Objective To investigate the possibility of survivin inhibition by lentiviral vector-mediated RNA interference and the influence on cell apoptosis in pancreatic cancer cell line.Methods The lentiviral vector of SiRNA targeted against survivin(LV-shRNA-survivin-1,LV-shRNA-survivin-2,LV-shRNA-survivin-3) was constructed and transfected into the packaging cells 293T,and then the supernatant with virus was collected to transfect SW1990 cells.Quantitative real-time fluorescent PCR and Western-blot were used to detect the expression of survivin.DAPI staining and detection of enzymatic activity of caspase 3/7 were employed to examine cell apoptosis.Results Three lentiviral vector-survivin-shRNA were constructed successfully.In the LV-shRNA-survivin-1 group,the survivin mRNA and protein expression inhibitory rate was 73.50% and 87.64% respectively;when compared to control group,the activity of caspase-3/7 increased significantly,which showed a 14.5-fold increase,and apoptosis increased 11.95%.Conclusions Lentiviral vector-mediad RNA interference targeted against survivin can effectively inhibit survivin expression and increase cell apoptosis significantly.

Objective To determine,the clinical significance of serum myocardial enzymes (Mb,cTNI, CK,CK-Mb,AST,LDH) in the classification of the disease severity of non-cardiogenic critically-ill patients. Compared with APACHEⅡscore concerned as the standard diagnosis of the critical ills,these biomarkers were investigated for the evaluation possibility of the degree and the prognosis of the critical ills.Method Patients admitted to our EICU were consecutively collected for the research from April to December in 2005 and the myocardial enzymes,and routine serum biochemical test and APACHEⅡscore were detected simultaneously.All the patients were classified to three groups according to the APACHEⅡscore (mild group,APACHEⅡ25) and two groups (survive group and death group) according to the prognosis.All the patients were followed up till recovery/discharge or death. Covariance,Wilcoxon and x~2 were used for the statistical analysis.Results The myocardial enzymes rose when the disease deteriorated and the APACHEⅡscore went up.AST,LDH,CK,CK-Mb,Mb were significantly different in the three groups according to the APACHEⅡscore (P

OBJECTIVETo explore the effect of half circle external fixation for the treatment of open tibial fractures.

METHODSFrom March 2005 to March 2011, 94 patients with open tibiofibula fractures were treated by closed manipulative or Kirschner-wire poking reduction with half circle groove external fixation including 63 males and 31 females with an average age of 39 years old ranging from 17 to 65 years old. Among these patients, 5 cases were cross shaped fractures, 19 were oblique form and spiral fractures, 70 cases were comminuted fractures. According to the fracture Gustilo classification, 49 cases were type IIIA, 45 were type IIIB. The incidence of wound infection, fracture healing time were observed. The function was evaluated according to the Johner-Wruhs standard.

RESULTSAll patients were followed up for 14 to 63 months (averaged 29 months). The wound healing time was from 16 to 39 weeks (means 21.4 weeks). No fracture nonunion, osteomyelitis and calf compartment syndrome occurred. The wounds of 81 cases were healed at the first period,deep wound infection occurred in 2 cases. According to the function Johner-Wruhs evaluation criteria:the result was excellent in 52 cases, good in 37 cases, fair in 5 cases.

CONCLUSIONClosed manipulative or Kirschner-wire poking reduction and half circle groove external fixation can reduce the infection rate of open tibiofibula fractures. For open tibiofibula comminuted fractures, after the half circle groove external fixation for 3 to 6 weeks, a middle Kirschner-wire was removed according to fracture end healing situation to make fixation dynamic and promote fracture healing.

Adolescent ; Adult ; Aged ; Bone Wires ; External Fixators ; Female ; Fracture Fixation ; instrumentation ; Fractures, Comminuted ; surgery ; Fractures, Open ; surgery ; Fractures, Ununited ; surgery ; Humans ; Male ; Middle Aged ; Tibial Fractures ; surgery ; Young Adult

