Objective To extract the loci of murine MHC gene and construct plasmids.Methods The RNA of mice was extracted and reversely transcribed into cDNA.By using nested PCR,the products were connected with T vector,cloned,and sequenced.Subsequently,the genes were digested by endonucleases,connected with expression vector,and sequenced again to choose the correct clones.Results After the nested PCR,the products were approved by sequencing.After being connected with the vectors,they were approved again by sequencing and the correct clones were chosen.Conclusion All of the loci of the MHC gene can be obtained by nested PCR.The plasmids from the correct clone can be used in the further experiments of transferring the gene to mitigate the transplantation rejection.

ObjectiveTo discuss the clinical experience and influence factors of airway complications after lung transplantation for end-stage lung diseases through reviewed 28 lung transplantation cases in our center.MethodsFrom August 2005 to December 2010,28 patients with end-stage lung diseases received lung transplantations consecutively in our center,in which 13 patients were bilateral-lung transplantation(BLT) and 15 patients were single-lung transplantation(SLT).The donor lungs were perfused with LPD solution antegrade and retrograde followed.During operation,the pulmonary artery pressure and flow rate were tested real time through the transesophageal echocardiography and Swans catheter.Postoperative care of patients was in respiratory intensive care unit,and immunosuppressive drugs were adjusted according to blood drug concentration.ResultsThere were no airway complications including anastomotic fistula or stenosis found in all patients.The mortality was 7.2％ in the early postoperative period ( 1-30 days).Cumulative survival rate was 94.1％ ％ at 1 year,76.2％ at 2 years,and 71.4％ at 3 years respectively.Four patients (14.3％) died in the postoperative 90 days.Three patients were reanastomosed pulmonary artery in operation because of stenosis detected by transesophageal echocardiography.After operation,three patients were reoperated,in which two were bleeding and one was pulmonary bulla and pneumothorax.All patients were followed from 1 year to 6.1 years after operation.The quality of life was improved significantly.ConclusionThe well improved technique of lung transplantation is helpful to reduce the operation related complications,decrease the early mortality post operation and play the important role in the effects of quality of life and long term survival rate.The intraoperative transesophageal echocardiography examination could detect the anomalous situation of vascular anastomosis.At the same time the patients should get benefits from the routine and close follow-up.

Objective To observe the changes of thoracic drainage volume after lung transplantation,and to explore the influencing factors and their relationship with the prognosis of lung transplantation.Methods This retrospective analysis included 46 consecutive lung transplantations in our hospital between 2005 and 2016.The volume of postoperative chest drainage was recorded and analyzed.Single factor analysis of the factors that may affect the drainage was done.The patients were divided into different groups according to different prognosis at different time points after operation,and drainage volume was compared among groups.All analyses were performed with SPPS,version 19.0.Results There were no operative deaths.The median thoracic drainage time was 9.3 days,the median total thoracic drainage volume was 4318 mL,and the average daily drainage volume was 487 mL.During the perioperative period,the use of cardiopulmonary bypass or extracorporeal membrane oxygenation increased significantly the volume of unilateral pleural drainage (P＜0.05).The average daily drainage volume and the average daily unilateral pleural drainage volume in the postoperative dead patients were significantly greater than those of survivals at 1 st month,3rd month and 1 st year after lung transplantation (P＜0.05).At 3rd month after transplantation,the total amount of postoperative thoracic drainage was significantly greater than that of survivals (P＜ 0.05).The average daily unilateral pleural drainage volume in dead patients after surgery was significantly greater than that of the survival patients 3 years after operation (P＜0.05).Conclusion Intraoperative hemostasis in patients with lung transplantation should receive considerable attention,cardiopulmonary bypass and extracorporeal membrane oxygenation should be used cautiously during perioperative period,and greater postoperative chest drainage predicated poor short-term prognosis.

