculated into nude mice respectively.30 days after inoculated,the mice of two groups were executed,and immunohistochemical(ICH) assay was used to evaluate the VEGF protein of mice tumor.RESULTS: siRNA down-regulated the Cripto mRNA in a dose and time dependent manner.Protein and mRNA of VEGF in transfected cells reduced in a dose and time dependent manner.Compared to control,the expression of VEGF protein from ICH assay was lowered significantly(P

Objective To determine the EC50 of dexmedetomidine hydrochloride ( DEX) which causes disappearance of explicit memory by process dissociation procedure (PDP). Methods Forty patients those who had senior middle school or higher educational background undergoing lower extremity surgery with grade ASA Ⅰ or Ⅱ, without hearing impairment, dysphasia,nervous system disorders,and having no drugs in the treatment of the central nervous system were included.PDP was applied to establish study table and record, and calculate performance of explicit memory and implicit memory. Memory performance was statistically compared with 0, 0 memory was considered to be statistically significant and disappearance, respectively.Sequential method was used for determination.According to explicit memory disappearance or not,target concentration of the next patient was adjusted (increase or decrease).DEX target concentration of the first patient was set to 4 ng?mL-1,and the ratio of target concentration between the adjacent patients was 1.2.If the explicit memory of the former patient disappeared,the target concentration of the next patient was decreased by 1 concentration gradient;if the explicit memory of the former patient did not disappear,the target concentration of the next patient was increased by 1 concentration gradient, and so forth. All the 40 patients were determined.The median effective dose (D1) and 95% confidence interval (CI) of DEX were calculated. Results The ED50 of DEX causing explicit memory disappearance was 5.23 ng?mL-1,and the 95% CI was 4.07-6.39 ng?mL-1. Conclusion In clinical,target concentration of dexmedetomidine hydrochloride 5.23 ng?mL-1 levels for sedation,can cause half of patients’ explicit memory disappear,so as to avoid intraoperative awareness.

Objective To evaluate the effect of propofol with different sedation depth on CERO 2 of elder patientsduring anesthesia. Method 60 case patients under cholecystotomywith laparoscope, ASA IorⅡgrading, 65-75 year old , 45-75 kg , were randomly divided into 3 groups ( n = 20 ) and were divided into group A (3 μg/mL), group B (4 μg/mL), group, C (5 μg/mL), according to TCI of propofol. TCI in different groups were modified after gereral anesthesia. Bloodgas was analyzed by blood samples taken from radial artery and Sjv ball, Da-jvO2 and CERO2 were calculated. Results The Da-jvO2 and CERO2 of group B and C were significantly lower thanthose of group A at T2,3, while CjvO2 were significantly higher thangroup A (P < 0.05). The Da-jvO2 and CERO2 of group B and C werenot significantly different (P > 0.05); NI value of group A in T1-3 was significantly higher thangroup B (P < 0.05), while. NI value of group B in T1-3 was significantly higher thangroup C (P < 0.05). Conclusion Propofol TCI 4 μg/mL, can improve cerebral oxygen metabolism of elder patients and decrease CERO2.

