Objective To construct the eukaryotic expression vector for the fusion gene comprising the genes encoding mouse anti-human cross-linked fibrin single-chain fragment variable (scFv) antibody and low-molecular weight single-chain urokinase. Methods The single peptide of recombinant human ipro-urokinase was ligated with the target fusion gene, and the same reading frame was guaranteed. The ligated compound was subsequently incorporated respectively into pcDNA3 and pMJK expression vectors by DNA recombination technique. Results and Conclusion The desired recombinant plasmids pcDNA3 and pMJK3 containing the signal peptide of recombinant human ipro-urokinase gene and the fusion gene were constructed,which lays the foundation for establishing of a cell line that may secrete the fusion protein.

Objective To construct the eukaryotic expression vector for the fusion gene comprising the genes encoding mouse anti-human cross-linked fibrin single-chain fragment variable (scFv) antibody and low-molecular weight single-chain urokinase. Methods The single peptide of recombinant human ipro-urokinase was ligated with the target fusion gene, and the same reading frame was guaranteed. The ligated compound was subsequently incorporated respectively into pcDNA3 and pMJK expression vectors by DNA recombination technique. Results and Conclusion The desired recombinant plasmids pcDNA3 and pMJK3 containing the signal peptide of recombinant human ipro-urokinase gene and the fusion gene were constructed,which lays the foundation for establishing of a cell line that may secrete the fusion protein.

Objective To evaluate the feasibility and safety of vaginal enlarged amputation of cervix to treat patients with cervical cancer of stage Ⅰ a1 and cervical intraepithelial neoplasia grade Ⅲ(CIN Ⅲ)who were unfit for conization surgery.Methods From July 2002 to May 2007,patients with cervical cancer at stage Ⅰ a1,diagnosed by pathology after loop electrosurgical excision procedure(LEEP),large area CIN Ⅲ(the area of lesion≥3/4 on colposcopy),CIN Ⅲ coexisted with vaginal intraepithelial neoplasia (VAIN)in the superior segment of vagina,CIN Ⅱ-Ⅲ recurrence or with residual lesion,positive margin after conization of cervix,who wanted to preserve fertility and(or)corpus uteri were selected to receive vaginal enlarged amputation of cervix.Results Forty-eight eases including 5 with cervical cancer in stage Ⅰ a1,38 with large area CIN Ⅲ(9 with gland involvement),2 with residual lesion and 2 with positive margin after LEEP,1 recurrence after cold knife conization,received the procedure successfully.The median age was 34 years(range 27-40),median operation time was 60 minutes(range 30-100),median blood loss was 40 ml(range 5-300),and median hospital stay was 10 days(range 7-17).After follow-up 1-39 months,no patient had postoperative complications and recurrence,and all patients resumed normal menstrual cycle and sexual life.Condusion Vaginal enlarged amputation of cervix appears to be a safe and feasible procedure for patients with cervical cancer at stage Ⅰ a1 and CIN Ⅲ who are unfit for conization surgery.

Objective To investigate the proportion and function of CD_4~+ CD_(25)~+ regulatory T cells (CD_4~+ CD_(25)~+ Tr)in unexplained recurrent spontaneous abortion(URSA).Methods(1)Proportion measurement:the proportion of CD_4~+ CD_(25)~+ Tr cells in peripheral blood was measured by double-label flow cytometric analysis.The samples were taken from 15 URSA women,15 normal non-pregnancy women and 13 normal pregnancy women.(2)Function measurement:CD_4~+ CD_(25)~+ Tr ceils and CD_4~+ CD_(25)~+ T ce]ls were extracted from peripheral blood lymphocytes by the microbeads separation.The purity of CD_4~+ CD_(25)~+ Tr cells and CD_4~+ CD_(25)~+ T cells was measured by flow cytometry.The growth inhibitory effect of CD_4~+ CD_(25)~+ Tr cells on CD_4~+ CD_(25)~+ T cells was assessed in vitro.Results The proportion of CD_4~+ CD_(25)~+ Tr cells was decreased significantly in URSA women(6.9?1.8)% than that in normal non-pregnancy women[(10.8?1.1)%] (P0.05).Conclusion The results suggest that decrease in proportion and function of CD_4~+ CD_(25)~+ Tr cells may be associated with URSA.

The optimization methods in radiation treatment planning are reviewed in this paper, including the physical and biological optimization models, the optimization for Gamma knife treatment planning, the optimization for intensity modulated radiation treatment planning and the optimization for intravascular brachytherapy treatment planning. The development trend of radiation treatment planning is also introduced in the paper.
Antineoplastic Protocols ; Humans ; Models, Biological ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; methods ; trends

OBJECTIVETo study the heterogeneity of polymerase gene (P gene) within hepatitis B virus (HBV) genotypes based on a systematic analysis of 202 HBV P genes, providing some useful references for further studies on the relationship among HBV genotypes, P gene mutations, replication and nucleoside analogues drug-resistance.

