Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Objective:Chimeric antigen receptor T-cell(CAR-T)immunotherapy has made major breakthroughs in the treatment of blood tu-mors.However,current CAR-T therapies face several limitations:they require autologous cells,involve a lengthy and costly production pro-cess,and use lentiviral transduction that carry risk of insertional carcinogenesis due to random integration.Therefore,there is an urgent need to develop a universal cost-effective cancer immunotherapy method generating CAR-T cells for in vivo cancer immunotherapy.Meth-ods:This study successfully established an exosome-mediated,T-cell targeted delivery system,demonstrating both precise design and func-tional efficacy for biomedical applications.To optimize CAR-T cell generation the transfection dose was adjusted,and the kinetics of CAR-T cell percentage were recorded.The cytotoxicity of the resulting CAR-T cells was evaluated in vitro by calcein-AM release.To test the tumor-killing in vivo of engineered exosomes,human PBMCs were injected into NPG mice via the tail vein to establish humanized mice,followed by intravenous injection of tumor cells to induce cancer.Results:To overcome the limitations of conditional autologous CAR-T cells,we de-veloped a T cell-targeted exosome system capable of specifically targeting human CD3+,CD4+,and CD8+T cells.CAR-T production was dose-dependent,with transfection efficiency reaching upto 97.8%at 106 particles/cell.Both in vitro cytotoxicity assays and in vivo animal experi-ments demonstrated that exosome-incubated CAR-T cells effectively eliminated CD19-positive Raji cells,highlighting their specificity and therapeutic potential in antigen-directed applications.Conclusions:We successfully established a CD8-targeting exosome delivery system for CAR-T cell production capable of transforming CD8+T cells into functional CAR-T cells,which showed significant tumor-killing ability in vitro and in mice.Compared with the traditional lentiviral vector for the preparation of CAR-T cells in vitro,in vivo-reprogrammed CAR-T cells us-ing our CD8-targeted exosome delivery system,with higher transfection efficiency,shorter production period,lower cost,and eliminated the risk of insertion carcinogenesis.This strategy promises to bring a new era of universal CAR-T medicine,which can improve cancer immuno-therapy and may hold promise as a therapeutic platform to treat various diseases.

Objective To observe the effect of transcutaneous electrical acupoint stimulation(TEAS)on circulation depression in patients underwent thoracoscopic radical resection of lung cancer under general anesthesia combined with thoracic paravertebral block(TPVB).Methods A total of 150 patients from Octomber 2021 to May 2022,58 males and 92 females,aged 19-64 years,BMI 18-30 kg/m2,ASA physical status Ⅰ or Ⅱ,underwent thoracoscopic radical resection of lung cancer under general anesthesia combined with TPVB were enrolled.According to random number table method,the patients were divided into two groups:the TEAS group and the control group,75 patients in each group.In the TEAS group,transcutaneous electrical acupoint stimulation was performed at Hegu,Neiguan,and Zusanli 30 minutes be-fore induction until the end of operation.In the control group,the electrodes were only connected at the same time point without electrical stimulation.HR,SBP,DBP,MAP,and BIS were recorded before stimu-lation(T0),10 minutes after TPVB(T1),the time of skin incision(T2),30 minutes after operation star-ted(T3),60 minutes after operation started(T4),the end of operation(T5),and 30 minutes after opera-tion(T6).The incidences of bradycardia,tachycardia,hypotension,and hypertension,and the usages of vasoactive drugs during operation were recorded.The dosages of propofol,sufentanil,and remifentanil in the operation were recorded.The VAS pain score 1,2,and 7 days after operation,the usages of analgesics used within 7 days after operation,postoperative adverse effects such as nausea and vomiting,dizziness,chest tightness,and shortness of breath,and the length of hospital stay were recorded.Results Compared with the control group,intraoperative infusion volume,incidence of hypotension,hypertension,and circulation depression,the usages of deoxyepinephrine,ephedrine,norepinephrine,and urapidil intraoperation,VAS pain scores 1 and 2 days after operation,and the usage of analgesics within 7 days after operation were sig-nificantly decreased(P＜0.05),length of hospital stay was significantly shortened(P＜0.05),SBP,DBP,and MAP were significantly increased at T1(P＜0.05),the dosagesof propofol,sufentanil,and remifentanil were significantly decreased in the TEAS group(P＜0.05).There were no significantly differ-ences of nausea and vomiting,dizziness,and shortness of breath between the two groups.Conclusion TEAS can improve the circulation depression,and reduce the incidences of intraoperative hypotension and hypertension,decrease the dosages of anesthetics and the rate of using vasoactive drugs during operation,improve early postoperative acute pain and shorten the length of hospital stay in patients undergoing thoraco-scopic radical resection of lung cancer under general anesthesia combined with TPVB.

