ObjectiveTo compare the clinical results of small surgical incision of bilateral spinous process and traditional open surgical incision in posterior single level lumbar interbody fusion,and assess the application value of the small surgical incision of bilateral spinous process in posterior single level lumbar interbody fusion.MethodsFrom December 2006 to June 2008,70 patients with lower lumbar vertebral diseases underwent single segment posterior lumbar interbody fusion.Patients were randomly divided into small surgical incision of bilateral spinous process group(Hereinafter referred to as small incision group) of 36 cases and 34 cases of conventional open group.Small incision group included 20 males and 16 females with an average age of 52.0 years.Traditional open group included 16 males and 18 females with an average age of 53.2 years.Two groups of operative time,blood loss,postoperative drainage,X-ray projection operation frequency,hospital stay,complication rate,creatine phosphokinase (CPK) level,multifidus cross-sectional area,postoperative low back pain visual analogue scale(VAS),Oswestry disability index(ODI),interbody fusion rate were compared,respectively.ResultsAll of 70 cases were followed up for 12-24 months (average,16).There were no statistically differences in the operation time,the number of X-ray projection,complication rate,and fusion rate between the two groups (P＞0.05),but there were significant differences in blood loss,postoperative drainage,the first day and the third day of postoperative the level of CPK,postoperative multifidus muscle cross-sectional area,postoperative low back pain VAS,hospital stay,and postoperative ODI between the two groups(P＜0.05).ConclusionSmall surgical incision of bilateral spinous process and traditional open surgical posterior lumbar interbody fusion were satisfied with the efficacy,but small surgical incision of bilateral spinous process with less trauma,shorter hospital stay,and rapid postoperative recovery.

Objective To investigate the relationship between interaction of peroxisome proliferators-activated receptor alpha (PPARα), cytochrome P450 oxysterol 7α-hydroxylase (CYP7B1) and estrogen receptor (ER) and intrahepatic cholestasis in pregnant rats. Methods Eighty clean SD pregnant rats were selected and divided into four groups randomly with 20 in each. Since the 13th day of pregnancy,rats in the control group was injected subcutaneously with refined vegetable oil 2.0 ml · kg-1 · d -1 , those in the low-dose, moderate-dose and high-dose groups received 17-α-ethynylestradiol (EE) 1.0 mg · kg-1 · d-1,1.25 mg · kg-1 · d-1 and 1.5 mg · kg-1 · d-1, respectively. All rats were sacrificed at the 21at day of pregnancy and maternal hepatic tissues were collected. The serum levels of alanine aminotransferase(ALT), aspartate transaminase (AST), total bile acid (TBA) and bilirubin (BIL) were determined by enzyme linked immunosorbent assay (ELISA). The mRNA expressions of PPARα, CYP7B1, Erα and Erβ in maternal rat livers were examined by real-time PCR. Results (1) Biochemical indicators: the serum levels of ALT,AST, TBA and BIL were significantly lower in the control group than in the rest 3 groups,respectively [ control group: (41.1 ± 2.8 ) U/L, (44.4 ± 3.6) U/L, (26.4 ± 5.6 ) μmol/L and( 2.8 ± 0.2)U/L;low-dose group: (48.2 ±3.4) U/L,(47.9 ±3.7) U/L,(36.4 ±4.2) μmol/L and (4.2 ±0.2) U/L;moderate-dose group: (70.4 ± 5.3 ) U/L, (68.4 ± 5.6) U/L, (64.3 ± 3.8 ) μmol/L and ( 6.2 ± 1.2)U/L; high-dose group: (72.4 ±7.6) U/L, (70.2 ±3.8) U/L, (72.4 ±7.8) μmol/L and (8.2 ±2.2)U/L, P<0.05], and those in the moderate or high-dose groups were higher than in the low-dose group (P<0.05). (2) mRNA expression of Erα and Erβ: the mRNA expression of Erα in pregnant rat livers increased in a dose-dependent manner, which were all significantly higher than that in the control group,respectively ( low-dose group: 0.76 ± 0.02 ); moderate -dose group: ( 0.99 ± 0.04; high-dose group:1.21 ±0.01 ;control group:0.65 ±0.01, P <0.05), but no difference was found among the 4 groups in the mRNA expression of Erβ ( P > 0.05 ). (3) mRNA expression of CYP7B1 and PPARα: the mRNA expression of CYP7B1 in pregnant rat livers increased from the low-dose group to the high-dose group, and were all higher than that of the control group ( low-dose group: 0.93 ± 0.01; moderate-dose group: 0.99 ±0.06; high-dose group: 1.22 ± 0.04; control group: 0.75 ± 0.02, P < 0.05 ). However, the mRNA expression of PPARα decreased from the low-dose group to the high-dose group, and were all lower than that of the control group (low-dose group: 0.83 ± 0.05; moderate-dose group: 0.71 ± 0.02; high-dose group:0.64 ± 0.03; control group: 1.35 ± 0. 05; P < 0.05 ) . Conclusions The down regulated mRNA expression of PPARα, caused by higher dose of estrogen, may increase the expression of CYP7B1 due to the ineffectiveness of the inhibition of PPARα on CYP7B1, which may further stimulate the Erα activity and then induce intrahepatic cholestasis. Abnormal expression of PPARα, CYP7B1 and ER may play a role in the pathogenesis of estrogen-induced intrahepatic cholestasis.

