[摘 要] 目的：探讨异位表达血红蛋白亚基（HBA/HBB）对缺氧条件下嵌合抗原受体T细胞（CAR-T细胞）功能障碍的改善作用及其对肿瘤细胞的杀伤效应。方法：全基因合成技术合成靶向HER2的CAR序列，构建共表达HBA或HBB的CAR慢病毒载体，包装慢病毒后感染人原代T淋巴细胞，制备异位表达HBA/HBB的CAR-T细胞，命名为HBA CAR-T和HBB CAR-T。采用缺氧探针检测小鼠实体瘤缺氧状态。通过流式细胞术检测瘤内CAR-T细胞占比、异位表达血红蛋白亚基的CAR-T细胞阳性率及CAR-T细胞的活性氧、凋亡水平。WB法检测HBA CAR-T和HBB CAR-T内相关血红蛋白亚基表达情况，采用细胞计数板计数检测细胞增殖水平，通过萤光素酶报告基因法检测CAR-T细胞对肿瘤细胞的杀伤能力，qPCR检测CAR-T细胞中缺氧诱导因子-1α（HIF-1α）表达水平，利用MitoXpress Intra试剂盒检测CAR-T细胞内氧气含量。结果：不同细胞构建的实体瘤模型均存在明显缺氧情况，且CAR-T细胞浸润水平与缺氧程度呈显著负相关（P < 0.000 1）。HBA CAR-T与HBB CAR-T构建成功（阳性率 > 60%），相应血红蛋白亚基可稳定表达。缺氧环境下HBA CAR-T和HBB CAR-T的ROS水平、凋亡水平显著下降，增殖、对肿瘤细胞的体外杀伤能力显著强于传统CAR-T细胞（均P < 0.05）。HBA CAR-T与HBB CAR-T内HIF-1α表达降低（均P < 0.001），且缺氧程度显著降低（均P < 0.001）。结论：异位表达血红蛋白亚基可改善缺氧条件下CAR-T细胞功能障碍并增强其对肿瘤细胞的体外杀伤作用。

Monogenic hereditary cerebral small vessel disease （CSVD） is a group of genetic disorders characterized by cerebrovascular lesions， including cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy， cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy， COL4A1 and COL4A2-related CSVD， and other rare types such as retinal vasculopathy with cerebral leukoencephalopathy and systemic manifestations and leukoencephalopathy with calcifications and cysts. These diseases are often caused by specific gene mutations and exhibit highly heterogeneous pathological mechanisms and clinical manifestations， including inflammatory response， abnormal gene expression， and microangiopathy. The advances in imaging findings and biomarkers have provided new methods for early diagnosis， while treatment strategies include stem cell therapy， immunotherapy，gene editing， and molecular targeted therapy. However， further studies are needed to develop individualized treatment regimens for different subtypes. This article reviews the important advances in the pathogenesis， clinical features， diagnostic methods， and treatment modalities of monogenic hereditary CSVD in recent years， in order to provide guidance for future research and clinical practice.

Objective To investigate the feasibility of a "pericardial lining" modified Bentall procedure for the treatment of patients with aortic root aneurysm. Methods This was a retrospective study that consecutively enrolled patients treated at the Affiliated Suzhou Hospital of Nanjing Medical University, The Affiliated Traditional Chinese Medicine Hospital of Southwest Medical University, and the First People&#39;s Hospital of Guangyuan from January 2023 to February 2024. Preoperative clinical data, imaging findings (including echocardiography and CT scans of the aortic root and the entire aorta), details of coronary artery management, surgical outcomes, and postoperative follow-up results were collected. All patients underwent the "pericardial lining" modified Bentall procedure: the aortic valve was replaced, and an autologous pericardial patch was divided into three equal leaflets based on the circumference of the aortic annulus measured by a valve sizer. These leaflets were then sutured to the aortic annulus. Fenestrations were created in two of the pericardial leaflets for anastomosis with the left and right coronary ostia. The pericardial leaflets were sutured to the wall of the aortic sinuses to form an integrated structure, thereby narrowing the sinus portion. A prosthetic vascular graft was anastomosed to the proximal and distal aorta, and no aortic root-to-right atrium shunt was created. Results A total of 5 patients, aged 37 to 68 years, were included. The preoperative Society of Thoracic Surgeons (STS) risk scores ranged from 2.8% to 3.9%. The diameter of the ascending aorta was 40-73 mm, the left ventricular end-diastolic diameter (LVEDD) was 45-71 mm, and the left ventricular ejection fraction (LVEF) was 47%-64%. Intraoperatively, the aortic cross-clamp time ranged from 85 to 180 min, and the cardiopulmonary bypass time ranged from 110 to 302 min. Postoperative follow-up echocardiography revealed that the ascending aortic diameter was 27-35 mm, LVEDD was 39-57 mm, and LVEF was 43%-61%. All surgeries were completed successfully with satisfactory immediate outcomes and no intraoperative complications. During the follow-up period, there was no mortality or reoperation. Conclusion For patients with aortic root aneurysm, the "pericardial lining" modified Bentall procedure yields satisfactory preliminary results, and the technique is demonstrated to be feasible.

