Objective To investigate the causes and preventive measures for failure and complications of minimally invasive percutaneous nephrolithotomy (mPCNL). Methods From January 2005 to October 2008,totally 353 cases of mPNCL were performed in our hospital. Among the cases,30 patients experienced failure of the surgery or postoperative complications. The data of the 30 patients were analyzed retrospectively in this study. Results Of the 30 cases,puncture failure occurred in 5 patients,while 2 of them were converted to open surgery,and the other 3 were treated by a second puncture successfully; Zebra wire extrusion was found in 5 cases,who were then cured by re-puncture or a second operation; in 3 patients,the wire or PCN tube was moved into the renal vein (2 cases) or the colon (1 case) without causing bleeding or intestinal fistula,the cases were cured afterwards by a second operation; 3 patients developed hydrothorax and then was cured by chest drainage; postoperative arteriovenous fistula was detected in 2 patients,who were cured by interventional therapy; in 5 days after the operation,one patient developed massive hemorrhage from the PCN tube when driving cars,and then recovered by clipping the PCN tube and hemostasis; another patient showed hemorrhage and infection due to extraction of the PCN tube,and open surgery was carried out to cure this patient. Conclusions Failure of puncture and Zebra wire extrusion are most common causes of failure of mPCNL. Whereas,post-mPCNL complications is often caused by unskillful surgeons or noncompliant patients.

OBJECTIVETo compare the effectiveness of percutaneous microwave ablation ( MWA) versus hepatectomy for multifocal hepatocellular carcinoma.

METHODSFrom August 2002 to March 2012, one hundred and twenty-two patients with multifocal hepatocellular carcinoma (diameters 1 to 7 cm, 2 to 4 lesions) were treated by either complete MWA or radical hepatectomy, and their clinical data were collected and analyzed.The patients were divided into MWA group (n = 50) and resection group (n = 72), and the resection group was matched by MWA group based on clinical parameters. The survival and complications in the two groups were compared.

RESULTSThe overall 1-, 3- and 5-year survival rates were 100.0%, 73.0% and 62.0%, respectively, in the MWA group, and 80.0%, 56.0%, and 41.0%, respectively, in the resection group (P < 0.05). The corresponding recurrence-free survival rates were 88.0%, 63.0%, and 52.0% in the MWA group, and 68.0%, 45.0%, and 36.0%, respectively, in the resection group (P< 0.05). The multivariate Cox regression analysis indicated that albumin level, performance status, treatment modality, and tumor size were independent prognostic factors.

CONCLUSIONCompared with hepatectomy, percutaneous microwave ablation is a minimally invasive and reproducible procedure, and can improve the survival in patients with multifocal hepatocellular carcinoma.

Carcinoma, Hepatocellular ; mortality ; radiotherapy ; surgery ; Catheter Ablation ; Genetic Engineering ; Hepatectomy ; mortality ; Humans ; Liver Neoplasms ; mortality ; radiotherapy ; surgery ; Microwaves ; therapeutic use ; Regression Analysis ; Survival Rate ; Treatment Outcome

Objective To observe the transfection efficiency and anti-fibrotic effect of miR-29b transfected by anti-TGF-β Ⅱ R ScFv/Ck/tP fusion protein (new vector) in hepatic stellate cell (HSC),and to provide a new vector in gene therapy for liver fibrosis.Methods The liposome vector,new vector,and lentiviral vector were used as transfection reagents to transfect miR-29b into HSC.Transfection efficiency was observed under fluorescence microscope and flow cytometry.Collagen α1 (Ⅰ) mRNA and protein expression in different groups were analyzed by real-time RT-PCR and Western Blot,respectively.Results Compared to the control,transfection efficiencies in lentiviral vector,new vector,and liposome vector groups were about 70％,58％,and 29％,respectively.Collagen α1 (Ⅰ) mRNA expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 70％,50％,and 38％,respectively ((t =6.316,P ＜0.01 ; t =4.082,P ＜0.01 ; t =3.014,P ＜0.05).Collagen α1(Ⅰ) protein expression in lentiviral vector,new vector,and liposome vector groups was decreased by about 59％,41％,and 27％,respectively (t =4.209,P ＜0.01; t =4.033,P ＜0.01; t =2.842,P ＜0.05).Conclusions The new vector constructed by us has a high transfection efficiency.MiR-29b transfected by the new vector has a good anti-liver fibrosis effect.