METHODS202 HBV complete sequences containing P genes were obtained from GenBank and were analysed using computer softwares.

RESULTSThere were some genotype-related characteristics of HBV P genes. As reverse transcriptase domain was concerned, there were more amino acid divergences in genotype C and D compared with these in genotype A. There were also amino acid substitutions in the A-F conserved regions of the reverse transcriptase domain within and between HBV genotypes.

CONCLUSIONSThere are divergences of P genes and amino acids within and between HBV genotypes, which should be considered when amino acid changes are analyzed whether they are proposed to be drug-resistance mutations or the results from quasispecies-selected. Moreover, these divergences may affect the antiviral effect of nucleoside analogues on HBV with different genotypes.

Amino Acid Sequence ; DNA, Viral ; genetics ; DNA-Directed DNA Polymerase ; genetics ; Gene Products, pol ; genetics ; Genetic Heterogeneity ; Genotype ; Hepatitis B virus ; genetics ; physiology ; Humans ; Molecular Sequence Data ; Mutation ; Phylogeny

OBJECTIVETo prepare and identify a polyclonal antibody (pAb) against (mouse) cysteinyl leukotriene receptor 1 (CysLT(1)) and to investigate the changes of CysLT(1) receptor expression in BV2 microglial cells after rotenone treatment.

METHODSRabbits were immunized with KLH-coupled CysLT(1) peptide to prepare the pAb. The titer of the pAb in rabbit plasma was detected by ELISA method, and the specificity of the pAb was tested by antigen blockade. After BV2 cells were treated with rotenone (0.01-1 μmol/L) for 24 h, the expression of CysLT(1) was determined by immunostaining, Western blotting and RT-PCR.

RESULTThe pAb showed a titer of 1/32728, and was not cross-reacted with antigens of CysLT(2) receptor and GPR17. Immunostaining, Western blotting and RT-PCR analysis showed the expression of CysLT(1) receptor in BV2 microglia. Rotenone at 1μmol/L significantly induced an increased expression of CysLT(1) receptor.

CONCLUSIONThe prepared CysLT(1) receptor polyclonal antibody has a high titer and high specificity to meet testing requirements of Western blotting and immunostaining; CysLT(1) is associated with rotenone-induced injury of BV2 microglial cells.

Animals ; Cells, Cultured ; Male ; Mice ; Microglia ; drug effects ; metabolism ; pathology ; Rabbits ; Receptors, Leukotriene ; immunology ; metabolism ; Rotenone ; pharmacology

OBJECTIVETo evaluated the safety and efficacy of hyperthermic intraperitoneal perfusion chemotherapy(HIPC) in the prevention and treatment of pseudomyxoma peritonei (PMP) recurrence after cytoreductive surgery(CRS).

METHODSStudies published in English before 2010 on HIPC after CRS for PMP were searched in PubMed database. Each study was carefully evaluated based on pre-determined criteria. Study results were comprehensively displayed in a form. A descriptive systematic review was performed.

RESULTSA total of 11 studies were included. The median survival time of patients in these studies ranged from 25.6 months to 156 months. The ranges of 1-year, 2-year, 3-year, 5-year, and 10-year survival rates were 72%-100%, 55%-96%, 59%-96%, 52%-96%, and 55%-96%, respectively. The overall complication rate ranged from 2%-15%, and the total perioperative mortality were from 0 to 7%.

CONCLUSIONHIPC after CRS is effective and safe for patients with PMP.

Chemotherapy, Cancer, Regional Perfusion ; methods ; Humans ; Peritoneal Neoplasms ; drug therapy ; surgery ; Postoperative Care ; Pseudomyxoma Peritonei ; drug therapy ; surgery ; Treatment Outcome

OBJECTIVETo evaluate the access to the frontal recess by identifying the agger nasi cell and uncinate process.

METHODSForty-seven patients (85 sides) who underwent endoscopic frontal sinus surgery in our department constituted the study population. Computed tomographic (CT) scans of the sinuses were obtained in coronal and axial views. The frontal ostium was identified by using agger nasi cell approach or identifying the uncinate process.

RESULTSThe frontal sinus ostium was identified in 100% of patients (85 sides). After an average follow-up of 9 months, 41 sides of 49 sides (84%) had endoscopically healed sinuses by using agger nasi cell approach. And 21 sides of 36 sides (81%) had endoscopically healed sinuses by identifying the uncinate process.

CONCLUSIONSThe agger nasi cell approach to the frontal recess gives an access and allows identification of the frontal ostium. In addition, it provides direct visualization with a 0 degree endoscope into the frontal recess.

