BACKGROUND:Now it has cooperation and facilitative effete between myogenic regulatory factors through a long time study. So, gene therapy of double genes of Myod1 and Myog can obtain better effect, and can provide a new way for preventing denervated skeletal muscle atrophy.
OBJECTIVE:To construct eukaryotic co-expression vector carrying Myod1 and Myog genes.
METHODS:Ful-length Myod1 gene and Myog gene cDNA were amplified by reverse transcription PCR, and then inserted into pVAX1 vector after digested to establish the recombined Myod1 and Myog eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and then identified with gene sequencing. The in vitro cultured 3T3 cel s were transfected with pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and the expressions of Myod1 and Myog genes in the 3T3 cel s were detected with western blot assay in order to identify whether the 3T3 cel s could express the target protein correctly.
RESULTS AND CONCLUSION:The sequencing results showed that the sequence length and base sequence of Myod1 and Myog cDNA in eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP were identical with the reported sequences. Myod1 and Myog protein band expressions were detected in 3T3 cel s by western blot after transient transfection. The pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, a eukaryotic co-expression vector of Myod1 gene and Myog gene is successful y constructed.

Objective Identification of anatomic structures are essential for totally thoracoscopic anatomic pulmonary segmentectomies, however sometimes the procedure are difficulty.This study was to assess whether three-dimensional computed tomography angiography(3D-CTA) could contribute to the preoperative arrangement of thoracoscopic complex segmentectomy.Methods Between September 2012 and August 2014, 29 patients were performed thoracoscopic complex segmentectomies under the guidance of preoperative 3D-CTA.The segmentectomies pattern were based on the nodules' diameter, location,and pathology.The targeted vessels and bronchus were marked in preoperative simulated segmentectomies.Results Of the 29 cases, 9 right upper lobe segmentectomies, 13 left upper segmentectomies, and 7 bibasilar segmentectomies were achieved, among which 6 subsegmentectomies were also inclued.The mean lesion diameter, operative time and intraoperative blood loss were(1.35 ± 0.80) cm, (190.53 ± 50.83) min, and (26.90 ± 32.24) ml respectively.Under the guidance of preoperative 3D-CTA , 8(27.5％) nodules were detected accurately, moreover 2(6.9％) aberrant arteries and 1 (3.4％) aberrant bronchus were observed.According to the marked vessels and bronchus preoperatively, 27 (93.1％) arteries, 25 (86.2％)veins,and 29(100％) bronchus were identified and dissected in the operation.Three cases converted to unplanned segmentectomies.No serious complications or death occurred.Conclusion 3D-CTA is an effective tool to enhance security and efficiency in thoracoscopic complex anatomical segmentectomy.

Objective To evaluate the safety and feasibility of totally thoracoscopic anatomic pulmonary segmentectomy (TTAS) for the treatment of the peripheral stage ⅠA non small cell lung cancer(NSCLC).Methods The study involved 50 consecutive patients undergoing totally thoracoscopic anatomic segmentectomy (TTAS) from September 2010 to November 2012 in the First People's Hospital affiliatied to Nanjing Medical University.The diameter of the tumors were less than 2 cm [(mean diameter(1.35 ±0.48) cm].All lymph node sampling of N1 and N2 were neglive,All patients received symtematic lymph node dissection.The pulmonary vessels were individually ligated,and the bronchi were closed using an endoscopic stapler.The intersegmental plane was identified using the demarcation between the resected(inflated) and preserved(collapsed) lungs.Staplers were used for intersegmental dissection.Results The mean operative time and intraoperative bleeding were (191.5 ± 50.4) min and (49.2 ± 54.6) ml respectively.The chest tube drainage duration was (3 ± 1) days.The number of stapler cartridges used for intersegmental division was 3.9 ±0.8.The mean number of lymph nodes and nodal stations dissected were 12.6 ± 2.8 and 6.0 ± 1.5 respectively.No mortality and complications were observed 30 days after the surgery.Further,no local recurrence or metastases were observed during follow-up.Conclusion Totally thoracoscopic anatomic segmentectomy(TTAS) is a feasible and safe technique.With systematic lymph node dissection,TTAS can be a reasonable therapeutic option for stage ⅠA NSCLC.