Objective To observe the effect of small interfering RNA(siRNA)expression plasmids targeting transforming growth factor p receptor(TαR)Ⅰ gene on the collagen synthesis of hepatic stellate cells(HSCs).Methods Three siRNA expression plasmids were designed and constructed according to TBR Ⅰ sequence.Then the plasmids were transfected into HSC-T6 using 1ipofectamine2000 reagent. The mRNA and protein expressions of TβR Ⅰ were analyzed by reverse transcription polymerase chain reaction(RT-PCR)and Western blot technique, respectively. The cell proliferation was detected using methylthiazo-lyldiphenyl-tetrazolium bromide(MTT)methods. Concentrations of haluronic acid and type Ⅲ pro-collagen in the supernatants were determined by radioimmunoassay. The data were analyzed using least significant difference(LSD).Results Three recombinant plasmids expressing siRNAs were successfully constructed and confirmed by restriction enzyme assay. Compared with the blank control,all the three recombinant plasmids could inhibit the expressions of TβR Ⅰ mRNA,of which plasmid expressing siRNA2 exhibited the strongest inhibitory effect(psiRNA1 group:t=7.354,P＜0.01;psiRNA2 group:t=9.214,P＜0.01;psiRNA3 group:t=5.967,P＜0.01).The expressions of TβR Ⅰ protein were also reduced by all the three recombinant plasmids,of which the plasmid expressing siRNA2 showed the strongest inhibitory effect(psiRNA1 group: t=6.324,P＜0.01;psiRNA2 group:t=8.741,P＜0.01;psiRNA3 group:t=4.128,P＜0.01).The proliferation activity and collagen synthesis of HSCs also decreased in all three HSC groups treated with recombinant plasmids, of which, again, plasmid expressing siRNA2 exhibited the strongest inhibitory effect. However, no significant change was observed in HSCs transfected with non-related siRNA. Conclusion Recombinant plasmids targeting TβR I can inhibit collagen synthesis, which suggests a novel target for gene therapy of liver fibrosis.

Objective To observe the effects of adiponectin on mRNA and protein expressions of connective tissue growth factor (CTGF) in hepatic stellate cells (HSCs) and the levels of procollagen type Ⅲ (PC Ⅲ) and hyaluronic acid (HA).Methods Cultured rat HSCs were treated with different concentrations of adiponectin.CTGF mRNA and protein expressions were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western Blot.The levels of PCⅢ and HA were detected by enzymelinked immunosorbent assay (ELISA).Results Compared with the control group,the result of RT-PCR showed that the four groups had different degrees of inhibitory effect,of which group D exhibited the strongest inhibitory effect.The absorbance ratio was 1.54 ±0.18,1.21 ±0.14,0.96 ±0.10,and 0.79 ± 0.08,respectively (t =2.42,P ＜0.05;t =2.73,P ＜0.05;t =3.28,P ＜0.01;t =4.67,P ＜0.01).Western Blot also indicated that four groups had different degrees of inhibitory effect,of which D group exhibited the strongest inhibitory effect.The ratio of integral absorption was 1.54 ± 0.18,1.21 ±0.14,0.96±0.10,and 0.79 ±0.08,respectively (t =2.84,P ＜0.01;t =4.05,P ＜0.01;t =6.25,P ＜0.01;t =9.72,P ＜0.01).The levels of PCⅢ and HA secreted in culture media were also decreased.It was significantly decreased with the concentration of adiponectin increased.Conclusion Adiponectin can inhibit the levels of PC Ⅲ and HA,which may be achieved through reducing CTGF mRNA and protein expressions.

