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.

0.05).However,the percentage of patients with cholecystolithiassis was 86.5% in IGC group,and 50.6% in GC group(P=0.000).Besides the percentage of IA stage in IGC group(29.7%) was relatively higher than that(9.0%)in GC group(P=0.03);the surgical resection rate of tumor in IGC group was 56.8% and 32.6% in GC group(P=0.01).Nevertheless,the percentage of advance stage in IGC group(43.2%) was relatively lower than that in GC group(74.2%)(P=0.001).The overall 1,3,and 5-year survival rate of IGC group was 70.0%,31.2% and 26.8% repectively,and the mean survival time was17 months(51?13);which were significantly higher than those in GC group,in which the 1,3,5-year survival rate was 27.0%,17.7% and 15.1% repectively and the mean survival time was(25?8),5 months(all P=0.006).Single factor analysis showed that the survival time in IGC patients was influenced by the TNM stage(P=0.000),pT-category(P=0.000),operation-category(P=0.008);however,postoperative pathological grade(P=0.080),age(P=0.188) and sex(P=0.234) had no influence on survival rate.According to multivariate analysis,pT-category(P=0.000)was an independent factor for the survival time of IGC.Conclusions Comparing with GC group,IGC has a higher percentage of cholecystolithiassis,IA tumor stage and surgical resection rate,and thus,it has relatively better progonosis.pT-category is the vital independent prognostic factor in IGC.If a patient in ICG has been misdiagnosed during the primary operation,the patient should be re-operated for radical excision as soon as possible,except when the tumor is in stage Tis or T1a.

Fanconi anaemia (FA) is an autosomal recessive inherited disorder caused by defects in hematopoietic stem cells. The clinical manifestations of FA are diverse and complicated. FA cells display high hypersensitivity to agents which produce interstrand DNA cross-links such as mitomycin C (MMC) or diepoxybutane (DEB). At least eight complementation groups with defects in eight genes (FANCA, FANCB, FANCC, FANCD(1), FANCD(2), FANCE, FANCF and FANCG) have been identified by gene analysis. Six genes (corresponding to subtypes A, C, D(2), E, F and G) have been coloned, and the encoded FA proteins interact in a common cellular pathway - "FA Pathway", through which modulate DNA repair. The progress of research on FA molecular mechanism provides gene therapy of FA with theory basis. FA cells transduced with the use of retrovirus carring the normal FA gene cDNA manifestate phenotypic correction of hypersensitivity to DNA cross-linking agents, such as MMC. In this review the clinical manifestations and gene composition of FA, and the functions of encoded FA proteins were summarized. The hematopoietic stem cell transplantation and gene therapy for FA patients were discussed.
Cell Cycle Proteins ; DNA-Binding Proteins ; Fanconi Anemia ; genetics ; metabolism ; therapy ; Fanconi Anemia Complementation Group C Protein ; Fanconi Anemia Complementation Group D2 Protein ; Fanconi Anemia Complementation Group Proteins ; Genetic Therapy ; Hematopoietic Stem Cell Transplantation ; Humans ; Mutation ; Nuclear Proteins ; genetics ; Proteins ; analysis ; genetics

Objective To evaluate the diagnostic role of multi-slice CT(MSCT)angiography for tumors of intra-abdominal testes.Methods MSCT angiography of 5 cases with seminoma of intra-abdominal testes proved by surgery and pathology was analyzed.All patients were male,and the age ranged from 33 to 59 years with the mean of 43.6 years.Results(1)The arterial-supply of tumors completely came from the testicular artery of the same side.The arteries went into tumors from the posterior side.(2)The tumors had comparatively large volume with complete or incomplete amicula.The tumor density of 3 cases was inhomogeneous.The degree of entity augmentation of 5 patients was about 14—25 HU.Conclusion MSCT angiography can reveal the tumor vessel and is essential for the diagnosis of tumors of intra-abdominal testes.

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 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

OBJECTIVETo investigate the feasibility of in vivo magnetic resonance imaging (MRI) tracking of transplanted adipose-derived stem cells (ADSCs) labeled with superparamagnetic iron oxide (SPIO) in rat heart.

METHODSADSCs were labeled with poly-L-lysine (PLL)-SPIO complexes. Intracellular iron uptake was identified by Prussian blue stain and transmission electromicroscopy. Trypan blue staining was used to test the viability of the labeled cells. In vitro MRI of labeled cells was performed. SPIO-labeled ADSCs were transplanted into normal rat hearts and were in vivo imaged with MRI. Image findings on MRI were correlated with histological findings of the rat hearts.

RESULTSThe labeling efficacy of ADSCs with PLL-SPIO was nearly 100%. Light microscopy revealed the SPIO particles were located in the cytoplasm of the ADSCs by Prussian blue staining. Transmission electromicroscopy revealed that the SPIO particles were located in the endosomes in the cytoplasm. There was no significantly deference in viability between labeled and unlabeled groups demonstrated by Trypan blue test (P > 0.05). MRI showed signal loss in gel mixed with labeled cells as compared with the unlabeled cells group and blank group. Signal void on rat hearts were demonstrated on MRI and were well correlated with histological findings where Prussian-blue-stain positive cells presented.

CONCLUSIONMRI can be used to in vivo track the transplanted ADSCs labeled with SPIO into rat hearts and facilitate to understand the conditions of the labeled cells in the transplanted areas.