Objective: To evaluate the incidence of postoperative venous thromboembolism (VTE) after thoracic surgery and its characteristic. Methods: This was a single-center, prospective cohort study. Patients undergoing major thoracic surgeries between July 2016 and March 2017 at Department of Thoracic Surgery, Beijing Chaoyang Hospital Affiliated to Capital Medical University were enrolled in this study. Besides the routine examination, all patients were screened for deep venous thrombosis (DVT) by using noninvasive duplex lower-extremity ultrasonography after surgery. CT pulmonary angiography (CTPA) was carried out if patients had one of the following conditions including typical symptoms of PE, high Caprini score (>9 points) or new diagnosed postoperative DVT. Caprini risk assessment model was used to detect high risk patients. No patients received any prophylaxis of VTE before surgery. Further data was analyzed for identifying the incidence of postoperative VTE. The t-test, χ² test or Wilcoxon rank-sum test was used to analyze the quantitative data and classification data, respectively. Results: Totally 345 patients who undergoing major thoracic surgery were enrolled in this study including 145 benign diseases and 200 malignant diseases.There were 207 male and 138 female, aging from 15 to 85 years. Surgery procedures included 285 lung surgeries, 27 esophagectomies, 22 mediastinal surgeries and 11 other procedures. The overall incidence of VTE was 13.9% (48 of 345) after major thoracic surgery including 39 patients with newly diagnosed DVT (81.2%), 1 patient with PE (2.1%) and 8 patients with DVT+ PE (16.7%). The median time of VTE detected was 4.5 days postoperative. There were 89.6% (43/48) VTE cases diagnosed in 1 week. The incidence of VTE was 9.0% in patients with benign diseases, while 17.5% in malignant diseases (χ²=5.112, P<0.05). The incidence of VTE in patients with pulmonary diseases was 12.6%, among that, in patients with lung cancer and benign lung diseases was 16.4% and 7.5 % (χ²=4.946, P<0.05), respectively. Regarding to Caprini risk assessment model, the incidence of VTE in low risk patients, moderate risk patients (Caprini score 5 to 8 points)and high risk patients(≥9 points)were 0(0/77), 15.2%(33/217) and 29.4%(15/51), respectively(Z=-12.166, P<0.05). In patients with lung cancer, 98.2% of patients were moderate risk or high risk; only 3 cases scored low risk. The incidence of VTE in moderate risk and high risk patients was 13.4%(18/134) and 32.1%(9/28), respectively, while it was 0(0/3) in low risk patients. Conclusion s The overall incidence of VTE after major thoracic surgeries is 13.9%, and the incidence of VTE after lung cancer surgeries was 16.4%. Most of the VTE cases occurr within one week after the surgery. Caprini risk assessment model can identify high risk patients effectively.

Objective     To investigate the surgical procedure selection, operation technique and safety of anatomic sublobar resection for pulmonary nodules. Methods     The clinical data of 242 patients with clinical stage ⅠA lung cancer who underwent anatomic sublobar resection in our hospital between 2017 and 2020 were retrospectively analyzed. There were 81 males and 161 females with a median age of 57.0 (50.0, 65.0) years. They were divided into 4 groups according to the surgical methods, including a segmentectomy group (n=148), a combined segmentectomy group (n=31), an enlarged segmentectomy group (n=43) and an anatomic wedge resection group (n=20). The preoperative CT data, operation related indexes and early postoperative outcomes of each group were summarized. Results     The median medical history of the patients was 4.0 months. The median maximum diameter of nodule on CT image was 1.1 cm, and the consolidation/tumor ratio (CTR) was ≤0.25 in 81.0% of the patients. A total of 240 patients were primary lung adenocarcinoma. The median operation time was 130.0 min, the median blood loss was 50.0 mL, the median chest drainage time was 3.0 d, and the hospitalization cost was (53.0±12.0) thousand yuan. The operation time of combined segmentectomy was longer than that of the segmentectomy group (P=0.001). The operation time (P=0.000), intraoperative blood loss (P=0.000), lymph nodes dissected (P=0.007) and cost of hospitalization (P=0.000) in the anatomic wedge resection group were shorter or less than those in the other three groups. There was no significant difference in the drainage time, total drainage volume, air leakage or postoperative hospital stay among the four groups (P>0.05). Conclusion     The combined application of segmentectomy and wedge resection technique provides a more flexible surgical option for the surgical treatment of early lung cancer with ground glass opacity as the main component.