Objective To isolate and culture splenic CD11clow CD45RBhigh dendritic cells (DC) derived from endotoxin tolerance (ET) mice and investigate its biological characterization.Methods Mice weighed 20 to 25 gram were completely randomized into two groups including ET group and control group with 6 each.ET mice were modeled by intraperitoneal injection of low-dose lipopolysaccharide (LPS) for several days (pretreated with LPS 0.1 μg/mouse for 5 d).Mice in control group were given the same volume of normal saline (NS).CD11clowCD45RBhighDC were isolated from spleen by magnetic activated cell sorting (MACS).The immunological phenotypes were detected by flow cytometry.The suppressive capacity of CD11clow CD45RBhigh DC was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay in allogenic mixed T cells reaction.The expressions of interleukin (IL)-10 and IL-12 produced by CD11clow CD45RBhigh DC were measured by enzyme-linked immunosorbent assay (ELISA).Statistical significance was analyzed through one-way analysis of variance (ANOVA).The homogeneity of variances was detected by Levene test.If variances were homogeneous,the least significant difference (LSD) test was used.If not,Dunnett T3 test was applied.Results The consistence of CD1 1 clow CD45RBhigh DC in control group was 30 ％,reaching the amount of (5.30±0.12) × 105/mouse ;In ET group,the percentage of CD11clow CD45RBhighDC achieved 80 ％ and the production was (1.20 ± 0.13) × 106/mouse the difference was statistically significant (t=3.23,P＜0.01).The cellar morphology in two groups showed no obvious difference.Compared to expression levels of all cell phenotypes (histocompatibility complex-Ⅱ,CD40 and CD80) in normal mice,the cell surface expression levels of CD11clowCD45RBhigh DC in ET mice were much lower.The difference in two groups was statistically significant.Splenic CD11clowCD45RBhighDC derived from ET mice with cell concentration of 1∶ 10,1∶50and 1∶100 had more obvious prohibitory effects on allogenic T cells (t1∶0 =1.36,P1∶10 ＜0.01,t1∶50 =2.49,P1∶50 ＜0.01,1∶100 =1.88,Pm00 ＜0.01).Secretion of IL-10 produced by CD11clowCD45RBhighDC of ET mice was significantly increased (t1∶0=13.63,P1∶10 ＜0.01,t1∶50 =13.45,P1∶50 ＜0.01,t1∶00 =9.31,P1∶00 ＜0.01),but the expression of IL-12 was lower (t1∶0 =2.62,P1∶0 =0.02,1∶∶50 =2.74,P1∶0=0.02,t1∶100 =2.99,P1∶100 =0.01).Conclusion Splenic CD11clow CD45RBhigh DC from ET mice have weaker ability of antigen presenting and allogeneic lymphocytes proliferation stimulating than those from normal mice.

Objective To explore the relations of the doctor-patient relationship to psychological security and stress.Methods Totally 430 operation patients were selected and followed-up.A survey was performed using the Security Questionnaire (SQ),the Patient Doctor Relationship Questionnaire 13-Item Version (PDRQ-13),the Hospital Stress Rating Scale (HSRS) Results The average score of SQ was lower than medium value in operation patients.The SQ scores of operation patients with emergency operation were 39.50±10.51,lower than those of selective operation which were 49.02±10.37 and limited operation which were 47.67±10.29 (P ＜ 0.05).The SQ scores of operation patients who didn't provided themselves were 37.33±10.10,lower than those of basically which were 40.10±10.83 and partly provided for themselves which were 43.50±10.51 (P＜ 0.05).The SQ scores of operation patients in severe group according severity of illness were 37.13±10.40,lower than those of them in mild group which were 39.19±10.24 and moderate group which were 41.38±10.01 (P＜ 0.01).The SQ scores of post-operation patients were 44.32±9.96,higher than post-operation patients which were 40.36±10.81 (P ＜ 0.01).The lack of comforts (LC),lack of information (LI) and risks of serious disease (RSD) scores of post-operation patients were 49.23±30.62,30.46±21.42,52.84±31.22,lower than those of post-operation patients which were 52.37±32.06,35.25±20.14,80.12±33.18 (P ＜ 0.05).The path analysis showed that the regression relationship between SQ scores and PDRQ scores,RSD scores was established (β=0.40,-0.26,P ＜ 0.01),and the PDRQ-13 scores was a mediator between the scores of LC,LI,RSD and the SQ scores (β=0.40,P＜0.01).The PDRQ-13 scores was a adjustor between the RSD scores and the SQ scores (β=-0.34,P ＜ 0.01).Conclusions The operation patients have a low level of the sense of security which is statistically significant with surgical types,the ability to care for themselves and severity of illness.The sense of security of post-operation patients were higher than those of pre-operation patients.The recognition of physician-patient relationship may be play a mediating and adjusting role between the sense of security and stress.The surgical stress affect patient security and physician-patient relationship.

Objective To observe the effect of sustained release type Ⅰ collagen-vascular endothelial growth factor (VEGF) on healing of bone-tendon junction injuries.Methods Partial patellectomy was conducted in 72 rabbits divided equally into control group,type Ⅰ collagen group,and collagen type Ⅰ-VEGF group.The scaffold was planted into the bone-tendon interface.Animals were sacrificed at 4,8 and 12 weeks.New bone formation into the patella-patella tendon surface was detected using X-ray films and histological observations.Quality of bone healing was assayed using biomechanical testing.Results At postoperative 4,8 and 12 weeks,X-ray films showed bone formation of type Ⅰ collagen group [(4.1 ± 0.4) mm2,(12.1 ± 0.5) mm2,(13.0 ± 1.2) mm2 respectively] and of collagen type Ⅰ-VEGF group [(3.8 ± 0.4) mm2,(11.0 ± 0.5) mm2,(13.1 ± 1.0) mm2 respectively] were more than that of control group [(2.1 ± 0.6) mm2,(4.1 ± 0.3) mm2,(6.6 ± 0.6) mm2 respectively] (P ＜ 0.05).Histology identified few new bone,massive fibrocyte accumulation and disrupted alignment of tendon fiber in control group,massive new bone formation,neat and orderly alignment of collagen fiber tissues and massive aggrecan expression at postoperative 4 and 8 weeks (fibrous cartage repair in largely) in collagen type Ⅰ-VEGF group,and massive new bone formation but worse alignment of tendon collagen fibers and less aggrecan expression (fibrous repair in largely) in type Ⅰ collagen group.Biomechanical test showed the ultimate tensile strength increased over time in all groups,with significantly higher value at 12 weeks than that at 4 and 8 weeks.At the same time point,ultimate tensile strength ranged in an order as follows:collagen type Ⅰ-VEGF group ＞ collagen type Ⅰ group ＞ control group (P ＜ 0.05).Conclusion Sustained release type Ⅰ collagen-VEGF can accelerate early healing of bone-tendon junction injury and improve the histological and mechanical properties.

