Background: The aim of this study is to investigate the specific pathway involved in human leukocyte antigen (HLA) sensitization using single-cell RNA-sequencing analysis and an allo-sensitized mouse model developed with an HLA.A2 transgenic mouse. Methods: For sensitization, wild-type C57BL/6 mouse received two skin grafts from C57BL/6-Tg(HLA-A2.1)1Enge/J mouse (allogeneic mouse, ALLO). For syngeneic control (SYN), skin grafts were transferred from C57BL/6 to C57BL/6. We performed single-cell RNA-sequencing analysis on splenocytes isolated from ALLO and SYN and compared the gene expression between them. Results: We generated 9,190 and 8,890 single-cell transcriptomes from ALLO and SYN, respectively. Five major cell types (B cells, T cells, natural killer cells, macrophages, and neutrophils) and their transcriptome data were annotated according to the representative differentially expressed genes of each cell cluster. The percentage of B cells was higher in ALLO than it was in SYN. Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated that the highly expressed genes in the B cells from ALLO were mainly associated with antigen processing and presentation pathways, allograft rejection, and the Th17 cell differentiation pathway. Upregulated genes in the T cells of ALLO were involved in the interleukin (IL)-17 signaling pathway. The ratio of Th17 cluster and Treg cluster was increased in the ALLO. On flow cytometry, the percentage of Th17 (IL-17+/CD4+ T) cells was higher and regulatory T cells (FOXP3+/CD4+ T) was lower in the ALLO compared to those in the SYN. Conclusion Our results indicate that not only the B cell lineage but also the Th17 cells and their cytokine (IL-17) are involved in the sensitization to HLA.

Objective:To summarize the effect of Chinese Children′s Cancer Group (CCCG) Wilms tumor (WT)-2015 protocol.Methods:This was a prospective study. CCCG-WT-2015 protocol was revised on the basis of the CCCG-WT-2009 protocol. Clinical data of 288 children diagnosed with newly diagnosed kidney neoplasms in fourteen pediatric centers between September 2015 to December 2018 were summarized. The age of onset, distribution of pathological subtypes, staging, curative effect and prognostic factors of these children were analyzed. Kaplan-Meier method was used for survival curve and Log-Rank method was used for univariate analysis.Results:Among 288 cases with kidney neoplasms, there were 261 cases of WT, including 254 cases (97.3%) with favorable histology (FH) WT and 7 cases (2.7%) with unfavorable histology WT (UFHWT). The 3 year events free survival (EFS) rate for FHWT and UFHWT were (88.9±2.1)% and (80.0±17.9)%, which were better than that in WT-2009 (81.2% and 71.7%). In the 96 cases of stage Ⅲ/Ⅳ FHWT with indications for radiotherapy, 76 cases received radiation, another 20 cases received M protocol chemotherapy (cyclophosphamide, etoposide, gentamycin, vincristine and adriamycin) instead of radiation. The 3 year EFS rate for these two groups were (84.7±4.3)% and (84.7±8.1)%(χ 2=0.015, P=0.902). There were 22 renal clear cell sarcoma and 5 malignant rhabdoid tumor, 3 year EFS rate of them was (94.4±5.4)% and (20.0±17.9)%. Univariate analysis was performed for age, gender, pathological type, stage, whether rupture occurred during operation, whether complete remission (CR) occurred at the end of treatment and radiotherapy. Pathological types (χ 2=44.329, P<0.01) and failure to achieve CR at the end of the treatment (χ 2=49.459, P<0.01) were independent factor for predicting survival. Conclusion:Compared with CCCG-WT-2009, treatment of renal tumors in CCCG-WT-2015 study yielded good survival outcome, which can be further applied.

ObjectiveTo assess the effects of constraint-induced movement therapy(CIMT) on sub-acute stroke patients with upper extremity motor dysfunction.Methods63 patients with hemiplegia after stroke were divided into two groups: control group(31 cases) and CIMT group(32 casese). The two groups received the regular rehabilitation training for 14 d. Then the control group went on the the regular rehabilitation training, while the CIMT group received CIMT for 14 d. All patients were assessed by the Action Research Arm Test(ARAT) and Fugl-Meyer(FMA) on the first day, the 15th day and the 30th day after the treatment.ResultsARAT score and FMA scores were higher in CIMT group than in the control group 30 d after the treatment and in CIMT group 15 days after the treatment(P<0.001).ConclusionCIMT is more effective to improve the upper limb motor function of the sub-acute stroke patients than the regular rehabilitation training.

