Objective To construct the specific small hairpin RNA(shRNA)plasmids for Formin-like 2(FMNL2)gene,and to observe its effect on proliferation of SW620.Methods Plasmids FMNL21 shRNA and plasmids FMNL22 shRNA complimentary to the sequence responsible for the function domain of human FMNL2 were constructed and transfected into SW620,respectively.The inhibition rates of FMNL2 expression were detected by re-al-time PCR after transfection with PGenesil-FMNL21 and PGenesil-FMNL22,respectively.The expression of FMNL2 was detected by real-time PCR at 12,24,48,72 and 96h after transfection.At 48 h and 72 h after transfection,the proliferation of SW620 was detected using MTT assay.Resuits The transfection rate in SW620 was(60.8±2.8)%for PGenesil-FMNL21,(58.7±2.9)%for PGenesil-FMNL22,and(62.6±1.7)%for PGenesil-HK(P＞0.05).The inhibition rate of FMNL2 expression after transfection with PGenesil-FMNL21 was 77.1%,which was the highest.The inhibition rates of FMNL2 expression were 13.5%,57.3%,80.3%,62.4%,and 33.2%at 12,24,48,72and 96h after transfection respectively.The proliferat capability of SW620 was obviously lower than that of control cells(P＜0.05).Conclusion Both plasmids FMNL21 shRNA and plasmids FMNL22 shRNA can inhibit the expression of FMNL2,of which Pgenesil-FMNL21 execs the greatest role.FMNL2 could be related to the proliferation of SW620.

Objective To analyze the relationship between the expression of formlin-like 2(FMNL2)and colorectal carcinoma metastasis through the detection of the expression of FMNL2 in colorectal carcinoma tissue and paracancerous tissue.Methods ①Immunohistochemistry was used to detect the expression of FMNL2 in 175 cases of paraffin wax specimens(including 30 cases of normal mucosa,25 cases of adenoma,75 cases of colorectal carcinoma and 45 cases of metastatic tumors).②Real time-PCR was used to detect FMNL2 mRNA expression in 32 cases of fresh specimens of carcinoma and 32 cases of paracancerous tissue.Results The positive detection rate of FMNL2 was higher in colorectal carcinoma tissue than in paracancerous tissue(P=0.003),higher in lymphatic metastasis than in primary carcinoma tissue(P=0.037),and was aslo higher in metastatic carcinoma tissue than in non metastatic carcinoma tissue(P=0.011).The expression of FMNL2 mRNA was not related to the infiltration and differentiation of carcinoma tissue but its concentration in metastatic carcinoma tissue was 2.5 times higher than in non metastatic carcinoma tissue.Conclusion FMNL2 gene may play an important role in the metastasis of colorectal carcinoma.

Objective To study the synergistic antitumor effects of quercetin and cisplatin in human prostate cancer PC3 cells and LNCaP cells.Methods Twelve h after PC3 cells or LNCaP cells were seeded,different dose of quercetin (10 μmol/L,20 μmol/L,40 μmol/L,80 μmol/L,160 μmol/L) or cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L),or quercetin (20 μmol/L) + cisplatin (0.01 μmol/L,0.05 μmol/L,0.1 μmol/L,1 μmol/L,10 μmol/L) were added for48 h and then the antiproliferative effects were detected with MTT assay.After incubated with quercetin (20 μmol/L) or cisplatin (0.05 μ mol/L),or quercetin (20 μ mol/L) + cisplatin (0.05 μmol/L) for48 h,cell cycle distribution and apoptosis of PC3 cells or LNCaP cells were detected by flow cytometer,PI and Annexin V staining.Protein expression was detected by Western blotting.Results After treatment with quercetin or cisplatin alone,the IC50 were (0.99 ± 0.13) μmol/L,(0.75 ± 0.09) μmol/L and (91.60 ± 6.10) μ mol/L,(72.90±4.70) μ mol/L for LNCaP cells or PC3 cells,respectively;The IC50 were (0.11±0.06)μ mol/L,(0.07±0.02) μmol/L for quercetin + cisplatin treatment (Compared with quercetin,P＜0.01 ;Compared with cisplatin,P＜0.05.After treatment with cisplatin or quercetin + cisplatin for 48 h,the S phase percent of LNCaP cells or PC3 cells were (22.4±2.7)％,(31.2±2.4)％ and (20.1±1.6)％,(31.0±2.5)％,respectively,(Compared with control,P＜0.05,however,treatment with quercetin alone has no significant difference (Compared with control,P＞0.05).After treatment with cisplatin or quercetin + cisplatin for 48 h,the apoptotic percent of LNCaP cells or PC3 cells were (14.8 ± 1.9) ％,(39.6 ± 3.1) ％ and (11.5± 1.2) ％,(34.1 ±3.3) ％,respectively,(compared with control,P ＜ 0.05,however,treatment with quercetin alone had no significant difference (compared with control,P＞0.05).After treatment with quercetin alone for 48 h,the activation of caspase-3,caspase-8 and caspase-9 were slightly increased,the expressions of Bax and p21 were up-regulated,the expressions of Bcl-2 and CDK2 were down-regulated.Furthermore,these effect of cisplatin and quercetin + cisplatin were significantly enhanced (compared with quercetin,P＜0.05;compared with quercetin,P＜0.01,respectively.Conclusions The combination modality with quercetin and cisplatin has a better treatment effect in vitro not only in androgen-dependent LNCaP cells but also in androgen-independent PC3 cells.