Objective: To study the mechanism of baicalin in inhibiting Mycobacterium tuberculosis（MTB）and to provide reference for drug-resistant tuberculosis treatment. Methods: Forty male Kunming mice were injected isoniazid-resistant MTB into their tail veins to build models of infection. They were evenly divided into MTB group,isophosiazone group,NF-κB inhibition group and baicalicin group according to treatment. The lung tissue and peripheral blood of the mice were collected on the 8th day after modeling. The morphological changes of the lungs were observed by HE staining. The number of MTB in lung tissue was detected by acid-fast staining and quantitative PCR. The number of macrophagein lung tissue was detected by immunohistochemistry. The expression of NF-κB and TLR4 in monocytes/macrophages were detected by flow cytometry. Results: The average weight of mice in the baicalicin group was significantly higher than that in the MTB group,the isophosiazone group and the NF-κBinhibition group（P<0.05）. The average fluorescence intensity of NF-κB and TLR4 in monocytes/macrophages in the baicalicin group were 448.21±30.61 and 401.01±34.58,which were significantly higher than those in the MTB group and the isophosiazone group（P<0.05）. Typical tuberculous chronic granulomatous lesions were observed in the MTB group,isophosiazone group and NF-κB inhibition group,except the baicalin group. The mean number of MTB and CD68+ macrophagesin lung tissue of mice in the baicalin group were significantly less than that in the MTB group,the isophosiazone group and the NF-κB inhibition group（P<0.05）. Conclusion Baicalin achieves an anti-tuberculosis effect by regulating the expression of NF-κB and TLR4 in macrophages,which can be weakened by adding NF-κB inhibitor.

Objective To investigate the correlation between occupational stress,uncertain endur-ance and job burnout of pre-hospital emergency nurses. Methods A total of 380 pre-hospital emergency nurses were recruited by stratified cluster sampling method.The occupational burnout scale(MBI)including emotional exhaustion(EE),depersonalization(D)and diminished personal accomplishment(PA),the In-tolerance of Uncertainty Scale(IUS),the Nursing Stressor Scale(NSS)including stresses associated with nurse expectations(NSS1),with family(NSS2),with interpersonal relationship(NSS3),with nature of work (NSS4),with patient(NSS5)and with work load(NSS6)were applied for all subjects.Results The pre-hospital emergency nurses who had worked less than 3 years got the higher EE score(33.58 ± 4.34)and higher PA score(26.98±4.31)than those who had worked 3 to 10 year(22.33±4.05,16.14±4.28,P<0.05) or longer than 10 years(21.50±4.36,16.81±4.36,P<0.05).Nurses got higher EE score(33.82±4.28,22.96 ±4.39,21.04±4.62,P<0.05),and higher PA scores(27.02±4.21,16.04±4.31,16.01±4.30,P<0.05)than the nurse practitioner or above levels.People whose monthly income were below 0.3 million got higher PA scores than those whose monthly income were 0.3 million or more(26.75±4.24,16.47±4.27,17.11±4.34,P<0.05).People who got degrees of bachelor or above had PA scores which was more than people who got de-grees under bachelor(26.33±4.36,17.24±4.28,P<0.05).Regression analysis showed that the multiplication score between IUS and NSS1 had the interaction effect with EE and D scores(β=0.33,0.33,P<0.05); the multiplication score between IUS and NSS5 had the interaction effect with EE and D scores(β=0.31,0.30, P<0.05)and the multiplication score between IUS and NSS3 had the interaction effect with D scores(β=0.34,P<0.05).Conclusion The pre-hospital emergency nurses'job burnout varies according to their length of service,job title,educational level and salary,and is also influenced by the combination of occupational stress and uncertain endurance.