Objective To isolate stable passage primary HIV-1 drug resistant strains and observe replication dynamics of the drug resistant isolates and evolvement tendency of the drug resistant mutations in vitro.Methods Peripheral blood mononuclear cells(PBMCs)from 15 AIDS patients receiving highly active antiretroviral therapy(HAART)were collected,and the primary HIV-1 stains were separated utilizing co-cultivated with PBMCs from normal people.HIV-1 pol genes from those strains were obtained by RT-PCR and sequenced.The drug resistant mutations were analyzed in the Stanford HIV Drug Resistance Database.Results Eight strong positive strains were isolated from 15 AIDS patients with viral loads higher than 1000 copies/ml,and two of them were drug resistant.Drug resistant mutations of the two strains were respectively K103N/K238T and M184V/K103N/Y181C/H221Y which show high-level resistance to NVP and 3TC/NVP,respectively.K103N,M184V,Y181C and H221Y exist stably in the environment without drug pressure,however,RT K238T reverted to K238.Conclusion Two drug resistant strains were successfully isolated in vitro without drug pressure.Strains with K103N shows superior fitness and can exist steadily.Strains with M184V and K103N/Y181C/H221Y can also replicate stably in vitro without drug pressure.NNRTI mutation K238T reproduces astatically,which suggests that RT 238 codon might revert gradually to wild genotype.

Objective To elucidate the molecular evolutional characteristics of HIV-1 non-nucleoside reverse transcriptase inhibitor (NNRTI) drug resistance-associated mutations in AIDS patients receiving highly active antiretroviral therapy (HAART).Methods Four AIDS patients receiving HAART with good adherence within a HlV-1 drug resistance cohort from a rural region in central China were selected,who possessed susceptible virus at the beginning of treatment and gradually came to produce resistance to NNRTIs during the process of antiretroviral therapy (ART),reverse transcriptase (RT) genes from each patient's peripheral blood samples (from 3 to 30 months after withdrawal) were cloned and sequenced in succession.Results To sequenced total 855 clones and obtained the HIV-1 NNRTI drug resistance-asseciated mutations patterns of the four patients: (1)G190A often appeared with F227 L and had the tendency of accumulating P236V during the process of treatmenL (2)Y188C always presented alone and sometimes it concured with P236V.(3) YI81C frequently concured with VI79D or KIO3N and the combination varies from patient to patient.(4)K103N often combined with Y181C or M230L Conclusions The molecular evolutional characteristics of HIV-1 NNRTI drug resistance-asseciated mutations in the 4 AIDS patients are summarized.They showed different pathways on HIV-1 NNRTI drug resistance-associated mutations and those mutations detected early tend to be predominant strains.

Objective To evaluate the safety and efficacy of basic anesthesia combined with local anesthesia in the preoperative localization of multiple pulmonary nodules. Methods The clinical data of patients who underwent preoperative localization for multiple pulmonary nodules resection under single-port thoracoscopy in Nanjing Brain Hospital from July 2023 to September 2023 were extracted. They were divided into a group A and a group B according to the localization method. The patients in the group A were localized under local anesthesia, and the patients in the group B were localized with basic anesthesia combined with local anesthesia. The basic clinical characteristics, localization success rate, incidence of localization complications, localization time, and pain score of the two groups were compared and analyzed. Results Finally, we included 200 patients with 100 patients in each group. There were 49 males and 51 females at age of 25-77 (50.94±14.29) years in the group A. There are 45 males and 55 females at age of 24-78 (48.25±14.04) years in the group B. The incidence of localization complications (4% vs. 13%, P=0.04), localization time [(19.90±8.66) min vs. (15.23±5.98) min, P<0.01], and pain score[ (2.01±2.09) vs. (3.29±2.54), P<0.01] in the group B were significantly lower than those in the group A, and the differences were statistically significant. The localization success rate of the group B was significantly higher than that of the group A (98% vs. 92%, P=0.04), and the difference was statistically significant.Conclusion Mobile CT combined with basic anesthesia for preoperative localization of multiple pulmonary nodules is highly safe, has a high success rate, and provides high patient comfort, making it a valuable approach for clinical promotion.