Objective:To investigate the application of three-dimensional image reconstruction combined with problem-based learning (PBL) in the teaching of physicians receiving continuing education in thoracic surgery.Methods:A total of 68 physicians who received continuing education in Department of Thoracic Surgery in our hospital were selected as research subjects, and they were divided into control group and observation group using a random number table, with 34 physicians in each group. The physicians in the control group received traditional teaching, while those in the observation group received three-dimensional image reconstruction combined with PBL teaching. A questionnaire survey, theoretical assessment, and assessment of practical skills were performed to evaluate the effect of teaching. SPSS 22.0 was used to perform the t-test. Results:Compared with the control group, the observation group had significantly higher degrees of satisfaction with each item of the questionnaire survey ( P<0.05). Compared with the control group, the observation group had significantly higher scores of theoretical assessment [(94.07±6.03) vs. (86.34±5.46), P<0.001] and the assessment of practical skills [(95.20±5.48) vs. (84.71±6.14), P<0.001]. Conclusion:The application of three-dimensional image reconstruction combined with PBL teaching can help to improve the comprehensive ability of physicians receiving continuing education.

Objective:To establish a candidate reference procedure for the enumeration of cell particles in urine and applied to the multi-center performance evaluation of an automated urine formed elements analyzer.Methods:According to the standardized mannual microscopic examination of fresh non-centrifuged urine samples and the recommended reference method for enumeration of cell particles in urine published by ISLH, we established a candidate reference procedure for the enumeration of cell particles in urine. From four class A tertiary hospitals′ clinical laboratories, three rigorous trained technicians per hospital tested the same specimen respectively using the reference procedure. Each specimen was repeatedly counted 5 times, obtaining the quantitative results of cell particles were obtained in urine. Four hospitals used the established candidate reference measurement procedure and the automated urine formed elements analyzer to detect 40 to 60 urine specimens from September 2020 to January 2021, and evaluate the established reference method, meanwhile evaluate the accuracy and consistency of the each count from automated urinalysis analyzer.Results:Using the candidate reference measurement procedures, the coefficient of variation of results derived from three trained technicians per hospital was less than 6.98% (red blood cells), 6.99% (white blood cells), 13.94% (epithelial cells) and met the quality requirements. The performance evaluation results of automated urine formed elements analyzer showed that the accuracy of red blood cells, white blood cells and epithelial cells met the requirements (bias≤4.98%) and was well consistent with the reference measurement procedure ( R2≥0.989). Conclusions:A candidate reference measurement procedure for the enumeration of urine cell particles was successfully established with satisfactory precision and accuracy. This procedure was applied to multicenter performance evaluation of an automated urine formed elements analyzer with good accuracy and consistency.

