Background Researches showed that CD4+CD25+ natural regulatory T cells (nTregs) play an important role in maintaining peripheral immune tolerance, while immunotherapy using in vitro-expanded induced regulatory T cells (iTregs) suppresses allograft rejection in multiple organ transplantation.The inducing method of iTregs still needs to be optimized.Furthermore,the effect of iTregs on grafts of keratoplasty is unclear.Objective This study was to investigate the inducing and expansion method of iTregs and explore its inhibitory effects on corneal allograft rejection.Methods Bone marrow-derived dendritic cells (BMDCs) were isolated from C57BL/6 mice femora and cultured.CD4+ CD25+ T cells and CD4+ CD25-T cells were isolated from mouse spleen and separated using flow cytometry.The CD4+CD25-T cells were divided into negative control group (CD4+CD25-T cells), CD3/ 28 antibody bead group (CD4+CD25-T cells+CD3/28 antibody bead) ,2.5 ng/ml transforming growth factor (TGF)-β1 induced group and 10.0 ng/ml TGF-β1 induced group.The iTregs was formed after induction of different concentrations of TGF-β1 and CD3/CD28 antibody bead (1 : 1).CD3/CD28 antibody bead (1 : 2) , interleukin-2 (IL-2) and TGF-β1 were used to expand iTregs.The phenotype and proliferation of iTregs were assayed by flow cytometry,and the inhibitory effect of iTregs on effector T cells (Teffs) was analyzed by mixed lymphocyte reaction.Allogenic keratoplasty model (C57BL/6→BALB/c) was build,and 0.1 ml iTregs or nTregs suspension or PBS was injected via posterior venous plexus of fellow eyes to assess the graft survival time.The use and care of the mice followed the ARVO statement.Results The CD4+CD25+ T cell proportions were (6±3)％ ,(91±4)％ ,(91±3)％ and (86± 6) ％ in the negative control group,CD3/CD28 antibody bead group, 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1induced group, showing significant increases in the CD3/CD28 antibody bead group, 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group compared with the negative control group (all at P＜0.01).The Foxp3+ T cell proportions of the CD3/CD28 antibody bead group,2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group were (1.18 ±0.20) ％ , (8.70± 1.80) ％ and (21.80±3.36) ％ , showing significant increases in the 2.5 ng/ml TGF-β1 induced group and 10.0 ng/ml TGF-β1 induced group compared with the CD3/CD28 antibody bead group (both at P＜0.01).Compared with the nTregs, the expression of CD69 was lower, and the expressions of PD-1 and Foxp3 were raised in the iTregs (all at P＜0.01).The proliferation of Teffs were decreased when cocultured with iTregs in comparison with nTregs at 1 : 1,1 : 2,1 : 4,1 : 8,1 : 16 Tregs/Teffs rations (all at P＜ 0.01).The survival time of mouse corneal grafts was 4 weeks with the permanent tolerance of 50％ in the iTregs injected group,which was superior to the 3 weeks survival time and 17％ permanent tolerance in the nTregs injected group(P＜0.05).Conclusions TGF-β1 can induce CD4+ CD25-T cells to form iTregs, which highly express Foxp3.iTregs show a stronger inhibitory effect on the growth of lymphocytes than nTregs, and therefore suppress the graft rejection after keratoplasty.