Adult ; Chronic Disease ; Endoscopy ; methods ; Female ; Frontal Sinus ; surgery ; Humans ; Male ; Middle Aged ; Nasal Cavity ; Nose ; surgery ; Paranasal Sinuses ; Sinusitis ; surgery

Objective To investigate the application value of limited liquid resuscitation combined with continuous renal replacement therapy (CRRT) in treatment of severe acute pancreatitis (SAP) companied with abdominal compartment syndrome (ACS).Methods The retrospective cohort study was adopted.The clinical data of 67 patients with SAP companied with ACS who were admitted to the Fuling Center Hospital of Chongqing from January 2005 to December 2014 were collected.Among 67 patients,33 receiving conventional liquid resuscitation between January 2005 and December 2010 were allocated into the control group and 34 receiving limited liquid resuscitation combined with CRRT between January 2011 and December 2014 were allocated into the observation group.Observation indicators included:(1) required fluid volume and time of negative fluid balance in the 2 groups;(2) changes of pathological and physiological indicators after treatment in the 2 groups;(3)outcomes and prognosis of patients in the 2 groups.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using t test.Count data were analyzed using the chi-square test.Repeated measurement data were analyzed by repeated measures ANOVA.Results (1) Required fluid volume and time of negative fluid balance in the 2 groups:required fluid volumes at 6,24,48 and 72 hours after resuscitation were (2 449±339) mL,(4 820±757) mL,(9 428± 1 272) mL,(13 127± 1 565) mL in the control group and (2 360± 314) mL,(4 582±530) mL,(8 564± 970) mL,(11 470± 1 253) mL in the observation group,respectively,with a statistically significant difference in required fluid volume between the 2 groups (F=13.035,P＜0.05) and in required fluid volume at 48 and 72 hours between the 2 groups (t=3.132,4.794,P＜0.05).Time of negative fluid balance in the observation group and control group was (4.3± 1.7)days and (6.4 ±1.8)days,respectively,showing a statistically significant difference between the 2 groups (t =4.913,P＜0.05).(2) Changes of pathological and physiological indicators in the 2 groups after treatment:time factors:from 0 h to 168 h postoperatively,APACHE Ⅱ score,C-reaction protein (CRP),D-dimer,IAP,Bla and oxygenation index were changed from 20.9±4.1 to 13.9±2.6,from (167±39)mg/L to (55±17) mg/L,from (1 652±1 544) μg/L to (993±500)μg/L,from (23.4±3.4)cmH2O (1 cmH2O=0.098 kPa) to (21.4±2.9)cmH2O,from (4.6±1.6) mmol/L to (1.4±0.5)mmol/L,from (189±27) mmHg (1 mmHg =0.133 kPa) to (152±23)mmHg in the control group,and chaged from 21.5±5.1 to 11.0±2.8,from (168±36)mg/L to (44±19)mg/L,from (1 634±1 525) μg/L to (578±350) μg/L,from (23.2±2.5)cmH2O to (17.4±2.6)cmH2O,from (4.5±1.6)mmol/L to (0.8±0.3)mmol/L,from (188±26)mmHg to (196±25)mmHg in the observation group,respectively,showing gradual decreasing with time and statistically significant differences between the 2 groups (F =186.415,581.118,34.618,212.416,262.272,207.645,P＜0.05).Treatment factors:there were no significant differences in changing trends of APACHE Ⅱ score,D-dimer and Bla between the 2 groups (F=3.499,2.350,3.516,P＞0.05),and there were significant differences in changing trends of CRP,IAP and oxygenation index between the 2 groups (F=4.009,15.276,14.959,P＜0.05).Interaction effect between time factors and treatment factors:there were obviously interaction effects between time factors and treatment factors in APACHE Ⅱ score,CRP,IAP and oxygenation index (F=4.890,4.912,23.874,28.615,P＜0.05) and no interaction effects between time factors and treatment factors in D-dimer and Bla (F=2.803,1.920,P＞0.05).(3) Outcomes and prognosis of patients in the 2 groups:numbers of patients with surgery,local complications and infection and duration of hospital stay were 11,16,14,(46±17)days in the control group and 4,6,6,(36±14) days in the observation group,respectively,with statistically significant differences between the 2 groups (x2=4.484,7.221,4.910,t =2.433,P＜0.05).Mortality and hospital expenses were 24.2％ (8/33),(33± 18) x 104 yuan in the control group and 8.8％ (3/34),(27± 14)× 104 yuan in the observation group,respectively,with no statistically significant difference between the 2 groups (x2 =2.901,t =1.283,P＞ 0.05).Conclusion Limited liquid resuscitation combined with CRRT can effectively control IAP of patients with SAP companied with ACS and improve oxygenation index,meanwhile,it can also reduce number of patients with surgery,infection and local complications and duration of hospital stay.