Objective: To analyze the influencing factors for hospitalization costs among stroke patients with different subtypes, so as to provide the reference for reducing the economic burden of patients. Methods: Data of patients with hemorrhagic or ischemic stroke who were discharged from hospitals in Nanshan District, Shenzhen City from January 1, 2016 to December 31, 2021 were collected through Hospital Information System. Hospitalization costs were analyzed between hemorrhagic and ischemic stroke patients, and factors affecting hospitalization costs among stroke patients with different subtypes were identified using a structural equation model. Results: A total of 10 298 patients with stroke were recruited, including 2 820 patients with hemorrhagic stroke (27.38%) and 7 478 patients with ischemic stroke (72.62%). The patients with hemorrhagic stroke had a median duration of hospital stay of 19.00 (interquartile range, 18.00) d, and a median hospitalization cost of 26 759.48 (interquartile range, 51 000.87) Yuan. The patients with ischemic stroke had a median duration of hospital stay of 12.00 (interquartile range, 10.00) d, and a median hospitalization cost of 12 199.87 (interquartile range, 13 290.20) Yuan. Structural equation model analysis showed that department of hospitalization, discharge status, ways of leaving hospital, surgery and hypertension had direct effects on hospitalization costs and indirect effects on hospitalization costs through duration of hospital stay among hemorrhagic stroke patients, and duration of hospital stay had the highest total effect (0.684), followed by surgery (0.632). Employment status, admission route, department of hospitalization, ways of leaving hospital, payment mode, surgery and dyslipidemia had direct effects on hospitalization costs and indirect effects on hospitalization costs through duration of hospital stay among ischemic stroke patients, and duration of hospital stay (0.746), surgery (0.424) and department of hospitalization (0.151) ranked the top three in total effects. Conclusion The hospitalization cost is relatively high among stroke patients in Nanshan District, and duration of hospital stay and surgery have great influence on hospitalization costs among stroke patients with different subtypes.

Objective: To investigate the trend in incidence of stroke in Nanshan District, Shenzhen City from 2010 to 2021, so as to provide the evidence for formulating the integrated strategy for stroke control. Methods: The data of stroke morbidity in Nanshan District from 2010 to 2021 were collected through Shenzhen Chronic Disease Prevention and Control Management System. The incidence of stroke was calculated, and standardized by the population of the Chinese Sixth National Census in 2010. The trends in stroke incidence were evaluated in Nanshan District from 2010 to 2021 using annual percent change (APC) and average annual percent change (AAPC), and gender-, age- and subtype-specific incidence of stroke was descriptively analyzed. Results: A total of 30 377 cases with stroke were reported in Nanshan District from 2010 to 2021, with a crude incidence rate of 190.45/105 and a standardized incidence rate of 405.65/105. The crude incidence rate of stroke appeared a tendency towards a rise in Nanshan District from 2010 to 2021 (APC=5.38%, t=4.678, P=0.001), and a higher crude incidence rate of stroke was seen among men than among women (227.57/105 vs. 148.40/105; χ2=1 309.580, P<0.001). The incidence of stroke appeared a tendency towards a rise with age (χ2trend =435.717, P<0.001), and there was a tendency towards a rise in stroke incidence among residents under 40 years of age (APC=2.89%, t=2.538, P=0.029). The crude incidence of ischemic stroke was 151.24/105, which was higher than that of hemorrhagic stroke (39.21/105) (χ2=10 521.000, P<0.001). Conclusions The incidence of stroke appeared a tendency towards a rise in Nanshan District from 2010 to 2021, with ischemic stroke as the predominant subtype of stroke. Males and middle-aged and elderly residents should be given a high priority for stroke prevention and treatment.

