Objective To determine the factors influencing the indication for tracheostomy following cervical spinal cord injury.Methods A retrospective study was performed to analyze the 118 patients who had been treated for cervical fracture/dislocation along with cervical spinal cord injury from July 2004 to June 2014 and whose abbreviated injury scale score (AISS) was lower than 3.They were 96 men and 22 women,19 to 68 years of age (average,45.2 years).The patients were divided into a tracheostomy group (n ＝ 28) and a non-tracheostomy group (n =90).The 2 groups were compared in terms of gender,age,presence or absence of complete spinal cord injury at admission,injured segment,injury mechanism,smoking history,injury severity score (ISS),motor AISS,systolic pressure at admission,hospital stay,and ICU stay to determine the factors influencing allocation of tracheostomy.Results Compared with the non-tracheostomy group,the tracheostomy group had a higher rate of complete spinal cord injury at admission,a higher rate of smoking,a higher ISS at admission,a lower motor AISS,and longer hospital and ICU stay,with statistically significant differences (P ＜ 0.05).There were no significant differences between the 2 groups in gender,age,injured segment,injury mechanism,or systolic pressure at admission (P ＞ 0.05).Increased severity of cervical spinal cord injury was associated with significantly decreased motor AISA,increased rate of tracheostomy and increased ISS (P ＜ 0.05).Conclusion The influencing factors for indication of tracheostomy after cervical spinal cord injury are complete cervical spinal cord injury irrespective of the level of injury,ISS,motor AISS,and history of smoking.

Cerebral vasospasm (CVS) is one of the serious complications of subarachnoid hemorrhage (SAH).Pathogenesis of CVS has not been fully clarified,and it may be associated with a variety of factors.With the development of molecular biology techniques,people have more understanding on SAH caused pathogenesis of CVS,and the research has also made considerable progress in the prevention of CVS.

Objective:After construction and identification of HBsAg gene recombinant backbone adenoviral vector,it is to prouduct HBsAg gene recombinant adenoviruses by packing PAd-Easy-1-HBs in 293 cells.Methods:The gene of interest was amplified from plasmid pEcob-6 PCR, the gene of interest which contained HBsAg gene was cloned into on adenoviral shuttle vector pAd-track-cmv. The pAd-track-cmv-HBs was linearized by digesting with restriction endonuclease Pme-1, and subsequently cotransformed into E.coli BJ-5183 cells with an adenoviral backbone vector pAd-Easy-1, Homologous recombinants were performed in bacterial cells. Finally, the linearized backbone adenoviral vector was transfected into adenoviruses packing cells lines,e.g. 293 cell by lipofectamine transfection. Transfections and viral productions can be minotored by green fluorescent protein(GFP). The expression of HBsAg in supermatant was investigated by ELISA. It was a certain HBsAg vaccine to amplify recombinant adenoviruses by repeating the infevtion cell to collect the viral supermatant.Results:GFP expression was visible by fluorescence microscopy after transfection. Adenoviral titer was monitored by GFP expression. GFP expression was visible after repeating the infection cell using the viral supernatant in more than 90 percent of the cells.The HBsAg also expressed in supermatant.Conclusion:HBsAg gene recombinant adenoviral backbone vector has been constructed successfully. HBsAg gene recombinant adenoviruses have been producted by packing in 293 cells. The study provides the possibility of further researches on the development of new anti-HBV vaccines.

Object In order to identify the medicine at the molecular level, the internal transcribed spacers (ITS) of Saussurea medusa Maxim and its easily confusable species were sequenced Methods The double stranded DNA was amplified using PCR systems 9 600 kits and sequenced on an ABI 377 automated sequencer from both directions Results The ITS sequences of S medusa of different populations showed no variation, but there existed distinct variation between S medusa and its confusable species Conclusion ITS sequences can be used for the molecular authentication between S medusa and its confusable species

The experience in the orthopedic teaching of the interns of medical science of law was explored.The features of the students of the medical science of law were analyzed.And related teaching project was established during the progress of orthopedic practice.Our experience emphasized on the knowledge teaching of medical ethics and medical disputes.

Objective To study the effect of stable expression of reconstructed human transforming growth factor-β3 ( hTGF-β3) on proliferation of precartilaginous stem cells ( PSCs) and their differentiation into cartilage chondrocytes.Methods After isolated and purified by immunological microbeads,PSCs of rats were transfected with pcDNA3.1 ( + )-hTGF-β3.MTT reduction assay ( MTT) and flow cytometry were performed to investigate the effect of transduction on proliferation and DNA synthesis.Biosynthesis of hTGF-β3 and expressions of cartilage associated genes and proteins were examined by qRT-PCR,immunohistology and Western blot.Results hTGF-β3 was expressed in PSCs stably.Compared with non-transfection group,PSCs' DNA synthesis level and proliferation rate were significantly increased after transfection.Quantitative real-time PCR and immunological investigation suggested up-regulated expression of specific genes and proteins of chondrocyte and increase of deposition of chondrocyte typical extracellular matrices proteoglycan and collagen type Ⅱ .Conclusions Gene enhanced PSCs can stably express hTGF-β3 protein to promote proliferation of PSCs and induce differentiation of PSCs in to chondrocytes,which provides a fresh approach to cartilage tissue engineering.