OBJECTIVE: To investigate the simultaneous inhibition of X-linked inhibitor of apoptosis protein (XIAP) and survivin expression on epithelial-mesenchymal transition (EMT) and invasiion of pancreatic cancer cells Panc-1, and its mechanism. METHODS: On the established human pancreatic cancer cells Panc-1-XS, the expression of XIAP and survivin was inhibited simultaneously. Cell invasion and migration were detected by Transwell chamber experiments and scratch test, and the expression of epithelial marker E-cadherin, mesenchymal markers Slug, phosphatase and tensin homolog deleted on chromosome ten (PTEN) and P-Akt protein was determined by Western blot. RESULTS: Cell invasion and migration of Panc-1-XS cells decreased significantly, accompanied by significantly upregulated protein expression of E-cadherin, and significantly declined protein expression of the Slug, indicating increased mesenchymal-epithelial conversion (MET); and increased protein expression of PTEN, and declined protein expression of P-Akt. CONCLUSION Simultaneously inhibiting the expression of XIAP and survivin can partially reverse EMT phenotype of pancreatic cancer Panc-1 cells, which then significantly reduces the cell invasion and migration of Panc-1 cell lines. This process may be regulated by PTEN/PI3K/Akt signaling pathway.
Antigens, CD ; Cadherins ; metabolism ; Cell Line, Tumor ; Cell Movement ; Epithelial-Mesenchymal Transition ; genetics ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; metabolism ; Pancreatic Neoplasms ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Snail Family Transcription Factors ; Survivin ; Transcription Factors ; metabolism ; X-Linked Inhibitor of Apoptosis Protein ; genetics ; metabolism

OBJECTIVETo compare clinical efficacy differences among electroacupuncture (EA), transcutaneous electrical nerve stimulation (TENS) therapy and acupoint massage (AM) on periarthritis of shoulder.

METHODSOne hundred and twenty cases were randomly divided into an EA group, a TENS group and an AM group, 40 cases in each one. The selection of acupoints in the three groups were all the same, which were Jianqian (Extra), Xiabai (LU 4), Jianyu (LI 15), Binao (LI 14), Jianliao (TE 14), Naohui (TE 13), Xiaohai (SI 8) and Houxi (SI 3). EA was applied in the EA group with dense-disperse wave for 30 min, TENS therapy with dense-disperse wave for 30 min in the TENS group and AM combined with mobilization manipulation of Tuina on the shoulder in the AM group. All of the treatment was given once a day, six times a week and totally 4 weeks were required. The score of visual analogue scale (VAS) and improvement of shoulder joint activities in the three groups before and after the treatment were observed.

RESULTSAfter the treatment, the VAS score and activities of shoulder joint were all improved in the three groups (all P < 0.001) and AM group had more obvious improvement than TENS group and EA group (all P < 0.05). The effective and curative rate was 61.50 (24/39) in the AM group, which was superior to 42.9% (15/35) in the EA group and 44.4% (16/36) in the TENS group (both P < 0.05). The effective and curative rate of shoulder joint activities in the AM group was 61.5% (24/39), which was superior to 48.6% (17/35) in the EA group and 44.4% (16/36) in the TENS group (both P < 0.05).

CONCLUSIONThe acupoint massage has obvious clinical efficacy on periarthritis of shoulder, which could effectively relieve the pain and improve activities of shoulder joint, and it is superior to EA and TENS therapy.

Acupuncture Points ; Adult ; Aged ; Electroacupuncture ; Female ; Humans ; Male ; Massage ; Middle Aged ; Periarthritis ; physiopathology ; therapy ; Range of Motion, Articular ; Shoulder Joint ; physiopathology ; Transcutaneous Electric Nerve Stimulation

Derris eriocarpa, a traditional Chinese medicine belonging to the family of Leguminosae, is widely distributed mainly over Yunnan, Guangxi and Guizhou of China. Modern pharmacological researches on this herb showed that it had extensive bioactivities, such as promoting urination, removing dampness and cough and reducing inspissated mucus and other biological activities. The extensive studies on the chemical constituents of this plant have resulted in the isolation of triterpenoids, steroids, fatty acid and others, but the flavone compounds haven't reported before. In our further research on the ethyl acetate of this plant, nine flavone compounds were obtained by column chromatography on silica gel, Sephadex LH-20, semi-prep HPLC, polyamide column chromatography and recrystallization for separation and purification. The structures were determined on the basis of extensive spectroscopic analysis, including MS, NMR experiments and comparison with spectroscopic data in the literature, respectively, as diosmetin (1), 3, 3'-di-O-methylquercetin (2), afromosin (3), 6, 3'-dihydroxy-7, 4'-dimethoxyisoflavone (4), odoratin (5), 7, 3'-dihydroxy-8, 4'-dimethoxyisoflavone (6), 6, 4'-dihydroxy-7, 3'-dimethoxyisoflavone (7), 5, 7, 4'-trihydroxy-3, 3', 5'-trimethoxyflavone (8), and alpinumisoflavone (9). All these compounds were isolated from Derris eriocarpa How for the first time. And the in vitro assays showed that compound 2 possessed moderate inhibitory activity against human cancer cells K562 and HEL.
Derris ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Humans ; K562 Cells