Objective    To investigate the predictive value of right atrial myocardial fibrosis in the prognosis of isolated tricuspid regurgitation surgery after left heart valve surgery. Methods    The patients who underwent tricuspid valvuloplasty by the same operator in Guangdong Provincial People&#39;s Hospital from April 2016 to August 2021 due to long-term isolated severe tricuspid regurgitation after left heart valve surgery were included in the study. According to the degree of right atrial myocardial fibrosis, the patients were divided into three groups: a mild group, a moderate group, and a severe group. The clinical data of these patients were compared and analyzed. Results    A total of 75 patients were enrolled, including 16 males and 59 females with an average age of 57.0±8.4 years. There were 30 patients in the mild group, 29 patients in the moderate group and 16 patients in the severe group. In terms of the preoperative data, there were statistical differences in cardiac function grade, right atrial diameter, tricuspid incompetence area among the three groups (P<0.05). In terms of the postoperative data, there were statistical differences among the three groups in the cardiopulmonary bypass time, mechanical ventilation time, ICU monitoring time, complication rate and mortality (P<0.05). Further pairwise comparison showed that, compared with the mild group, the severe group had longer mechanical ventilation time (P=0.024), longer ICU monitoring time (P=0.003) and higher incidence of postoperative complications (P=0.024), while the moderate group had no statistical difference in all aspects (P>0.05); compared with the moderate group, the severe group had longer ICU monitoring time (P=0.021) and higher incidence of complications (P=0.006). Conclusion    The early outcome of tricuspid valvuloplasty in patients with isolated tricuspid regurgitation after left heart valve surgery with severe right atrial myocardial fibrosis is worse than that in the patients with mild and moderate fibrosis, suggesting that the degree of myocardial fibrosis in the right atrium can be a predictor of the effect of tricuspid regurgitation surgery and a judgement indicator of the surgery timing.

Abstract OBJECTIVE To quickly qualitatively analyze the chemical components in Sanzi powder compound prescription by HPLC-Q-Exactive-MS/MS. METHODS SHIMADZU GIST C18 column(4.6 mm×150 mm, 5 μm) was used, using 0.1% formic acid water-methanol as mobile phase with gradient elution, the flow rate was 0.5 mL·min-1, the column temperature was 30 ℃. Under positive and negative ion modes, the primary and secondary mass spectrometry information of Sanzi powder was scanned. Qualitative attribution of each component in Sanzi powder was carried out based on the mass spectrometry information, molecular formula, and retention time of molecular ion peaks and fragment ions analyzed using total ion flow diagrams, combined with the molecular formula and structural formula searched in the Chemspider database and references. RESULTS Based on the analysis of the mass spectrometry cleavage rules and references of various components, 95 possible chemical components were preliminarily inferred, including 39 phenolic acids, 20 tannins, 9 organic acid esters, 5 monoterpenoids, 12 iridoids, 8 triterpenes and 2 flavonoids. Among them, 57 components were from Chebulae Fructus, 30 were from Gardeniae Fructus, 10 were from Toosendan Fructus, and among these compounds, rutin from Chebulae Fructus, Gardeniae Fructus and Toosendan Fructus. CONCLUSION The HPLC-Q-Exactive-MS/MS detection method has good separation and high sensitivity, and can quickly and efficiently infer various components in Sanzi powder. It establishes a fast and efficient analytical method for identifying the chemical components in Sanzi powder.

Objective: To investigate the prevalence of current smoking and smoking cessation, and identify the influencing factors among hypertensive patients in Nanshan District, Shenzhen City, so as to provide insights into smoking control among hypertensive patients. Methods: The demographic features, life style, status of smoking and smoking cessation in hypertensive patients were collected from 69 community health centers in Nanshan District from 2017 to 2019. The gender- and age-specific prevalence of smoking and smoking cessation was estimated among hypertensive patients, and the factors affecting hypertensive patients&#39; smoking and smoking cessation were identified using a multivariable logistic regression model. Results: Totally 4 385 patients with hypertension were enrolled, with a mean age of (58.41±11.19) years, and the participants included 2 265 men (51.65%) and 2 120 women (48.35%). There were 724 current smokers, and the prevalence of current smoking was 16.51%, with 29.71% prevalence in males and 2.41% in females. There were 424 hypertensive patients quitting smoking, and the prevalence of smoking cessation was 36.93%, with 38.20% prevalence in males. Multivariable logistic regression analysis identified male ( OR=12.885, 95%CI: 9.567-17.354 ) and drinking ( OR=2.567, 95%CI: 2.118-3.111 ) as facilitating factors for current smoking among hypertensive patients, and increasing age (OR=0.723, 95%CI: 0.642-0.815) and high exercise frequency (OR=0.881, 95%CI: 0.817-0.950) as barrier factors, while male ( OR=7.309, 95%CI: 3.304-16.165), increasing age ( OR=1.381, 95%CI: 1.120-1.703 ), unmarried ( OR=1.819, 95%CI: 1.329-2.490 ), divorced ( OR=7.837, 95%CI: 1.254-48.975 ), retired ( OR=1.545, 95%CI: 1.095-2.180 ), unemployed (OR=1.801, 95%CI: 1.057-3.066), and high exercise frequency ( OR=1.221, 95%CI: 1.096-1.360 ) were identified as facilitating factors for smoking cessation among hypertensive patients and widowed ( OR=0.285, 95%CI: 0.089-0.906 ), high educational level ( OR=0.766, 95%CI: 0.619-0.949 ), and drinking ( OR=0.488, 95%CI: 0.368-0.647 ) as barrier factors. Conclusions The prevalence of smoking is lower, and the prevalence of smoking cessation is higher among hypertensive patients than among general populations in Nanshan District. Young and middle-aged, employed, widowed men with a high educational level are key populations for tobacco control, and alcohol consumption control and intensified exercises are important measures to reduce the prevalence of smoking and improve the prevalence of smoking cessation.