Objective To analyze the independent risk factors affecting complications of preoperative CT-guided Hookwire localization of pulmonary nodules, and establish and validate a nomogram risk prediction model. Methods Clinical data of patients who underwent thoracoscopic lung surgery with preoperative CT-guided Hookwire localization at the Department of Thoracic Surgery, Affiliated Nanjing Brain Hospital, Nanjing Medical University from January 2023 to October 2023 were collected. Patients were divided into a complication group and a non-complication group according to whether they had complications. The clinical data of the two groups were compared by univariate analysis and multivariate binary logistic regression analysis to determine the independent risk factors causing complications during localization, and a nomogram prediction model was established. The discrimination of the model was evaluated by receiver operating characteristic (ROC) curve, and the consistency between predicted events and actual results was evaluated by calibration curve. Results A total of 300 patients were included, including 143 males and 157 females, aged 24-68 (46.00±22.81) years. Univariate analysis showed that there were statistically significant differences in age, number and location of nodules, preoperative anxiety score, history of chronic obstructive pulmonary disease (COPD), number of needle adjustments, pain score, and distance between the tip of the localization needle and the visceral pleura between the two groups (P<0.05). Multivariate binary logistic regression analysis suggested that pain score [OR=1.253, 95%CI (1.094, 1.434), P=0.001], age [OR=1.020, 95%CI (1.000, 1.042), P=0.049], history of COPD [OR=3.281, 95%CI (1.751, 6.146), P<0.001], number of nodules [OR=1.667, 95%CI (1.221, 2.274), P=0.001], preoperative anxiety score [OR=1.061, 95%CI (1.031, 1.092), P<0.001], number of needle adjustments [OR=1.832, 95%CI (1.263, 2.658), P=0.001], and distance between the needle tip and the visceral pleura [OR=1.759, 95%CI (1.373, 2.254), P<0.001] were associated with localization complications. The area under the ROC curve for the modeling group was 0.825, and that for the validation group was 0.845. Hosmer-Lemeshow test showed that there was no statistically significant difference between the ideal curve of the model fitting curve and that of the modeling group and internal validation group, indicating good goodness of fit (χ2=6.488, P=0.593). Conclusion Advanced age, multiple nodules, preoperative anxiety, history of COPD, multiple needle adjustments, severe pain during localization, and long distance between the tip of the localization needle and the visceral pleura are independent risk factors for complications of lung nodule localization, and the prediction model based on these factors has good predictive performance.

Objective To explore the feasibility and accuracy of using indocyanine green fluorescence (ICGF) to identify the intersegmental plane after ligation of the target pulmonary vein during thoracoscopic segmentectomy. Methods From December 2022 to June 2023, the patients with pulmonary nodules undergoing video-assisted thoracoscopic anatomical segmentectomy with intersegmental plane displayed using ICGF after ligation of the target pulmonary vein by the same medical team in our hospital were collected. Preoperative three-dimensional reconstruction was used to identify the target segment where the pulmonary nodule was located and the anatomical structure of the arteries, veins, and bronchi in the target segment. The intersegmental plane was first determined by the inflation-deflation method after the target pulmonary vein was ligated during the operation. During the waiting period, the target artery and bronchus could be separated but not cut off. The inflation-deflation boundary was marked by electrocoagulation, and then ICGF was injected via peripheral vein to identify the intersegmental plane again, and the consistency of the two intersegmental planes was finally evaluated. Results Finally 32 patients were collected, including 14 males and 18 females, with an average age of 58.69±11.84 years, ranging from 25 to 76 years. The intersegmental plane determined by inflation-deflation method was basically consistent with ICGF method in all patients. All the 32 patients successfully completed uniportal thoracoscopic segmentectomy without ICGF-related complications or perioperative death. The average operation time was 98.59±20.72 min, the average intraoperative blood loss was 45.31±35.65 mL, and the average postoperative chest tube duration was 3.50±1.16 days. The average postoperative hospital stay was 4.66±1.29 days, and the average tumor margin width was 26.96±5.86 mm. Conclusion The ICGF can safely and accurately identify the intersegmental plane by target pulmonary venous preferential ligation in thoracoscopic segmentectomy, which is a useful exploration and important supplement to the simplified thoracoscopic anatomical segmentectomy.