1.Construction of recombinant retroviral vector of short interfering RNAs specific for macrophage migration inhibitory factor (MIF) and establishment of stable HeLa cell line with a persistent knockdown of MIF
Bo DAI ; Dingzhang XIAO ; Xiyong YU
Chinese Journal of Pathophysiology 2000;0(11):-
AIM:To construct recombinant retroviral vector of short interfering RNAs (siRNA) specific for macrophage migration inhibitory factor (MIF) and to establish the stable knockdown of MIF cell line of mammalian cells by transfecting the recombinant retroviral vectors. METHODS: We synthesized oligo-nucleotides for MIF in vitro, and cloned them into retroviral vector pSuper.retro. Subsequently the plasmids were sequenced and digested to identify the construction of the recombinant retroviral vectors. The vectors RNAi were transfected into packing cell line PHOENIX, which was selected by puromycin later. HeLa cell line was infected by the virus supernatant of stable PHOENIX cell lines, and the stable HeLa cell line showed significantly to silence MIF was established by selecting with puromycin. We also compare the characters of HeLa-pSuper-mock to HeLa-pSuper-MIF cells by using migration assay, adhesion assay, soft agar assay and FACS analysis of the cell-cycle progression. RESULTS: The recombinant retroviral vectors were constructed successfully. The HeLa cell line infected by the supernatant containing the retrovirus of package PHOENIX cells was persistent knockdown of MIF confirmed by Western blotting. Knockdown of MIF in HeLa cells inhibited the migration and adhesion, and decreased the clone formation. FACS analysis revealed that knockdown of MIF arrested HeLa cells in G0/G1 phase. CONCLUSION: We establish the stable HeLa cell line with a persistent knockdown of MIF. Our current studies reveal that MIF is necessary for HeLa cell migration and anchorage-independent growth.
2.Application of uniportal video-assisted thoracoscopic surgery in the treatment of tuberculous destroyed lung
Yuhui JIANG ; Qibin LIU ; Li YAO ; Xiyong DAI
Chinese Journal of Surgery 2024;62(5):432-437
Objective:To examine the efficacy of uniportal video-assisted thoracoscopic surgery in the treatment of tuberculous destroyed lung.Methods:This is a retrospective case series study. The clinical data of 33 patients with tuberculous destroyed lung who had received uniportal video-assisted thoracoscopic pulmonary resection from June 2020 to May 2022 in Department of Surgery, Wuhan Pulmonary Hospital were retrospectively analyzed. There were 13 males and 20 females, aged (47.5±16.2) years (range: 19 to 68 years). The course of the disease was from 15 days to 8 years. All 33 cases had pleural adhesions, including 30 cases with total pleural adhesions and atresia. There were 21 cases of calcification of the thoracic lymph node, 17 cases of aspergillus infection, 4 cases of drug-resistant tuberculosis. The surgical incision was located at the midline of the fifth intercostal axilla, length 4 to 5 cm. The principle of separating pleural adhesions was easy first and difficult later, and then appropriate procedures were selected to resect the diseased lung based on the exploration situation. There were 12 cases that underwent superior lobectomy, 11 cases that underwent superior lobectomy and dorsal segmentectomy, 3 cases that underwent inferior lobectomy, 3 cases that underwent pneumonectomy, 2 cases that underwent middle and inferior lobectomy, and 1 case that underwent superior lobectomy, dorsal segmentectomy and basal segment wedgectomy. The surgical techniques, perioperative evaluation and treatment, management of complications, and the outcome were summarized.Results:Six cases were converted to thoracoscope assisted small incision or thoracotomy. For 27 cases who successfully underwent uniportal VATS, the operation time was (238.7±76.8) minutes (range: 60 to 420 minutes), the intraoperative bleeding was (400.4±315.9) ml (range: 50 to 1 200 ml). The duration of postoperative drainage was (12.7±8.3) days (range: 3 to 42 days). The postoperative hospital stay was (15.2±7.9) days (range: 6 to 43 days). Persistent postoperative pulmonary leakage occurred in 12 cases. There were 2 cases of active thoracic bleeding, one of which was cured with conservative treatment. The other case underwent secondary operation. One case of bronchopleural fistula was cured after continuous thoracic drainage to control infection and implantation of one-way bronchial valve through a fiberoptic bronchoscope.Conclusion:For selected patients with tuberculous destroyed lung, choosing the reasonable surgical procedures and techniques, the uniportal VATS could reduce surgical trauma.