Objective To study the clinicopathologic features,immunophenotype,diagnosis and differential diagnosis,prognosis and treatment of solid papillary carcinoma(SPC)of breast.Methods A retrospective study was performed to analyse the clinicalpathologic features and immunophenotype of 52 case of SPC,with or without invasion. The cytokeratins(CK),myoepithelial markers,neuroendocrine markers,proliferation marker Ki -67,estrogen receptor (ER),progesterone receptor(PR),HER2,ect were detected by immunohistochemical staining,and HER2 was detected by FISH.All patients were followed up.Results All the patients were females with a mean age of (66.76 ±10.19)years. The incidence of SPC among all the breast cancers in this department was 0.99%.The presenting symptoms was a painless mass with or without nipple discharge in the central of breast.All of them accepted surgical resection,13 cases were administrated with chemotherapy after surgery,6 cases were radiotherapy.Metastasis was observed four in 34 cases who had undergone axillar lymph node dissection.General observation tumor was substantive and nodular lesions,may be accompanied by cyst formation,microscopy tumor for swelling,substantive nodules within only fine fiber vascular axis,and lacked obvious nipples and screen structure.The cells were lack of atypia,adhesive in cells nest,35 patients were accompanied with mucus secretion.Immunohistochemical study show that the tumor cells experssed CK8 /18,ER and PR,but not basal cell cytokertin CK5 /6 and 34βE12.Positivity for CK5 /6,34βE12 and P63 were 5.78%, 9.62% and 7.69% in the myoepithelial layers of axis of papillary,and were 21.15%,38.46% and 19.23% in cell nests and tumor peripheral myoepithelial,respectively.Positivity for CgA,Syn,NSE,CD56 were 82.69%,88.46%, 94.23% and 90.38%,respectively.The staining of HER2 oncoprotein was most negative,and 23 cases of FISH detection were all negative.The average positive index of Ki -67 was 4.28%.47 cases received follow -up of patients(90.38%),only 1 case death,1 case recurrence the rest were no tumor recurrence or metastasis,disease -free survival.Conclusion SPC of breast,a rare pathologic type,usually occurs in old females.The diagnosis depends on pathological characteristics and its unique immune phenotypic characteristics,is closely associated with mucinous carcinoma and neuroendocrine carcinoma,and the treatment is given priority tosurgery with a relatively inert biological behaviour.

Objective To study the role of urinary kidney injury molecule-1 (KIM-1) in pediatric Henoch-Sch?nlein purpura (HSP). Methods Urinary levels of KIM-1 were examined using ELISA in 48 children with HSP including 23 HSPN children (HSPN group) and 25 non-HSPN children (HSP group), and 20 healthy children. The levels of urinary creatinine and 24-hour urine protein were also detected. The results were analyzed and compared among groups. Results The ratio of urinary KIM-1/creatinine (Cr) in HSPN children was signiifcantly higher than that in the other two groups (P<0.05). There was no signiifcant difference in the ratio of urinary KIM-1/Cr between HSP group and the control group (P>0.05). The ratio of urinary KIM-1/Cr had no correlation with 24-hour urine protein in all HSP children (r=0.239, P=0.590). Conclusions Urinary KIM-1 may play a role in the pathogenesis of pediatric HSPN.

Objective To explore the feasibility and safety of Mckeown-type minimally invasive esophagectomy in para-recurrent laryngeal nerve lymphadenectomy. Methods A total of 163 cases underwent minimally invasive McKeown resection for esophageal carcinoma in the Affiliated Tumor Hospital of Xinjiang Medical University were retrospectively analyzed. Patients were divided into routine treatment group (n=63), right recurrent nerve lymph node dissection group (right group, n=53) and bilateral recurrent laryngeal nerve lymph node dissection group (bilateral group, n=47) according to the operation modes. The postoperative pathology, operation time, intraoperative bleeding volume, postoperative hospitalization time, number of lymph nodes and pneumonia, anastomotic fistula, laryngeal nerve injury and other complications were compared between three groups of patients. Pathological conditions were consisted of the location of tumor, the degree of differentiation, T stage and pathological stage. Postoperative complications were followed up for 6 months. Results There were no significant differences in tumor location, pathological differentiation degree, T stage and pathologic stage between three groups. The amount of bleeding was more in the routine group than that of the right group and the bilateral group (P<0.05). There was no significant difference in the amount of bleeding between the right group and the bilateral group. The total lymph nodes and thoracic lymph nodes were increased in order in routine group, the right group and the bilateral group, and there was significant difference between three groups(P<0.05). There were no significant differences in other operation data and complications between three groups. Conclusion McKeown-type minimally invasive esophagectomy shows good feasibility and safety for para-recurrent laryngeal nerve lymphadenectomy .

