Objective: To investigate the influence of transfection of TCRVJ37.1 gene in cytotoxicity of PBMC of healthy donor to breast cancer cell line MCF-7/S. Methods :pcDN*31vp'1 packaged by lipofectamine were transfected to healthy donor's PBMC,Flow Cytometer Analysis and MTT Colorimetric Assay were used respectively to test the expression of pcDNA3.1VB7.1 gene and the cytotoxicity of PBMC before and after gene transfection of healthy donor to breast cancer cell line MCF-7/S. Results: The expression of TCRVJ37.1 gene after transfection was obviously higher.There was distinguish differences of cytotoxicity before and after gene transfection. Conclusion:The modified healthy donor's PBMC by TCR gene could have stronger cytotoxicity to breast cancer cell line.

Objective To discuss the cause and treatment strategy for open laryngotracheal trauma.Methods The clinical data of 38 cases with open laryngotracheal trauma admitted from 1998 to 2012 were collected and analyzed.Of them,one patient died in emergency department despite energetic resuscitation,37 were hospitalized and given advanced treatment after life support measures in emergency department.They were treated with tracheotomy before or during operation.Of them,32 patients were regularly followed up for imaging studies.Result Except 1 patient died of exsanguination from carotid artery rupture,the other 37 survived after successful treatment.Twenty patients were treated with debridement and suture of the wounds and laryngoplasty (20/37,54.1％),8 patients were operated with laryngoplasty plus intraluminal stents implanted (8/37,21.6％) ; the rest 9 patients (9/37,24.3％) were separately given placement of nickel-titanium shape memory alloy stent (n =2),laryngofissure with rubber gloves throat models implanted and laryngoplasty (n =3),linear silicone tube implanted and laryngoplasty with trans-cervical approach (n =1) and tracheoesophageal fistula neoplasty with laryngoplasty and laryngofissure (n =3).The operated patients were followed-up for 0.5 to 3 years after discharge.Of them,35 patients (35/37,94.6％) had successful decannulation,breathing smoothly and swallowing normally,23 patients (23/37,62.2％) had almost normal pronunciation,12 patients (12/37,32.4％) had different degrees in hoarseness of voice,and 2 patients (2/37,5.4％) were referred to other hospital because of failure in extubation with severe stenosis of laryngotrachea.Conclusions When the patients with open laryngotracheal trauma were treated,the essential strategy was to prevent shock,hemorrhage,and asphyxia.In the case of patent respiratory tract and stable vital signs,laryngotracheal reconstruction should be carried out as soon as possible to prevent complications,thereby obtaining good therapeutic effect.

ObjectiveTo investigate the cause,diagnosis and treatment measures of neonatal pneumothorax.MethodsThe clinical data of 98 cases with neonatal pneumothorax in our hospital from Mar 2005 to Mar 2010 were reviewed retrospectively.ResultsThere were 27 premature neonates and 71 full-term neonates in the 98 cases.Ten cases were spontaneous pneumothorax and 88 cases were non spontaneous pneumothorax.After the treatnent of primary disease,pleural aspiration,chest drainage and mechanical ventilation,88 cases were cured,8 cases were improved and 2 cases died.ConclusionNeonatal pneumothorax is related with cesarean section,improper use of endotracheal intubation,meconium aspiration,premature hyaline membrane disease using pulmonary surfactant and ventilator therapy.Most pneumothorax is due to lung disease.The prevention of preterm birth and asphyxia can reduce the incidence of neonatal pneumothorax.

Objective To evaluate the therapeutic effects of endoscopic biliary double stents for advanced malignant hilar biliary obstruction.Methods From January 2007 to December 2010,double stents was attempted in 28 patients (15 men and 13 women,median age 66.4 years (44-88 years),including 9 with Bismuth Ⅱ,8 with type Ⅲa,5 with type Ⅲb and 6 with type Ⅳ.A total of 23 consecutive patients ( 11 men and 12 women,median age 65.8 years (42-83 years) with malignant hilar obstruction undergoing a therapy with single stent were recruited as the control group,including 7 with Bismuth Ⅱ,5 with Ⅲa,6 with Ⅲlb and 5 with Ⅳ.The rates of successful drainage,complications,mean survival time of patients and the average duration of biliary stent patency were compared between the two groups.Results Successful rate of cannulation was both 100％ in the two groups.Successful rate of drainage and complications of double stent group were 96.4％ (27/28) and 17.9％ (5/28),and these two variables of single stent group were 87.0％ (20/23) and 13.0％ (3/23),which were not significantly different (P ＞0.05).23 patients (82.1％ ) in double stent and 19 ( 82.6％ ) in single stent group were followed up.The average duration of stent patency and mean survival time of double stent group were ( 129 ±48.5) d and ( 187 ±94.5) d,which were superior to those of the single stent group,i.e.( 102 ±37.8) d and ( 103 ±98.5) d.Conclusion Double stenting is an effective therapy for malignant hilar obstruction of Bismuth Ⅱ and above.It is superior to single stent method in the mean duration of patency and mean survival time.

