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 assess the effect of different postoperative body positions on respiratory func-tion of patients undergoing shoulder arthroscopy under general anesthesia with interscalene brachial plex-us block.Methods Seventy patients undergoing arthroscopic repair of the supraspinatus tendon were ran-domly allocated to a supine position group and a group with a 30° head elevation.Patients in both groups were in a horizontal position before anesthesia,and diaphragm movement was measured under B-ultrasound as baseline.Promptly after removal of the endotracheal tube upon arrival at the anesthe-sia recovery room,the supine group was conducted blood gas analysis,and monitored heart rate,res-piration,non-invasive arterial blood pressure and blood oxygen saturation,wearing a mask for oxygen supplementation,while the group with a 30° head elevation was given identical nursing procedures af-ter the bed's head was elevated 30°.Then right after entering the anesthesia recovery room,as well as 5 and 15 minutes after deoxygenation,the oxygen partial pressure(PaO2),carbon dioxide partial pressure(PaCO2)and percutaneous oxygen saturation(SpO2)were recorded and compared between the two groups.Moreover,the diaphragmatic movements during deep breathing before nerve block and pri-or to discharge from the anesthesia recovery room were measured,and such adverse reactions as hypox-emia,labored breathing and glossoptosis were observed.Results Compared with the supine position group,a significant increase in the average PaO2 and SpO2 15 minutes after deoxygenation,diaphrag-matic movement during deep breathing and comfort was observed in the group with a 30° head eleva-tion,but a significant decrease in the incidence of postoperative laboured breathing and glossoptosis.However,no significant differences were found between the two groups in the incidence of hypoxemia and the average PaO2,PaCO2 and SpO2 immediately after entering the anesthesia recovery room and 5 minutes after deoxygenation.Conclusion Elevating the head of the bed by 30° enhances respiratory function in patients undergoing shoulder surgery with general anesthesia intubation and intermuscular groove brachial plexus block,compared to the supine position.Although no significant reduction in hy-poxemia incidence is observed,there is a notable relief of adverse reactions such as laboured breath-ing and glossoptosis,which makes patients feel more comfortable.Therefore,such posture care is wor-thy of application in clinical practice.

ObjectiveTo investigate the antiviral effect of Menispermi Rhizoma total alkaloids and its relationship with the type Ⅰ interferon （IFN-Ⅰ） signaling pathway. MethodThe effects of Menispermi Rhizoma total alkaloids on the intracellular replication of influenza A virus （H1N1）， vesicular stomatitis virus （VSV）， and cerebral myocarditis virus （EMCV） were detected by fluorescent inverted microscope， flow cytometry， Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）， and Western blot. A mouse model infected with H1N1 was constructed， and the mice were divided into a control group， H1N1 model group， Menispermi Rhizoma total alkaloids groups （10， 20， 30 mg·kg^-1）， and oseltamivir group （40 mg·kg^-1）， so as to study the effects on the weight and survival rate of infected mice. Real-time PCR was used to detect the activation effect of Menispermi Rhizoma total alkaloids on the IFN-Ⅰ pathway in cells， and the relationship between the antiviral effect of Menispermi Rhizoma total alkaloids in IFNAR1 knockout A549 cells （IFNAR1^-/--A549） and IFN-Ⅰ pathway was detected. ResultCompared with the control group， the virus proliferated significantly in the model group （P<0.01）. Compared with the model group， Menispermi Rhizoma total alkaloids could significantly inhibit the replication of H1N1， VSV， and EMCV in vitro （P<0.01）， inhibit the weight loss of the mice infected with the H1N1 in vivo， and improve the survival rate of mice （P<0.05）. In addition， Menispermi Rhizoma total alkaloids activated the IFN-I pathway and relied on this pathway to exert the function of antiviral infection. ConclusionMenispermi Rhizoma total alkaloids exert antiviral effects in vivo and in vitro by activating the IFN-Ⅰ pathway.