BACKGROUND:Laminoplasty and laminectomy are the most commonly used treatment methods for multilevel cervical spondylotic myelopathy, which is more common in elderly patients. C5 nerve root palsy is the common postoprative complication after posterior cervical repair. OBJECTIVE:To compare the incidence of C5 nerve root palsy after laminoplasty with mini-titanium plate fixation and laminectomy with internal fixation in repair of multilevel cervical spondylotic myelopathy. METHODS:Total y 134 patients with multilevel cervical spondylotic myelopathy from August 2010 to December 2014 were enrol ed, and then divided into laminoplasty group (n=45) and laminectomy group (n=89) owing to different ways of repair. Patients in the laminoplasty group were treated with laminoplasty with mini-titanium plate fixation, and patients in the laminectomy group were treated with laminectomy with internal fixation. C5 nerve root palsy condition after repair was recorded and evaluated. The cervical lordosis angle (Cobb angle) and cervical curvature index were compared. The Japanese Orthopaedic Association score was used for neurological assessment. RESULTS AND CONCLUSION:Al patients in both groups were fol owed up for more than 6 months. There were no significant differences in cervical lordotic angle and cervical curvature index at the first week before and after the treatment between these two groups (P>0.05). The Japanese Orthopaedic Association scores of patients after 6 months of treatment were significantly improved compared with that before treatment in these two groups (P<0.05). There were 2 cases of C5 nerve root palsy after the treatment of laminoplasty, the occurrence rate was 4%(2/45);there were 10 cases of C5 nerve root palsy after the treatment of laminectomy, the occurrence rate was 11%(10/89);there was significant difference between these two groups (P<0.05). These results suggest that compared with laminectomy fixation, the incidence of C5 nerve root palsy was lower after the laminoplasty with mini-trianium plate fixation, which can be widely used in decompression repair treatment of multilevel cervical spondylotic myelopathy.

Objective:To explore the clinical efficacy and surgical indications of Y type osteotomy in the treatment of post-tuberculous thoracolumbar severe angular kyphosis.Methods:From March 2012 to June 2018, 36 patients with post-tuberculous thoracolumbar severe angular kyphosis were treated with Y type osteotomy, including 22 males and 14 females, aged 23.6±5.7 years (range, 7-57 years). The parietal vertebrae of kyphosis were located in the upper thoracic vertebra in 3 cases, the thoracic vertebra in 11 cases, the thoracolumbar segment in 17 cases, and the lumbar vertebra in 5 cases. The Cobb angle of kyphosis before the operation was 92.8°±23.3° (range, 60°-147°). The visual analogue scale (VAS), American Spinal Injury Association (ASIA) neurological function grade, and Kirkaldy-Willis function score were used to evaluate the clinical effect. The imaging evaluation indexes were interbody kyphosis angle and spinal bone fusion.Results:The operation was successful in all the 36 patients. The operation time was 210 ±25.9 min (range, 180-270 min), the intraoperative blood loss was 520 ±110 ml (range, 400-800 ml), and the postoperative follow-up time was 26.38±1.75 months (range, 22-30 months). The postoperative kyphosis Cobb angle was corrected to 16.5°±7.7° (range, 5°-35°), which was significantly improved compared with that before operation( t=25.438, P<0.01), and the correction rate was 82.2%. At the last follow-up, the kyphosis angle was 16.5°±7.1° (range, 6°-32°), which was not significantly different from that after the operation. The preoperative VAS score was 7.3±1.8 (range, 3-9), and the postoperative VAS score was 2.4±0.8 (range, 1-3), while the improvement rate was 67.1%. At the last follow-up, it was 1.1±0.6 (range, 0-2), and the improvement rate was 85.0%. According to the Kirkaldy-Willis functional score, the results were excellent in 25 cases, good in 8 cases, and fair in 3 cases at the last follow-up, with an excellent and good rate of 91.7%. Before the operation, 9 cases were accompanied by neurological dysfunction (ASIA grade: grade C in 2 cases, grade D in 7 cases). At the last follow-up, all the 9 patients recovered to grade E. During the operation, the electrophysiological nerve monitoring was abnormal in 2 patients, and the awakening test was negative in 1 case. In another patient, neuroelectrophysiological monitoring after posterior column osteotomy showed a decrease in bilateral sensory and motor function. There was no compression around the spinal cord in the osteotomy area, so the operating bed was gradually folded and partially restored to kyphosis and temporarily fixed with double rods. Neuroelectrophysiological monitoring suggested the recovery of nerve function. The awakening test showed that the nerve function of both lower limbs recovered close to the preoperative state, and further osteotomy and internal fixation was performed 2 weeks later. The nerve function of both lower limbs returned to normal after 3 months. After the operation, one patient's muscle strength of the lower limbs decreased from grade 5 to grade 3, and the sensory function was normal. After symptomatic support treatment such as neurotrophic drugs, it returned to normal 2 weeks later. 1 case developed delayed neurological dysfunction 1 year after the operation. Neurotrophic drugs and rehabilitation treatment improved it. The sinus of the incision was formed in one case 3 months after the operation and healed after debridement and suture. Conclusion:Y typeosteotomyis a safe and effective method for patients with post-tuberculous thoracolumbar severe angular kyphosis. Compared with traditional osteotomy, anterior support bone grafting can be avoided, and spinal shortening can be reduced.

