OBJECTIVE To investigate the effect of neferine （NEF） on mitophagy in Parkinson’s disease （PD） cells by regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/UNC-51-like kinase 1 （ULK1） signaling pathway， and explore the mechanism of this drug to improve PD. METHODS SH-SY5Y cells were treated with 100 μmol/L 1-methyl-4-phenylpyridinium ion （MPP+） for 24 h to construct a PD cell model. PD model cells were divided into model group （PD group）， NEF low-， medium- and high-concentration groups （NEF-L， NEF-M， NEF-H group， 2.5， 5.0， 10.0 μmol/L）， and high concentration of NEF+AMPK inhibitor group （NEF-H+Compound C group， 10.0 μmol/L NEF+50 μmol/L Compound C）. The cells treated without MPP+ and NEF were used as the control group. The ultrastructure of the cells in each group was observed； the amount of autophagosomes， survival rate， apoptosis rate， mitochondrial membrane potential， and the protein expressions of Caspase-3， microtubule-associated protein 1 light chain 3 （LC3） and Beclin-1， as well as the phosphorylation levels of mTOR， AMPK and ULK1 were detected. RESULTS Compared with PD group， the amount of autophagosomes in NEF-L， NEF-M and NEF-H groups was increased， and membrane potential was increased； survival rate， LC3- Ⅱ/LC3-Ⅰ， protein expression of Beclin-1， and protein phosphorylation levels of AMPK and ULK1 were significantly increased or up-regulated； the apoptotic rate， protein expressions of Caspase-3 and p62， and protein phosphorylation level of mTOR were significantly decreased or down-regulated， and the above improvements were in a dose-dependent manner （P＜0.05）. Compound C could significantly reverse the above improvement effect of high concentration of NEF （P＜0.05）. CONCLUSIONS NEF can promote mitophagy and inhibit apoptosis of PD model cells by up-regulating protein phosphorylation levels of AMPK and ULK1， and down-regulating protein phosphorylation level of mTOR， thus playing a protective role in nerve cells.

OBJECTIVE To investigate the effect of neferine （NEF） on mitophagy in Parkinson’s disease （PD） cells by regulating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/UNC-51-like kinase 1 （ULK1） signaling pathway， and explore the mechanism of this drug to improve PD. METHODS SH-SY5Y cells were treated with 100 μmol/L 1-methyl-4-phenylpyridinium ion （MPP+） for 24 h to construct a PD cell model. PD model cells were divided into model group （PD group）， NEF low-， medium- and high-concentration groups （NEF-L， NEF-M， NEF-H group， 2.5， 5.0， 10.0 μmol/L）， and high concentration of NEF+AMPK inhibitor group （NEF-H+Compound C group， 10.0 μmol/L NEF+50 μmol/L Compound C）. The cells treated without MPP+ and NEF were used as the control group. The ultrastructure of the cells in each group was observed； the amount of autophagosomes， survival rate， apoptosis rate， mitochondrial membrane potential， and the protein expressions of Caspase-3， microtubule-associated protein 1 light chain 3 （LC3） and Beclin-1， as well as the phosphorylation levels of mTOR， AMPK and ULK1 were detected. RESULTS Compared with PD group， the amount of autophagosomes in NEF-L， NEF-M and NEF-H groups was increased， and membrane potential was increased； survival rate， LC3- Ⅱ/LC3-Ⅰ， protein expression of Beclin-1， and protein phosphorylation levels of AMPK and ULK1 were significantly increased or up-regulated； the apoptotic rate， protein expressions of Caspase-3 and p62， and protein phosphorylation level of mTOR were significantly decreased or down-regulated， and the above improvements were in a dose-dependent manner （P＜0.05）. Compound C could significantly reverse the above improvement effect of high concentration of NEF （P＜0.05）. CONCLUSIONS NEF can promote mitophagy and inhibit apoptosis of PD model cells by up-regulating protein phosphorylation levels of AMPK and ULK1， and down-regulating protein phosphorylation level of mTOR， thus playing a protective role in nerve cells.

