ObjectiveTostudytheinhibitioneffectof c-mycASODN(antisense oligodeoxynucleotide) and 5-FU (5-fluorouracil) on the expression of c-myc gene and the proliferation of human hepatomacellsHEPG-2. MethodsAfter treatedbyliposomemediatedc-mycantisense phosphorothioate oligodeoxynucleotide (APSODN) and 5-FU, the growth inhibition rate was detected by MTT assay, the expression of c-myc mRNA was detected by RT-PCR and immunohistocehemical methods HEPG-2cells. The cell cycle was analyzed by flow cytometric analysis. The morphological changes were observed by fluorescence staining and cellular genome electrophoresis. ResultsAfter sealing c-myc gene with ASODN,the growth of cells was repressed and the effect was time-dependent and dose-dependent ( P = 0. 02 ). The ability of proliferation decreased, the expression of c-myc gene was inhibited on transcription and translation levels; 5-FU can induce apoptosis of hepatoma cells HEPG-2 dramatically with the dose of 10 μ mol/L, when treated by both c-myc ASODN and 5-FU, HEPG-2 cells was induced apoptosis in a cooperative style ( P =0. 01 ).ConclusionsThe liposome mediated c-myc (APSODN) and 5-FU can inhibit the proliferation of HEPG-2 cells by inhibiting the expression of c-myc gene and can induce apoptosis of hepatoma cells HEPG-2 in a cooperative style. c-myc ( APSODN ) can increase the sensitivity of 5-FU to hepatoma cells and decrease the effective concentration of 5-FU.

BACKGROUND:Ribbed anatomic femoral stem meets the biomechanical features of the femur, and has achieved satisfactory clinical efficacy with good initial stability, rapid bone growth and low stress shielding.
OBJECTIVE:To evaluate the design features and clinical results of the cementless anatomic Ribbed femoral stem (Ribbed stem) in total hip arthroplasty.
METHODWe retrospectively analyzed 52 patients (52 hips) with hip joint disease who were treated with Ribbed stem in total hip arthroplasty between March 2010 and March 2012. There were 20 males and 32 females. The mean age was 59 years (range 22-78 years). The mean fol ow-up was 3.1 years (range 2-4 years). The preoperative and postoperative Harris hip scores, the rate of postoperative thigh pain, the incidence of intraoperative femoral fracture, wound healing, dislocation and revision were recorded. Radiographic evaluation was used to evaluate the rate of stem fil , subsidence of femoral stem, periprosthetic radiolucent line, bone ingrowth, stress shielding, and osteolysis.
RESULTS AND CONCLUSION:The incidence of intraoperative femoral fracture was 6%. Al incisions were healed. There were no cases of hip dislocation and revision;the rate of thigh pain was 6%, no patient had thigh pain after 1 year postoperatively. The mean preoperative Harris hip score was 48 points, which improved to a mean of 96 points at the final fol ow-up. Postoperative X-ray showed that al patients had a satisfactory femoral fil in both planes and al of the femoral stems were wel-fixed at the final fol ow-up. The average fil ing rate was 91%, 88%and 86%by normotopia imaging, while 88%, 85%and 81%by lateral imaging, at the metaphysic, middle and distal ends respectively. At the final fol ow-up, 49 hips (92%) showed stable bone ingrowth, 3 hips showed stable fiber ingrowth. No osteolysis around the components at the femur or acetabulum was observed. 5 (10%) hips appeared the subsidence of femoral stems, which were<2 mm. The stress shielding phenomenon occurred in al patients, including first-degree in 31 hips, second-degree in 19 hips, third-degree in 2 hips, and fourth-degree in no case. Experimental findings indicate that, Ribbed stem with a rational design can achieve satisfactory clinical outcomes with good initial stability, rapid bone growth and low stress shielding.

Objective To develop a one-step PCR assay for rapid discrimination of six Brucella species and some intraspecific biovars.Methods Using 6 pairs of primers in one-step PCR to differentiate six classical Brucella species and some biovar in ordinary PCR instrument.The tested strains including 27 reference strains of six Brucella species and 239 Brucella strains were estimated by the PCR assay and biological identification methods.Results The six Brucella species could be precisely differentiated by the onestep PCR assay from the tested strains.Five biovars and vaccine strain of B.suis species could be determined,and biovars 1,3,4 and biovars 2,5,6,7,9 of B.abortus species could be identified at the level of their biovar,moreover,biovars 1,2 and 3,and vaccine strain Rev 1 of B.melitensis species were also discriminated at the biovar and strain level.The accurate rates of the biological identification method and the PCR assay were 98.33％ and 100％ respectively.Conclusion One-step PCR assay was a rapid,specific,and low cost method for identification of Brucella species and discriminating biovars in ordinary PCR instrument.

