Objective There are complex pathophysiological changes in each component of cerebral tissue after cerebral ischemia-reperfusion injury，and routine neuroprotective strategies fail to exert an effective therapeutic effect in clinical practice. However， the proposal of neurovascular unit（NVU） provides a new conceptual framework for a more comprehensive understanding of the pathophysiological process for interactions between each component of cerebral tissue after cerebral ischemia-reperfusion injury， laying a theoretical foundation for investigating new targets for clinical treatment. Methods A novel NVU in vitro model was established in Transwell co-culture plate using the neurons， astrocytes， and brain microvascular endothelial cells （BMECs） derived from the cortex of Sprague-Dawley rats， and MTT assay was used to measure cell viability in the modified and classical NVU models after OGD/R injury for 1 hour. Results An in vitro NVU model with the three-layer structure of neurons， astrocytes， and BMECs from the top to the bottom was successfully established using Transwell co-culture plate. After OGD/R injury for 1 hour， the neurons in the modified in vitro NVU model showed a significantly higher viability than those in the classical NVU model （P<0.05）， while there were no significant differences in the viability of astrocytes and BMECs between the two models （P>0.05）. Conclusion The modified in vitro NVU model is more suitable for the research on ischemic cerebrovascular disease.

Objective:To investigate the relationship between CXCL6 expression and prostate cancer,as well as its effects on the malignant progression of PC-3 cells and the underlying mechanisms.Methods:Multiple databases were used to analyze CXCL6 expression in prostate cancer.And survival analysis,GO,and KEGG enrichment analysis were conducted.According to the concentration of CXCL6,PC-3 cells were divided into 0,100 and 200 ng/ml CXCL6 groups.The effects of up-regulated CXCL6 expression on PC-3 cell proliferation,migration,apoptosis,cycle,and related genes were investigated using CCK-8,EdU assay,scratch test,flow cytometry,and prostate cancer-targeted PCR array.Results:Database analysis revealed that CXCL6 was significantly down-regulated in prostate cancer(P＜0.05),and patients with low CXCL6 expression had a worse prognosis(P＜0.05).In prostate cancer,proteins associated with CXCL6 expression included CXCL1 and CXCL2,and related genes included MIR325 and RASSF6.GO analysis indicated that differentially expressed genes(DEGs)of CXCL6 were primarily enriched in processes such as response to chemokines and the tubular endoplasmic reticulum network(P＜0.05).KEGG analysis showed that DEGs were mainly enriched in signaling pathways such as the Hippo signaling pathway(P＜0.05).CCK-8,EdU assay,scratch test,and flow cytometry results demonstrated that compared with the 0 ng/ml CXCL6 group,the 100 and 200 ng/ml CXCL6 groups exhibited significantly reduced proliferation and migration rates(P＜0.05),significantly increased apoptosis rate(P＜0.05).And the proportion of S-phase cells in the 200 ng/ml CXCL6 group was lower compared to the 0 ng/ml CXCL6 group(P＜0.05).PCR array results showed that compared with the 0 ng/ml CXCL6 group,the expression of genes such as CDH1 and HAL was increased in the 200 ng/ml CXCL6 group,while the expression of genes such as CD5L and EDNRB was decreased.Conclusion:CXCL6 plays a significant role in the malignant progression of PC-3 cells,and its mechanism may be associated with the methylation of promoters of genes such as RASSF1,RARB,and CDH1.