Objective: To analyze the impact of parental sports concept and behavior on physical activity in preschool children, so as to provide a foundation for future guidance on fostering childrens physical activity within the family context. Methods: From November to December 2020, a clustered convenience sampling method was employed to conduct surveys, and a total of 283 children were selectal from one kindergarten each in Beijing, Shenyang, and Xian. Participating children wore ActiGraph GT9X accelerometers continuously for one week to collect data on different intensity levels of physical activity. Physical Activity afterschool Questionnaire for Preschooler (P-PAQ) was utilized to assess parental sports concept and behavior. The gender differences in physical activity level and physical activity compliance rate were analyzed by using ttest, Mann-Whitney U test, and Chisquare test; and the relationship between parental exercise concepts and behaviors and physical activity of preschool children was analyzed using Spearman rank correlation analysis and multiple linear regression analysis. Results: Parental sports concept was significantly positively correlated with average daily moderatetovigorous physical activity (MVPA) and total physical activity (TPA) in children (r=0.12-0.16, P<0.05). Parental sports behavior was significantly positively correlated with childrens average daily TPA (r=0.25, P<0.05). Multiple linear regression revealed that parental sports concept was positively correlated with average daily MVPA and TPA in both boys and girls (B=0.65-0.83), while parental sports behavior only was positively correlated with boys average daily MVPA and TPA (B=0.24-0.25)(P<0.05). Conclusions Parental sports concept and behavior can impact physical activity levels in preschool children, exhibiting gender differences. Future guidance on physical activity in family upbringing should consider both parental sports concept and behavior, and pay attention to the influence of childrens gender.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in China. Currently, hepatectomy, radiofrequency ablation, interventional therapy, and so on are widely used in the treatment of liver cancer; however the 5-year survival rate of patients is still very poor. The onset of liver cancer is insidious, and most patients do not have the opportunity to undergo surgery when they are initially diagnosed. In addition, patients with advanced liver cancer lack effective treatment, which is mainly due to chronic inflammation of the liver. Immunotherapy has great potential advantages in the treatment of liver cancer, so it has attracted more and more researchers' attention. Importantly, with the development of biomedical science into the era of precision medicine, more precise and effective immunotherapy strategies for HCC treatment have been explored, thus bringing new hope to HCC patients. This article reviews the research status and progress of liver cancer immunotherapy in the context of precision medicine.

Objective To analyze the differences of clinical manifestations and organ damage between patients with severe fever with thrombocytopenia syndrome(SFTS)and patients with tsutsugamushi disease,and to investigate the prognostic factors of SFTS.Methods The research was performed on 49 patients with SFTS and 16 patients with tsutsugamushi disease who visited the First Affiliated Hospital of Anhui Medical University from October 2014 to June 2017.The general information of patients including region,age,gender and clinical manifestations were evaluated.Blood routine,liver and kidney function,myocardial enzyme levels,lipase,amylase,electrolytes,C-reactive protein,procalcitonin,prothrombin time(PT)and activated partial thromboplastin time(APTT)were continuously monitored during the course of disease.T test was used for continuous variables of normal distribution,and non-parametric test was used for variables of non-normal distribution.Chi-square test was used for categorical variables.Results The mean age of SFTS patients was 62.1±15.5(ranging from 17 to 87 years)and the mean age of tsutsugamushi patients was 56.1±9.2(ranging from 47 to 73 years).There was no significant difference between the two groups(t=1.47,P=0.147).There were 25 males(51%)in SFTS patients and 8 males(50%)in tsutsugamushi disease patients.There was no significant difference between the two groups(x2=0.005,P=0.943).The incidences of headache,vomiting,superficial lymphadenectasis,disturbance of consciousness,proteinuria,hematuria,pulmonary infection,multiple organ dysfunction and acute pancreatitis in SFTS patients were all significantly higher than those in tsutsugamushi disease patients(x2=8.82,4.38,8.71,11.17,7.88,5.56,4.35,9.43,and 8.13,respectively,P <0.05 or 0.01).The counts of leukocytes(Z=2.73),neutrophils(Z=2.46),lymphocytes(Z=3.15),platelets(Z=4.25),albumin(Z=2.65)and sodium ion(t=2.10)in SFTS patients were all significantly lower than those in patients with tsutsugamushi disease(P <0.05 or 0.01).The levels of aspartate aminotransferase(Z=2.94),lactate dehydrogenase(Z=3.42),creatine kinase(CK)(Z=2.88),amylase(Z=2.11),lipase(Z=2.82),creatinine(Z=2.07)and urea nitrogen(Z=2.50)in fatal SFTS patients were all significantly higher than those in patients with tsutsugamushi disease(P <0.05 or 0.01).Among 49 SFTS patients,16 patients died and 33 patients recovered finally.The age(t=3.33),platelet count(Z=2.55),alanine aminotransferase(ALT)(Z=2.10),aspartate aminotransferase(AST)(Z=2.22),lactate dehydrogenase(Z=2.26),CK(Z=3.50),CK-MB(Z=3.10),creatinine(Z=2.17),urea nitrogen(Z=2.36),and sodium(t=2.65)between the two subgroups had significant differences(P <0.05 or 0.01).Conclusions SFTS is more severe and has high mortality,while tsutsugamushi disease has a better prognosis.Early differential diagnosis and early rational treatment are important to reduce the mortality of patients with SFTS.