hToracoscopic segmentectomy is technically much more meticulous than lobectomy, due to the com-plicated anotomical variations of segmental bronchi and vessels. Preoperative three-dimensional computed tomography bron-chography and angiography, 3D-CTBA) could reveal the anatomical structures and variations of the segmental bronchi/vessels and locate the pulmonary nodules, which is helpful for surgery planning. Preoperative nodule localization is of vital importance for thoracoscopic segmentectomy. Techniques involved in this procedure include dissection of the targeted arteries, bronchus and intra-segmental veins, retention of the inter-segmental veins, identiifcation of the inter-segmental boarder with the inlfa-tion-delfation method and seperation of intra-segmental pulmonary tissues by electrotome and/or endoscopic staplers. hTe incision margin for malignant nodules should be at least 2 cm or the diameter of the tumor. Meanwhile, sampling of N1 and N2 station lymph nodes and intraoperative frozen section is also necessary. hTe complication rate of thoracoscopic segmentectomy is comparatively low. hTe anatomic relationship between pulmonary segments and lobes is that a lobe consists of several ir-regular cone-shaped segments with the inter-segmental veins lies between the segments. Our center has explored a method to separate pulmonary segments from the lobe on the basis of cone-shaped principle, and we named it“Cone-shaped Segmentec-tomy”. hTis technique could precisely decide and dissect the targeted bronchi and vessels, and anatomically separate the inter-segmental boarder, which ultimately achieve a completely anatomical segmentectomy.

Objective:To investigate the predictive value of Graeb score for the outcome of high-grade aneurysmal subarachnoid hemorrhage (aSAH) patients with intraventricular hemorrhage (IVH).Methods:Consecutive high-grade aSAH patients with IVH admitted to the No. 1 People's Hospital of Jiujiang from January 2012 to March 2020 were enrolled retrospectively. High-grade aSAH was defined as grade Ⅳ to Ⅴ according to the World Federation of Neurological Surgeons (WFNS) scale. The outcome of patients was evaluated by the modified Rankin Scale (mRS) at 3 months after discharge. A score of ≤2 was defined as a good outcome and a score of >2 were defined as a poor outcome. Multivariate logistic regression model was used to evaluate the correlation between Graeb score and clinical outcome, and the receiver operating characteristic (ROC) curve was used to determine the predictive value of Graeb score for clinical outcome. Results:A total of 86 high-grade aSAH patients with IVH were enrolled. Aneurysm treatment: craniotomy clipping in 42 patients (48.8%), intravascular embolization in 21 (24.4%), and conservative treatment in 23 (26.7%). Twenty-nine patients (33.7%) had a good outcome and 57 (66.3%) had a poor outcome. Multivariate logistic regression analysis showed that the Graeb score >6 (odds ratio [ OR] 26.360, 95% confidence interval [ CI] 4.106-169.235; P<0.001), the modified Fisher grade 3-4 ( OR 11.674, 95% CI 1.540-88.512; P=0.017) and complicated with chronic hydrocephalus ( OR 21.236, 95% CI 2.883-156.431; P=0.003) were the independent risk factors for the poor outcome. ROC curve analysis showed that the area under the curve of the Graeb score predicting for poor outcome was 0.843 (95% CI 0.760-0.926; P<0.001), the best cut-off value was 6.5, and the corresponding sensitivity and specificity were 71.9% and 86.2%, respectively. Conclusion:The Graeb score is an independent influencing factor affecting the clinical outcome of high-grade aSAH patients with IVH. Graeb score >6.5 had higher predictive value for the poor outcome in such patients.

Objective To investigate the effect of hyperbaric oxygen preconditioning (HBO) on activation of extracellular signal-regulated kinase (ERK) and death-associated protein kinase 1 (DAPK1)in ischemia-like treated neurons.Methods (1) Cultured mouse primary neurons were allocated into control,ischemia-like condition treated and HBO preconditioning groups.Neurons in ischemia-like condition treated group were treated with 95％ NO2+5％ CO2 for 30 min,and cells in the HBO preconditioning group were pretreated with 98％ O2+2％ CO2 for 2 h,followed by ischemia-like condition treatment.At the end ofthe treatment,the growth of neurons was measured by MTT assay and expression of phosphorylated extracellular signal-regulated kinase (p-ERK) was detected by Western blotting.Neurons were transfected with ERK over-expression vector and pretreated with HBO and ischemia-like condition,and then,the expressions of activated caspase-3 (cleavage caspase-3) and DAPK1 in neurons were analyzed by Western blotting.The interactions of ERK with DAPK1 were also detected by irnmunoprecipitation.(2) Forty-five BALB/c mice were randomized into control,ischemia treated and HBO preconditioning groups.After finishing the treatments,the interactions of ERK with DAPK1 in hippocampal neurons were detected by immunoprecipitation.Results As compared with control group,cells in ischemia-like condition group showed a reduction of cell survivals,while cells in HBO preconditioning group exhibited an inhibition effect on ischemia-like condition-induced cell survival reduction.The expressions of p-ERK were increased by ischemia-like condition,while decreased by HBO preconditioning,with significant difference (P＜0.05).Moreover,over-expression of ERK promoted level of cleavage caspase-3 in neurons; however,DAPK1 expressions were not affected by ischemia-like condition or HBO preconditioning.The interactions of ERK with DAPK1 were attenuated by HBO preconditioning as compared with those by ischemia-like condition treatment.Conclusion HBO preconditioning inhibits ERK activation and disturbs the interaction between ERK and DAPK1,which blocks cell apoptosis and thus induces ischemic tolerance.