Objective To study the clinicopathologic features,immunophenotype,diagnosis and differential diagnosis of solid papillary carcinoma (SPC) of breast.Methods A retrospective study was performed to analyse the clinicopathologic features and immunophenotype of 31 cases of SPC with or without invasion.The cytokeratins (CK),myoepithelial markers,neuroendocrine markers,proliferation marker Ki-67,estrogen receptor (ER),progesterone receptor (PR),HER2,ect were detected by immunohistochemical staining,and HER2 was detected by FISH.All patients were followed up.Results All the patients were females with a mean age of (67.0±12.5) years.The presenting symptom was a painless mass with or without nipple discharge.In 19 cases who had undergone axillar lymph node dissection,only one metastasis was observed.General observation showed that tumors had substantive and nodular lesions,accompanied by cyst formation.Microscopy examination showed that tumors had swell and substantive nodules with only fine fiber vascular axis,and lacked obvious nipples and screen structure.The cells were lack of atypia,adhesive in cells nest.20 patients were accompanied by mucus secretion.Immunohistochemical study showed that the tumor cells expressed CK8/18,ER,PR,but didn' t express basal cell CK5/6,34β12.The positive rates of CK5/6,34β12,p63 were 3.23 ％ (1/31),9.68 ％ (3/31),6.45 ％ (2/31) in the myoepithelial layers of axis of papillary,and were 19.25 ％ (6/31),38.71％ (12/31),16.13 ％ (5/31) in peripheral cell nests and tumors,respectively.The positive rates of CgA,Syn,NSE,Cd56 were 83.87 ％ (26/31),87.10 ％ (27/31),93.55 ％ (29/31),90.32 ％ (28/31),respectively.The staining of HER2 oncoprotein was negative mostly,and the FISH detection outcome of 15 cases were all negative.The average positive index of Ki-67 was 4.12 ％.28 cases (90.23 ％) received follow-up,only 1 patient died,and the rest patients were without tumor recurrence or metastasis.Conclusions SPC of breast,a rare pathologic type,usually occurs in old females.The diagnosis depends on pathological characteristics and its unique immune phenotypic characteristics.SPC of breast has close relationship with mucinous carcinoma and neuroendocrine carcinoma,with a relatively inert biological behavior.

Objective To detect the expression of Livin,an apoptosis-inhibiting protein,and Smac,an apoptosis-promoting protein,in basal cell carcinoma (BCC) lesions.Methods Skin specimens were obtained from the lesions of 80 patients with BCC and normal skin of 30 human controls.Immunohistochemical SP method was used to detect the pmtein expression of Livin and Smac in these specimens.Chi-square test was conducted to compare the expression rate of Livin and Smac protein between the lesional and control specimens.The relationship between the protein expression of Livin and Smac in BCC was analyzed by Spearman correlation coefficients.Results The expression rate of Livin protein was significandy higher (77.50％ vs.3.33％,x2 =49.04,P ＜ 0.001),while that of Smac protein was statistically lower (46.25％ vs.100％,x2 =26.47,P ＜ 0.001),in BCC than in the control specimens.No significant difference was observed in the expression rate of Livin or Smac protein between nodular ulcerative and pigmented BCC specimens (75.41％ vs.80.00％,x2 =0.001,P ＞ 0.05; 47.54％ vs.40.00％,x2 =0.28,P＞ 0.05) or between nodulocystic and pigmented BCC specimens (73.58％ vs.80.00％,x2 =0.03,P ＞ 0.05; 45.28％ vs.40.00％,x2 =0.13,P ＞ 0.05).There was a negative relationship between the protein expression of Livin and Smac in BCC lesions (r =-0.432,P ＜ 0.01).Conclusion The upregulated expression of Livin and downregulated expression of Smac may be invoved in the occurrence and development of BCC.

Objective To explore the assessment system of humanistic communication in the grad-uation examination of pediatric residents. Methods A humanistic communication assessment program and an evaluation form were designed and put into practice in the graduation examination of pediatric residents in 2016 and 2017 in Beijing Children's Hospital affiliated to Capital Medical University and the scores were collected and analyzed to compare the differences in pediatric residents who have undergone system-atic physician-patient communication scene training and who didn't. With this analysis, the assessment system was further improved. Results Of all the 5 dimensions, communication content, nonverbal skills, and communication effects and the comparison of their respective scores were statistically significant. The scores of the residents who have undergone systematic physician-patient communication scene training are much better than those who didn't. Conclusion This assessment system focuses on the investigation of ability. From the design of examination questions and assessment scenes, to the evaluation of students per- formances, only when candidates have mastered the skills of communication can they do well in examina-tions and achieve good results.