Objective This meta-analysis aims to identify consistent results of vulnerable brain regions through the existing resting-state fMRI studies, thus exploring the changes of neural spontaneous brain activity in type 2 diabetes mellitus (T2DM) patients. Methods A systematic analysis of studies on brain resting-state changes in T2DM patients using ALFF, fALFF and ReHo analysis. The meta-analysis was based on the activation likelihood estimation method, using the software of Ginger ALE 2.3. Results Ten studies from 7 references (188 T2DM patients and 170 healthy controls) were included. Based on the analysis of ALFF and ReHo data, this meta-analysis identi fied the robust reduction of resting-state spontaneous brain activity in T2DM patients, including the left lingual gyrus of occipital lobe, right cerebellum posterior lobe,left postcentral gyrus and right insula(cluster size= 800,488,368,256 mm3,P<0.05 after FDR correction),while no increased spontaneous brain activation was found in any regions.The meta-analysis from ReHo studies showed reduced resting-state spontaneous brain activity in the left lingual gyrus of occipital lobe,left postcentral gyrus,right insula and posterior cingulum/lingual gyrus(cluster size=832,368,280,232 mm3,P<0.05 after FDR correction),while no increased spontaneous brain activation was found in any regions. Conclusion This meta-analysis study using the activation likelihood estimation method demonstrated that the resting-state spontaneous brain anomalies in T2DM patients might contribute to exploring machenism underlying diabetic encephalopathy.

Objective To observe expression level of serum vascular endothelial growth factor-C (VEGF-C),VEGF-C receptor-2 and VEGF-C receptor-3 in patients with acute leukemia (AL) and to explore its clinical significance.Methods Enzyme-linked immuno sorbent assay (ELISA) was used to detect the serum expression levels of VEGF-C,VEGFR-2,and VEGFR-3 of 51 patients diagnosed with acute leukemia,43 patients under medical treatment and 16 healthy blood donors.Results (1) Serum VEGF-C,VEGFR-2,and VEGFR-3 expression levels in AL patients were significantly higher than those in normal control group.(2) Serum VEGF-C and VEGFR-2 expression levels in complete remission (CR) group significantly declined after treatment.Serum VEGF-C and VEGFR-2 expression levels in non-complete remission (NR) group slightly declined after treatment but no significant difference was found (P＞0.05).(3) No significant difference was found in serum VEGFR-3 expression levels both in CR group and NR group after treatment (P＞0.05).(4) Serum VEGF-C,VEGFR-2,and VEGFR-3 expression levels in NR group were significantly higher than those in CR group before treatment (P＜0.08).Conclusions Observing serum expression level of VEGF-C,VEGFR-2,and VEGFR-3 of AL patients may be helpful in monitoring curative effects and prognosis of acute leukemia.