OBJECTIVE: To evaluate the cost-effectiveness of 3 therapeutic schemes in the treatment of chronic Hepatitis B with YMDD mutation.METHODS: 90 patients were randomly assigned into three groups: the patients in Group A were assigned to receive Adefovir Dipivoxil in combination with Lamivudine for 12 weeks followed by administration of Adefovir alone for 36 weeks;Group B received Adefovir in combination with Lamivudine for 48 weeks,while Group C received Entecavir alone for 48 weeks.The cost-effectiveness of the 3 groups were analyzed.RESULTS: In the three groups(A,B,and C),the cost-effectiveness ratios for HBV DNA negative-conversion ratewere 12 685.6,15 481.3,and 31 462.2,respectively and the incremental cost-effectiveness ratios of Group B and Group C were 29 501.5 and 106 907.8 respectively as against Group A.The cost-effectiveness ratios of the three Groups(A,B,and C) for serum alanine aminotransferase normalization rate were 12 685.6,14 284.9,31 462.2 respectively,and the incremental cost-effectiveness ratios of Group B and Group C were 19 618.5 and 106 907.8 respectively as against Group A.The cost-effectiveness ratios of the three Groups(A,B,and C) for HBeAg/HBeAb seroconversion rate were 76 227.9,93 120.3,and 209 664.0 respectively,and the incremental cost-effectiveness ratios of Group B and Group C were 178 350.0 and 1 267 621.4 respectively as against Group A.In Group A,1 case showed rtA181V mutation and another one showed rtN236T mutation after 48-week treatment.CONCLUSION: Group B(ADV in combination with LAM) is more cost-effective in the treatment of chronic hepatitis B with YMDD mutation.

Objective To analyze the quality of life (QOL) of mothers of children with cerebral palsy and it's influencing factors.Methods The QOL of 123 children with cerebral palsy and their mothers was assessed a 36-item short form health survey (SF-36) and basic questionnaires combined with interviews.The factors influencing QOL were analyzed with t-tests and one way analysis of variance,and multiple regression analysis was used to determine the main influencing factors.Results Scores on all of the SF-36 items were much lower for mothers of children with cerebral palsy than for mothers of normal children.Average scores in the domains of emotional health,general health and vitality were the lowest (55.28,60.49 and 65.26 respectively).Correlation analysis showed that general health,physical function and role emotions were positively correlated with the child's age.All domains except pain were negatively correlated with the child's condition.Social functioning was positively correlated with the mother's age.The general health,role emotional,social function,and mental health scores were all negatively correlated with the mother's education level.Univariate analysis showed that the child's age and condition,the mother's education level and occupation,and the father's occupation all significantly predicted the mother's QOL.Multivariate regression analysis confirmed that the child's age and the father's occupation had significant predictive power.Conclusions The QOL of the mother of a child with cerebral palsy is mainly influenced by the child's age and condition,the mother's education level and occupation,and the father's occupation.Comprehensive and effective measures should be taken to improve the QOL of mothers of children with cerebral palsy and to improve the efficacy of rehabilitation for children with cerebral palsy.

Objective To investigate the killing effect of ethacrynic acid (EA) on lung cancer A549 cells derived spheres and explore the underlying mechanism.Methods A549 spheres were cultured in serum-free medium,and the protein expression of CD133,SOX2,EpCAM and ABCG2 was detected by Western blotting.MTT assay was used to evaluate the cell viability of A549 spheres and A549 cells after treated by 1,2,5,10 and 20 mg/mL cisplatin (DDP) for 48 h.The activity of glutathione S-transferase (GST) was measured by colorimetric method after A549 spheres were treated with 10,50,100 and 200 μmol/L EA,respectively.Flow cytometry,Western blotting,real-time PCR and luciferase assay were used to analyze the levels of cellular reactive oxygen species (ROS),formation of A549 spheres,mRNA and protein expression levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin upon 200 μmol/L EA treated cells for 48 h.A549 sphere was infected with β-catenin adenovirus for 24 h,followed by 200 μmol/L EA treatment (in presence or absence of 5 mg/mL DDP) for 24 h.The expression of β-catenin,Sox2 and ABCG2 at mRNA and protein levels was detected by real-time PCR and Western blotting,and cell growth of A549 spheres was evaluated by MTT assay.Results The A549 spheres,with high expression of tumor stem cells markers CD133,SOX2,EpCAM and drug resistance related molecule ABCG2,and resistance to DDP at different doses,were successfully derived.After 200 μmol/L EA had treated A549 sphere for 48 h,the levels of ROS were significantly increased (P ＜ 0.05),and the mRNA and protein levels of β-catenin,Sox2 and ABCG2,and promoter activity of β-catenin were notably decreased (P ＜ 0.05).The treatment of 200 μmol/L EA enhanced the inhibitory effect on proliferation and the promoting effect on apoptosis in A549 spheres induced by 5 mg/mL DDP (P ＜ 0.05).Up-regulation of β-catenin by adenoviral infection partly reversed the effects of 200 μmol/L EA on suppressing the expression levels of β-catenin,Sox2 and ABCG2,compared to the spheres infected with blank adenovirus.Additionally,β-catenin over-expression significantly remitted the inhibitory effect of 200 μmol/L EA and 5 mg/mL DDP on the proliferation in A549 spheres.Conclusion EA exerts inhibitory effect on the proliferation and stemness of A549 spheres through suppressing GST activity and β-catenin expression,and then promotes cell apoptosis.EA might be a novel drug in treatment of lung cancer and cancer stem cells.