Objective    To compare the clinical effects of segmentectomy and lobectomy for ≤2 cm lung adenocarcinoma with micropapillary and solid subtype negative by intraoperative frozen sections. Methods    The patients with adenocarcinoma who received segmentectomy or lobectomy in multicenter from June 2020 to March 2021 were included. They were divided into two groups according to a random number table, including a segmentectomy group (n=119, 44 males and 75 females with an average age of 56.6±8.9 years) and a lobectomy group (n=115, 43 males and 72 females with an average of 56.2±9.5 years). The clinical data of the patients were analyzed. Results    There was no significant difference in the baseline data between the two groups (P>0.05). No perioperative death was found. There was no statistical difference in the operation time (111.2±30.0 min vs. 107.3±34.3 min), blood loss (54.2±83.5 mL vs. 40.0±16.4 mL), drainage duration (2.8±0.6 d vs. 2.6±0.6 d), hospital stay time (3.9±2.3 d vs. 3.7±1.1 d) or pathology staging (P>0.05) between the two groups. The postoperative pulmonary function analysis revealed that the mean decreased values of forced vital capacity and forced expiratory volume in one second percent predicted in the segmentectomy group were significantly better than those in the lobectomy group (0.2±0.3 L vs. 0.4±0.3 L, P=0.005; 0.3%±8.1% vs. 2.9%±7.4%, P=0.041). Conclusion    Segmentectomy is effective in protecting lungs function, which is expected to improve life quality of patients.

AIM:To compare the effectiveness and safety of intravitreal injection of conbercept(IVC)and intravitreal injection of ranibizumab(IVR)for type 1 retinopathy of prematurity(ROP).

METHORDS: A retrospective case series observation was carried out. Clinical data of patients with type 1 ROP treated with IVC(46 cases, 91 eyes)or IVR(55 cases, 109 eyes)from August 2018 to January 2020 in Xijing Hospital were collected. Regression, progression, recurrence, retreatment of ROP, the proportion of adverse outcomes and ocular and systemic complications were all analyzed.

RESULTS:All the 101 infants(200 eyes)with type 1 ROP treated with intravitreal injection of anti-vascular endothelial growth factor(VEGF)were enrolled, in which 20 eyes with aggressive posterior ROP(AP-ROP), 86 eyes with threshold ROP, and 94 eyes with type 1 pre-threshold ROP were included. There were no statistical differences in the basic conditions and the severity of ROP between the two groups before treatment(P>0.05). There was no difference in the primary cure rate between IVC and IVR groups(93.4% vs 87.2%, P>0.05). A total of 6 eyes(6.6%)developed ROP recurrence in the IVC group, in which 2 eyes received a second IVC, and 4 eyes were treated with laser photocoagulation(LP). In the IVR counterpart, 11 eyes(10.1%)developed ROP recurrence, among them 4 eyes underwent another IVR, and 11 eyes were treated with LP. The recurrence interval was 11.7±4.13wk and 9.82±4.02wk in the IVC and IVR groups. All these results of recurrence showed no significant statistical difference between these two groups(P>0.05). There was no progression of ROP appeared in IVC group. The progression of ROP developed in 3 eyes after initial ranibizumab injection, among them retinal fibrous proliferation and hemorrhage treated with LP in 2 eyes, and tractional retinal detachment treated with vitrectomy in 1 eye. No drug or injection related side effect was observed in infants of both groups. There were some adverse prognosis, such as temporal retinal folds, smaller angle between upper and lower temporal retinal vessel trunks caused by retinal vascular traction in 3 eyes in IVR group during the long-term follow up.

CONCLUSION: Both IVC and IVR are effective and safety choices for the treatment of ROP. There was no significant difference between the first cure rate and the recurrence rate. Some of the most severe cases treated by IVR were at risk for progression or poor prognosis that need to be followed up for a long time.