Objective To investigate the efficacy, safety and tolerance of bosentan, a dual endothelin receptor antagonist, in Chinese patients with idiopathic pulmonary arterial hypertension (IPAH).Methods Totally 79 IPAH patients (hemodynamic criteria confirmed by right heart catheterization) were included in this open-label, prospective multicenter study. Patients received 62. 5 mg of bosentan twice daily for the first 4 weeks, and then up-titrated to 125 mg twice daily for another 12 weeks. The primary end point was the change in exercise capacity showed by six-minute walk distance (6MWD) from baseline to 16 weeks. Secondary end points included the change in World Health Organization (WHO) functional class,Borg dyspnoea scale and systolic pulmonary artery pressure measured by echocardiography. Results The 6MWD increased from (343. 7 ± 93.7) meters at baseline to (397.5 ± 104. 4) meters after 16 weeks ( P ＜0. 01 ), WHO functional class and Borg dyspnoea scale were also significantly improved after 16 weeks therapy compared to baseline levels (all P ＜0. 01 ). Furthermore, the systolic pulmonary artery pressure was significantly decreased from (97.8±25.2) mm Hg (1 mm Hg=0. 133 kPa) to (92.8 ±29.5) mm Hg (P ＜0. 05) after 16 weeks bosentan treatment. There was no patient withdrawal from this study for safety consideration. Conclusion Bosentan therapy is well tolerated and can improve the exercise capacity and WHO functional class in Chinese IPAH patients.

Objective To evaluate interventional therapy for biliary stricture (BS) after orthotopic liver transplantation (OLT). Methods The efficacy of interventional therapy for BS after OLT from Oct 2003 to Jan 2006 was analyzed retrospectively. Fifty-three patients received 107 times of interventional therapy through endoscopic retrograde cholangiography ( ERC) which included 68 nasobiliary catheter placements,26 biliary balloon dilatations and stent placements and 13 ERC. Nine patients received 11 times of interventional therapy through percutaneous transhepatic cholangiography ( PTC) including 2 PTC, 7 percutaneous drainages,3 biliary balloon dilatations and 1 biliary stent replacement. One patient received bile drainage through T tube. Results The success rate of ERC was 88. 8% (95/107) , that of nasobiliary catheter placement 94% (64/68) , biliary stent placement 88. 5% (23/26). The success rate of PTC was 81. 8% (9/11) , that of percutaneous drainage was 100% (7/7) , biliary stent replacement 100% (1/1). The curative rate of interventional therapy for 53 patients with BS was 28. 3% (15/53) ,the improvement rate was 41. 5% (22/53). The curative rate of interventional therapy for anastomotic, extrahepatic, intrahepatic hilar and diffuse BS was respectively 66. 7% (4/6)、66. 7% (10/15)、50% (1/2)、0 (0/7) and 0 (0/22). Conclusions The efficacy of interventional therapy for BS after OLT was not satisfactory. The result relates to the type of BS, for anastomotic, extrahepatic and solitary intrahepatic BS this therapy was effective, while that for hilar and diffuse BS the prognosis was poor.

OBJECTIVEInhaled glucocorticosteroids (ICS) remains the first line controller medication for chronic airway inflammation in asthma till now. If the impact of allergen could not be eliminated, how would the improvement of airway inflammation be achieved with inhaled glucocorticosteroids therapy? What was its effect on airway remodeling? In this study, an animal model of asthma was established and the effects of budesonide on airway allergic inflammation and extracellular matrix (ECM) deposition in sensitized guinea pigs with repeated exposure to allergen were investigated.

METHODSThirty-two male Hartley guinea pigs were randomly divided into four groups with 8 in each group: (A) Group of repeated exposure to ovalbumin (OVA), (B) Group of repeated exposure to OVA plus budesonide (BUD) intervention, (C) Group of stopping repeated exposure to OVA plus stopping BUD intervention, (D) Control group. At 24 h after the last OVA challenge (8 weeks after the first OVA challenge), bronchoalveolar lavage fluid (BALF) was collected from each animal. Total and differential leukocyte counts in BALF was performed on cell suspension smear stained with May-Grünwald-Giemsa (MGG) method. The upper lobe of right lung was removed and regularly fixed, then paraffin embedded lung tissues sections were prepared. The count of eosinophils infiltrated in the airway wall was performed on H&E stained lung tissue sections with LEICA Q500IW computerized image analysis system. Fibronectin and collagen type III (Col-III) deposited in the airway wall were detected by immunohistochemical staining on the paraffin embedded lung tissues sections. The intensity of positive reaction of fibronectin or Col-III deposited in the airway wall was analyzed with LEICA Q500IW computerized image analysis system.