Objective:To investigate the relationship between CXCL6 expression and prostate cancer,as well as its effects on the malignant progression of PC-3 cells and the underlying mechanisms.Methods:Multiple databases were used to analyze CXCL6 expression in prostate cancer.And survival analysis,GO,and KEGG enrichment analysis were conducted.According to the concentration of CXCL6,PC-3 cells were divided into 0,100 and 200 ng/ml CXCL6 groups.The effects of up-regulated CXCL6 expression on PC-3 cell proliferation,migration,apoptosis,cycle,and related genes were investigated using CCK-8,EdU assay,scratch test,flow cytometry,and prostate cancer-targeted PCR array.Results:Database analysis revealed that CXCL6 was significantly down-regulated in prostate cancer(P＜0.05),and patients with low CXCL6 expression had a worse prognosis(P＜0.05).In prostate cancer,proteins associated with CXCL6 expression included CXCL1 and CXCL2,and related genes included MIR325 and RASSF6.GO analysis indicated that differentially expressed genes(DEGs)of CXCL6 were primarily enriched in processes such as response to chemokines and the tubular endoplasmic reticulum network(P＜0.05).KEGG analysis showed that DEGs were mainly enriched in signaling pathways such as the Hippo signaling pathway(P＜0.05).CCK-8,EdU assay,scratch test,and flow cytometry results demonstrated that compared with the 0 ng/ml CXCL6 group,the 100 and 200 ng/ml CXCL6 groups exhibited significantly reduced proliferation and migration rates(P＜0.05),significantly increased apoptosis rate(P＜0.05).And the proportion of S-phase cells in the 200 ng/ml CXCL6 group was lower compared to the 0 ng/ml CXCL6 group(P＜0.05).PCR array results showed that compared with the 0 ng/ml CXCL6 group,the expression of genes such as CDH1 and HAL was increased in the 200 ng/ml CXCL6 group,while the expression of genes such as CD5L and EDNRB was decreased.Conclusion:CXCL6 plays a significant role in the malignant progression of PC-3 cells,and its mechanism may be associated with the methylation of promoters of genes such as RASSF1,RARB,and CDH1.

Minimal residual disease (MRD), a crucial biomarker for assessing efficacy and predicting recurrence, refers to residual tumor cells remaining in the body of patients with hematological malignancies who achieved complete remission after treatment. This study aimed to conduct a retrospective analysis of the clinical diagnosis, treatment, and MRD monitoring of a pediatric patient with multiple acute B-lymphocytic leukemia relapses, alongside a review of relevant literature. In this case, Ig rearrangement based on next-generation sequencing (NGS) was more accurate in assessing the MRD level, compared with the traditional method of MRD detection, indicating the risk of earlier relapse and guided interventions in time. Additionally, NGS-MRD detected clonal evolution, providing new ideas to further investigate the intrinsic factors of disease development.

Objective To investigate the effects and possible mechanisms of the growth differentia-tion factor 15(GDF-15)/glial-derived neurotrophic factor receptor alpha-like(GFRAL)pathway on the progression of atherosclerosis in ApoE-/-mice.Methods Eight 8-week-old male ApoE-/-mice were randomly divided into control group and rGDF-15 group.The mice in the control group received an injection of phosphate-buffered saline via tail vein once a week after 4 weeks of high-fat diet feeding,and those in the rGDF-15 group received an injection of recombinant GDF-15(0.05 mg/kg)via tail vein once a week after 4 weeks of high-fat diet feeding.The mice were fed with high-fat diet for another 8 weeks,the body weight was monitored during this period.After 12 weeks'feeding,the mice were euthanized.Another 4 normal mice(at the same age,20 weeks old)were subjected and served as normal control group.The levels of fasting blood glucose,blood lip-ids,cortisol,and aldosterone were compared among the three groups.Oil red O staining was used to evaluate plaque size in the aorta,and immunohistochemistry was employed to assess the expression of GDF-15 and GFRAL in the brain tissue.Results The serum level of GDF-15 was higher in the rGDF-15 group than in the control group(52.59±2.90 ng/ml vs 20.09±1.27 ng/ml,P＜0.01).The weight of mice was significantly lower in the rGDF-15 group than the con-trol group during Week 11(28.60±0.22 g vs 29.47±0.25 g,P＜0.01)and 12(28.98±0.22 g vs 30.35±0.13 g,P＜0.01).The rGDF-15 group had a statistically lower level of triglycerides(0.22±0.02 mmol/L vs 0.38±0.09 mmol/L,P＜0.05),lighter plaque burden[(22.22±2.58)％vs(31.61±3.51)％,P＜0.01],and enhanced expression levels of GDF-15 and GFRAL in the brain tissue(0.088±0.007 vs 0.030±0.006,0.031±0.003 vs 0.010±0.001,P＜0.01).The levels of cor-tisol and aldosterone in the control group and rGDF-15 group were significantly higher than those in the normal group(P＜0.01).The aldosterone level in the rGDF-15 group was significantly re-duced compared to the control group(22.013.67 mg/ml vs 87.29±8.63 mg/ml,P＜0.01).Conclusion GDF-15 may regulate body weight and triglyceride and aldosterone levels through GFRAL,and then affect the progression of atherosclerosis.