Adipocytes ; cytology ; Animals ; Cell Differentiation ; Contrast Media ; administration & dosage ; Dextrans ; administration & dosage ; Feasibility Studies ; Image Enhancement ; methods ; Magnetic Resonance Imaging ; methods ; Magnetite Nanoparticles ; administration & dosage ; Male ; Myocardium ; cytology ; pathology ; Rats ; Rats, Wistar ; Stem Cell Transplantation ; methods ; Stem Cells ; cytology

To investigate the function of exogenous unsaturated fatty acids in hyposmotic membrane stretch enhancement of muscarinic current (ICCh) in antral circular smooth muscle cells of guinea pig, we recorded the membrane current with the conventional whole cell patch-clamp technique. I(CCh) elicited by 50 micromol/L carbachol (CCh) at the holding potential of 20 mV under isosmotic condition was taken as control. Hyposmotic membrane stretch increased I(CCh) to 226.0+/-21.0%. When the cells were pretreated with 5 micromol/L arachidonic acid (AA), linoleic acid (LA) or oleic acid (OA), I(CCh)was inhibited to 3.8+/-0.6%, 35.2+/-0.8% and 66.6+/-0.6% respectively. Hyposmotic membrane stretch increased I(CCh) to 106.0+/-2.5%, 173.2+/-6.8% and 222.1+/-11.0% of the control respectively. Five micromol/L AA inhibited hyposmotic membrane stretch-enhanced I(CCh) by 51.2+/-3.8%, while the control I(CCh) under isosmotic condition was inhibited by 96.2+/-1.6%. The results suggest that unsaturated fatty acids inhibited I(CCh) and the inhibitory effect is more significant when the unsaturation degree is increased. However, the unsaturated fatty acids are not involved in the increase of I(CCh) induced by hyposmotic membrane stretch.
Animals ; Fatty Acids, Unsaturated ; pharmacology ; Guinea Pigs ; Membrane Potentials ; drug effects ; Myocytes, Smooth Muscle ; cytology ; physiology ; Osmotic Pressure ; Patch-Clamp Techniques ; Pyloric Antrum ; cytology ; physiology ; Receptors, Muscarinic ; physiology ; Sodium Chloride

Objective To discuss the microsurgical resection of intracranial cystic tumors.Methods From August 2003 to August 2010, 47 patients who had been diagnosed by imaging with intracranial cystic tumor received microsurgical resection. The tumors were confirmed by pathological examination postoperation.There were 22 cases of cystic glioma,8 cases of cystic meningioma,3 cases of malignant cystic meningioma,4 cases of cystic ependymoma,4 cases of angioreticuloma and 6 cases of cystic metastatic tumor.When the skull was minimally opened according to the location of the tumor,the hydatid fluid was absorbed before separation and resection of the tumor with its capsule under endoscopy. Results Total resection was performed in 31 cases (65.96％) and sub-total resection in 16cases (34.04％).No operative death occurred.Neural dysfunction was observed not long postoperation in 6 cases (12.77％),including one case of trigeminal nerve injury,one case of abducent nerve injury,one case of facial nerve dysfunction,one case of glossopharyngeal nerve injury,2 cases of paralysis and 2cases of hydrocephalus.Forty-one patients were followed up for 0.5 to 6 years.Eight patients (17.02％)had recurrence and 2 died. Conclusion Microneurosurgery can improve the total resection rate of intracranial cystic tumors with minimal invasion to neural functions.

Background There is a growing interest in studying the relationship between intrinsic resistance and biofilms resistance to drugs. However, the relationship still remains unclear in the macroscopic bacterial growth. Our study is to illuminate the change of bacterial drug resistance of gyrA mutant and active efflux pump during the development of Pseudomonas aeruginosa (P. aeruginosa) biofilms. Methods The strains of type Ⅱ topoisomerase gene mutant (gyrA mutant) and multidrug resistance (MDR) efflux pump were clinical isolates and detected by polymerase chain reaction (PCR). The process of bacterial biofilms development was observed by scanning electron microscope. Triparental mating experiments were performed to transfer report gene of green fluorescent protein (GFP) into P. aeruginosa biofilms strains and followed by analysis of bacterial survival rate between intrinsic resistance and biofilms resistance.Results The fluorescent strains with pGFPuv could develop mature biofilms on Teflon surface. Before a period of 72 hours, the survival rate of biofilms bacteria and intrinsic resistance strains in ciprofloxacin solution was significantly different (P<0.05). The survival number of intrinsic resistance strains (gyrA mutation and active efflux pump) was illustriously higher than biofilm strain in the initial stage of biofilms development. After 72 hours incubation, there was no clearly difference between mutants and biofilms strains in the survival rate (P>0.05). The carbonyl cyanide m-chlorophenylhydrazone and azithromycin could significantly reduce the drug resistance of biofilm strains and efflux pump strains.Conclusions In the development of P. aeruginosa biofilms, the strains of gyrA mutation and MDR efflux could be conferred with new level of drug resistance. When co-cultured mutated strains with biofilm strains, biofilms may play a major role in bacterial resistance. But after 72 hours incubation (a mature biofilms had been developed), there was no clearly difference between the number of mutant strains and biofilm strains.