Objective·To investigate the immunotherapeutic effects of exosomes derived from leukemia cells with high expression of B7 molecules(co-stimulatory molecules CD80 and CD86)in tumor-bearing mice.Methods·L1210 leukemia cells were transfected with lentiviral vectors containing the CD80 and CD86 genes,and thereby the leukemia cells with high expression of CD80 and CD86 molecules were obtained.Exosomes LEX-8086 with high expression of CD80 and CD86 molecules were then isolated from the culture supernatant.The biological characteristics of LEX-8086 were analyzed by using transmission electron microscopy(TEM)and Western blotting.A DBA/2-leukemia tumor-bearing mouse model was established.The tumor-bearing mice were divided into LEX-8086 group and PBS group.The LEX-8086 group received intravenous injections of 500 μL LEX-8086(150 μg/mL)via the tail vein 7 d and 12 d after tumor cell inoculation,while the PBS group received injections of equal volumes of PBS at the same time points.Differences in body weight,tumor volume,tumor mass,and survival of the mice between the two groups were observed.Flow cytometry was used to analyze the changes in the immune cells in spleens and lymph nodes of the mice in both groups.Western blotting and real-time fluorescent quantitative PCR(qPCR)were used to detect the expression of markers for M1 and M2 macrophages in the spleens of the mice in both groups.Results·The exosomes LEX-8086 were isolated from the supernatant of L1210 cells overexpressing CD80 and CD86 molecules.TEM revealed that LEX-8086 exhibited a disc-shaped vesicular structure with a diameter of approximately 100 nm.Western blotting demonstrated the expression of exosome-specific markers.In the animal experiments,compared with the PBS group,the body weight of the LEX-8086 group showed no significant change,while both the tumor volume and the relative tumor mass ratio decreased significantly,and the survival was significantly prolonged(all P<0.05).Flow cytometry results indicated that the proportions of natural killer(NK)cells and CD4+T cells in the lymph nodes and spleens of the LEX-8086 group significantly increased(all P<0.05);as for CD8+T cells,they only increased in the lymph modes(P=0.012).Moreover,flow cytometry results showed that the proportion of M1 macrophages in the spleens of mice in the LEX-8086 group was significantly elevated(P=0.003),while the proportion of M2 macrophages remained unchanged.Western blotting and qPCR also revealed that the mRNA and protein expression levels of M1 macrophage markers interleukin-6 and tumor necrosis factor-α in the spleens of mice in the LEX-8086 group increased significantly(all P<0.05).Conclusion·Exosomes derived from leukemia cells with high expression of co-stimulatory molecules CD80 and CD86 can induce a therapeutic anti-leukemic effect,which may be achieved by increasing the proportions of NK cells,M1 macrophages,CD4+T cells,and CD8+T cells.