Objective: To improve the understanding of thyrotropin-secreting adenoma in multiple endocrine neoplasia type 1(MEN1) through analyzing the clinical diagnosis and treatment process, as well as outcomes in one case of this disorder. Methods: The clinical manifestations, biochemical and hormone levels, imaging presentations, medical and surgical treatments, and post-operational pathologic findings in the process of diagnosis and treatment of a patient with thyrotropin-secreting adenoma in MEN1 were analyzed. The next generation sequencing followed by Sanger method was used for analyzing MEN1 and related genes. The results were evaluated with online PolyPhen2 and PROVEAN for variation hazard. Results: One 19-year old male patient was diagnosed with hyperthyroidism due to thyrotoxicosis and high level of thyroid hormones(THs) with measurable TSH(2.78 mIU/L) and negative thyrotropin receptor antibody(TRAb). Meanwhile, primary hyperparathyroidism was suggested by hypercalcemia, hypophosphatemia, and elevated intact parathyroid hormone(PTH) level, all the parameters were returned to normal after surgical resection of the mass which was below the left thyroid lobe indicated by ultrasound and ^99mTc scan. Thyrotoxicosis remained in spite of one year treatment with antithyroid drug, thyrotropinoma was then suspected, and subsequent MRI scan found a macroadenoma at right pituitary. TSH and THs returned to normal 1 month after transsphenoidal removal of the adenoma. As expected, immunohistochemical staining revealed TSH positive. In addition, a pancreatic mass was found by both CT and MRI scan, which was considered as a silent neuroendocrine tumor. Gene analysis revealed a missense mutation of MEN1 as c. 415C>T and p. His139Tyr(H139Y), which was predicted highly hazard. Only five cases of thyrotropinoma in MEN1 were previously reported. Conclusion Thyrotropinoma should be cautiously identified from hyperthyroidism to avoid misdiagnosis and mistreatment, and it should keep in mind that thyrotropinoma may be associated with MEN1 though it would be very rare.

Objective     To study the correlation of preoperative hemoglobin amount with venous thromboembolism (VTE) after surgical treatment of bronchiectasis and the clinical significance. Methods     A retrospective study was performed on patients with bronchiectasis who underwent surgical treatment in our center from June 2017 to November 2021. The differences in blood parameters between the VTE patients and non-VTE patients were compared. The relationship between preoperative hemoglobin and VTE was confirmed by quartile grouping and receiver operating characteristic (ROC) curve. Results     A total of 122 patients were enrolled, including 50 males and 72 females, with a mean age of 52.52±12.29 years. The overall incidence of VTE after bronchiectasis was 9.02% (11/122). Preoperative hemoglobin amount (OR=0.923, 95%CI 0.870-0.980, P=0.008) and D-dimer amount (OR=1.734, 95%CI 1.087-2.766, P=0.021) were independent influencing factors for VTE after bronchiectasis. The incidence of VTE after bronchiectasis decreased gradually with the increase of preoperative hemoglobin amount. The area under the ROC curve (AUC) of postoperative D-dimer alone was 0.757, whereas the AUC of postoperative D-dimer combined with preoperative hemoglobin amount was 0.878. Conclusion     Low preoperative hemoglobin is an independent risk factor for postoperative VTE. Postoperative D-dimer combined with preoperative hemoglobin amount has a better predictive performance compared with postoperative D-dimer alone for postoperative VTE.

Metabolic-associated fatty liver disease (MAFLD), which is previously known as non-alcoholic fatty liver disease (NAFLD), represents a major health concern worldwide with limited therapy. Here, we provide evidence that ferroptosis, a novel form of regulated cell death characterized by iron-driven lipid peroxidation, was comprehensively activated in liver tissues from MAFLD patients. The canonical-GPX4 (cGPX4), which is the most important negative controller of ferroptosis, is downregulated at protein but not mRNA level. Interestingly, a non-canonical GPX4 transcript-variant is induced (inducible-GPX4, iGPX4) in MAFLD condition. The high fat-fructose/sucrose diet (HFFD) and methionine/choline-deficient diet (MCD)-induced MAFLD pathologies, including hepatocellular ballooning, steatohepatitis and fibrosis, were attenuated and aggravated, respectively, in cGPX4-and iGPX4-knockin mice. cGPX4 and iGPX4 isoforms also displayed opposing effects on oxidative stress and ferroptosis in hepatocytes. Knockdown of iGPX4 by siRNA alleviated lipid stress, ferroptosis and cell injury. Mechanistically, the triggered iGPX4 interacts with cGPX4 to facilitate the transformation of cGPX4 from enzymatic-active monomer to enzymatic-inactive oligomers upon lipid stress, and thus promotes ferroptosis. Co-immunoprecipitation and nano LC-MS/MS analyses confirmed the interaction between iGPX4 and cGPX4. Our results reveal a detrimental role of non-canonical GPX4 isoform in ferroptosis, and indicate selectively targeting iGPX4 may be a promising therapeutic strategy for MAFLD.