Purpose　The aim is to establish fermentation process of new type recombinant human necrosis factor (nrhTNF)α.Methods　The optimized host cell,culture medium and induction time were deternined according to the growth and expression specificity of nrhTNF on test tube and flask shaker. Then fed-batch culture was carried out on 5 L automatic fermentor.Results　The expression of nrhTNF in fermentor could keep in a higher level of about 50% of total bacterial proteins when fermented to around A600nm 30，with a fermentation time of 12 h to 13 h.Conclusion　A short cycle,high expression and stabilized fermentation process was established.

Objective To explore the effect of the 3rd generation Gamma nail in the treatment of intertrochanteric fractures.Methods 32 cases with intertrochanteric fractures were treated with the third generation Gamma nail internal fixation.Results Mter follow-up,the excellent and good rate was 87.6％.Conclusion The third generation Gamma nail treatment is a good internal fixation for intertrochanteric fracture of femur.

Objective To investigate the therapeutic effect of the complicated fracture of tibial plateau treated by open reduction and internal fixation. Methods Surgical treatment in 68 cases with complicated fractures of the tibial plateau were analyzed retrospectively, T type plate, L type buttress plate, double plates, anatomical plate,locking plate and less invasive stabization system were adopted for different fractures,iliac or artificial bone grafted in the reconstruction of bone defect after the depression replaced. The time of operation and the effect of functional exercise in the postoperative patients to recover the function of the knees were analyzed at the same time. Results 52 patients of 68 cases were followed-up. Based on Rasmussen rating system,the recovery of knee joint function were excellent in 42cases(67%) ,good in 12 cases(19%) ,fair in 6 cases(10%) and poor in 2 cases(4%) ,the excellent-and-good rate was 86%. Conclusion Surgical treatment of the complicated fractures of the tibial plateau depended on the kind of fracture, therapeutic principles by stages. Moreover, bone grafte after the depression replaced in bone defect was neeassary,good internal fixtion oould meet functional exercise,soft tissue injuries like ligament and meniscus should be well treated,follow the principles of early exercise and delay weightbearing.

Objective To construct a recombinant bacillus Calmette-Guérin(BCG) vaccines based on different tandem repeats of MUC1 and GM-CSF, rBCG-MVNTR1/4/8-CSF, and to observe the ability of three recombinant BCG vaccines in the inhibition of breast cancer. Methods After MUC1 variable-number tandem repeats (MVNTR1/4/8) were cloned in a stepwise manner, the E. coli-Mycobacteria shuttle expression vector pDE22-MVNTR1/4/8-CSF were constructed by fusing MVNTR1/4/8 and GM-CSF, and then used to transform competent BCG by electroporation after identification by restriction endonuclease digestion analysis and DNA sequencing. A novel breast cancer vaccines, rBCG-MVNTR1/4/8-CSF was constructed. The expression of fused MVNTR1/4/8-CSF protiens was analyzed by SDS-PAGE and Western blot. The ability of rBCG vaccines inhibiting the growth of breast cancer was observed in hu-PBL-SCID mice. The specific T cell responses in mice were assessed by immunohistochemistry. Results The expression of recombinant MVNTR1/4/8-CSF fusion proteins were detected by SDS-PAGE and Western Blot in rBCG-MVNTR1/4/8-CSF vaccines, respectively. Tumor incidence in mice prophylactic immunized with rBCG-MVNTR4-CSF (37.5%) or rBCG-MVNTR8-CSF (25%) significantly decreased compared with PBS and BCG-pDE22 control ( 100% ) at 42 days after tumor implantation ( P ＜ 0. 05 ). MCF-7 tumor growth inhibition in rBCG- MVNTR4/8-CSF-immunized mice was more significant than that in controls ( P ＜0. 01 ).The inhibition effect of three rBCG vaccines on breast rumor growth appeared to rise with increase of numbers of the tandem repeats of MUC1. Survival rate was 75% of mice in the rBCG-MVNTR4-CSF-treated group and 87. 5% of mice in the rBCG-MVNTR8-CSF-treated groups at 70 days after tumor implantation; however,survival rate was only 12. 5% in control group( P ＜0. 05). The CD4-positive and CD8-positive lymphocytes were detected only in rBCG-MVNTR4/8-CSF-immunized mice. Conclusions rBCG-MVNTR4/8-CSF immunization inhibits breast cancer growth in mice.