Minimal residual disease (MRD), a crucial biomarker for assessing efficacy and predicting recurrence, refers to residual tumor cells remaining in the body of patients with hematological malignancies who achieved complete remission after treatment. This study aimed to conduct a retrospective analysis of the clinical diagnosis, treatment, and MRD monitoring of a pediatric patient with multiple acute B-lymphocytic leukemia relapses, alongside a review of relevant literature. In this case, Ig rearrangement based on next-generation sequencing (NGS) was more accurate in assessing the MRD level, compared with the traditional method of MRD detection, indicating the risk of earlier relapse and guided interventions in time. Additionally, NGS-MRD detected clonal evolution, providing new ideas to further investigate the intrinsic factors of disease development.

Objective:To investigate the correlation between serum cystatin C (CysC) and clinical and pathological features of IgA nephropathy.Methods:Four hundred and twenty-one cases of primary IgA nephropathy diagnosed by renal biopsy in Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from January 2010 to January 2021 were retrospectively analyzed. According to the serum CysC level at the time of renal biopsy, the patients were divided into high serum CysC group and normal serum CysC group, and the clinical data and pathological indices of the patients were compared. Spearman correlation analysis was used to analyze the correlation between estimated glomerular filtration rate (eGFR) and serum CysC. The clinicopathological factors related to the serum CysC level were analyzed by multiple linear regression. The area under the receiver operator characteristic curve (AUC) was used to evaluate the ability of serum CysC level to predict related pathological injury.Results:The age, prevalence of hypertension, serum creatinine, urea and uric acid levels of high serum CysC group were significantly higher than those of normal serum CysC group, while the eGFR level was significantly lower than that of normal serum CysC group ( P<0.05). Spearman correlation analysis showed that serum CysC was negatively correlated with eGFR ( r=-0.744, P<0.001). In terms of pathological injury, the degree of renal tubular atrophy and renal interstitial fibrosis (T) and renal arteriole wall thickening (A) in high serum CysC group were more serious than those in normal serum CysC group ( P<0.05). Multiple linear regression analysis showed that the prevalence of hypertension, serum creatinine, urea, uric acid, T and A were correlated with serum CysC levels (standard regression coefficient β=0.048, 0.299, 0.260, 0.134, 0.195, 0.068, respectively, P<0.05). After adding serum CysC on the basis of clinical features, the prediction efficiency of renal tubular atrophy and renal interstitial fibrosis was higher (AUC were 0.829 [95% CI 0.787-0.870], 0.847 [95% CI 0.808-0.886], P<0.05). Conclusions:Patients with older age, hypertension, poor renal function and severe pathological damage are more likely to have elevated serum CysC levels. Serum CysC was related to the prevalence of hypertension, creatinine, urea, uric acid, T and A. Combined with serum CysC level can effectively improve the ability prediction of T.