[Objective]To compare the differences of neuromyelitis optica spectrum disorder(NMOSD)and multiple sclerosis (MS) on pregnancy ,and analyze the mutual impact of pregnancy on the diseases.[Methods]Prospectively collected clinical information of 235 NMOSD patients and 125 MS patients ,including the annualized relapse rate (ARR),the Expanded Disability Status Scale(EDSS)and pregnancy outcomes. 70 NMOSD patients and 30 MS patients were screened out as information patients. The ARR and EDSS score in two groups were compared during the year before pregnancy,during pregnancy and after 1 year postpartum, respectively. 50 cases of normal pregnant women for the same period as the control group ,then to compared the difference of three groups on pregnancy outcomes.[Results]Attacks occurring during pregnancy or one year after childbirth/abortion in NMOSD and MS were 53.25%(41/77)and 20.00%(7/35)(P=0.001);The ARR during the first 3 months postpartum periods of NMOSD and MS group(2.65,2.51)was significantly higher than during the year before pregnancy(0.27,0.49,P < 0.001)and during pregnancy (0.32,0.2,P<0.001);The EDSS score of two groups increased after 1 year postpartum(3.06 ± 2.16,2.19 ± 1.28)than that during the year before pregnancy(1.58 ± 0.48,1.92 ± 1.29,P < 0.001)and during pregnancy(1.92 ± 1.35,1.67 ± 0.70,P < 0.001). There was no difference on ARR and EDSS score between NMOSD and MS group. NMOSD ,MS and normal control group had no dif?ference on pregnancy outcomes and neonatal weight.[Conclusions]Compared with MS,the attack of NMOSD had more closer relation?ship with pregnancy;both NMOSD and MS would increase the risk of disease relapsing and disability after pregnancy;the diseases had no effect on pregnancy outcomes.

Object To establish the method for quality control of Fushengkang Capsule. Methods Semen Livistonae Chinensis, Fructus Camptothecae Acuminatae and Rhizoma Curcumae in this capsule were identified by TLC. The content of camptothecine was determined by HPLC. Condition of HPLC was: Dikma Diamonsil C 18 column (250 mm? 4 6 mm, 5 ?m) the mobile phase was a mixture of methanol water (55∶45), the flow rate was 0 8 mL/min, and detection wavelength at 254 nm. Results The spots of TLC were clear and concentrative. There was a good linear relationship for camptothecine within the range of 0 050 6 - 0 253 0 ?g. The average recovery was 96 90%, RSD=1 74% (n=6). Conclusion This method is sensitive, accurate, reproducible and exclusive. It can be used for quality control of Fushengkang Capsule

Objective To investigate the expression of HIF-1α and P-gp protein in pancreatic carcinoma and determine their clinicopathological significance and the correlation between the expression of HIF-1α, P-gp and the clinical prognosis. Method In samples from 74 cases of pancreatic carcinoma and 10 healthy individuals, the expression of HIF-1α and P-gp were detected by immunohistochemical method. Results The positive expression rate of HIF-1α and P-gp was 75.7% and 86.5%,respectively, which were remarkably higher than that in normal pancreatic tissue (P＜0.05). There was a positive correlation between the expression of HIF-1α and that of P-gp. The aberrant expression of HIF-1α and P-gp was associated with lymph node metastasis but not the location, size, clinical stages and nerve invasion of the tumor. Patients with high intensity of HIF-1α and P-gp expression showed a significantly lower median survival time than those with low intensity expression.Conclusions The expression of HIF-1α and P-gp is up-regulated in pancreatic carcinoma and there is a positive correlation between them. The expression of HIF-1α and P-gp might be related to the lymph node metastasis and poor prognosis.