Objective:To explore the clinical effects of unilateral secondary puncture percutaneous vertebroplasty (PVP) in the treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fractures (ASOTLF).Methods:A retrospective case-control study was conducted to analyze the clinical data of 193 patients with type ⅡA ASOTLF who had been admitted to Department of Spine Surgery, Honghui Hospital from February 2016 to October 2018. They were 71 males and 122 females, aged from 65 to 90 years [average, (73.9±4.3) years]. The segments injured were T10 in 21 cases, T11 in 27 cases, T12 in 44 cases, L1 in 48 cases, L2 in 29 cases, L3 in 14 cases, and L4 in 10 cases. Of them, 85 received unilateral secondary puncture PVP (observation group) and 108 did not (control group). The clinical effects were evaluated by comparing between the 2 groups the operation time, bone cement injection volume, intraoperative blood loss, hospital stay, and visual analogue scale (VAS) for back pain, spinal Oswestry disability index (ODI), anterior height of the injured vertebral body (AH) and kyphosis angle (KA) of the injured vertebra before operation, at 3 days after operation and the last follow-up. The bone cement leakage and fracture of adjacent vertebral body were observed.Results:All patients were followed up for 12 to 24 months (average, 15.8 months). There was no significant difference in the preoperative general data between the 2 groups, showing they were comparable ( P>0.05). The operation time and bone cement injection volume [(36.2±1.4) min and (5.5±0.7) mL] in the observation group were significantly longer or more than those in the control group [(32.3±1.7) min and (4.0±0.7) mL] ( P<0.05). There was no significant difference in the hospital stay or intraoperative blood loss between the 2 groups ( P>0.05). The VAS, ODI, AH and KA at 3 days after operation and the last follow-up were significantly improved compared with those before operation in both groups ( P<0.05). There was no significant difference in VAS, ODI, AH or KA between the 2 groups before operation or at 3 days after operation ( P>0.05). However, the VAS, ODI, AH and KA at the last follow-up in the observation group [(2.2±0.8) points, 19.2%±5.8%, (2.90±0.21) cm, and 12.2°±1.5°] were better than those in the control group [(3.1±0.9) points, 22.8%±5.3%, (2.41±0.15) cm, and 13.3°±1.2°]. There was no significant difference between the 2 groups in the incidence of postoperative bone cement leakage or that of adjacent vertebral fracture ( P>0.05). Conclusions:In the treatment of type ⅡA ASOTLF, unilateral secondary puncture PVP can result in satisfactory clinical effects, because it effectively promotes dispersion of bone cement and prevents re-collapse of the vertebra operated but does not increase the risks of bone cement leakage and adjacent vertebral fracture.

Objective:To explore the treatment options for patients with osteoporotic thoracolumbar injury classification and severity score (OTLICS) of 4 points following acute symptomatic osteoporotic vertebral compression fracture (OVCF) .Methods:A retrospective case-control study was conducted to analyze the clinical data of 108 patients with acute symptomatic OVCF admitted to Honghui Hospital, Xi'an Jiaotong University Faculty of Medicine from February 2016 to February 2018. There were 21 males and 87 females, aged 55-92 years [(71.6±5.3)years]. All patients had OTLICS score of 4 points. Of all, 76 patients were treated operatively (surgery group) and 32 patients were treated non-operatively (non-surgery group). The visual analogue scale (VAS), Oswestry disability index (ODI) and quality of life score (SF-36) were compared between the two groups before treatment, at 1 week, 3 months, 6 months after treatment, and at the last follow-up. The recovery of the injured vertebrae before and after treatment (Cobb angle of the injured vertebra, relative height of the anterior edge of the injured vertebrae), adjacent vertebral body fractures, and complications were compared between the two groups.Results:All patients were followed up for 12-18 months [(13.4±4.2)months]. At 1 week, 3 months and 6 months after treatment, the VAS in surgery group [(3.5±1.0)points, (2.1±0.6)points, (2.0±0.4)points] was better than that in non-surgery group [(6.4±1.7)points, (5.3±1.5)points, (3.3±0.6)points] ( P<0.05), the ODI in surgery group [(45.8±10.3)points, (35.3±8.6)points, (26.5±7.1)points] was better than that in non-surgery group [(65.7±12.1)points, (58.3±10.7)points, (45.6±9.3)points] ( P<0.05), the SF-36 in surgery group [(82.8±1.4)points, (85.0±1.5)points, (88.0±1.3)points] was better than that in non-surgery group [(80.4±1.3)points, (81.5±1.4)points, (85.6±1.1)points] ( P<0.05). At the last follow-up, there were no significant differences in VAS, ODI and SF-36 between the two groups ( P>0.05). At 3 months, 6 months and the last follow-up, the Cobb angle in surgery group [(18.3±3.9)°, (17.5±3.0)°, (17.8±1.6)°] was better than that in non-surgery group [(22.4±2.2)°, (22.5±1.7)°, (22.1±1.3)°] ( P<0.05), and the relative height of the anterior edge of the injured vertebra in surgery group [(75.4±8.6)%, (76.6±8.6)%, (75.2±8.3)%] was better than that in non-surgery group [(63.5±7.6)%, (65.2±7.4)%, (62.8±7.2)%] ( P<0.05). The adjacent vertebral fractures occurred in 6 patients (8%) in surgery group, and in 2 patients (6%)in non-surgey group ( P>0.05). Postoperative complications occurred in 8 patients (11%) in surgery group, and occurred in 6 patients (19%) in non-surgery group ( P>0.05). Conclusion:When applying the OTLICS scoring system to treat acute symptomatic OVCF, patients with OTLICS score of 4 points should be promptly treated with operatively, which can quick relieve pain, correct deformities promote functional recovery, and improve quality of life.