We constructed prokaryotic recombinant expression vector of P61 gene from Nocardia brasiliensis,expressing P61 protein with biological activity in E.coli,and lay a foundation for further studies related to P61.P61 gene was synthesized and cloned into an expression vector pET-30a(+).The recombinant vector was transformed into Escherichia coli BL21 and induced with IPTG.The production was analyzed with Western blot and the catalase activity of P61 was tested with Catalase Assay Kit.The protein of P61was successfully expressed in E.coli with solubility and high catalase activity,and could be identified by anti-N.brasiliensis sera from mice.The prokaryotic expression plasmid of protein P61 was constructed successfully and can be expressed efficiently in E.coli BL21 cells with higher catalase.

Objective To evaluate the mid-and long-term clinic results of ceramic-on-ceramic joint in total hip arthroplasty (THA) in younger patients.Methods A retrospective review was made on 72 younger patients who had THA with ceramic-on-ceramic bearings due to femoral head necrosis or inborn hip dysplasia between March 2001 and March 2011.There were 32 females and 40 males,at a mean age of (35.2 ± 7.6)years (range,17-45 years).Patients were evaluated with Harris hip score and University of California,Los Angeles (UCLA) activity score.Radiological findings were recorded including component loosening,ostelysis,and ceramic bearing related complications.Results Duration of followup was (8.5 ±2.4) years (range,5-12 months).At the final follow-up,Harris hip score was increased to (93.8 ±4.3)points (range,85-100 points) compared to preoperative (46.1 ± 12.3) points (range,27-70 points) and UCLA activity score was increased to (7.1 ± 1.4)points (range,4-10 points) compared to preoperative (4.0 ± 1.2) points (range,2-6 points) (P ＜ 0.05).At the final follow-up,no osteolysis or prosthesis loosening occurred,but there were ceramic sandwich liner fractures in two patients (two hips),dislocation in one patient (one hip) and squeaking in one patient (one hip).After revision due to the ceramics break,the 8-year survival rate was 97％ (95％ CI91.9-100.0).Conclusion Ceramicon-ceramic joint in THA satisfies the activity demand in younger patients,with excellent mid-and longterm clinical results.

BACKGROUND:Femoral neck fracture is mainly fixed by three inverted triangle cannulated screws. Scholars have proposed to add a cannulated screw to enhance the fixation strength of femoral neck fracture of Pauwels III type based on three cannulated screw fixation, but the stability is not verified. OBJECTIVE:To analyze the biomechanical stability and stress of the three and four cannulated screws for the treatment of the Pauwels III femoral neck fractures. METHODS:The CT imaging results of the fourth generation of artificial bone sawbones were imported into the Mimics software wherein a three-dimensional finite element model of the proximal femur was prepared and introduced in the 3-matic software. Models of middlesegment of femoral neck with Pauwels III fractures were established. Cannulated screw models were established with UG 8.0 software and introduced in the fractures models. Finally, finite element models of Pauwels III femoral neck fractures fixed with three and four screws were established. In the same condition, an axial load of 411 N was applied on the femoral head with Abaqus software. The displacement of two markers of the broken ends and internal fixation system Von Mises stress distribution were compared between the two models. RESULTS AND CONCLUSION:(1) The displacement was 0.42 mm in three screws model, and 0.17 mm in the four screws model. (2) Von Mises stress peak was 547 MPa and 27.8 MPa in both models. The peak value was lower in models of fourscrews than that of three screws. Stress concentration position was at the fracture site in both models. The stress range of models of four screws was more extensive and scattered. (3) Finite element analysis results demonstrated that four-screw implantation for Pauwels III femoral neck fractures had strong anti-shearing force and biomechanical stability. Clinical advantages need further clinical comparative study.

OBJECTIVE:To optimize the formulation of ferulic acid ligustrazine (FATM) solid lipid nanoparticles (FATM-SLN),and conduct the quality evaluation. METHODS:Emulsification ultrasonic method was used to prepare FATM-SLN. Using particle size and entrapment efficiency as indexes,amount of glyceryl monostearate,egg yolk lecithin (PC),poloxamer 188 (P188),and sodium stearate as factors,single factor test and orthogonal test were used to optimize the formulation;and verifica-tion test was conducted. The appearance morphology,distribution of particle size,Zeta potential,stability and in vitro release de-gree of prepared FATM-SLN were investigated. RESULTS:The optimal formulation was as follows as FATM of 10 mg,glyceryl monostearate of 300 mg,PC of 200 mg,P188 of 200 mg,sodium stearate of 10 mg,and purified water of 20 mL. The prepared FATM-SLN showed spherical solid particles,appearance morphology was round,distribution of particle size was 40-800 nm,parti-cle size was 106.23 nm,polydispersity index was 0.254,Zeta potential was -34.8 mV,entrapment efficiency was 73.32%,drug loading was 1.20%;the appearance had no obvious changes within 10 d in 4 ℃(RSD<2%). The drug-release in 0.5-1 h was the fastest,the cumulative release degree reached to 60.47%;it tended to be stable after 8 h,the cumulative release degree reached to 93.46%,and drugs were basically released completely. CONCLUSIONS:FATM-SLN formulation is successfully optimized,and the prepared FATM-SLN has small particle size,high entrapment efficiency and good stability.

