1.Surgical treatment of unstable fractures of distal radius
Tao LI ; Qizhi SONG ; Shuhua YANG
Chinese Journal of Trauma 2010;26(7):602-605
Objective To explore the surgical methods for unstable fractures of distal radius and evaluate the clinical results. Methods A retrospective analysis was conducted on 87 patients with unstable fractures of distal radius treated surgically from May 2001 to June 2008. According to AO classification and comminution severity, three methods including plate fixation, external fixation and Kirschner' s pins and plate fixation plus external fixation were employed accordingly. Results Of all, 78 patients were followed up for average 18 months. The result of Cooney's assessment of the wrist was excellent in 44 patients, good in 21, fair in 11 and poor in 2, with excellence rate of 83%. Conclusions The surgical methods should be selected based on different types of unstable distal radius fractures. In the meantime, maximal recovery of the relative length of the radius and ulna, formation of articular surface, volar tilting angle and ulnar inclination angle and reasonable rehabilitation exercises are helpful for the functional recovery of the wrist to a largest extent.
2.Construction of targeting short hairpin RNA plasmid vector expressing TERT gene
Yang SONG ; Tao XU ; Mingkun YANG ; Weibin SHENG
Chinese Journal of Tissue Engineering Research 2014;(7):1057-1062
BACKGROUND:Telomerase reverse transcriptase (TERT) plays an important role in telomerase activation.
OBJECTIVE:To construct the targeting short hairpin RNA plasmid vector expressing TERT gene from astrocytes by using pLentilox3.7.U6.
METHODS:By using two sequences from TERT gene, we synthetized sense and antisense strand template sequences of RNA interference molecular in vitro, and then obtained the complementary strands through annealing procedure. We connected the strands with pLentilox3.7.U6 that was sequenced and transfected into the Escherichia coli. In the end, we tested its effect of reducing the TERT gene expressing by using cultured astrocytes from rat spinal cord in vitro through western blot and immunofluorescence technique.
RESULTS AND CONCLUSION:Western blot and immunofluorescence assay showed that, compared with the control group, the interference groups had a lower TERT expression in astrocytes. The targeting short hairpin RNA plasmid vector expressing TERT gene is useful to reduce the TERT gene expression. The targeting short hairpin RNA plasmid vector expressing TERT gene is valid for us to do the further test learning the mechanism of astrocytes in spinal cord injury.
3.The expression of matrix metalloproteinase-9 and the pathological changes of vascular smooth muscle cell in patients with acute ascending aortic dissection
Tao WEI ; Tao HONG ; Zhaohui HU ; Kai SONG ; Jiayu ZHENG ; Shouguo YANG ; Zhaohua YANG ; Chunsheng WANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2010;26(3):176-179
Objective Ascending aortic dissection(AAD),for which the pathogenesis remains unknown,is life-threatening.Matrix metalloproteinase-9(MMP-9)and the pathological changes of vascular smooth muscle cells(VSMCs)have been reported to have roles the pathogenesis.The study examined the expression of matrix metalloproteinase-9(MMP-9)and the pathological changes of,VSMCs in patients with AAD.Methods AAD samples were taken from 35 patients(disease group)in acute phase during aortic replacement operation for AAD and control samples were corresponding part of ascending aorta(control group,n=21)collected from the donor hearts for transplantation.Transmission electron microscepe,hematoxylin-eosin(H-E)staining.Mallory staining were used for observing the pathological changes of VSMCs and matrix in the affected aortic wall.The immunohistochemicai staining of MMP-9 was carried out in both groups and semi-quantified by staining intensity analysis.The affected patients were further grouped according to the diameter of dissected aorta as with a AAD of <55 mm or with a AAD of≥55 mm.The associations of clinical factors,such as smoking status,hypertensive disease and aneurysm diameter,with the expression of MMP-9 were analyzed.Results Increased synthetic function of VSMCs with decreased density,disrupted elastic fibers and fibrosis in the dissected aortic wall were observed in the disease group,but not in the control group.MMP-9 was scarcely expressed in the aortic wall of the patients in the control group,though it was notably expressed in the VSMCs of disease group.Both subgroups presented more MMP-9 than the control group(both P<0.001).In the disease group,sub-group with a AAD diameter of ≥55 mm presented more MMP-9 than that with a diameter of <55 mm(P<0.05).MMP-9 expression was positively correlated with a history of hypertension(P<0.01)or a great aneurysm diameter(P<0.05).MMP-9 expression was not associated with age,smoking status or other clinical factors.Conclusion Increased secretion of VSMCs and the expression of MMP-9 induced by elevated blood pressure may lead to the destruction of matrix proteins.The resulting fibrosis of the aortic wall would decrease the tensile strength of the wall.When the fibrotic aortic wall dilated further,the increased expression of MMP-9 would aggravate the damage to the wall.It can be speculated that acute AAD would occur as a result of partial tearing of the aortic intima.