3.Application of uniportal video-assisted thoracoscopic surgery in the treatment of tuberculous destroyed lung
Yuhui JIANG ; Qibin LIU ; Li YAO ; Xiyong DAI
Chinese Journal of Surgery 2024;62(5):432-437
Objective:To examine the efficacy of uniportal video-assisted thoracoscopic surgery in the treatment of tuberculous destroyed lung.Methods:This is a retrospective case series study. The clinical data of 33 patients with tuberculous destroyed lung who had received uniportal video-assisted thoracoscopic pulmonary resection from June 2020 to May 2022 in Department of Surgery, Wuhan Pulmonary Hospital were retrospectively analyzed. There were 13 males and 20 females, aged (47.5±16.2) years (range: 19 to 68 years). The course of the disease was from 15 days to 8 years. All 33 cases had pleural adhesions, including 30 cases with total pleural adhesions and atresia. There were 21 cases of calcification of the thoracic lymph node, 17 cases of aspergillus infection, 4 cases of drug-resistant tuberculosis. The surgical incision was located at the midline of the fifth intercostal axilla, length 4 to 5 cm. The principle of separating pleural adhesions was easy first and difficult later, and then appropriate procedures were selected to resect the diseased lung based on the exploration situation. There were 12 cases that underwent superior lobectomy, 11 cases that underwent superior lobectomy and dorsal segmentectomy, 3 cases that underwent inferior lobectomy, 3 cases that underwent pneumonectomy, 2 cases that underwent middle and inferior lobectomy, and 1 case that underwent superior lobectomy, dorsal segmentectomy and basal segment wedgectomy. The surgical techniques, perioperative evaluation and treatment, management of complications, and the outcome were summarized.Results:Six cases were converted to thoracoscope assisted small incision or thoracotomy. For 27 cases who successfully underwent uniportal VATS, the operation time was (238.7±76.8) minutes (range: 60 to 420 minutes), the intraoperative bleeding was (400.4±315.9) ml (range: 50 to 1 200 ml). The duration of postoperative drainage was (12.7±8.3) days (range: 3 to 42 days). The postoperative hospital stay was (15.2±7.9) days (range: 6 to 43 days). Persistent postoperative pulmonary leakage occurred in 12 cases. There were 2 cases of active thoracic bleeding, one of which was cured with conservative treatment. The other case underwent secondary operation. One case of bronchopleural fistula was cured after continuous thoracic drainage to control infection and implantation of one-way bronchial valve through a fiberoptic bronchoscope.Conclusion:For selected patients with tuberculous destroyed lung, choosing the reasonable surgical procedures and techniques, the uniportal VATS could reduce surgical trauma.
4.Curative effect analysis of decortication in the treatment of durg-resistant tuberculous empyema
Yuhui JIANG ; Lei SHEN ; Xiyong DAI ; Jian SHENG ; Xiaoyu LIU
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(8):491-495
Objective:To investigate the risk and efficacy of the decortication in treatment of drug-resistant tuberculous empyema.Methods:A retrospective analysis was conducted on the 146 patients with tuberculous empyema, who underwent decortication in Wuhan Pulmonary Hospital from March 2016 to November 2018, according to the drug-susceptibility testing of the pleural effusion or tissue specimens, the patients had been divided into drug-resistant group(29 cases) and control group(117 cases), compared the clinical datas such as operation time, intraoperative blood loss, postoperative drainage duration, pulmonary reexpansion duration, postoperative complications and curative ratio between both groups, and the factors that influence the risk and efficacy of surgery had been analyzed.Results:The operative time, intraoperative blood loss, postoperative drainage duration and postoperative complications in the drug-resistant group were larger than those in the control group, and the differences were statistically significant ( P<0.05). There was no significant difference in pulmonary reexpansion duration and curative ratio in the two groups( P>0.05). Conclusion:Decortication is safety, effective and feasible in the treatment of drug-resistant tuberculous empyema, although the operation is difficult and risky.