The influence of short hairpin RNA (shRNA)-mediated osteopontin (OPN) gene silencing on the proliferation and invasion of human renal cancer ACHN cells was investigated. Four types of OPN shRNA recombinant plasmids were constructed and RT-PCR assays were used to screen the most highly functional shRNA recombinant plasmids, which were transferred into the cultured ACHN cells by Lipofectamine 2000. The cells transfected by shRNA expression vectors (ACHN/OPN) were visualized under an inverted microscope and screened by G418. Untreated cells (ACHN) and cells transfected by mock vectors (ACHN/Vect) were used as control groups. The expression levels of OPN mRNA and protein were detected by real-time PCR and Western blot respectively. The cell cycle and ratios of apoptotic cells were assessed by flow cytometry. MTT method was used for drawing the growth curve and observing cell proliferation in vitro. The abilities of migration and invasion in three groups were measured by Transwell chamber test. The expression levels of matrix metalloproteinase (MMP)-2 and MMP-9 in three groups were examined by Western blot. Our results showed that the recombinant plasmid could be successfully transferred into ACHN cells by LipofectamineTM 2000. Compared with untreated cells, the expression levels of OPN mRNA and protein in ACHN/OPN cells were decreased by 59.68% and 76.42%, respectively (P<0.05), ACHN/OPN cells were blocked in S phase and apoptotic ratio increased significantly (P<0.05), however, no significant differences were found between ACHN/Vect and ACHN. Recombinant plasmid significantly attenuated expression levels of MMP-2 and MMP-9 proteins and suppressed the proliferation, migration, and invasion of ACHN cells. This study suggested that OPN may play an important role in the growth and invasion of human renal cancer ACHN cells, and these processes are correlated with the activations of MMP-2 and MMP-9. Our data provided preliminary experimental evidence for the feasibility of RNA interference technology in gene therapy of human renal cancer.

This study examined the construction of eukaryotic expression plasmid of human transforming growth factor-β3 (hTGF-β3) and its inducing effect on the differentiation of precartilaginous stem cells (PSCs) into chondroblasts. hTGF-β3 gene was amplified by using polymerase chain reaction (PCR) and then inserted into the eukaryotic expression plasmid pcDNA3.1 to construct the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3. Rat PSCs were isolated and purified by employing an immunomagnetic cell sorting system. pcDNA3.1(+)-hTGF-β3 was transfected into purified PSCs with the use of linear polyamines. The expression of TGF-β3 and cartilage-specific extracellular matrix (ECM) components was detected after transfection by real-time quantitative PCR, ELISA, immunochemistry and Western blotting, respectively. The results showed that the eukaryotic expression plasmid pcDNA3.1(+)-hTGF-β3 was successfully established as identified by enzyme digestion and DNA sequencing. Real-time quantitative PCR and ELISA revealed that hTGF-β3 was strongly expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs. Real-time quantitative PCR, immunochemistry and Western blotting showed that the cartilage-specific ECM markers, i.e., cartilage oligomeric matrix protein (COMP), Aggrecan, collagen type X and II were intensely expressed in the pcDNA3.1(+)-hTGF-β3-transfected cells. It was concluded that hTGF-β3 could be stably expressed in pcDNA3.1(+)-hTGF-β3-transfected PSCs and induce the differentiation of PSCs into chondroblasts.

Objective To comparatively study the differences of 18F‐FDG imaging agent by three kinds of different intravenous injection method for conducting PET / CT examination in aspects of the puncture success rate ,residual amount of drug injection and staff ray exposure time and their significance .Methods 240 patients with PET /CT examination were randomly divided into the group A ,B and C ,80 cases in each group .The drug injection adopted the traditional direct injection ,indwelling catheter injection and scalp venous needle connecting syringe(indwelling bubbles) .The puncture success rate ,drug residues and staff contacting radio‐pharmaceuticals time were compared among 3 groups .The obtained relevant data were statistically analyzed .Results The puncture success rate in the scalp venous needle connecting syringe (indwelling bubbles) and the indwelling catheter injection was higher than that in the traditional direct injection and the staff contacting radiopharmaceuticals time was significantly decreased ,the differ‐ences among them were statistically significant(P< 0 .01) ;the radioactive drugs residue in the scalp venous needle connecting syr‐inge was significantly decreased than that in other two kinds of injection method ,the difference was statistically significant (P <0 .01) .Conclusion The injection method of scalp intravenous needle connecting syringe (indwelling bubbles) significantly increases the puncture success rate ,reduces the radioactive drug residue ,at the same time decreases the staff radiation exposure time ,this method has the advantage in the radionuclide injection .

To explore the experience in teaching of interns in orthopedic,we put emphasis on the knowledge teaching of medical ethics and medical dispute in addition to the teaching of expertise of orthopedic.