4.Construction and appraisal of recombinant adeno-associated virus vector for expressing shRNA targeting CIP2A mRNA
Xue YANG ; Tao SONG ; Jie TAO ; Hao SUN ; Wei YANG ; Qingguang LIU
Journal of Xi'an Jiaotong University(Medical Sciences) 2015;(6):743-748
Objective To construct the recombinant adeno-associated virus vector carrying cancerous inhibitor of PP2A (CIP2A)short hairpin RNA (shRNA)for preparation of high-titer viruses.Methods The small hairpin RNA of CIP2A (CIP2A shRNA)was designed,synthesized and cloned into pDC31 6-EGFP-U6 plasmid which was double digested by Bam HⅠ and Hin dⅢ.The resultant plasmid pDC31 6-EGFP-shRNA was confirmed and served as template to appraise primers.EGFP-CIP2A shRNA sequence was amplified by PCR,double digested with Eco RⅠand Sal Ⅰ and ligated to pSNAV2.0 plasmid digested with the same enzyme pair.pSNAV2.0-EGFP-CIP2A shRNA plasmid DNA was prepared,purified,identified and transfected into BHK-21 cells.BHK-21 cells expressing CIP2A shRNA (BHK-21/CIP2A-shRNA ) were obtained and subsequently infected with VGTC’s proprietary AAV packaging system to package the rAAV2-CIP2A shRNA.After purification,the functional and infectious virus was obtained and the titer of virus was detected.Real-time PCR and Western blot methods were used to detect the expression of CIP2A after infection with HepG2 cells,and the empty viral vector rAAV2-EGFP was used as control. Results A recombinant adeno-associated virus-2 vector carrying CIP2A shRNA was constructed successfully.The presence of the target sequence in the vector was confirmed by double enzyme digestion and sequencing.By transfecting the pSNAV2.0-EGFP-CIP2A shRNA plasmid into BHK-21 cells,BHK-21/CIP2A shRNA cells were infected with helper virus HSV1-rc/ΔUL2 to package the rAAV2-CIP2A shRNA to obtain a functional and infectious virus.The titer of the recombinant virus was 0.25×10 1 2 v.g./mL.The expression of CIP2A mRNA and screening value of 1×10 5 MOI effected HepG2 cells.Conclusion A high-titer recombinant adeno-associated virus-2 vector carrying CIP2A shRNA has been constructed successfully.
6.Electrochemical Deoxyribonucleic Acid Biosensor Based on Multi-walled Carbon Nanotubes/Ag-TiO_2 Composite Film for Label-free Phosphinothricin Acetyltransferase Gene Detection by Electrochemical Impedance Spectroscopy
Na ZHOU ; Tao YANG ; Kui JIAO ; Caixia SONG
Chinese Journal of Analytical Chemistry 2010;38(3):301-306
A highly sensitive electrochemical deoxyribonucleic acid(DNA) biosensor based on multi-walled carbon nanotubes(MWNT)/Ag-TiO_2 composite film was developed. The solution containing Ag-TiO_2-MWNT composite was casted on the carbon paste electrode surface to form a robust film, which combine the advantages of the good biocompatibility of Ag-TiO_2 naocomposite and the fine conductivity, as well as the large active surface area of carbon nanotubes. The composite could greatly improve the immobilization capacity of the probe DNA. The morphologies and electrochemistry of the nanocomposite film were investigated by scanning electron microscopy and electrochemical techniques including electrochemical impedance spectroscopy and cyclic voltammetry, respectively. DNA hybridization events were monitored by a label-free method of electrochemical impedance spectroscopy. This label-free electrochemical impedance DNA biosensor showed high sensitivity and selectivity for phosphinothricin acetyltransferase gene sequence assay. The multicomponents films also displayed a high stability during repeated regeneration and hybridization process.
7.siRNA lentiviral vectors carrying telomerase reverse transcriptase gene hasten astrocytes apoptosis
Liang LIANG ; Tao XU ; Yang SONG ; Weibin SHENG
Chinese Journal of Tissue Engineering Research 2015;(11):1707-1711
BACKGROUND:Telomerase reverse transcriptase plays an important role in telomerase activation, and lentiviral vectors carrying human telomerase reverse transcriptase that can inhibit astrocyte expression are rarely reported to have effects on spinal cord injury. OBJECTIVE:To transfect rat astrocytes with telomerase reverse transcriptase gene lentiviral vectors and to observe the effect of telomerase reverse transcriptase gene lentiviral vectors on apoptosis of astrocytes. METHODS: Astrocytes from rats were subject to primary culture and subculture, and then transfected with siRNA lentiviral vector carrying telomerase reverse transcriptase gene (siRNA transfection group), simple lentiviral vector (lentiviral vector group) and nothing (blank group), respectively. Then, the transfection efficiency and apoptosis in different time periods after transfection were determined. RESULTS AND CONCLUSION:The transfection efficiency was up to 85%-90% after siRNA lentiviral vector and simple lentiviral vector transfection. The apoptosis rate of astrocytes in the siRNA transfection group was 50%-60% at 24-48 hours after transfection, but there was no significant change in the other two groups. These findings suggest that siRNA lentiviral vectors carrying telomerase reverse transcriptase gene can accelerate astrocytes apoptosis.