Objective To explore the effectiveness and safety of endoscopic papillary balloon dilation( EPBD ) for the removal of common bile duct stones. Methods Three hundred and twenty consecutive patients with common bile duct stones on endoscopic retrograde cholangiopancreatography (ERCP) who met all eligibility criteria were randomly assigned endoscopic sphincterotomy (EST) or EPBD.Complications were classified by an expert panel unaware of treatment allocation and outcome. Results After a single ERCP, all stones were removed from 112 patients ( 70% ) assigned EST and 104 ( 65% )assigned EPBD. Mechanical lithotripsy was used to fragment stones in 36 (22. 5% ) EPBD procedures and 32 ( 20. 0% ) EST procedures. Early complications occurred in 5.6% EST patients and 8.1% EPBD patients. No patient died. Gallstone disease recurrence, which is a long-term complication, is 7. 5% ( 12/160) in EST patients and 2.5% (4/160) in EPBD patients, P ＜0. 05. Conclusions The success rate of EPBD was similar to that of EST. We found no evidence of previously suggested higher risk of pancreatitis with EPBD, and suggest that EPBD is preferred in patients who are not suitable for EST, such as those with high risk of bleeding. This procedure is a valuable alternative to EST in patients with bile duct stones.

Objective To summarize the nursing experience during removing the upper gastrointestinal foreign bodies in special patients by painless endoscopy. Method Retrospective analysis was done to investigate the clinical records on endoscopy for removing the upper gastrointestinal foreign bodies in 69 special patients. Result The foreign bodies in 67 patients were removed by endoscopy successfully, without severe complications such as bleeding and perforation; one patient was removed with duodenum lateral telescope; one patient turned for sugery . Conclusion Sufficient preoperative preparation and skilled surgical nursing cooperation are promising for the successful removal of upper gastrointestinal foreign bodies in special patients.

Objective To study the value of three-dimensional pseudo-continuous arterial spin labeling(3D pCASL) for estimating collateral flow after middle cerebral artery(MCA) stenosis.Methods 3D pCASL was performed in 28 cases of negative diffusion weighted imaging(DWI) patients with MCA stenosis for estimating collateral vessels formation.The post-labeling delay(PLD) of 3D pCASL was set to 1.5 s and 2.5 s.The cerebral blood flow (CBF) of 3D pCASL with two PLD was calculated by AW workstation respectively.Nineteen normal persons were regarded as the control group.The 95% confidence interval of the ratio of left/right side of CBF of control group was set as reference value.The characteristic of the ratio of stenosed/contralateral side of CBF was analyzed.The characteristics of collateral flow was investigated with digital subtraction angiography(DSA).Results Abnormal perfusion was found in 28 patiens.Two types of abnormal perfusion were found, including typeⅠ: rCBF(PLD=1.5 s) decreased,rCBF(PLD=2.5 s) was normal (n=16);typeⅡ:rCBF(PLD=1.5 s) decreased,rCBF(PLD=2.5s) decreased (n=12).On brain DSA, abundant of collateral flow supply was found in 16 patients with typeⅠabnormal perfusion,whereas deficiency of collateral flow supply was found in 12 patients with typeⅡ.Conclusion 3D pCASL with two PLDs may provide important reference value for estimating collateral flow after middle cerebral artery stenosis.

OBJECTIVETo investigate clinical effects of calcaneal fracture with closed reduction and minimally invasive plate fixation assisted with bidirectional distractor distraction.