Objective·To investigate the immunotherapeutic effects of exosomes derived from leukemia cells with high expression of B7 molecules(co-stimulatory molecules CD80 and CD86)in tumor-bearing mice.Methods·L1210 leukemia cells were transfected with lentiviral vectors containing the CD80 and CD86 genes,and thereby the leukemia cells with high expression of CD80 and CD86 molecules were obtained.Exosomes LEX-8086 with high expression of CD80 and CD86 molecules were then isolated from the culture supernatant.The biological characteristics of LEX-8086 were analyzed by using transmission electron microscopy(TEM)and Western blotting.A DBA/2-leukemia tumor-bearing mouse model was established.The tumor-bearing mice were divided into LEX-8086 group and PBS group.The LEX-8086 group received intravenous injections of 500 μL LEX-8086(150 μg/mL)via the tail vein 7 d and 12 d after tumor cell inoculation,while the PBS group received injections of equal volumes of PBS at the same time points.Differences in body weight,tumor volume,tumor mass,and survival of the mice between the two groups were observed.Flow cytometry was used to analyze the changes in the immune cells in spleens and lymph nodes of the mice in both groups.Western blotting and real-time fluorescent quantitative PCR(qPCR)were used to detect the expression of markers for M1 and M2 macrophages in the spleens of the mice in both groups.Results·The exosomes LEX-8086 were isolated from the supernatant of L1210 cells overexpressing CD80 and CD86 molecules.TEM revealed that LEX-8086 exhibited a disc-shaped vesicular structure with a diameter of approximately 100 nm.Western blotting demonstrated the expression of exosome-specific markers.In the animal experiments,compared with the PBS group,the body weight of the LEX-8086 group showed no significant change,while both the tumor volume and the relative tumor mass ratio decreased significantly,and the survival was significantly prolonged(all P<0.05).Flow cytometry results indicated that the proportions of natural killer(NK)cells and CD4+T cells in the lymph nodes and spleens of the LEX-8086 group significantly increased(all P<0.05);as for CD8+T cells,they only increased in the lymph modes(P=0.012).Moreover,flow cytometry results showed that the proportion of M1 macrophages in the spleens of mice in the LEX-8086 group was significantly elevated(P=0.003),while the proportion of M2 macrophages remained unchanged.Western blotting and qPCR also revealed that the mRNA and protein expression levels of M1 macrophage markers interleukin-6 and tumor necrosis factor-α in the spleens of mice in the LEX-8086 group increased significantly(all P<0.05).Conclusion·Exosomes derived from leukemia cells with high expression of co-stimulatory molecules CD80 and CD86 can induce a therapeutic anti-leukemic effect,which may be achieved by increasing the proportions of NK cells,M1 macrophages,CD4+T cells,and CD8+T cells.

Objective:To compare the clinical outcomes of endovascular therapy in acute stroke patients with anterior circulation tandem occlusions caused by atherosclerosis or dissection.Methods:A retrospective cohort study. A total of 98 patients with anterior circulation tandem lesions undergoing endovascular therapy in the Wuhan NO.1 Hospital (March 2016 to March 2022) were analyzed. Median age was 64(55,71) years old, and 82.7% (81/98 cases) were males. According to the lesion etiology, the patients were divided into atherosclerosis and dissection groups. The differences in clinical outcomes between the two groups were investigated, including favorable 90-day functional outcome (modified Rankin Scale score of 0-2), successful reperfusion (modified Thrombolysis in Cerebrovascular Infarction score of 2b-3), symptomatic intracranial hemorrhage, stroke-associated pneumonia, 90-day all-cause mortality, and average hospitalization days. Logistic regression analysis was used to adjust for potential confounders affecting functional outcomes in both groups, and to determine odds ratios and 95% confidence intervals.Results:Seventy-one patients were grouped into the atherosclerotic cause and 27 into the dissection cause cohorts. The rate of favorable 90-day functional outcome was 43.7% (31/71 cases) in the atherosclerosis group versus 55.6% (15/27 cases) in the dissection group (adjusted odds ratio=1.339; 95% confidence interval, 0.374-4.798; P=0.654). No significant differences were found in other clinical outcomes between the two groups (all P>0.05). Conclusion:The clinical prognosis of patients with tandem lesions caused by atherosclerotic stenosis or artery dissection was similar after endovascular therapy. Future studies are still needed to verify our results.