8.Tracing rat nerve tracts with 7.0T Manganese-enhanced MRI
Song WEN ; Gejun GAO ; Tao YANG ; Feng DAI ; Zhongbao TAN
Chinese Journal of Medical Imaging Technology 2010;26(1):29-32
Objective To investigate the effect of manganese-enhanced MRI (MEMRI) at 7.0T for tracing nerve tracts in rat brain in vivo. Methods With brain stereotactic apparatus, 0.4 μl Mncl_2 with aqueous solution of 1 mol/L was injected into the right somatosensory cortex of 9 SD rats. MR scan was performed for tracing corticospinal tracts and other coherent nerve tracts pre-, and 24, 48, 72 h, 7 days post-injection with 7.0T micro-MRI system, respectively. Results Corticospinal tracts were showed in intact after Mn~(2+) administration from somatosensory cortex, thalamus, cerebral peduncle to pons at the time point of 24, 48, 72 h and 7 days, while the best tdisplaying was achieved at 24-48 h after Mn~(2+) administration. Simultaneously a small quantity of Mn~(2+) reached the opposite somatosensory cortex through the corpus callosum. Conclusion MEMRI for tracing rat nerve tracts can be showed clearly with 7.0T MRI. The location of manganese-enhanced corticospinal tracts in agreement with the rat brain atlas in stereotaxic is in agreement with that Paxinos' published. MEMRI can display the relationship between the two sides of hemisphere, and may play an important role in investigating the brain function and nerve plasticity after nerve injury in vivo.
9.Clinical therapy for blood culture-negative infective endocarditis
Jianguo YANG ; Daqing HU ; Laichun SONG ; Liang TAO ; Xueguo FENG
Clinical Medicine of China 2015;(3):215-218
Objective To summarize the clinical treatment experience of blood culture-negative infective endocarditis,and to explore the surgery chance and therapeutic strategy of blood culture-negative endocarditis. Methods One hundred and sixty-six patients who were diagnosed blood culture-negative endocarditis in the Aisa Heart Hospital of Wuhan from Jul. 2008 to Dec. 2012 were recruited in the study. Broad-spectrum antibiotics including cefuroxime axetil and levofloxacin were used before the result of blood culture,and sensitive antibiotics were selected to control patient's condition when getting the result of blood culture. But broad-spectrum antibiotics were continuously used to the blood culture-negative endocarditis until stable condition. When the conditions were stable,active preparation before surgery should be carried out. Thoroughly clear the vegetation and protect the cardiorespiratory function during operating. Kata-step antibiotics were used to control patient's condition until normal temperature,as well as the number of leukocytes decreased,blood sedimentation normalized and C-reaction protein decreased. Then,the narrow-spectrum antibiotics were selected including cephalosporin until discharged from hospital,and continued treatment of antibiotics for 4 - 6 weeks. Results Five patients died after the operation,including 1 case died of low cardiac output syndrome,2 cases died of multiple organ failure,1 case died of septicemia and the 1 case died of cerebral embolism. All the other patients discharged from hospital successfully. Conclusion The patients with blood culture-negative IE should be controlled rapidly. The duration and dose of antibiotics should be enough. Active operative preparation should be taken and then surgery timely. Thus,the hospital mortality could decrease and prompt the long-term outcome.
10.Thyroid Imaging Reporting and Data System in Identifying Benign and Malignant Thyroid Nodules
Xueqin HOU ; Xue FAN ; Lingfei YANG ; Jian DONG ; Tao SONG
Chinese Journal of Medical Imaging 2015;(7):489-493
PurposeAs a main method to detect thyroid nodules, ultrasonography seems to have a rather low accuracy in differentiating benign and malignant ones. The present study aims to explore the diagnostic value of thyroid imaging reporting and data system (TI-RADS) in identifying benign and malignant thyroid nodules.Materials and Methods A total of 168 patients with thyroid nodules conifrmed pathologically (with 251 suspicious nodules) underwent ultrasonography and were further grouped into category 3-5 according to TI-RADS classiifcation standard. The results were retrospectively compared with histopathological ifndings. The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS classiifcation in the diagnosis of benign and malignant thyroid nodules were calculated, and the differences in ultrasonic features between benign and malignant thyroid nodules were also compared.Results The surgical and pathological findings showed that 94 nodules were benign and 157 were sinister; TI-RADS regarded 93 nodules were benign and 185 were malignant. The difference of the two means of identification was significant (χ2=149.6,P<0.01). The sensitivity, speciifcity, accuracy, positive predictive value and negative predictive value of TI-RADS were 91.2% (144/157), 85.1% (80/94), 89.3% (224/251), 91.2% (144/158) and 86.0% (80/93), respectively. The ultrasonic manifestations of benign and malignant nodules were signiifcantly different with regard to aspect ratio, echogenicity, shape, calciifcation inside the nodule (χ2=8.7-121.4,P<0.01).Conclusion TI-RADS classiifcation standards have a high accuracy in the differentiation of benign and malignant thyroid nodules as a means of ultrasound examination, and thus can serve as an important guiding method in clinical diagnosis and treatment.