Objective To summarize and analyze the perioperative complications of irreversible electroporation (IRE) ablation in treating tumors at different locations and to discuss their managements. Methods A total of 200 patients with tumors, including pancreatic tumor (n=71), liver tumor (n=64) and other tumors (n =65), were enrolled in this study. All patients received IRE ablation treatment. The perioperative complications were recorded and the data were statistically analyzed. The causes of severe complications and the treatment of complications were discussed. Results None of the patients died during the course of IRE ablation procedure. Severe postoperative complications occurred in the patients with pancreatic tumor or liver tumor, including duodenal artery bleeding in 3 patients with pancreatic tumor, which occurred at 10 days, 11 days and 15 days after IRE ablation respectively, and gastrointestinal bleeding (n =1) and biliary septic shock (n=1) in 2 patients with liver tumor, which occurred at 9 days after IRE ablation, the clinical symptoms were controlled after interventional embolization and/or vascular ligation together with anti-infective therapy. All minor complications were relieved after symptomatic treatment within 14 days. Conclusion IRE ablation has less systemic inflammatory response, and both the intraoperative and postoperative adverse reactions can be easily controlled, besides, IRE ablation has higher clinical safety. Although IRE ablation procedure may damage the internal or peripheral vessels of the pancreatic tumor, this severe complications can be effectively avoided if proper measures are adopted based on the causes of complications. (J Intervent Radiol, 2018, 27: 223-227)

Objective This study applied " modified inflation-deflation" to identify the intersegmental plane and explored a method to remove pulmonary segments from the lobe on the basis of cone-shaped principle."Cone-shaped" Segmentectomy includes anatomical separation of the hilum(conical tip) and the intersegmental plane(undissociative conical surface).The feasibility was evaluated.Methods We retrospectively reviewed 146 patients with peripheric pulmonary nodules who underwent cone-shaped segmentectomy between September 2013 and August 2016.Under the guidance of preoperative 3D-CTBA (three-dimensional computed tomography bronchography and angiography)the targeted bronchi and arteries were dissected.The intersegmental plane was identified by " modified inflation-deflation" method.The inflation-deflation interface was anatomically separated from hilum to distal region along the intersegmental veins,while leaving 1-2 cm in thickness from far end.The residual intersegmental parenchyma was then dissected using endo staplers.Mean folhow-up was 20.4 months.Results Sixty two single segmentectomies,38 combined segmentectomies,43 subsegmentectomies and 3 subsubsegmentectonies were performed.Twelve diagnoses of benign nodule,15 diagnoses of atypical adenomatous hyperplasia,6 diagnoses of metastatic carcinona and 113 diagnosis of primary puhmonary cancer were confirmed pathologically." Modified inflation-deflation" spent (12.5 ±6.4)minutes to show the clear inflation-deflation line.All the targeted bronchi and arteries were dissected precisely and the intersegmental veins were preserved.All the preserved adjacent segments kept approximately original shape after inflated.The tumor size was(1.4 ±0.7)cm,the surgical margin width was(2.5 ± 1.6)cm.The depth of the separated intersegmental plane was(4.2 ±1.7)cm,and the width of the plane was(6.5 ± 2.2)cm.The operative time was(158.5 ± 42.6)min,the volume of hemorrhage was(25.2 ± 15.7) ml,the postoperative hospital stay was (5.2 ± 2.2) days.The total incidence of complications was 5.5％ (8/146).No death within 30 days occurred.Among 113 primary pulmonary cancers,there were 35 T0 N0M0(adenocarcinoma in situ) and 78 T1a No M0 non-small-cell lung cancers.One patient had local recurrence,and no death occurred during follow-up period.Conclusion Modified inflation-deflation can show clear inflation-deflation line to identify the intersegmental plane.Thoracscopic cone-shaped segmentectomy can achieve a complete anatomical segmentectomy.