BACKGROUND: Previous studies demonstrated that construction of eoexpression plasmid containing multiple genes on the same vector could improve transfection and expression rates.OBJECTIVE: To construct eukaryotic expression plasmid pcDNA3.1 (+)-MUC1 -GM-CSF by human polymorphic epithclial mucin (MUC 1) and granulocyte macrophage colony stimulating factor.(GM-CSF) and to observe expression of recombinant plasmid in COS-7 cells.DESIGN,TIME AND SETTING: Gene recombination design,which was carried out in the Animal Central Laboratory,the Fourth Military University of Chinese PLA from September 2005 to December 2006.MATERIALS: Eukaryotic expression vector pcDNA3.1 (+) was presented by Pro.Taylor-Papadimitriou;pGEM-3zf()-GM-CSF plasmid,COS-7 cells,pUCI 8 vector,and E.coli DH5α were made in the center; female BALB/c mice were provided by Experimental Animal Center of the Fourth Military University of Chinese PLA.METHODS: Signal peptide was synthesized with encoded MUCI gene sections to obtain repeated sequence coneatemer after renaturation.Next,the accepted concatemer was cloned with GM-CSF following enzyme identification and sequencing analysis,and then they were put in eukaryotic expression vector pcDNA3.1(+) to construct eukaryotic coexpression plasmid pcDNA3.1 (+)-MUCI -GM-CSF in order to transform COS-7 cells.MAIN OUTCOME MEASURES: Gene expression was detected by indirect immunofluorescence and enzyme linked immunosorbent assay (ELISA).RESULTS: Enzyme identification and sequencing analysis showed that recombinant plasmid contained a fusion gene encompassing human MUC1 repeated sequence concatemer and GM-CSF; moreover,MUC1 expression was detected in COS-7 cells,while recombinant plasmid could induce the production of anti-GM-CSF antibody.CONCLUSION: The recombination between human MUC1 repeated sequence concatemer and GM-CSF gene successfully constructs eukaryotic coexpression plasmid.

Objective To compare the clinical effect of arthroscopic single-row and suture-bridge double-row fixation of full-thickness rotator cuff tear.Methods A retrospective study was conducted on 40 patients with full-thickness rotator cuff tear treated by single-row and suture-bridge double-row fixation between May 2010 and May 2012.Single-row rotator cuff repair was performed in 18 patients and doublerow repair in 22 patients.The two techniques were compared in aspects of complications,recovery time to daily life,time to achieve satisfaction score,re-tear rate,pre-and post-operative VAS,University of California at Los Angeles Shoulder Scores (UCLA),and American Shoulder and Elbow Surgeons (ASES).Results Mean period of follow-up was 15.6 months (range,13-24 months).Both treatment were effective in improving VAS,UCLA,and ASES,but the differences were insignificant (P ＞ 0.05).Recovery time to daily life,time to achieve satisfaction score,and large rotator cuff re-tear rate were 69.2 ± 16.6)d,(196.4 ±24.6)d,and 4/5 respectively in single-row fixation,with significant differences from (54.1 ± 13.3)d,(165.0 ±20.6)d,and 1/8 in suture-bridge double-row fixation (P ＜0.05).Conclusion Both treatments are effective for rotator cuff reconstruction,but suture-bridge double-row fixation provides earlier recovery to daily life,shorter time to achieve satisfaction score,and lower incidence of large rotator cuff retear.

Aim To express the fusion protein hTNFα -hPgnK5 in E.Coli and to identify its activity. Methods The hTNFα -hPgnK5 gene express vector was constructed and the fusion protein was expressed in E.coli. The activity of fusion protein to inhibit the proliferation of tumor cells or angioendothelial cells in vitro was determined by MTT. Results Fusion protein hTNFα -hPgnK5 possessed immunoactivity in Western blot analysis and could inhibit proliferation of tumor cells or angioendothelial cells in vitro test. Conclusion The hTNFα gene and hPgnk5 gene can be coexpressed in prokaryotic system and expression products can inhibit the proliferation of tumor cells or angioendothelial cells in vitro.

ObjectiveTo investigate the expression of MUC1 in thyroid cancer and benign thyroid diseases (nodular goiter and adenoma) and its clinical significance. Methods The expression level of MUC1 was examined by immunohistochemical analysis with MUC1 monoclonal antibody in 68 samples of thyroid carcinoma and in 18 samples of benign diseases and 10 normal thyroid tissues. Results Immunohistochemical analysis showed positive expression of MUC1 in 51 out of 68 cases of thyroid cancer, in 4 out of 18 cases of benign thyroid diseases and 1 out of 10 normal control thyroid tissue, with a difference statistically significant between the malignancy and benign and normal control (? 2=17.20,P0.05).Conclusion The over-expression of MUC1 in thyroid cancer may act as diagnostic markers of thyroid carcinoma.