BACKGROUND: Long-term remote ischemic conditioning (RIC) has been proven to be beneficial in multiple diseases, such as cerebral and cardiovascular diseases. However, the hyperacute and acute effects of a single RIC stimulus are still not clear. Quantitative proteomic analyses of plasma proteins following RIC application have been conducted in preclinical and clinical studies but exhibit high heterogeneity in results due to wide variations in experimental setups and sampling procedures. Hence, this study aimed to explore the immediate effects of RIC on plasma proteome in healthy young adults to exclude confounding factors of disease entity, such as medications and gender. METHODS: Young healthy male participants were enrolled after a systematic physical examination and 6-month lifestyle observation. Individual RIC sessions included five cycles of alternative ischemia and reperfusion, each lasting for 5 min in bilateral forearms. Blood samples were collected at baseline, 5 min after RIC, and 2 h after RIC, and then samples were processed for proteomic analysis using liquid chromatography-tandem mass spectrometry method. RESULTS: Proteins related to lipid metabolism (e.g., Apolipoprotein F), coagulation factors (hepatocyte growth factor activator preproprotein), members of complement cascades (mannan-binding lectin serine protease 1 isoform 2 precursor), and inflammatory responses (carboxypeptidase N catalytic chain precursor) were differentially altered at their serum levels following the RIC intervention. The most enriched pathways were protein glycosylation and complement/coagulation cascades. CONCLUSIONS One-time RIC stimulus may induce instant cellular responses like anti-inflammation, coagulation, and fibrinolysis balancing, and lipid metabolism regulation which are protective in different perspectives. Protective effects of single RIC in hyperacute and acute phases may be exploited in clinical emergency settings due to apparently beneficial alterations in plasma proteome profile. Furthermore, the beneficial effects of long-term (repeated) RIC interventions in preventing chronic cardiovascular diseases among general populations can also be expected based on our study findings.
Young Adult ; Humans ; Male ; Proteome ; Cardiovascular Diseases ; Proteomics ; Ischemia ; Blood Coagulation

No case of moyamoya syndrome with bilateral posterior cerebral artery (PCA) occlusion has been reported in China so far as this disease is extremely rare. The case shown in this article is a middle-aged women who has a history of atrial fibrillation, hypertension and type 2 diabetes acutely attacked by this syndrome. The main clinical manifestations included binocular blindness, right limb weakness. Imaging findings showed bilateral acute cerebral infarction in the parietal occipital lobe, bilateral anterior cerebral artery and middle cerebral artery smoke angiogenesis, bilateral PCA occlusion with distal smoke angiogenesis. Considering the medical history of the patient, the cause of the disease was diagnosed as embolic stroke of undetermined source. The patient′s consciousness has been recovered and the limb weakness has been improved after active symptomatic treatment. However, the blindness did not see any improvements. This case report aims to improve clinicians′ understanding of bilateral PCA embolization in patients with moyamoya syndrome so the occurrence of cerebral infarction can be effectively prevented.

【Objective】 To analyze the yield, specificity and detection time of red blood cell(RBC)alloimmunization in 104 588 inpatients. 【Methods】 The clinical information of patients who underwent at least one antibody screening in our hospital from November 2017 to December 2019 was retrospectively analyzed. The demographic characteristics, transfusion history, pregnancy history and antibody screening results of patients were collected. The RBC alloantibody yield, specificity and detection time were analyzed, and differences of transfusion units and frequency between patients with and without alloimmunization were compared. 【Results】 Eight hundred cases of alloantibodies with clinical significance were detected in blood samples of 723 patients, with a positive rate of 0.7% (723/104 588). The incidence rate of alloimmunization in females was higher than that in males (0.9% vs 0.5%, P<0.05). Rh alloantibodies accounted for 76.4%(611/800), of which 61.4%(375/611)were anti-E. Transfusion units and frequency of patients with alloimmunity were higher than those without(median: 6.0 vs 4.0, P<0.05; 4.0 vs 2.0, P<0.05, respectively). And 67.5% of RBC alloantibodies were detected within 6 months, with the median (IQR) detection time of 97.0 (22.5-247.0) days. 【Conclusion】 Routine antibody screening should be performed before transfusion in order to reduce the occurrence of adverse reactions, and Rh typing transfusion with compatible crossmatch should be performed if necessary.

