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.

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.

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.

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.

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.