Objective To explore the clinical outcomes of locking plate fixation of intertrochanteric fractures in patients with deformed proximal femur.Methods A retrospective review was made of the 25 patients with intertrochanteric fracture and deformed proximal femur who had been treated from January 2009 to July 2015.They were 17 men and 8 women,from 22 to 76 years of age (mean,45.2 years).The proximal femoral deformities were postpoliomyelitis syndrome in 6 cases,fibrous dysplasia in 4,malunion in 13,and hip varus caused by implant breakage following intramedullary nailing in 2.By AO classification,there were 4 cases of type 31-A2.1,3 ones of type 31-A2.2,16 ones of type 31-A3.1,and 2 ones of type 31-A3.2.All the patients were treated by open reduction plus locking plate fixation.At the final follow-up,function of the affected hip was evaluated by the Hip Disability and Osteoarthritis Outcome Score (HOOS),and quality of life of the patients was evaluated using the SF-36 health survey questionnaire.Results The 25 patients were effectively followed up for 12 to 24 months (mean,17.4 months).No infection,pneumonia,fat embolism,or deep venous thrombosis of lower limb was observed.The fractures healed without nonunion or refracture after a mean time of 5.9 months (range,from 3 to 6 months) in the 25 patients.By the HOOS evaluation at the final follow-ups,the 25 patients scored,on average,83.7± 15.6 in pain,55.6± 14.1 in symptoms,53.6 ± 9.5 in daily living function,52.7 ± 8.9 in sports and recreational activities and 62.4 ± 12.3 in quality of life;by SF-36 evaluation,they had a mean total score of 71.2 ± 13.8.Both the HOOS and SF-36 scores were not significantly different from the pre-injury values (P ＞ 0.05).Conclusion Locking plate fixation is a simple and effective way for the intertrochanteric fractures complicated with deformed proximal femur because intramedullary nailing is difficult or infeasible.

G mutation of SLC26A4, the parents and the second child were carriers of the same mutation, while the fetus had a wild-type form. Conclusion It is feasible to identify deafness related genes by screening for GJB2 and SLC26A4 mutation, thus providing correct prenatal diagnosis and avoiding deaf delivery of baby.

Objective To evaluate the clinical application of our self-designed ultradistal locking tool in the intramedullary nailing for tibial fractures.Methods From January 2014 to May 2016,175 patients with tibial fracture were treated at our department.They were 119 men and 56 women,from 19 to 73 years of age (average,46.3 years).They were divided into 2 groups according to the different targeting devices used in the intramedullary nailing.Conventional locking tools were used in the 83 patients from January 2014 to January 2015 and our self-designed new ultradistal locking tools in the 92 patients from February 2015 to May 2016.The 2 groups were compared in terms of operation time,frequency of intraoperative fluoroscopy,and successful rate of one-time locking.Results There were no significant differences between the 2 groups in general clinical data(P ＞ 0.05),showing similarities of the 2 groups.The operation time(59.8 ±4.3 min),frequency of intraoperative fluoroscopy(11.0 ± 2.1 times),and rate of one-time successful locking[94.4％ (238/252)] in the ultradistal locking group were significantly better than those in the conventional locking group [73.6 ± 5.3 min,23.0 ± 3.8 times and 85.7％ (180/210),respectively] (P ＜ 0.05).Conclusions Our new ultradistal locking tools are superior to the conventional ones in that they lead to shorter operation time,less intraoperative fluoroscopy and higher successful rate of one-time locking.Additionally,the new locking tools are easy to handle and incur no extra costs.

Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.

OBJECTIVETo construct a human source vector containing minidystrophin-EGFP fusion gene and investigate its expression in Cos-7 cells.

METHODSThe recombinant human source vector named pHrnDysG was constructed with PCR-clone methods. Three fragments of dystrophin gene were PCR amplified from normal human dystrophin gene cDNA (GenBank NM04006). These three fragments were ligated to generate a minidystrophin gene. The enhanced green fluorescent protein (EGFP) gene was fused to the C terminal of the minidystrophin gene, and then the pHrnDysG was finally obtained by cloning the fusion gene to pHrneo. Fluorescence microscope and RT-PCR were used to detect the expression of minidystrophin-EGFP fusion gene after the recombinant construct was transfected into Cos-7 cells by lipofectamine.

RESULTSRestrictive enzyme digestion analysis and sequencing confirmed that pHrnDysG vector was constructed successfully. After the recombinant pHrnDysG was transfected to Cos-7 cells, RT-PCR demonstrated that the fusion gene was successfully transcribed, and the green fluorescence was observed at the cell membrane.

CONCLUSIONThe minidystrophin-EGFP fusion gene mediated by pHrneo vector could express in Cos-7 cells and its products' localization in the cell membrane was the same as that of full length dystrophin. These results suggested that the recombinant human source vector pHrnDysG might be potentially used in studies on the gene therapy of Duchenne muscular dystrophy.