Objective To investigate the expression level of soluble Programmed cell death ligand2 (sPD-L2) in the peripheral blood of patients with Brucella infection and their correlation with helper T cells (Th) subsets. Methods Data from eighty-three patients with confirmed Brucella infection, including 37 patients with acute phase Brucella infection and 46 patients with chronic phase Brucella infection, collected from the People's Hospital of Xinjiang Uygur Autonomous Region between January 2019 and December 2021, were analyzed. Fifty healthy individuals undergoing regular medical check-ups during the same period were included as the Healthy Control (HC) group. Flow cytometry was used to detect the percentage of Th1 and Th2 cells as CD4+ T cells in the peripheral blood of the three groups of samples. Quantitative real-time fluorescence PCR (qRT-PCR) was performed to measure the mRNA expression levels of Th1 and Th2 cell-associated transcription factors T-bet and GATA-3 in the peripheral blood of the three samples. The expression levels of cytokines sPD-L2, IFN-γ, and IL-4 in the serum of the three groups of samples were measured by the cytometric bead array (CBA) technique. Results Th1 cells in the acute phase brucellosis infection group (45.33±4.96)%, mRNA expression levels of T-bet (1.91±0.41), and IFN-γ expression levels (74.42±13.95) pg/mL were significantly higher in the acute phase Brucella disease group compared to the chronic phase group (21.78±4.42)%, (0.65±0.24), and (10.64±3.04) pg/mL, as well as the HC group (28.87±6.67)%, (1.12±0.25), and (7.48±2.92) pg/mL, respectively (P<0.05). Th2 cells (59.80±10.09)%, GATA-3 mRNA expression level (1.50±0.44), and IL-4 level (8.76±2.06) pg/mL were significantly higher in the chronic phase brucellosis infection group compared to the acute phase brucellosis patient group (40.61±9.32)%, (1.02±0.23), and (2.08±0.50) pg/mL, as well as the HC group (46.06±8.84)%, (1.18±0.28), and (2.70±0.75) pg/mL, respectively (P<0.05). Compared to the HC group (91.61±11.44) pg/mL, the sPD-L2 levels were significantly higher in the acute phase brucellosis patient group (156.77±24.69) pg/mL and slightly higher in the chronic phase brucellosis patient group (110.99±29.44) pg/mL, with a statistically significant difference between the acute and chronic phase brucellosis patient groups (P<0.05). Correlation analysis of the brucellosis patient group showed that sPD-L2 levels were positively correlated with the percentage of Th1 cells (r=0.540, P<0.01) and the levels of IFN-γ (r=0.692, P<0.01), and negatively correlated with the percentage of Th2 cells (r=-0.565, P<0.01) and the levels of IL-4 (r=-0.528, P<0.01). Conclusions In patients with acute brucellosis, sPD-L2 levels are significantly increased, which may resist brucellosis by promoting Th1 cells to secrete more pro-inflammatory factors to defend against Brucella infection. In chronic Brucella disease patients, the tilting of immune response types from Th1 to Th2 may be due to a decrease in sPD-L2 expression, which reduces the inhibitory effect of Th2, leading to incomplete clearance of Brucella and thus immune escape. Brucella may affect the conversion of Th1 to Th2 through the sPD-L2 pathway, exerting negative regulatory effects and resulting in difficulty in complete Brucella clearance, thus turning into a chronic phase.