Objective:To investigate the clinical efficacy of pentoxifylline sequential therapy combined with rasagiline and levodopa in the treatment of elderly patients with symptoms of Parkinson disease (PD), and the influence on hemorheology and serum Toll-like receptor 4 (TLR4) signaling pathway downstream related inflammatory factors.Methods:A prospective study method was used to select 90 elderly patients with PD with fluctuating symptoms who were admitted to the Eighth People′s Hospital of Hebei Province from June 2021 to October 2022 as research objects. The patients were divided into observation group and control group with 45 cases in each group according to random number table method. The observation group was treated with pentoxifylline sequential therapy combined with rasagiline and levodopa. The control group was treated with rasagiline combined with levodopa. The clinical efficacy of the two groups was compared. The unified Parkinson disease rating scale (UPDRS), Montreal cognitive assessment scale (MoCA), Berg balance scale (BBS) and 39-item Parkinson disease quality of life questionnaire (PDQ-39) were scored before and after treatment. Hemorheology indexes and serum levels of related inflammatory factors downstream of TLR4 signaling pathway, including whole blood high viscosity (HBV), whole blood low shear viscosity (LBV), plasma viscosity (PV), fibrinogen (FIB); TLR4, interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF) -α, were detected before and after treatment in both groups. The adverse reactions of the two groups were compared.Results:The total effective rate in observation group was significantly higher than that in control group: 93.33% (42/45) vs. 77.78% (35/45), and there was statistical difference ( P<0.05). After treatment, the UPDRS mental activity and emotional disorders, daily living ability, motor function, motor complications scores and PDQ-39 score of the two groups were significantly lower than before treatment, the MoCA and BBS scores were significantly higher than before treatment, and the improvement was more significant in the observation group, there were statistical differences ( P<0.05). After treatment, HBV, LBV, PV and FIB in the observation group were significantly decreased compared with those before treatment: (6.52 ± 0.92) mPa·s vs. (7.25 ± 1.24) mPa·s, (11.45 ± 1.24) mPa·s vs. (14.13 ± 1.64) mPa·s, (1.55 ± 0.17) mPa·s vs. (1.88 ± 0.22) mPa·s, (3.25 ± 0.47) g/L vs. (3.82 ± 0.52) g/L, and there were statistical differences ( P<0.05). There were no significant differences in hemorheology indexes of control group before and after treatment ( P>0.05). After treatment, serum levels of TLR4, IL-1β, IL-6 and TNF-α in both groups were significantly decreased compared with those before treatment, and the indexes in observation group were significantly lower than those in control group: (2.07 ± 0.18) ng/L vs. (2.58 ± 0.21) ng/L, (1.42 ± 0.17) ng/L vs. (2.28 ± 0.25) ng/L, (1.56 ± 0.22) ng/L vs. (2.42 ± 0.28) ng/L, (46.31 ± 3.17) ng/L vs. (54.34 ± 3.65) ng/L, and the differences were statistically significant ( P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:Pentoxifylline sequential therapy combined with rasagiline and levodopa can effectively improve the hemorheology of elderly patients with PD accompanied by symptom fluctuations, reduce the levels of related inflammatory factors downstream of serum TLR4 signaling pathway, and improve clinical efficacy.

Objective To understand the proficiency level and sources of Traditional Chinese Medicine(TCM)health education among hemodialysis patients,in order to provide the basis for promoting the application of TCM nursing in hemodialysis patients.Methods Utilizing a convenience sampling approach,we conducted a survey from April to May 2023 involving hemodialysis patients from 7 tertiary TCM hospitals across Beijing,Hebei Province,Sichuan Province,and other regions.Custom-made questionnaires were utilized,gathering general information,respons-es conceming the level of mastery on TCM health education,and their primary sources of such knowledge.Results A total of 413 questionnaires were disseminated,and 392 proved valid,yielding a valid response rate of 94.92％.The score of patient's knowledge on TCM health education averaged(8.25±3.73),and an average score rate of 48.53％was obtained.In the patients undergoing hemodialysis,a relatively larger number of people possess knowl-edge of TCM health knowledge about relieving symptoms(50.8％～55.9％)and traditional Chinese daily life routines(56.4％～90.1％).However,fewer people are aware of how to use the five-tone therapy method for emotional inter-vention(40.8％)and principle of taking restorative Traditional Chinese Medicine(24.5％～36.7％),the dietary prin-ciples of"nurturing yang in spring and summer,and nurturing yin in autumn and winter"(14.3％),and theories of TCM kidney functions(9.9％).Approximately 23.0％of the patients sourced their TCM health education knowledge from new media platforms.Compared with new media method alone,patients who utilized only traditional education-al platforms(P=0.020),and those who combined both new media and traditional methods(P=0.018)demonstrated higher proficiency in TCM health education.Conclusion Hemodialysis caregivers are urged to develop a TCM health education framework that emphasizes emotional well-being and traditional wellness concepts,thereby fostering patient-centric TCM health ideologies.Hemodialysis education personnel are encouraged to leverage new media whilst ensuring education quality and effective outcomes.