Objective To investigate feedback regulation of close-loop muscle relaxant injection system on accuracy of cisatracurium besilate usage. Methods Two hundred patients undergoing laparoscopic cholecystectomy surgery, aged 20 to 40 years old, at ASA Ⅰ or Ⅱ, were randomly divided into two groups:control group and treatment group (n=100 each group).In the control group, the patients received injection of cisatracurium besilate with closed-loop muscle relaxant injection system at 1.5-2.0 μg·kg-1 ·min-1 , until 30 min before the end of surgery;if the muscle relaxant level could not meet the requirement of the operation, extra 0.05 mg·kg-1 was added.The treatment group was adopted closed-loop muscle relaxant monitoring under negative feedback regulation of infusion cisatracurium, and the close-loop control parameters were set to: drug was added when TOF was 8%, and injection speed was 2. 5 μg · kg-1 · min-1 , maintaining speed was 0. 33 μg · kg-1 · min-1 , the stimulus current for monitoring muscle relaxant was 60 mA , and the pulse width was 200μs.The Cooper score, cisatracurium dosage, and muscle recovery index, TOFr75 and TOFr90 of the two groups were compared. Prediction probability ( Pk ) of NI on awakening period of eye opening and directional force recovery of the two groups were detected, and regression equation was established to predict ED50 and ED95 related NI . Results Cooper score was significantly higher in the treatment group than in the control group ( P<0. 01 ) . Muscle recovery index, TOFr75 , TOFr90 , and cisatracurium dosage per unit time and body mass were significantly lower in the treatment group than in the control group(P<0.01). Pk of NI on awakening period of eye opening and directional force recovery of the two groups were higher than 0.5; and Pk of the treatment group were significantly higher than those of the control group ( P<0.01) . Regression equation predicted that ED95 was significantly lower in the treatment group than in the control group ( P<0.01) , while the ED50 between the two groups has no significant difference ( P>0.05) . Conclusion The accuracy of closed loop muscle relaxant injection system is higher than that of the traditional method, it provides better muscle relaxation effect for tracheal intubation, reduces recovery time, increases the Pk of NI on patient awakening.

Objective To investigate the correlation between the ratio change of circulating myeloid-derived suppressor cells (MDSCs) and cellular immune function in healthy volunteers,chronic gastritis patients,gastric intraepithelial neoplasia patients and gastric cancer patients.Methods From February 2011 to July 2011,129 peripheral blood samples were collected,including 32 healthy volunteers,48 chronic gastritis patients,27 gastric intraepithelial neoplasia patients and 22 gastric cancer patients.The percentages of peripheral blood MDSC,T lymphocyte subsets and regulatory T cells (Treg) were determined by flow cytometry.The data were analyzed by one way analysis of variance,pearson and spearman correlation.Results The percentages of circulating MDSC,CD8+ T lymphocyte and Treg were highest in gastric cancer patients (9.63％±3.24％,10.03％ ± 1.26％,69.45％±3.42％) and lowest in healthy volunteers (0.92％±0.33％,4.12％ ±0.99％,32.35％ ±4.83％).Those of gastric intraepithelial neoplasia patients (5.13％ ± 1.30％,7.54％ ± 0.79％,53.26％±4.30％) were lower than gastric cancer patients but higher than chronic gastritis patients (2.76％ ±0.64％,6.28％ ±0.61％,42.37％ ±4.02％).The differences among each groups were statistically significant (F=24.85,20.88,37.84,all P＜0.05).However,the percentage of circulating CD4+T lymphocyte was highest in healthy volunteers (65.10％±4.10％),55.15％ ± 4.00％ in chronic gastritis group,42.23％ ± 3.91％ in gastric intraepithelial neoplasia group,and lowest in gastric cancer group (26.84％ ± 3.69％).The differences among each groups were statistically significant (F=46.80,P＜0.05).A significant correlation between circulating MDSC and TNM stages of gastric cancer was also observed (r=0.856,P＜0.01).The percentage of circulating MDSC was positively correlated with Treg percentage (r =0.862,P ＜ 0.01),and negatively correlated with CD4+/CD8+ ratio (r=-0.768,P＜0.01).Conclusion The increase of MDSC percentage in peripheral blood is correlated with human cellular immune function,which might play an important role in the tumor immune evasion during the development of gastric cancer.