Objective:To explore the effects of foraging exercise (FE) on depressive-like behaviors and expression of transforming growth factor-β1 (TGF-β1) in hippocampus of rats with ischemic stroke after chronic stress.Methods:The right middle cerebral artery occlusion (MCAO) model was used in 30 male adult clean grade SD rats by suture method.According to the body weight, rats were evenly divided into stroke group ( n=10) and chronic unpredictable mild stimulation (CUMS) group ( n=20). Rats of CUMS group received stress induction 1 week after operation and lasted for 3 weeks. Then, according to random number generator of SPSS 24.0 software, the depression rats were divided into post-stroke depression (PSD) group( n=10) and FE groups ( n=10). The FE group received free FE intervention for 4 weeks. Body weight, water maze test, novelty inhibition feeding test (NSFT) and sucrose preference test (SPT) were performed at the end of the 1st, 4th and 8th week, respectively. The expression of TGF-β1 in hippocampus was detected by Immunohistochemistry (IHC) and Western blot (WB), and the levels of TGF-β1 and TNF-α in serum were detected by ELISA. SPSS 24.0 software was used for statistical analysis. The behavioral data were compared by two factor repeated measurement analysis of variance. One way ANOVA was used for comparison among groups, and LSD test was used for further pairwise comparison. Results:(1) The interaction between group and time had statistical significance on body weight, latency and food intake of NSFT and sucrose preference index(SPI) ( F=2.936-12.098, all P<0.05). After 4 weeks, compared with the stroke group((343.80±19.34)g, (12.10±6.97)s, (0.75±0.09)%), the body weight((307.80±17.23)g, (305.30±24.39)g), and SPI((0.52±0.06)%, (0.53±0.07)%) of PSD group and FE group were lower and the NSFT latency((21.70±7.02)s, (22.40±0.84)s) was longer (all P<0.05). After 8 weeks, SPI in FE group was higher than that in PSD group ( P=0.045). There were significant differences in body weight of three groups, NSFT latency and SPI of PSD group and FE group, and food intake of stroke and FE group ( F=8.478-196.548, all P<0.05). There was no interaction between group and time in the water maze test. Main effect of time ( P=0.034) and main effect of group ( P<0.01) had statistical significance on escape latency. The escape latency after 4 weeks was longer than that after 1 week ( P=0.003). The latency of PSD group was longer than that of stroke group ( P=0.005), and latency of FE group was shorter than that of the PSD group ( P<0.01). The main effect of group had statistical significance in the number of crossing quadrant ( P<0.01). The number of crossing quadrant of FE group was less than that of PSD group ( P<0.01). (2) Immunohistoche mistry staining showed that compared with the stroke group, the expression of TGF-β1 was down-regulated in 3 areas of hippocampus of PSD group (CA1, CA3 and DG) ( t=5.449-9.353, all P<0.01). Compared with stroke group, the expression of TGF-β1 of CA1 ( t=7.433, P<0.01) in FE group was down-regulated, but was up-regulated in CA3 ( t=3.342, P<0.05) of FE group. Compared with the PSD group, the expression of TGF-β1 was up-regulated in CA3 and DG of FE group ( t=7.811, 8.790, both P<0.01). (3) Western blot results: Compared with stroke group, the expression of TGF-β1 in hippocampus of PSD group was down-regulated ( t=3.255, P<0.01). Compared with the PSD group, the expression of TGF-β1 in hippocampus of FE group was up-regulated ( t=2.906, P<0.05). (4) ELISA detection showed that compared with the stroke group, the levels of TGF-β1 decreased ( t=2.224, P<0.05), but TNF-α increased ( t=6.127, P<0.01) in PSD group.Compared with the PSD group, the expression of TGF-β1 in FE group increased significantly ( t=4.417, P<0.01). Conclusion:Foraging exercise can improve the depressive behavior symptoms of ischemic stroke rats after chronic stress, and its mechanism may be related to the increasing expression of TGF-β1, which can alleviate the inflammatory reaction in hippocampus.

Objective:To develop an online interactive cytopathological training program, and to evaluate it for improving the cytopathological diagnostic ability of endoscopists in endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) of pancreas.Methods:A total of 5 500 cytopathological images were collected from 194 patients with pancreatic solid mass who underwent EUS-FNA in Nanjing Drum Tower Hospital from August 2018 to August 2019. The cell type in each cytopathological picture was labeled by senior cellular pathologists, which was used to build a learning and testing platform for online interactive cytopathological training. Five endoscopists without cytopathological background were invited to participate in this training. Sensitivity, specificity, positive predictive value and negative predictive value of endoscopists in differential diagnosis of cancer and non-cancer before and after training were compared to evaluate the effect of the online interactive cytopathological training program on improving the ability of endoscopists in diagnosis of cytopathology.Results:A cytopathological training platform for endoscopists to learn and take online test was successfully built. Before training, sensitivity, specificity, positive predictive value, negative predictive value and accuracy of diagnosis of cancer and non-cancer for endoscopists were 0.55 (95% CI: 0.53-0.58), 0.32 (95% CI: 0.30-0.35), 0.43 (95% CI: 0.41-0.45), 0.44 (95% CI: 0.41-0.47) and 0.43 (95% CI: 0.42-0.45), respectively. After training, the above indicators were 0.96 (95% CI: 0.95-0.97), 0.70 (95% CI: 0.68-0.73), 0.74 (95% CI: 0.72-0.76), 0.95 (95% CI: 0.94-0.96) and 0.81 (95% CI: 0.80-0.83), respectively, which were significantly improved compared with those before ( P<0.001). Conclusion:The online interactive cytopathological training program can improve the understanding and diagnostic ability of endoscopists in pancreatic cytopathology, help to implement rapid on-site evaluation in the process of EUS-FNA, and improve the diagnostic efficiency of EUS-FNA.

Objective　To explore the relevant clinical phenotype of different diseases relating to hot cross bun (HCB) sign and to determine the correlation between neuroimaging and clinical features.Methods　Data of one patient with HCB syndrome in the Department of Neurology of our hospital were collected.The published cases of HCB syndrome were systematically reviewed.According to the etiology,14 cases were analyzed.The clinical symptoms,Magnetic resonance imaging (MRI) features,Laboratory examination treatment and prognosis and so forth were retrospectively analyzed.Results　Published cases through the system references,classify disease and its clinical heterogeneity were reviewed.We found that HCB sign associated with 9 kinds of disease,including nutrition metabolic disease,inflammatory demyelinating disease,nervous system degenerative disease,secondary to suspected vasculitis Parkinson’s syndrome,tumor paraneoplastic disease testicular cancer is caused by the death of infectious diseases,genetic correlation disease related diseases,stroke and poisoning.Conclusion　This study shows that the basic disease spectrum of HCB syndrome may be more extensive than before.According to different clinical phenotypes,imaging features and basic diseases,the occurrence of HCB may be involved in different pathogenesis.