OBJECTIVE To investigate the intervention effect and mechanism of Wuwei baogan pill on mice with non- alcoholic fatty liver disease （NAFLD）. METHODS The mice were given high-fat and high-sugar diet for 19 weeks to induce NAFLD model. The model mice were randomly grouped into model group， positive control group （polyene phosphatidylcholine capsules， 23.30 mg/kg）， Wuwei baogan pill low-dose， medium-dose and high-dose groups （0.11， 0.23， 0.45 g/kg）， with 8 mice in each group； the normal group was additionally set up without modeling. Administration groups were given relevant medicine intragastrically， and model group and normal group were given constant volume of normal saline， once a day， for consecutive 4 weeks. After the last administration， glucose metabolism （including fasting blood glucose， fasting insulin， insulin resistance index）， liver function [liver index， alanine aminotransferase （ALT）， aspartate aminotransferase （AST），liver tissue pathological score]， lipid metabolism [triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， and high- density lipoprotein cholesterol （HDL-C）] were measured； the pathological morphology of liver tissue， as well as fibrosis， lipid droplet formation， and glycogen synthesis were observed； the levels of free fatty acid （FFA） in serum and inflammatory factors in liver tissue [tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） and IL-1β] were detected； the expressions of insulin receptor substrate/phosphoinositide 3-kinase/protein kinase B/ glycogen synthase kinase 3β （IRS/PI3K/AKT/GSK3β） signaling pathway-related protein in liver tissue were investigated. RESULTS After intervention with high-dose Wuwei baogan pill， liver index of NAFLD mice， serum levels of ALT， AST， FFA， TC， TG and LDL-C， the levels of TNF-α， IL-6 and IL-1β in liver tissue， fasting blood glucose， fasting insulin， insulin resistance index， liver tissue pathological score， proportions of fibrotic staining area and lipid droplet staining area all significantly decreased （P＜0.05）； the level of HDL-C， proportion of glycogen staining area， the phosphorylation of IRS1， PI3K， AKT and GSK3β protein increased significantly （P＜0.05）； the degree of liver cell necrosis and steatosis was reduced， and the fibrotic lesions were alleviated. The above indexes of mice were improved in Wuwei baogan pill low-dose and medium-dose groups， but there was no statistically significant difference in some indexes. CONCLUSIONS Wuwei baogan pill can regulate lipid and glucose metabolism disorders in the liver of NAFLD mice， and improve liver injury， the mechanism of which may be associated with the activation of IRS/PI3K/AKT/GSK3β signaling pathway.

OBJECTIVE To investigate the intervention effect and mechanism of Wuwei baogan pill on mice with non- alcoholic fatty liver disease （NAFLD）. METHODS The mice were given high-fat and high-sugar diet for 19 weeks to induce NAFLD model. The model mice were randomly grouped into model group， positive control group （polyene phosphatidylcholine capsules， 23.30 mg/kg）， Wuwei baogan pill low-dose， medium-dose and high-dose groups （0.11， 0.23， 0.45 g/kg）， with 8 mice in each group； the normal group was additionally set up without modeling. Administration groups were given relevant medicine intragastrically， and model group and normal group were given constant volume of normal saline， once a day， for consecutive 4 weeks. After the last administration， glucose metabolism （including fasting blood glucose， fasting insulin， insulin resistance index）， liver function [liver index， alanine aminotransferase （ALT）， aspartate aminotransferase （AST），liver tissue pathological score]， lipid metabolism [triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， and high- density lipoprotein cholesterol （HDL-C）] were measured； the pathological morphology of liver tissue， as well as fibrosis， lipid droplet formation， and glycogen synthesis were observed； the levels of free fatty acid （FFA） in serum and inflammatory factors in liver tissue [tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6） and IL-1β] were detected； the expressions of insulin receptor substrate/phosphoinositide 3-kinase/protein kinase B/ glycogen synthase kinase 3β （IRS/PI3K/AKT/GSK3β） signaling pathway-related protein in liver tissue were investigated. RESULTS After intervention with high-dose Wuwei baogan pill， liver index of NAFLD mice， serum levels of ALT， AST， FFA， TC， TG and LDL-C， the levels of TNF-α， IL-6 and IL-1β in liver tissue， fasting blood glucose， fasting insulin， insulin resistance index， liver tissue pathological score， proportions of fibrotic staining area and lipid droplet staining area all significantly decreased （P＜0.05）； the level of HDL-C， proportion of glycogen staining area， the phosphorylation of IRS1， PI3K， AKT and GSK3β protein increased significantly （P＜0.05）； the degree of liver cell necrosis and steatosis was reduced， and the fibrotic lesions were alleviated. The above indexes of mice were improved in Wuwei baogan pill low-dose and medium-dose groups， but there was no statistically significant difference in some indexes. CONCLUSIONS Wuwei baogan pill can regulate lipid and glucose metabolism disorders in the liver of NAFLD mice， and improve liver injury， the mechanism of which may be associated with the activation of IRS/PI3K/AKT/GSK3β signaling pathway.