Gene co-expression network (GCN) mining identifies gene modules with highly correlated expression profiles across samples/conditions. It enables researchers to discover latent gene/molecule interactions, identify novel gene functions, and extract molecular features from certain disease/condition groups, thus helping to identify disease bio-markers. However, there lacks an easy-to-use tool package for users to mine GCN modules that are relatively small in size with tightly connected genes that can be convenient for downstream gene set enrichment analysis, as well as modules that may share common members. To address this need, we developed an online GCN mining tool package:TSUNAMI (Tools SUite for Network Analysis and MIning). TSUNAMI incorporates our state-of-the-art lmQCM algorithm to mine GCN modules for both public and user-input data (microarray, RNA-seq, or any other numerical omics data), and then performs downstream gene set enrichment analysis for the identified modules. It has several features and advantages:1) a user-friendly interface and real-time co-expression network mining through a web server;2) direct access and search of NCBI Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases, as well as user-input gene ex-pression matrices for GCN module mining; 3) multiple co-expression analysis tools to choose from, all of which are highly flexible in regards to parameter selection options;4) identified GCN modules are summarized to eigengenes, which are convenient for users to check their correlation with other clinical traits;5) integrated downstream Enrichr enrichment analysis and links to other gene set enrichment tools;and 6) visualization of gene loci by Circos plot in any step of the process. The web service is freely accessible through URL:https://biolearns.medicine.iu.edu/. Source code is available at https://github.com/huangzhii/TSUNAMI/.

Objective To evaluate the expression differences of C-reactive protein(CRP),procalcitonin(PCT)and cytokines[interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),interleukin(IL)-10,IL-6,IL-4,IL-2,IL-17 A,and IFN-γ/IL-10]in human immunodeficiency virus(HIV)infection,tuberculosis(TB)infec-tion and HIV/TB co-infection in order to provide evidence for early clinical diagnosis of HIV/TB co-infection.Methods Medical records and clinical information of 103 patients with HIV(HIV group),42 patients with TB(TB group)and 67 patients with HIV/TB co-infection(HIV/TB group)were collected from January 2021 to September 2022.CRP,PCT and cytokines were detected in all patients,and their expression differ-ences among the three groups were analyzed.Receiver operating characteristic(ROC)curves were drawn and screening ability was evaluated.Results Compared with HIV group and TB group,CRP,PCT,IFN-γ,IL-10,IL-6 levels and IFN-γ/IL-10 in HIV/TB group were increased,with statistical significance(P＜0.001).Bon-ferroni correction results showed that CRP,PCT,IFN-γ,IL-6 levels and IFN-γ/IL-10 were significantly differ-ent between HIV and HIV/TB groups(all P＜0.001).ROC curve analysis showed that the area under the curve of CRP,PCT,IFN-γ,IL-6 and IFN-γ/IL-10 combined detection was 0.826,which was higher than that of single detection.Conclusion CRP,PCT,IFN-γ,IL-6,IFN-γ/IL-10 combined detection can help the early screening of HIV infection and HIV/TB co-infection.

OBJECTIVE: To establish a method for detecting triphenyl phosphate( TPP) in the workplace air by gas chromatography. METHODS: TPP in the air of workplace was collected with glass fiber filter paper,desorbed with ether,separated by HP-5 gas chromatographic column,and detected by flame photometric detector. RESULTS: This method has good linear range of 12. 50-800. 00 mg/L,with the correlation coefficient of 0. 999 9. The detection limit was 0. 78 mg/L,and the minimum detectable concentration was 0. 09 mg/m3(sample volume was 45 L). Desorption efficiency was 97. 2%-99. 4%; standard recovery rate was 99. 5%-100. 3%. The within-run relative standard deviation( RSD) was 2. 7%-3. 4%and the between-run RSD was 1. 4%-3. 2%. The sampling efficiency was 99. 6%-100. 0%. The samples could be stored at room temperature for at least 14 days. CONCLUSION: The method is simple,accurate and highly sensitive for detecting TPP in workplace air.