Objective:To explore the clinical effect of percutaneous vertebroplasty (PVP) at stage I in treatment of acute multi-segment osteoporotic vertebral compression fracture (OVCF) in the elderly with over 3 vertebrae operated.Methods:A retrospective case-control study was conducted to analyze the clinical data of 105 aged patients with acute multi-segment OVCF admitted to Honghui Hospital, Xi'an Jiaotong University from October 2015 to February 2019. There were 27 males and 78 females, aged 65-92 years [(73.0±14.5)years]. All patients received stage I multi-segmental PVP and standard anti-osteoporosis treatment. There were more than 3 operative vertebral segments in 30 patients (observation group) and less than or equal to 3 operative vertebral segments in 75 patients (control group). The operation time, intraoperative fluoroscopy frequency, cement injection volume, cement leakage rate and incidence of recurrent vertebral fractures were compared between the two groups. The visual analogue scale (VAS), Oswestry disability index (ODI) and activity of daily living (ADL) score were compared before operation, 1 day, 1 month after operation and at the last follow-up.Results:All patients were followed up for 11-13 months [(12.5±1.8)months]. The observation group showed operation time of (71.2±12.2)minutes, intraoperative fluoroscopy frequency of (38.8±6.4)times and cement injection volume of (20.2±4.6)ml, more than those in control group [(52.3±10.6)minutes, (25.4±5.3)times, (12.3±4.3)ml] ( P<0.05). There was no significant difference in cement leakage rate and incidence of recurrent vertebral fractures between the two groups ( P>0.05). No complications such as infection, nerve damage or cement implantation syndrome occurred. Before operation, 1 day after operation, 1 month after operation and at the last follow-up, the VAS in observation group [(7.6±0.7)points, (3.0±0.8)points, (2.3±0.7)points, (2.2±0.6)points] showed no significant difference from those in control group [(7.4±0.5)points, (2.9±0.4)points, (2.1±0.5)points, (2.0±0.5)points], the ODI in observation group [(74.6±3.3)%, (36.8±4.6)%, (29.7±4.0)%, (24.0±3.6)%] did not differ from those in control group [(73.8±1.0)%, (35.1±0.9)%, (28.4±2.2)%, (22.8±0.9)%], the ADL score in observation group [(34.5±5.0)points, (54.5±3.8)points, (73.7±3.9)points] were similar with those in control group [(36.2±3.4)points, (56.8±4.7)points, (75.3±5.3)points, (81.3±4.5)points] (all P>0.05). The postoperative VAS, ODI and ADL score in both groups were significantly improved in comparison with preoperation ( P<0.05). Conclusion:For acute multi-segment OVCF in the elderly with over 3 or not more than 3 the vertebrae operated, PVP at stage I has the same advantages in early pain relief and improvement of motor function and quality of life.

Objective:To compare the clinical efficacy of percutaneous vertebroplasty (PVP) and non-surgical treatment of patients with type I fracture according to the acute symptomatic osteoporotic thoracolumbar fracture (ASOTLF) classification.Methods:A retrospective case-control study was used to analyze the clinical data of 115 patients with ASOTLF admitted to Honghui Hospital of Xi'an Jiaotong University from January 2015 to December 2018. There were 48 males and 67 females, aged 65-92 years [(75.3±8.5)years]. According to clinical symptoms and imaging characteristics, all patients were identified with type I fracture according to the ASOTLF classification. Injury segments were at T 6 to T 10 in 10 patients, at T 11 in 15, at T 12 in 26, at L 1 in 34, ay L 2 in 18, at L 3 in 7, and at L 4 in 5. A total of 73 patients received PVP combined with anti-osteoporosis treatment (surgery group), and 42 patients received non-surgery combined anti-osteoporosis treatment (non-surgery group). Before treatment, at 1 day, 1 month, 3 months, 6 months after treatment, and at the last follow-up, the visual analogue scale (VAS) was used to assess the pain, the Roland Morris Disability (RMD) score to assess the spinal function, and the Quality of Life Questionnaire of the European Foundation for Osteoporosis (QUALEFFO) score to