Chimeric antigen receptor T (CAR-T) cell has achieved excellent efficacy in hematological tumors, especially for lymphoma. Many products have been approved to market all over the world, and 2 products targeting CD19 have been approved to treat relapsed and refractory large B-cell lymphoma in China. The current experiences of using CAR-T cells come from previous clinical studies. How to use CAR-T cells in a standardized and rationalized way is still a challenge faced by our clinicians. Based on the CAR-T cell treatment experiences from Peking University Cancer Hospital and the latest research progresses in CAR-T in China and abroad, this article will elaborate on patient screening, peripheral blood mononuclear cell collection, bridging treatment, lymphocyte depletion chemotherapy, CAR-T cell infusion, the monitoring and treatment of adverse events after infusion, and long-term follow-up after infusion, in order to guide clinicians to better use CAR-T cell and to bring maximum benefits to patients.

Objective:To investigate the risk factors for hemorrhagic transformation (HT) in patients with acute posterior circulation ischemic stroke (PCIS) and its impact on outcomes.Methods:From July 2016 to October 2019, patients admitted to the Department of Neurology, the People's Hospital of Zhengzhou and diagnosed as PCIS were enrolled retrospectively. Their demography, clinical data, laboratory and imaging findings were collected. HT was defined as no intracranial hemorrhage detected by the first head CT/MRI after onset, and intracranial hemorrhage was found during head CT/MRI reexamination within 10 d after onset. Symptomatic HT was defined as intracranial hemorrhage indicated by imaging reexamination and the National Institutes of Health Stroke Scale (NIHSS) score was higher than the baseline. The outcome was evaluated by the modified Rankin Scale at 3 months after onset, and >2 were defined as poor outcome. Multivariate logistic regression analysis was used to identify the independent risk factors for HT, symptomatic HT, and poor outcomes. Results:A total of 242 patients with PCIS were enrolled. Their age was 68.02±12.0 years, and 111 were females (45.9%). The baseline median NIHSS score was 5.9 (interquartile range: 3.1-8.8). HT occurred in 19 patients (7.9%), and 14 of them (73.7%) were symptomatic HT. Follow-up at 3 months showed that 74 patients (30.58%) had poor outcomes, of which 12 died. Multivariate logistic regression analysis showed that higher baseline systolic blood pressure (odds ratio [ OR] 1.076, 95% confidence interval [ CI] 1.021-1.135, P=0.006; OR 1.161, 95% CI 1.087-1.240, P<0.001) and larger infarct volume ( OR 31.293, 95% CI 4.542-215.592, P<0.001; OR 2.084, 95% CI 1.414-3.073, P<0.001) were the independent risk factors for HT and symptomatic HT. The higher NIHSS score ( OR 1.511, 95% CI 1.307-1.746; P<0.001), diabetes mellitus ( OR 2.041, 95% CI 1.054-3.952; P=0.034) and symptomatic HT ( OR 4.514, 95% CI 1.458-13.979; P=0.009) were the independent risk factors for poor outcomes. Conclusions:HT is rare in patients with PCIS. Higher baseline systolic blood pressure and larger infarct volume are the independent risk factors for HT in patients with PCIS. Higher baseline NIHSS scores, diabetes mellitus, and symptomatic HT are the independent risk factors for poor outcomes in patients with PCIS.

Objective:To investigate the intra-observer reproducibility of Ki-67　　　assessment in breast cancers using three methods based on digital slide.Methods:Thirty cases of invasive breast cancer tissues were immunostained for Ki-67 by automatic stainer, and then scanned into digital pathological slides. Ki-67 positive index was measured individually by three pathologists using size-set visual assessment of hot spot (SSVAHS), size-set semi-automatic counting of hot spot(SSSACHS), and size-set automatic counting of hot spot (SSACHS), respectively, and repeated for 10 times. Intraclass correlation coefficient (ICC) of each assessment method was calculated, and the intraobserver reliability was classified as excellent, good, fair and poor according to ICC.Results:The ICC by 3 pathologists using SSVAHS was 0.832, 0.843 and 0.826, respectively, The ICC using SSSACHS was 0.926,0.938,0.929, and the ICC using SSACHS was 0.964, 0.971 and 0.968.The intraobserver reliability level of all three methods was excellent.Conclusion:The three methods of Ki-67 assessment achieve satisfactory intraobserver reproducibility, and the order of reproducibility from high to low is SSACHS, SSSACHS, and SSVAHS.