5.Uniportal thoracoscopic decortication in the treatment of chronic tuberculous empyema
Yuhui JIANG ; Xiaoyu LIU ; Chao GAO ; Jian SHENG ; Li YAO ; Xiyong DAI
Chinese Journal of Thoracic and Cardiovascular Surgery 2022;38(1):18-22
Objective:To investigate the surgical procedure and outcome of uniportal thoracoscopic decortication in the treatment of chronic tuberculous empyema.Methods:From March 2019 to December 2019, the clinical data of 53 patients with chronic tuberculous empyema, who underwent uniportal thoracoscopic decortication in the Surgical Department of Wuhan Pulmonary Hospital were retrospectively analyzed. There were 40 males and 13 females. Age ranged from 16 to 69, averaged 36 years old.36 cases were on the right side and 17 cases on the left side, 38 cases were complicated with pulmonary tuberculosis. All cases had been diagnosed as tuberculous empyema by pathogeny and pathology test in preoperative or postoperative, and received tuberculosis management treatment between 2 and 12 months prior to surgery.The surgical procedure and clinical efficacy of uniportal thoracoscopic decortication were expounded in treatment of chronic tuberculous empyema.Results:Among the 53 patients, 49(92.45%) cases underwent uniportal thoracoscopic decortication, and 4(7.55%) cases changed to thoracotomy. The duration of surgery was 100-370 min, mean(234.53±56.06)min. Intraoperative hemorrhage was 50-1 400 ml, median value 300(175.0, 402.5)ml. Catheter retention time was 3-22 days, median value 8(6.00, 11.25)days. The incidence of surgical complications was 1.89%(1/53), the cure rate was 92.45%(49/53).Conclusion:If the perioperative evaluation and treatment are appropriate, and the operator is experienced, the uniportal thoracoscopic decortication is feasible, safe and effective in the therapy of chronic tuberculous empyema.
6.Analysis of the expression levels and clinical diagnostic value of miR-455 and miR-383 in serum of non-small cell lung cancer patients
Chao GAO ; Xiyong DAI ; Yuhui JIANG ; Xiaoyu LIU ; Jing LI ; Hao XIONG
Journal of Clinical Surgery 2024;32(6):607-610
Objective To detect and analyze the expression levels of serum microRNA(miR)-455 and miR-383 in patients with non-small cell lung cancer(NSCLC),and to explore their clinical diagnostic value.Methods A total of 98 NSCLC patients in our hospital from March 2020 to January 2023 were regarded as the NSCLC group,98 patients with benign lung diseases admitted during the same period were regarded as the benign lung disease group,and another 98 healthy volunteers who underwent physical examination in our hospital during the same period were collected as the health group.Real-time fluorescence quantitative PCR(RT-qPCR)method was applied to detect the expression levels of miR-455 and miR-383 in serum.Pearson was applied to analyze the correlation between serum miR-455 expression level and miR-383 level in NSCLC patients;multivariate Logistic regression and ROC curve was applied to analyze the influencing factors and diagnostic value of NSCLC occurrence.Results The expression levels of miR-455 and miR-383 in the serum of the healthy group,benign lung disease group,and NSCLC group decreased sequentially.Pearson analysis showed there was a positive correlation between serum miR-455 expression level and miR-383 level in NSCLC patients(r=0.582,P<0.05).Logistic analysis found that low expression levels of miR-455 and miR-383 were risk factors for the occurrence of NSCLC(P<0.05).ROC curve analysis showed that the AUC of NSCLC diagnosed by the combination of miR-455 and miR-383 was obviously higher than that diagnosed by miR-455 alone(Z=3.604,P=0.000)and miR-383 alone(Z=2.594,P=0.010).Conclusion The relative expression levels of miR-455 and miR-383 in serum of NSCLC patients are obviously down-regulated.The combined detection of the two has high diagnostic value for NSCLC.