evaluate the quality of life. The bone mineral density was compared between groups before treatment and at 1 year after treatment. The complications were observed as well. Results:All patients were followed up for 12-18 months [(13.2±4.6)months]. At 1 day, 1 month, and 3 months after treatment, the VAS in surgery group [(3.9±0.6)points, (3.3±0.6)points, (2.9±0.3)points] was significantly lower than that in non-surgery group [(6.0±0.7)points, (5.0±0.8)points, (4.2±1.0)points, respectively] (all P<0.05); the RMD score in surgery group [(15.2±0.7)points, (12.6±0.7)points, (10.6±0.7)points] was significantly lower than that in non-surgery group [(16.4±0.8)points, (14.6±0.8)points, (12.7±0.6)points, respectively] (all P<0.05). At 1 day and 1 month after treatment, the QUALEFFO score in surgery group [(46.0±1.1)points, (41.4±0.8)points] was lower than that in non-surgery group [(50.3±0.8)points, (44.7±1.2)points] (all P<0.05). There was no statistically significant difference between the two groups at other time points of the above indicators ( P>0.05). At 1 year after treatment, the bone mineral density in surgery group was (-3.0±0.9)SD, and was (-2.8±1.1)SD in non-surgery group ( P>0.05). There was no significant difference in the incidence of complications between surgery group [37%(27/73)] and non-surgery group [33%(14/42)] ( P>0.05). Conclusion:For patients with ASOTLF type I fracture, PVP and non-surgical treatment has similar effects in anti-osteoporosis and occurrence of complications, but the PVP is superior to non-surgical treatment in early pain relief, spinal function improvement and quality of life.

Objective:To investigate the difference of functional connectivity(FC) between the habenula and other brain regions in the patients with first-episode current depression (fMDD) and remitted depression (rMDD).Methods:Thirty-five patients with first-episode current depression (fMDD), 35 patients with remitted depression (rMDD) and 30 healthy controls (HC) matched with gender, age and education years were scanned by resting-state fMRI. Hamilton Depression Scale (HAMD 17) was used to assess the severity of the patients. After preprocessing, seed-based FC analysis was performed on the habenula. Pearson correlation analysis was performed between the FC values and HAMD 17 and duration of disease. Results:(1) Compared with the HC group, FC decreased between the habenula and left middle occipital gyrus(l-MOG) in fMDD group (x, y, z=-48, -84, 3, t=-4.335, P<0.05), while FC increased between the habenula and left supramarginal gyrus (x, y, z=-66, -30, 36, t=4.876, P<0.05), left superior frontal gyrus(l-SFG) (x, y, z=-6, -33, 51, t=4.402, P<0.05), left inferior parietal lobe(l-IPL)( t=3.300, P<0.05) and right inferior parietal lobe(r-IPL) ( t=3.557, P<0.05) in fMDD group. Compared with the HC, FC decreased between the habenula and l-MOG (x, y, z=-48, -84, 3, t=-4.321, P<0.05) and left thalamus (x, y, z=-9, 3, 3, t=-3.971, P<0.05) in rMDD group, while FC increased between the habenula and left middle temporal gyrus(l-MTG)( x, y, z=-48, -39, 9, t=4.062, P<0.05), left supramarginal gyrus (x, y, z=-51, -15, 45, t=2.906, P<0.05), l-SFG (x, y, z=-24, -21, 39, t=3.044, P<0.05), l-IPL ( t=2.880, P<0.05) and r-IPL ( t=3.512, P<0.05) in rMDD group. (2) FC decreased in fMDD group between the habenula and right orbitofrontal cortex(r-OFC) ( t=-3.899, P<0.05) and l-MTG ( t=-4.023, P<0.05) than rMDD group, while FC increased between the habenula and left supramarginal gyrus ( t=4.157, P<0.05), l-SFG( t=3.327, P<0.05), left thalamus ( t=3.316, P<0.05) and l-IPL ( t=3.909, P<0.05) than rMDD group. (3)There was no significantly correlation between the HAMD 17 and the FC value changes among the different regions, and was marginal significantly correlation between duration of disease and the FC values from the habenula and l-MOG( r=0.328, P=0.063). Conclusion:Both fMDD and rMDD show abnormal FC between the habenula and default mode network, visual network and reward network, which may be related to the pathogenesis of depression. The FC of the habenula in rMDD still had not recovered to normal.