Animals ; COS Cells ; Cercopithecus aethiops ; Dystrophin ; genetics ; metabolism ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Models, Genetic ; Recombinant Fusion Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

Animals ; Cadherins ; metabolism ; Catenins ; metabolism ; Estradiol ; pharmacology ; Estrogen Receptor Modulators ; pharmacology ; Female ; Ovariectomy ; Rats ; Uterus ; drug effects ; metabolism

OBJECTIVETo explore the role of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in myocardial ischemia/reperfusion injury (MI/RI) by observing the dynamic expression changes at mRNA and protein levels early after myocardial ischemia/reperfusion (I/ R).

METHODSThe Wistar rats were randomly divided into Sham and I/R group (n = 42), and killed according to different reperfusion time (1, 2, 4, 6, 12, 24 h and 7 d). Structural and morphous changes of myocytes were observed under optical microscope. The mRNA and protein levels of TLR2 and TLR4 were detected using real-time PCR (RT-PCR). Monocyte chemokine protein-1 (MCP-1) and interleukine-6 (IL-6) mRNA levels were measured by reverse transcriptase-polymerase chain reaction (rt-PCR).

RESULTS(1) With the extension of reperfusion time, the myocardial infarct size increased smoothly, and reached the plateau at 4 h, then stayed in the platform. After reperfusion for 7 d, the ventricular had been remodeled. (2) At the beginning of reperfusion, myocardial structure showed no significant change in Sham group, but had different degrees of injury in I/R group. In rats of the group reperfused for 7 d the left ventricular remodeling could be visible. (3) Compared to sham group,TIR2, TLR4, MCP-1, IL-6 mRNA level were increased in myocardium in I/R group. TLR2 and TLR4 both peaked at 4 h of reperfusion, IL6 peaked at 6 h, followed by a gradually decrease. TLR4 and IL-6 mRNA levels rose again at 7 d. MCP-1 level in I/R group remained fairly with sham group at the beginning of reperfusion, and markedly elevated at 7 d.

CONCLUSIONExpression of TLRs mRNA in myocardium during early after myocardial ischemia/reperfusion increased rapidly and activated TLRs might play an important role in MI/RI through promoting the generation of inflammatory factors. At the late reperfusion, TLRs levels raise again and the expression of inflammatory factors increase once again, Those may probably affect the remodeling of ventricular, and injure myocardial structure and function.

Animals ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Interleukin-6 ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Wistar ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

Human ribosomal DNA (hrDNA) targeting vector pHr is a homologous recombinant plasmid for human genome which developed in the State Key Laboratory of Medical Genetics. pHr was used to construct a recombinant plasmid pHr-CMG expressing mda-7/GFP fusion gene and its efficacy in the hepatocellular carcinoma cell line Bel-7402 was investigated. The expression of mda-7/GFP fusion gene was detected by fluorescent microscope, RT-PCR and Western blotting, and its function was detected by cell-cycle analyses, MTT assay and Hoechst33258 staining. The results demonstrated that pHr-CMG vector could express MDA-7/GFP fusion protein effectually and the mda-7 gene could induce cell apoptosis and proliferation suppression in Bel-7402 cell line, which might be caused by the G2/M cell cycle arrest. These results also suggested that human ribosomal DNA targeting vector system and the pHr-CMG vector may be applied in further gene therapy researches for hepatocellular carcinoma.

In order to study the tumor suppression effect of p53 with CMV enhancer and hTERT promoter mediated by human-derived vector pHrn in liver cancer cell Bel-7402, report plasmid pchEGFP, tumor suppressor plasmids pchp53Arg and pchp53Pro were constructed by inserting expression cassette CMVe+hTERTp+EGFP, CMVe+hTERTp+p53Arg and CMVe+hTERTp+p53Pro into pHrn respectively. 24 h after cell transfection by lipofectamine 2000, GFP expression pattern was analyzed through fluorescence microscope and flow cytometry; RT-PCR and Western blot were taken to study the p53 expression pattern. The cell apoptosis by Hoechst 33258 and Annexin V-FITC/PI staining was also studied. Results show that the expression of GFP and p53 protein in Bel-7402were detected, but apparent cell apoptosis could not be found. The recombinant p53 mediated by human-derived vector could express in Bel-7402, but no significant tumor suppression effect was detected, which might result from the down regulation effect of the wild type p53 on hTERT promoter.

Objective To develop a sinusoidal alternating magnetic field therapy system in order to overcome the disadvantages of the single output frequency and the low effective value of the output magnetic field strength of the alternating magnetic field therapy system in the market,of which the frequency and magnetic density both were continuously adjustable. Methods Multi winding Helmholtz coil was used as the magnetic field generator.On the basis of inverter technology,bipolar equivalent area method considering dead zone and variable speed integral incremental PID control algorithm were used to achieve the accuracy control of magnetic frequency and density in the coil.The accuracy of the resulting waveform and the accuracy of the magnetic field strength was verified by simulation calculation and system current and magnetic field strength test.Results The magnetic field treatment system gained high performance,total harmonic distortion (THD)of sine wave met the requirements of international standards.The obtained magnetic density was as expected of the simulation and calculation. Conclusion The device provides continuously adjustable magnetic field,which has a positive effect on the research for the medical staff, and technical references are provided to the research of magnetic field therapy system.