[摘 要] 目的：探讨在靶向HER2的CAR的CD3ζ链胞内区引入YRHQ基序对CAR-T细胞的特异性杀伤活性及免疫记忆形成的影响。方法：通过DNA合成获得包含靶向HER2的编码抗原受体H28ζ或H28ζ(YRHQ)的DNA片段，通过慢病毒载体将不同CAR的DNA片段分别转导健康人外周血T细胞，制备靶向HER2的H28ζ-CAR-T及H28ζ(YRHQ)-CAR-T细胞。扩增过程中对不同CAR-T细胞进行计数，FCM检测CAR的表达率。将CAR-T细胞分别与HER2阳性的SKOV3、MDA-MB-453或HER2阴性的MCF-7细胞共培养，LDH释放法检测其杀伤活性，ELISA法检测IL-2、IFN-γ和颗粒酶B的水平，WB法检测STAT3磷酸化水平及免疫检查点分子TIM-3和PD-1的表达，通过FCM检测CCR7、CD45RO的表达，分析CAR-T细胞的表型。结果：H28ζ-CAR-T和H28ζ(YRHQ)-CAR-T细胞扩增能力较好，体外培养7 d时扩增4~5倍。H28ζ-CAR和H28ζ(YRHQ)-CAR表达率分别为（33.3±2.85）%和（28.30±3.2）%。H28ξ(YRHQ)-CAR-T细胞的杀伤活性较H28ζ-CAR-T细胞更高（P<0.05）。经HER2抗原刺激后，与T细胞或H28z-CAR-T细胞比较，H28ξ(YRHQ)-CAR-T细胞的STAT3磷酸化水平较H28ξ-CAR-T细胞明显升高（P<0.01）；而两者间PD-1和TIM-3的表达无明显差异。未经抗原刺激的CAR-T细胞CCR7和CD45RO表达与正常T细胞比较差异无统计学意义（均P>0.05），与SKOV3细胞共培养后，与T细胞或H28z-CAR-T细胞比较，H28ξ(YRHQ)-CAR-T细胞中TEM细胞比例明显增加、TCM细胞比例明显减少（均P<0.05）。结论：在CD3胞内区引入YRHQ基序可在一定程度上提高CAR-T细胞的杀伤潜力。

Objective: To assess the effects of acute exposure to electronic cigarette ( e-cigarette ) on leukocyte and total protein levels in bronchoalveolar lavage fluid ( BALF ) and pulmonary surfactant protein expression in a mouse model, so as to provide insights into the elucidation of the mechanism underlying the damages to the respiratory system caused by e-cigarette. Methods: Twenty-one C57BL/6N female mice were randomly divided into the blank control group, the solvent control group and the nicotine group. Mice in the solvent control group and the nicotine group were exposed to the solvent aerosol or e-cigarette aerosol containing 25 mg/mL nicotine for 3 hours daily, while mice in the blank control group were bred in clean air. Following 3-day exposure, mouse BALF and lung specimens were collected. The cell morphology was observed using microscopy following Wright-Giemsa staining and the leukocyte count was estimated in BALF, while the total protein expression was quantified using bicinchoninic acid ( BCA ) assay. In addition, the mRNA expression of pulmonary surfactant protein genes was detected in mouse lung specimens using quantitative real-time PCR ( qPCR ) assay. Results: All mice in three groups grew well without obvious abnormality or death seen. Wright-Giemsa staining showed a higher number of mononuclear macrophages in mouse BALF in the nicotine group than in the blank control group and the solvent control group. The leukocyte counts were ( 2.00±0.77 )×107, ( 1.79±0.99 )×107 and ( 4.00±1.35 )×107 cells/L ( F=9.199, P=0.002 ), and the total protein levels were ( 0.16±0.03 ), ( 0.12±0.02 ) and ( 0.16±0.04 ) mg/mL in mouse BALF in the blank control group, solvent control group and nicotine group ( F=3.610, P=0.048 ), and the relative mRNA expression of pulmonary surfactant protein B (SP-B) and SP-D was 1.00±0.14, 0.82±0.12 and 0.74±0.07 ( F=5.491, P=0.028 ), and 1.00±0.06, 0.90±0.02 and 0.71±0.15 in mouse lung specimens, respectively ( F=10.460, P=0.005 ). The leukocyte count was significantly higher in the nicotine group than in the blank control group and solvent control group (P=0.007, 0.003), and the total protein content was higher in the nicotine group than in the solvent control group ( P=0.060 ), while the relative SP-B mRNA expression was lower in the nicotine group than in the blank control group ( P=0.025 ), and the relative SP-D mRNA expression was lower in the nicotine group than in the blank control group and solvent control group ( P=0.004, 0.041 ). Conclusion Acute exposure to e-cigarette results in elevated intrapulmonary inflammatory responses, pulmonary capillary barrier impairment and reduced pulmonary surfactant protein expression.