Objective To explore the diagnosis value and the mechanism of triggering receptor expressed on myeloid cells-1 (TREM-1) in early liver damage of severe acute pancreatitis with secondary infection. Methods Twenty-four male SD rats were randomly divided into the control group, the severe acute pancreatitis (SAP) group and the secondary infection of SAP (SISAP) group.The animal model was established by intraperitoneal injection of L- arginine and E. coli. After 24 hours, the serum levels of amylase, glutamate aminotransferase (ALT), aspartate aminotransferase (AST), tumor necrosis factor (TNF)-α, C-reactive protein (CRP) and the variation of TREM-1 expression were tested. The blood and peritoneal fluid samples were collected for bacterial culture.Part of the pancreas and liver tissue were taken for histopathological score under microscope. The expression of TREM-1 at mRNA and protein level in liver tissue was detected through Real-time PCR and Western Blot. Results The histological score of pancreas and liver, serum amylase, ALT and AST were significantly higher in the SAP and SISAP groups than those in C group (P<0. 05), and higher in SISAP group than in SAP group (P<0. 05). The CRP and TNF-a expression in SAP and SISAP groups were higher then those in control group, while there was no significant difference between the two groups (P=0. 262 and 0. 359 , respectively). The positive ratio of bacterial culture in blood and peritoneal fluid was 0(0/8), 12. 5% (1/8), and 100% (8/8) in control group, SAP group and SISAP group respectively. The expression of TREM-lmRNA in liver was 2. 10 ± 0. 33 in SAP group and 4. 58+ 1. 00 in SISAP group, which were significantly higher than that in control group (1. 00,P<0. 05) , and the expression of TREM-1 mRNA in SISAP group was higher than that in SAP group (P < 0.05). The expression of TREM-1 at protein level was higher in SISAP group,significantly stronger than that in control and SAP group. Conclusions TREM-1 may play an important role in the early liver damage caused by severe acute pancreatitis.

Objective To detect the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in the early secondary infection of acute necrotizing pancreatitis (ANP) and to probe its diagnostic value for early infection. Methods Twenty-four male SD rats were randomly divided into the control (C) group, the ANP group and the secondary infection of ANP (SIANP) group. The constructions of the models were achieved through intraperitoneal injection of L-arginine and E. coli. After 24 hours, the blood and peritoneal fluid samples were collected for bacterial culture, and the serum levels of amylase, CRP, TNF-α and TREM-1 were detected. The pathological changes in the pancreas were observed. The expression of TREM-1 mRNA and TREM-1 protein in pancreatic tissue was detected by Real-time PCR and Western Blot. Results The histological score of pancreas, and serum amylase in ANP group and SIANP group were significantly higher than those in C group; the positive rate of bacterial culture of blood and peritoneal fluid in SIANP group was 100% , which suggested the model was successfully established. CRP and TNF-a levels in SIANP group were (8.7 ±3.1)mg/L and (185.7 ± 10.9) mg/L, which were not significantly different from that in ANP group [( 16.5 ±3.6) , ( 176.0 ± 18.6) mg/L]. The serum level of TREM-1, expression of TREM-1 mRNA and TREM-1 protein in pancreatic tissue was (9.3 ±0.9) ng/ml, 14.84 ± 3.45, 316.2 ± 59.2, which were significantly higher than those in ANP group [ (5.5 ±0.3)ng/ml, 4.51 ±1.44, 188.6 ±42.4, P <0.05]. Conclusions TREM-1 has diagnostic value for early secondary infection of ANP.

Objective To investigate the effect of miR-200 a mimic on transforming growth factor β1-mediated acti-vation and collagen secretion of rat pancreatic stellate cells .Methods PSCs were isolated and cultured from pan-creatic tissue and identified by desmin , GFAP and α-SMA immunofluorescence staining .PSCs of 2nd generation were divided into control group , TGF-β1 group, TGF-β1+miR-NC group and TGF-β1+miR-200a mimic group.α-SMA and collagen Ⅰ protein were measured by Western blot and immunofluorescence staining .The mRNA ofα-SMA and collagen Ⅰ and the expression of miR-200a were detected by quantitative real-time PCR.Results TGF-β1 stimulates the activation of PSCs and promote collagen synthesis in time-dependment manner ( P<0.05 ) . After transfection of the mimic , treating with the same concentration of TGF-β1, the expressions of protein and mR-NA of both α-SMA and collagen Ⅰ decreases significantly ( P<0.01 ) .Conclusions Over-expression of miR-200 a significantly attenuates α-SMA activity and further affects the collagen synthesis of TGF-β1-dependent activa-tion of PSCs.The mechanisms are potentially related to the biological effects of TGF-β1.