RESULTSThe count of eosinophils in BALF (x 10(5)/ml) of group A and B were higher than that of group C and D (35.70 +/- 25.22, 11.49 +/- 5.51 vs. 1.00 +/- 0.90, 1.02 +/- 0.78, P < 0.01), the difference between group A and B, group B and C was significant. The count of eosinophils infiltrated at each level of airway wall in group A and B were higher than that of group C and D (large airway: 6.95 +/- 2.28, 1.54 +/- 1.09 vs. 0.76 +/- 0.45, 0.88 +/- 0.25; medial airway: 9.22 +/- 3.89, 3.99 +/- 2.3 vs. 1.25 +/- 1.20, 0.64 +/- 0.36; small airway: 11.56 +/- 4.02, 2.67 +/- 1.15 vs. 1.32 +/- 0.83, 0.43 +/- 0.24, P < 0.01), the difference between group A and B, group B and C was significant. The gray values of fibronectin deposited in medial and small airway of group A and B were lower than those of group C and D (medial airway 122 +/- 22, 174 +/- 23 vs. 219 +/- 34, 229 +/- 20; small airway 135 +/- 29, 165 +/- 41 vs. 236 +/- 20, 220 +/- 16, P < 0.05), the difference between group A and B, group B and C was significant. The gray values of Col-III deposited in medial and small airway of group A and B were lower than those of group C and D (medial airway 153 +/- 21, 174 +/- 22 vs. 189 +/- 14, 200 +/- 18; small airway 133 +/- 23, 176 +/- 20 vs. 191 +/- 14, 198 +/- 20, P < 0.05), the difference between group A and B was significant.

CONCLUSIONInhaled budesonide could partially inhibit allergic inflammation and ECM deposition in airway wall in guinea pig chronic asthma model with repeated exposure to allergen. Early inhaled budesonide combined with avoidance of OVA exposure could completely inhibit allergic inflammation and ECM deposition. These results suggest that the inhibitory effect on airway allergic inflammation and airway remodeling of inhaled glucocorticosteroids would be limited when the allergen factor could not be avoided.

Administration, Inhalation ; Airway Remodeling ; drug effects ; immunology ; Allergens ; administration & dosage ; immunology ; Animals ; Asthma ; chemically induced ; drug therapy ; immunology ; Bronchitis, Chronic ; chemically induced ; drug therapy ; immunology ; Bronchoalveolar Lavage Fluid ; immunology ; Budesonide ; administration & dosage ; pharmacology ; Collagen Type III ; metabolism ; Disease Models, Animal ; Eosinophils ; immunology ; Extracellular Matrix ; immunology ; Fibronectins ; metabolism ; Glucocorticoids ; administration & dosage ; pharmacology ; Guinea Pigs ; Immunohistochemistry ; Lung ; drug effects ; immunology ; Male ; Ovalbumin ; administration & dosage ; immunology

Objective:To explore the effect of half circle external fixation for the treatment of open tibial fractures. Meth-ods: From March 2005 to March 2011,94 patients with open tibiofibula fractures were treated by closed manipulative or Kirschner wire poking reduction with half circle groove external fixation including 63 males and 31 females with an average age of 39 years old ranging from 17 to 65 years old. Among these patients ,5 cases were cross shaped fractures ,19 were oblique form and spiral fractures,70 cases were comminuted fractures. According to the fracture Gustilo classification,49 cases were typeⅢA,45 were typeⅢB. The incidence of wound infection,fracture healing time were observed. The function was evaluated according to the Johner Wruhs standard. Results:All patients were followed up for 14 to 63 months (averaged 29 months). The wound healing time was from 16 to 39 weeks (means 21.4 weeks). No fracture nonunion,osteomyelitis and calf compart-ment syndrome occurred. The wounds of 81 cases were healed at the first period ,deep wound infection occurred in 2 cases. According to the function Johner-Wruhs evaluation criteria:the result was excellent in 52 cases,good in 37 cases,fair in 5 cases. Conclusion:Closed manipulative or Kirschner wire poking reduction and half circle groove external fixation can reduce the infection rate of open tibiofibula fractures. For open tibiofibula comminuted fractures ,after the half circle groove external fixation for 3 to 6 weeks,a middle Kirschner wire was removed according to fracture end healing situation to make fixation dy-namic and promote fracture healing.