Objective: To investigate the impact of hepatoprotective drugs on the performance of APRI for diagnosing liver fibrosis in chronic hepatitis B (CHB). Methods: Patients with CHB who underwent a percutaneous liver biopsy were recruited and divided into hepatoprotective drugs using group and hepatoprotective drugs free group. Grouping was carried out according to the types of commonly used hepatoprotective drugs, and controls were matched for each groups by stage of liver fibrosis and age from hepatoprotective drugs free group. The performance of APRI for diagnosing significant fibrosis and cirrhosis was evaluated and compared between each experimental groups and control groups using receiver operating characteristic (ROC) curves. Results: A total of 1447 patients were enrolled, including 60 using glycyrrhizin, 54 using silymarin, 63 using traditional Chinese medicine (TCM) containing schisandra, and 113 using more than one hepatoprotective drug. In patients using glycyrrhizin, the sensitivity of APRI to predict significant fibrosis was higher than its control group. In patients using more than one hepatoprotective drug, the sensitivity of APRI to exclude significant fibrosis was higher than its control group; the area under ROC curve of APRI to diagnose significant fibrosis, the specificity of APRI to exclude and predict significant fibrosis were all lower than its control group. In patients using glycyrrhizin and patients using more than one hepatoprotective drug, the specificity of APRI to exclude cirrhosis were both lower than their control groups. In patients using silymarin and patients using TCM, the performance of APRI to diagnose significant fibrosis and cirrhosis was comparable with their control groups. Conclusions Glycyrrhizin and combination of hepatoprotective drugs would have significant impact on the performance of APRI for diagnosing liver fibrosis in CHB, silymarin and TCM containing schisandra would have less impact.

Objective: To investigate the role of HBsAg status and content in neonatal venous blood to predict HBV mother-to-children transmission. Methods: The study candidates from a prospective study about HBV mother-to-children transmission blocking who were hepatitis B surface antigen (HBsAg) positivity, hepatitis B e antigen (HBeAg) positivity, and HBV DNA levels >10⁵ IU/ml.All of their infants were enrolled.200 IU of hepatitis B immunoglobulin (HBIG)was injected within 6 hours after birth, and 200 IU HBIG was voluntarily selected 1 month after birth.All infants according to 0-1-6 month standard procedure were given 10 or 20 μg of hepatitis B vaccine. Pregnancy women before birth, and infants at the time of birth, 1-month and 7-month after birth, venous blood was tested for HBV virus and serological markers to assess the association with success of mother-to-children transmission blocking. Results: 530 pregnant women and 530 neonates were enrolled. 60.75% at birth and 86.02% at birth for one month children were HBsAg-negative. The successful transmission in HBsAg-negative neonates was 100.00%. According to the receiver operating characteristic curve, the AUC of HBsAg content≥0.35 IU/ml at birth predicted to block failure was 0.979. The sensitivity was 85.60%, and the specificity was 100.00%. The AUC of HBsAg content≥0.18 IU/ ml at one month after birth predicted to block failure was 0.988, the sensitivity was 89.40%, and the specificity was 100.00%. Conclusions The HBsAg level in venous blood at birth and 1 month after birth can predict the failure of HBV mother-to-children transmission, and the neonates with HBsAg positivity in venous blood are a high-risk group that may block failure.

Objective: To investigate the predictors of death after endovascular mechanical thrombectomy (EMT) in patients with acute vertebrobasilar occlusive stroke (VBOS). Methods: Patients with acute VBOS treated with EMT in Wuhan No. 1 Hospital were enrolled retrospectively. The demographic and clinical data were collected. According to whether the patients died at 90 d after procedure, they were divided into survival group and death group. The demographic and clinical data were compared between the two groups. Multivariate logistic regression analysis was used to determine the independent risk factors for death at 90 d after EMT. Results: A total of 47 patients were enrolled. The median age was 62 years, 34 were males (72.3%), the median baseline National Institutes of Health Stroke Scale (NIHSS) score was 16, 42 patients (89.4%) had recanalization (modified Thrombolysis in Cerebral Infarction[mTICI] 2b/3 grade), and 12 (25.5%) died within 90 d after procedure. Univariate analysis showed that the baseline NIHSS score (26 [21-28]vs. 12 [5-23]; Z=-3.165, P=0.002), percentage of neutrophil (81.61% ±11.82% vs. 72.20% ±12.09%; t=-2.137, P=0.033), neutrophil/lymphocyte ratio (10.54±7.17 vs. 4.98±3.57; t=-2.393, P=0.017), and incidence of sICH (25.0% vs. 2.9%; χ²=5.627, P=0.018) in the death group were significantly higher than those in the survival group, while the percentage of lymphocyte (12.00%±9.04% vs. 20.67%±10.39%; t=-2.429, P=0.015) was significantly lower than that of the survival group. Multivariate logistic regression analysis showed that high baseline NIHSS score (odds ratio [OR] 1.243, 95% confidence interval [CI] 1.046-1.318; P=0.038), high neutrophil/lymphocyte ratio (OR 1.278, 95% CI 1.002-1.630; P=0.049) and symptomatic intracranial hemorrhage (OR 5.088, 95% CI 1.065-38.718; P=0.046) were the independent predictors for death. Conclusion High baseline NIHSS score, high neutrophil/lymphocyte ratio and symptomatic intracranial hemorrhage are the independent predictors for death within 90 d after EMT in patients with acute VBOS.