Objective To understand the prevalence of autism spectrum disorders (ASD) in children aged 0-6 years old and influencing factors in Hainan province.Methods A total of 37 862 children aged 0-6 years were selected from 18 counties in Hainan province for a screening by using questionnaire of "warning signs in child development",then field diagnosis was made,and general descriptive statistic analysis was conducted.The prevalence of ASD and related factors were analyzed with x2 test and unconditional logistic regression model.Results Among 37 862 children aged 0-6 years,235 were diagnosed with ASD,the prevalence of ASD was 0.62％ (0.99％ in boys,0.17％ in girls),the differences was significant (x2=101.91,P=0.000).The prevalence of ASD increased with age (x2=288.62,P=0.000).The prevalence of ASD was significantly higher in urban area than in other areas (x2=114.77,P=0.000).Factors such as full term pregnancy or not,neonatal asphyxia,father' s characteristics,father' s habit of chewing areca or smoking,mother' s general mood,and mother' s induced abortion history were the influencing factors for ASD.Conclusion The prevalence of ASD in children aged 0-6 years was high in Hainan and was influenced by genetic factors,pregnancy and delivery process,parents unhealthy habit before and during pregnancy and other factors.

Objective: To evaluate the efficacy and safety of maintenance therapy with reduced dose of rhTPO in the patients with primary immune thrombocytopenia (ITP) who attained stable platelet (PLT) counts after daily administration of rhTPO. Methods: Treatment was started with a daily administration of rhTPO (300 U/kg) for 2 consecutive weeks. Patients who attained stable PLT≥50×10⁹/L were enrolled to maintenance therapy starting with every other day administration of rhTPO, then adjusted dose interval to maintain platelet count (30-100) ×10⁹/L. Results: A total of 91 eligible patients were enrolled. Fourteen patients discontinued the study due to noncompliance (12/14) and investigator decision (2/14) . Among 77 patients who completed the study, 38 patients with the administration of rhTPO at every other day or less could maintain PLT≥30×10⁹/L for 12 weeks. The percentage of patients with a platelet response (PLT≥30×10⁹/L) at 4^th week, 8^th week and 12^th week of maintain therapy was 92.6% (63/68) , 82.7% (43/52) and 85.0% (34/40) , respectively. Median platelet counts remained in the range of (70-124) ×10⁹/L. The overall incidence of rhTPO-related adverse events was 7.7%. All the adverse events were generally mild. Conclusion Extending the dose interval of rhTPO is feasible to maintain stable platelet count in the patients with ITP, but the optimal dose interval is uncertain and might vary with individuals.

Objective To discuss the maximum tolerated dose of oxaliplatin based on 5-fluorouracil derivative in patients with locally advanced rectal cancer who underwent neoadjuvant chemoradiotherapy.Methods From Mar 2015 to Oct 2015,15 locally advanced rectal cancer patients (T3,T4/N +) who received intensity modulated radiotherapy and concurrent chemotherapy with capecitabine and oxaliplatin were enrolled in this study.The prescription dose was 50.6 Gy for gross tumor volume(GTV) and 41.8 Gy for clinical tumor volume(CTV) in 22 fractions within 30 d with concomitant boost.There were four dose-level groups of oxaliplatin as 100,110,120 and 130 mg/m2 tri-weekly and fixed capecitabine dose (825 mg/m2 bid d1-5 per week).The first 12 patients were randomly assigned into 4 groups.For the 130 mg/m3 group,another 3 patients were enrolled because of dose-limiting toxicity (DLT).Treatment related toxicities and response rates were evaluated.Results The most common adverse events(AE) were radiation enteritis,skin reactions,nausea,fatigue,urinary system AE and bone marrow suppression.There was a trend of increase by the dose level of oxaliplatin for toxicities.Groups 100,110 and 120 mg/m2 had none DLT,while group 130 mg/m2 had 1 patient for grade 3 thrombopenia and 1 patient for grade 3 nausea.Postoperative pathology showed that all patients achieved tumor downstaging,among which 0,1,2,3 cases achieved complete remission of the four groups,respectively.Conclusions The combination regimen of capecitabine and oxaliplatin is safe and effective according to the preliminary results.The maximum tolerated dose of oxaliplatin was 130 mg/m2 tri-weekly.