Objective To assess the prognosis and effect on renal function of pediatric urolithiasis caused by melamine-contaminated milk powder (PUMMP) in a long-term follow-up.Methods One hundred and two of 8335 children (≤ 6-year-old) with history of consuming melamine-contaminated milk powder screened in our hospital were followed up for eighteen months after diagnosis. Urinary system ultrasonography, urinalysis, urinary microprotein profiles [microalbumin (ALBU), immunoglobulin G (IgG), and N-acetyl-β-D-glucosidase (NAG)], urinary melamine and cyanuric acid were examined in the first visit and at the end of follow-up. Results Follow-up was completed in 91 children and the stone was excreted in 82 children (90.1%).Stones less than 5 mm in diameter were most vulnerable to discharge, and stones larger than 10 mm could not be expelled without interventions. At the end of follow-up, no melamine or cyanuric acid was found in the urine samples of 74 patients. Urinalysis showed that incidences of proteinuria, microscopic hematuria and leukocyturia were 0%, 5.1% and 2.0%, which were significant different from those in the first visit (Pproteinutria=0.123, Phemnatuna=0.038 and Pleukocyhuris=0.005).Urinary microprotein profiles revealed that some children whose urinalysis was normal still presented glomerular and renal tubular injury and the abnormal rates were 8.8% and 12.1%respectively. The glomerular injury was mainly related to persistent stone, male and younger.Conclusions 90.1% of children with PUMMP passes urinary stones at the end of follow-up.Stone size is the major risk factor of discharge. No melamine or cyanuric acid is found in the urine of children. After eighteen months, glomerular and renal tubular injury is still found in some patients. Further follow-up is necessary.

Objective To investigate the expression of monocyte chemoattractant protein-1(MCP-1)and CCR2 in peripheral blood and pleural fluid of tuberculous pleurisy patients.Methods Flow cytometry was used to detect the expression of CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients;ELISA was used to detect the content of MCP-1 in serum and pleural fluid.Results MCP-1 in surem and pleural fluid and CCR2 in the peripheral blood mononuclear cell and pleural fluid cell of tuberculous pleurisy patients was significantly higher than that of the healthy normal controls[MCP-1:(340.8±220.8)and(9.0±3.8)ng/L,P＜0.01;CCR2(18.2±10.1)%and(6.9±3.5)%,P＜0.05];Both MCP-1 and CCR2 were detected in pleural fluid and both of them were positivley correlated(r=0.227,P＜0.05).Conclusion The expression of MCP-1 and CCR2 in peripheral blood of tuberculous pleurisy patient are significantly elevated which are significance molecule participating in the pathogenesis of tuberculous pleurisy.

Objective To evaluate the feasibility and the clinical value of MRI in the diagnosis of small-bowel disease. Methods Sixty-three patients with suspected small-bowel diseases and 3 volunteers without signs of small bowel disease underwent MRI examination. Thirty-one patients whose diagnoses were confirmed by pathology or clinical results were categorized into two groups (neoplastic and normeoplastic). The conspicuity of bowel wall, the sensitivity of MRI in detecting small-bowel lesions, and the accuracy rate of diagnosis were calculated. The average bowel wall thickness between the two groups was assessed by using Wilcoxon signed-rank test. Enlarged mesenteric lymph nodes, mesenteric infiltration, and small-bowel stenosis were analyzed by using Fisher's exact test in each group respectively. Results MRI examinations of all 66 subjects were successfully performed. Images were rated on a continuous 4-peint scale. Sixty-two cases (93.9%) were scored as 2 or 3. The diagnoses of 31 patients (neoplastic group (n = 10) and nonneoplastic group (n = 21) were confirmed by pathology or clinical results. The sensitivity, accuracy of MRI in identifying small bowel diseases were 100% (31/31) and 77.4% (24/31) respectively. The average bowel wall thickness of the two groups was 23 mm(7.0-65.0 mm) and 5 mm(2.0-35.0 mm) respectively, and there was a statistically significant difference between the two groups (Z = - 2.949, P < 0.01). Enlarged lymph nodes in mesentery were found in 7 cases in neoplastic group and 4 cases in nonneoplastic group, and there was a statistically significant difference between the two group (P < 0.05). Small-bowel stenosis was depicted in 10 cases in both groups and there was a statistically significant difference between the two groups (P <0.01). The mesenteric infiltration sign was seen in 5 cases and 17 cases respectively, and showed no significant difference between the two groups (P > 0.05). Conclusion MRI can depict the location and extension of the small-bowel disease accurately and it is an effective method in the diagnosis of small-bowel disease.