Objective To investigate the effects of high glucose and lysophosphatidylcholine (LPC) on the immune function of in vitro cultured macrophages during Nocardia farcinica infection. Meth-ods RAW264.7 macrophages were cultured in vitro under different conditions as follows: routine culture (control group),50 mmol/L glucose (high glucose group),10 mg/L LPC(LPC groupⅠ),25 mg/L LPC (LPC groupⅡ) and 50 mmol/L glucose+25 mg/L LPC(high glucose and LPC group). The activity of mac-rophages in each group was tested after 6,12,24 and 36 h of culture. After 24 h of culture, macrophages were collected from every group and co-cultured with Nocardia farcinica. Dynamic phagocytosis rates were detected at 1,2,3,4,5 and 6 h after co-culture. Toxic effects of Nocardia farcinica on macrophages and concentrations of IL-10 and TNF-α were measured at 1,3 and 6 h after co-culture. Results Macrophages in all four experimental groups showed decreased activity as compared with those in the control group (P<0.01). Phagocytosis of Nocardia farcinica by macrophages was also reduced by high glucose and LPC. Phagocytosis rates of high glucose group and LPC groupⅡ at 1 and 2 h,LPC groupⅠat 1,2 and 3 h,and high glucose and LPC group at 1,2,3 and 4 h after co-culture were significantly lower than that of the con-trol group (P<0.05 or P<0.01). Compared with the control group, significantly reduced toxic effects on macrophages caused by Nocardia farcinica was observed in the experimental groups (P<0.05 or P<0.01). Compared with the control group,LPC groupsⅠand Ⅱ and high glucose and LPC group had decreased se-cretion of IL-10 at 3 h,and high glucose group and LPC groupⅠhad decreased secretion of TNF-α at 1 h(P<0.05). Conclusion Culture macrophages under the conditions of high glucose and LPC would reduce their activity and impair their ability to phagocytose Nocardia farcinica. Moreover, high glucose and LPC might have impacts on the toxic effects of Nocardia farcinica on macrophages and the secretion of IL-10 and TNF-α.

Objective: To describe the general and histological features of the full-length superficial fascia of the circumferential upper limb. Methods: Fresh frozen arm specimens were dissected, and then MRI imaging in vivo, enhanced CT angiography and HE histological staining were used to describe the characteristics of the full-length superficial fascia of the circumferential arm and its relationship with important blood vessels. Results: The four typical structures of the superficial fascia of the arm were divided into subcutaneous superficial fat, membrane-like substance, deep fat and deep fascia from superficial to deep. The thickness and stratification, fusion degree and histological characteristics of the superficial fascia of these four layers were obviously different in different levels and regions of the arm. MRI confirmed that the total thickness of superficial fascia gradually decreased from shoulder to wrist. Venography showed that the cephalic vein ran below the second layer of superficial fascia and above the deep fascia. The basilic vein originated from the dorsal vein network of the hand and always lied below the second layer of membranous material until the basilic vein penetrates below the deep fascia of the upper arm. Conclusions The deep understanding of the circumferential full-length of superficial fascia structure of the upper limb provides an important theoretical basis for improving the surgical safety and fine operation for the Dynamic Arm Circumferential Liposuction.

Objective To construct a mutant strain of Nocardia farcinica ( N. farcinica ) IFM10152 with mammalian cell entry 4A gene (mce4A) deletion and to analyze the function of that gene dur-ing infection. -ethods The mutant strain of N. farcinica was constructed through in-frame deletion without antibiotic labeling and verified by PCR and sequencing analysis. To analyze the function of mce4A gene in the interaction between N. farcinica and host cells, in vitro growth experiment, macrophage killing experi-ment using THP-1 ( a human leukemia mononuclear cell line) as the model and adhesion and invasion exper-iments using HeLa cells ( cervical cancer epithelial cells) were carried out. Results The mutant strain with mce4A gene deletion was successfully constructed and named △mce4A. No significant difference in growth rate was observed between the mutant and the wild-type strains. After knocking out the mce4A gene, the ability of N. farcinica to resist macrophage killing was obviously weakened as well as its ability to adhere and invade. Conclusions The mutant strain of N. farcinica with mce4A gene deletion was successfully construc-ted. The mce4A gene might play an important role in the adhesion and invasion of N. farcinica to host cells and its survival in macrophages.