In view of the current teaching status of Traditional Rehabilitation Methodology for rehabilitation medicine specialty in higher medical colleges and universities,combining with the knowledge of some acupoints taught in this course,we put forward some suggestions for the teaching of this course.In teaching process of acupoints knowledge,while emphasizing the importance of its anatomical location,we should also attach importance to elaborating the name of acupoints and the rich cultural contents contained in the naming process.At the same time,paying attention to induction learning and finding memory rules,combining theoretical knowledge with classmates' hands-on practice,and encouraging students to participate early in clinical practice activities,we can develop students' practical work ability.

Objective:To preliminarily study the characteristics of ventricular synchronicity in patients of idiopathic pulmonary arterial hypertension (IPAH) with different heart function.
Methods:Our research included in 2 groups:IPAH group, n=40 patients intermittently collected in our hospital from 2010-10 to 2014-12 and Control group, n=23 normal subjects from the same period of time. Tissue Doppler echocardiography was conducted to compare interventricular systolic and diastolic time to peak (Ts and Te), their differences (Te-Ts) and (Te-Ts) of left ventricle (LV), right ventricle (RV), interventricular septum (IVS) between 2 groups.
Results:Ts and Te of RV were longer than Ts and Te of LV and IVS in both groups, P<0.05. Compared with Control group, by heart function reducing, IPAH group showed different Ts and Te of LV and IVS, P<0.05 and Ts was gradually prolonged, Te was gradually shorter;while Te of RV became longer firs tand then became shorter thereafter. In IPAH group, the patients with heart function grade IV had the longer Ts of LV and IVS than those with heart functionII, III;the patients with heart function IV had the shorter Te of LV, RV and IVS than those with heart function II, III, all P<0.05. In IPAH group with different heart function, Te between RV-IVS and RV-LV were different from Control group, all P<0.05;Te between LV-IVS in heart function IV patients had the largest difference from Control group, P<0.05. Compared with Control group, IPAH group had gradually decreased Te-Ts of LV and IVS by heart function reducing asgrade II>III>IV, all differences had statistic meaning.
Conclusion:IPAH patients with different right heart function may have interventricular systolic and diastolic de-synchronicity;by reduced heart function, interventricular Te-Ts could be specifically shortened.