Objective: To investigate the incidence and pathogen distribution of ventilator-associated pneumonia (VAP) among preterm infants admitted to level Ⅲ neonatal intensive care units (NICU) in China. Method: A prospective study was conducted in 25 level Ⅲ NICU, enrolling all preterm infants <34 weeks gestational age admitted to the participating NICU within the first 7 days of life from May 2015 to April 2016. Chi-square test, t test and Mann-Whitney U test were used for statistical analysis. Result: A total of 7 918 patients were enrolled, within whom 4 623(58.4%) were males. The birth weight was (1 639±415) g and the gestational age was (31.4±2.0) weeks; 4 654(58.8%) infants required non-invasive mechanical ventilation and 2 154(27.2%) required intubation. Of all the mechanically ventilated patients, VAP occurred in 95 patients. The overall VAP rate was 7.0 episodes per 1 000 ventilator days, varying from 0 to 34.4 episodes per 1 000 ventilator days in different centers. The incidence of VAP was 9.6 and 6.0 per 1 000 ventilator days in children′s hospitals and maternity-infant hospitals respectively, without significant differences (t=1.002, P=0.327). Gram-negative bacilli (76 strains, 91.6%) were the primary VAP microorganisms, mainly Acinetobacter baumannii (24 strains, 28.9%), Klebsiella pneumonia (23 strains, 27.7%), and Pseudomonas aeruginosa (10 strains, 12.0%). Conclusion The incidence of VAP in China is similar to that in developed counties, with substantial variability in different NICU settings. More efforts are needed to monitor and evaluate the preventable factors associated with VAP and conduct interventions that could effectively reduce the occurrence of VAP.

Objective To investigate the efficacy and prognostic risk factors of ALL-R-2003 protocol in the treatment of relapsed childhood relapsed acute lymphoblastic leukemia (ALL) in single center. Methods A retrospective study of clinical data of 51 children with relapsed ALL from January 2004 to December 2014 was performed by using SPSS version 19.0 statistical software for statistical analysis. Results The median age at initial diagnosis of 51 patients was 5.5 years (range, 0.8-13.4 years). The median time from initial diagnosis to relapse was 25 months (range, 3-68 months) and follow-up time was 39 months (range, 3-116 months). The relapse rate in the standard-risk, intermediate-risk and the high-risk groups were 27.5 % (14/51), 29.4 %(15/51) and 43.1 % (22/51), respectively. The probability of 3-year overall survival (pOS) after relapse was (18.8±5.9)%and the probability of event free survival (pEFS) was (16.2±5.8)%. The 3-year pOS in very early relapse, early relapse and late relapse were 0, (11.7 ±7.7) % and (51.7 ±14.8) %, respectively (P= 0.000). There was no statistical difference in survival rate of different immunophenotype groups and sites of relapse (P> 0.05). The 3-year pOS of group S1, S2, S3, S4 were (50.0±35.4) %, (39.9±1.3) %, (10.0±9.5) % and 0, respectively (P=0.000). The 3-year pOS of bcr-abl and MLL gene positive groups were (25.0±21.7) %and 0, respectively, with no statistically significance compared with the negtive group [(24.1±12.0)%] (P>0.05). The 3-year pOS rates of children with bone marrow transplantation and without transplantation were (40.0 ±15.5) %and (13.0 ±5.9) % respectively (P= 0.038). Conclusions The children who in high risk group at initial diagnose are easily to meet earlier relapse and poorer prognosis. The survival period after relapse of bcr-abl or MLL gene positive cases is very short. Bone marrow transplantation can improve survival rate. Risk group at initial diagnose, relapse time and transplantation are the main factors influencing prognosis, and the relapse time and transplantation are the independent prognostic factors for relapsed childhood ALL.