7.Application of single-direction lobectomy in the treatment of complicated pulmonary tuberculosis: A retrospective cohort study
Yuhui JIANG ; Lei SHEN ; Xiyong DAI
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(12):1607-1611
Objective To explore the value of the single-direction lobectomy in the treatment of complicated pulmonary tuberculosis. Methods A retrospective analysis was performed on 88 patients with complicated pulmonary tuberculosis who received lobectomy in our hospital from 2017 to 2019. There were 64 males and 24 females, with an average age of 21-70 (47.67±13.39) years. According to the surgical procedure, patients who received single-direction lobectomy were divided into a single-direction group (n=32), and those who received traditional lobectomy were divided into a control group (n=56). Results The two groups had no statistical differences in gender, age, primary disease and complications, lesion morphology, clinical symptoms, operative site, interlobitis adhesion or hilar mediastinal lymph node calcification (P>0.05). Operation time [210.0 (180.0, 315.0) min vs. 300.0 (240.0, 320.0) min], intraoperative blood loss [200.0 (100.0, 337.5) mL vs. 325.0 (200.0, 600.0) mL], postoperative lung air leak time [3.0 (2.0, 5.0) d vs. 9.0 (6.8, 12.0) d] and the postoperative hospital stay [11.5 (8.0, 14.8) d vs. 18.0 (14.0, 22.0) d] of the single-direction group were less or shorter than those of the control group (P<0.05). There was no statistical difference between the single-direction group and the control group in the incidence of surgical complications [1 patient (3.12%) vs. 10 patients (17.86%)] or the cure rate [32 patients (100.00%) vs. 54 patients (96.43%)]. Conclusion The single-direction lobectomy can reduce lung injury and bleeding, shorten the duration of operation and accelerate the postoperative recovery in patients with complicated pulmonary tuberculosis, which has certain advantages compared with traditional lobectomy.
8.Clinical comparison of pulmonary lobectomy in patients with massive hemoptysis of pulmonary tuberculosis after bronchial artery embolization
JIANG Yuhui ; SHEN Lei ; DAI Xiyong ; SHENG Jian ; LIU Xiaoyu
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2019;26(12):1190-1193
Objective To compare the clinical data of pulmonary lobectomy in patients with massive hemoptysis of pulmonary tuberculosis after bronchial artery embolization in the short and long term, so as to provide a reference for clinical choices of appropriate operation time. Methods A retrospective analysis was conducted on 33 patients with massive hemoptysis of pulmonary tuberculosis, who had received pulmonary lobectomy after bronchial artery embolization in Wuhan Pulmonary Hospital from January 2015 to November 2017, including 29 males and 4 females aged of 23-66 (52.64±9.70) years. According to the time interval between bronchial artery embolization and lobectomy, the patients were divided into a short-term group (<2 weeks, 14 patients) and a long-term group (>1 month, 19 patients). The clinical data, such as operation time, intraoperative blood loss, postoperative extubation time and serious postoperative complications, were observed in the two groups for statistical analysis. Results The operative time (297.13±75.69 min vs. 231.32±67.57 min, P=0.013), intraoperative blood loss (685.74±325.51 mL vs. 355.83±259.11 mL, P=0.002), postoperative extubation time (14.07±5.24 d vs. 8.90±3.57 d, P=0.003) of the short-term group were all higher than those in the long-term group. Conclusion For the patients with massive hemoptysis of pulmonary tuberculosis, who had surgical indications and no risk of early rebleeding after bronchial artery embolization, pulmonary lobectomy should be performed late until the patient's physical condition and the primary disease was stable.