Objective: To investigate the efficacy of 200IU hepatitis B immunoglobulin (HBIG) injection at 1 month after birth to interrupt the mother-to-children transmission (MTCT) of hepatitis B virus (HBV). Methods: Infants born to mothers who were hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) positive, with HBV DNA load ≥1.0×10⁶ IU/ml and who did not receive antiviral drug treatment during pregnancy, were randomly divided into 2 groups. Infants in the control group were treated with standard immunoprophylaxis: 200 IU HBIG and 10 μg recombinant hepatitis B vaccine injection within 2 h after birth and a vaccine booster at 1 and 6 months after birth. For infants in the HBIG group the standard immunoprophylaxis and an additional 200 IU HBIG were administered at 1 month. HBsAg, the antibody to HBsAg (anti-HBs), and HBV DNA load were measured at birth and after 7 months. later.Immunoprophylaxis failure was defined as the presence of HBV DNA and HBsAg positivity or the presence of HBV DNA and HBsAg negativity at 7 months. Results: In this prospective cohort study, of the 280 infants enrolled, 14 infants (HBIG/control: 6/8) were lost to follow-up and 266 subjects (HBIG/control: 134/132) completed the 7-month study. The log₁₀HBV DNA load of mothers in the HBIG group and control group were (7.31±0.66) log₁₀IU/ml and (7.32±0.74) log₁₀IU/ml, respectively (P=0.92). The MTCT rate of the two groups was similar (5.97% vs. 7.58%, P=0.63). At 7 months, the HBsAg positive rate and the level of anti-HBs in the two groups were 94.03%(126/134)vs. 91.67% (121/132) and 623.60±412.93 mIU/mL vs. 620.38±399.10 mIU/ml, respectively with no significant difference (P=0.48 and P=0.95, respectively). The log₁₀ HBV DNA load of mothers in immunoprophylaxis failure group and success group was similar (P=0.09). The number of infants who were serum HBsAg positive and HBV DNA positive at birth in the immunoprophylaxis failure group were higher than those in the success group (100% and 100% vs. 35.89% and 31.85%, P<0.01, respectively). The serum HBsAg levels in infants at birth was the only independent relevant factor for HBV MTCT, with risk rates of 11.18 (95% Confidence interval (CI), 1.23-101.88), 352.00 (95%CI, 15.82-7833.20), and 968.00 (95%CI 81.35-11519.19) for HBsAg levels of 0.05-< 1, 1-< 10, and ≥ 10 IU/ml, respectively, compared to infants with HBsAg levels < 0.05 IU/ml. Conclusions Administering 200IU HBIG injection at 1 month did not reduce the risk of HBV MTCT.

ObjectiveTo explore the value of conventional echocardiography and transthoracic real-time three-dimensional echocardiography (RT3DE) in diagnosis of Ebstein anomaly.MethodsWe investigated the morphology and function of right ventricle (RV) as well as the structure, morphology and regurgitation of tricuspid valve in 61 adult patients with Ebstein anomaly before surgery by using conventional echocardiography and transthoracic RT3DE. Twenty normal adults were enrolled as control group. ResultsThe surface of the tricuspid leaflets, the morphology of the tricuspid annulus as well as the three-dimensional structure of the tricuspid valve were displayed stereoscopically by RT3DE. Complete RV volume data could be acquired in 32 patients of Ebstein anomaly. The apex or part of RV could not be contained in the remaining 29 patients. Thirty-four (55.7%) patients with Ebstein anomaly had severe tricuspid regurgitation,
16 (26.2%) patients had moderate to severe regurgitation, and 11 (18.1%) patients had moderate regurgitation. Compared with the normal adults, patients of Ebstein anomaly showed higher RV end-diastolic volume (EDV), end-systolic volume (ESV), end-diastolic volume index (EDVI), end-systolic volume index (ESVI), stroke volume (SV), RV anterior-posterior diameter (RV), tricuspid valve annular transverse diameter (TV-R), and lower RV ejection fraction(EF) [(273.5±77.7) mlvs (74.3±15.9) ml, (187.1±96.8) mlvs (31.1±9.2) ml, (177.4±53.6) ml/m2vs (43.4±8.2) ml/m2, (121.7±65.5) ml/m2vs (18.4±5.1) ml/m2, (95.9±20.2) ml vs (43.6±8.8) ml, (48.1±13.3) ml/m2vs (19.0±1.9) ml/m2, (56.4±8.9) ml/m2vs (28.5±4.3) ml/m2, (38.3±12.8) %vs (59.3±5.1) %, allP<0.05). The tricuspid regurgitant orifice flow cross-sectional area (EROA) were correlated positively with RV anterior-posterior diameter (r=0.691), ratio of RV and LV anterior-posterior diameter (RV/LV) (r=0.6471).ConclusionTransthoracic RT3DE is a feasible method in addition to conventional two-dimensional echocardiography in evaluation of tricuspid valve mophology and function, as well as RV volume and EF in adult patients with Ebstein anomaly.