METHODSFrom September 2015 to October 2016, 11 male patients(13 feet) with calcaneal fractures treated with bidirectional distractor distraction assisted with minimally invasive plate fixation were retrospectively studied. They were aged from 24 to 57 years old with an average of 36.4 years old;8 feet were type IIand 5 feet were type III according to Sanders classification. Postoperative incision, fracture healing, Böhler angle, Gissane angle were observed and Maryland scoring system was used to evaluate clinical effects.

RESULTSAll fractures healed well without incision inflammation and incision disunion. All patients were followed up from 12 to 15 months with an average of 13.5 months. Böhler angle were improved from (9.6±7.3)° before operation to (20.2±4.6) ° at 1 year after operation, and had statistical meaning; Gissane angle increased from (92.7 ±8.5)° before operation to (121.7 ±7.6) ° at 1 year after operation. Maryland score at 1 year after operation was 88.79±8.25, and 11 feet got excellent results and 2 feet moderate.

CONCLUSIONSBidirectional distractor distraction assisted with minimally invasive plate fixation could effectively fix calcaneal fractures, reduce postoperative complications, and get satisfied results of postoperative images and functional recovery. It is one of effective methods for treating Sanders II and III calcaneal fractures.

Objective To investigate the role of UDP-glucuronosyltransferase 1A (UGT1A),nuclear factor erythroid-2-related factor 2 (Nrf2) and kelch-like ECH-associated protein 1 ( Keapl ) in the tumorigenesis of colonic carcinoma.Methods The expressions of UGT1 A,Nrf2 and Keapl were detected in normal colonic mucosa(24 cases),adenoma tissue (30 cases) and adenocarcinoma tissue (77 cases) by immunohistochemistry,and the relationship between their expressions and the clinical pathological characteristics was analyzed.Results The positive rates of UGT1 A in normal colonic mucosa,adenoma and adenocarcinoma tissue were 83.3％ ( 20/24),80.0％ ( 24/30 ) and 53.2％ ( 41/77 ),respectively.The positive rate of UGT1A in adenocarcinoma was lower than those in colonic mucosa and adenoma ( all P ＜0.05 ).On the contrary,the positive rates of Nrf2 in adenoma [70.0％ (21/30) ] and adenocarcinoma tissue [ 87.0％ (67/77) ] were higher than that in normal colonic tissue [ 41.7％ (10/24),all P =0.000 ].The positive rates of Keapl in normal colonic mucosa,adenoma and adenocarcinoma tissue were 54.2％ ( 13/24),70.0％ (21/30) and 61.0％ (47/77),respectively ( normal colonic tissue vs adenocarcinoma tissue,P =0.040 ; adenoma vs adenocarcinoma,P =0.002 ).There was no correlation between the expression of UGT1 A,Nrf2 and the clinicopathologic features of colon carcinoma,while the differences of Keapl positive rates in the various degrees of tumor differentiation [ moderately-well differentiated vs poorly differentiated:70.0％ (35/50) vs 44.4％ (12/27) ] and invasion [T1-T2 vs T3-T4:78.8％ (26/33) vs 47.7％ (21/44) ]were statistically significant (all P ＜ 0.05 ).Conclusion The decreased expression of UGT1A and the dysregulation of Nrf2/Keapl system may play a role in colonic tumorigenesis.

To evaluate the effect of components in Guanxin Ⅱ prescription on the pharmacokinetic profiles of paeoniflorin and ferulic acid. METHODS: Drug concentrations of rat plasmas after intravenous injection of paronia pall (PPE) or ferulic acid (FA) extract solution, as well as oral administration of PPE and FA solution, and different kinds of decoctions based on Guanxin Ⅱ prescription were determined by an HPLC system. NONMEM (nonlinear mixed-effect modeling) method was used to analyze the population pharmacokinetics of PF and FA. RESULTS: A two-compartment model with first order degradation in absorption phase, and an ordinary two-compartment model were adequately describe PF and FA pharmacokinetic profiles, respectively. The mean of PF population parameters, CL1, V1, CL2, V2, Ka0, and Ka1, were 0.509 L/h, 0.104 L, 0.113 L/h, 0.123 L, 0.135 /h, and 0.0135 /h, respectively, while the typical values of CL1, V1, CL2, V2, Ka1, and F in FA model were 0.295 L/h, 0.025 L, 0.0331 L/h, 0.0518 L, 0.110 /h, and 0.40, respectively. Inter-individual variabilities were estimated and dose formulation (DF) was identified as a significant covariate in the model. CONCLUSION: The results indicate that the pharmacokinetic behaviors of index components in Guanxin Ⅱ prescription can be influenced by different dose formulations administrated in rats.