Condensation of acrylonitrile and aryl acetonitrile: construction of α-amino-ß-cyano cyclohexene skeletons.

A representative condensation of acrylonitrile and aryl acetonitrile has been reported for the synthesis of α-amino-ß-cyano cyclohexene. The reaction was carried out mildly in an open environment at room temperature. The scope and versatility of the method have been demonstrated with 20 examples, containing highly active ethynyl groups. Further applications for 4-aminopyrimidine compounds were performed. A mechanism was proposed, involving Michael additions between acrylonitrile and aryl acetonitriles as well as intramolecular condensation.

Study on mechanisms of Th17/Treg imbalance in patients with cystic echinococcosis based on miRNA expression profiles / 中国血吸虫病防治杂志

OBJECTIVE@#To investigate the serum microRNA (miRNA) expression and examine the impact of miRNA expression profiles on T helper type 17 (Th17)/regulatory T cells (Treg) imbalance among patients with cystic echinococcosis, so as to provide insights into the illustration of the mechanisms underlying chronic Echinococcus granulosus infections, and long-term pathogenesis.@*METHODS@#Total RNA was extracted from the sera of cystic echinococcosis patients and healthy controls, and subjected to high-throughput sequencing with the Illumina sequencing platform. Known miRNAs were annotated and new miRNAs were predicted using the miRBase database and the miRDeep2 tool, and differentially expressed miRNAs were identified. The target genes of differentially expressed miRNAs were predicted using the software miRanda and TargetScan, and the intersection was selected for Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. Among the differentially expressed miRNAs with the 20 highest fold changes, miRNAs that targeted genes relating to key transcription factors RORC and FOXP3 that determine the production of Th17 and Treg cells or their important regulatory pathways (PI3K-Akt and mTOR pathways) were matched.@*RESULTS@#A total of 53 differentially expressed miRNAs were screened in sera of cystic echinococcosis patients and healthy controls, including 47 up-regulated miRNAs and 6 down-regulated miRNAs. GO enrichment analysis showed that these differentially expressed miRNA were involved DNA transcription and translation, cell components, cell morphology, neurodevelopment and metabolic decomposition, and KEGG pathway analysis showed that the differentially expressed miRNA were mainly involved in MAPK, PI3K-Akt and mTOR signaling pathways. Among the differentially expressed miRNAs with the 20 highest fold changes, there were 3 miRNAs that had a potential for target regulation of RORC, and 15 miRNAs that had a potential to target the PI3K-Akt and mTOR signaling pathways.@*CONCLUSIONS@#Significant changes are found in serum miRNA expression profiles among patients with E. granulosus infections, and differentially expressed miRNAs may lead to Th17/Treg imbalance through targeting the key transcription factors of Th17/Treg or PI3K-Akt and mTOR pathways, which facilitates the long-term parasitism of E. granulosus in hosts and causes a chronic disease.

Soluble egg antigen of Schistosoma japonicum induces macrophage apoptosis in mice / 中国血吸虫病防治杂志

OBJECTIVE@#To investigate the dynamic changes of macrophage numbers and apoptosis during Schistosoma japonicum infection, and to investigate the possible mechanisms of macrophage apoptosis induced by S. japonicum soluble egg antigen (SEA).@*METHODS@#C57BL/6 mice at ages of 6~8 weeks were randomly divided into 4 groups, including three experimental groups and a normal control group. Each mouse in the experimental groups was infected with (12 ± 1) cercariae of S. japonicum via the abdominal skin, and all mice in an experimental group were sacrificed 3, 5, 8 weeks post-infection, respectively, while mice in the control group were not infected with S. japonicum cercariae and sacrificed on the day of S. japonicum infection in the experimental group. Mouse liver specimens and peritoneal exudation cells were sampled in each group, and the dynamic changes of macrophage numbers and apoptosis were detected. Mouse peritoneal macrophages were isolated, purified and treated with S. japonicum SEA, PBS and ovalbumin (OVA) in vitro, and the macrophage apoptosis was detected using flow cytometry. The mRNA and protein expression of BCL-2 protein family members were determined in macrophages using real-time quantitative PCR (qP-CR) and Western blotting assays, and the activation of caspase 3 was determined using flow cytometry and Western blotting. In addition, macrophages were in vitro treated with S. japonicum SEA in presence of a caspase inhibitor, H2O2 or N-acetyl-L-cysteine, and the apoptosis of macrophages was detected using flow cytometry.@*RESULTS@#The total macrophage numbers continued to increase in mouse liver [(0.873 ± 0.106) × 106, (2.737 ± 0.460) × 106 and (3.107 ± 0.367) × 106 cells, respectively; F = 81.900, P < 0.01] and peritoneal specimens [(5.282 ± 1.136) × 105, (7.500 ± 1.200) × 105 and (12.800 ± 0.800) × 105 cells, respectively; F = 55.720, P < 0.01] 3, 5 and 8 weeks post-infection with S. japonicum, and the numbers of apoptotic macrophages also continued to increase in mouse liver [(0.092 ± 0.018) × 106, (0.186 ± 0.025) × 106 and (0.173 ± 0.0270) × 106 cells; F = 57.780, P < 0.01] and peritoneal specimens [(0.335 ± 0.022) × 105, (0.771 ± 0.099) × 105 and (1.094 ± 0.051) × 105 cells; F = 49.460, P < 0.01] 3, 5 and 8 weeks post-infection with S. japonicum. The apoptotic rate of SEA-treated macrophages [(24.330 ± 0.784)%] was significantly higher than that of PBS-[(18.500 ± 1.077)%] and OVA-treated macrophages [(18.900 ± 1.350)%] (both P values < 0.01). There were no significant differences in the mRNA or protein expression of Bcl-2 [Bcl - 2 mRNA expression: (1.662 ± 0.943) vs. (1.000 ± 0.000), t = 1.215, P > 0.05; BCL protein expression: (0.068 ± 0.004) vs. (0.070 ± 0.005), t = 0.699, P > 0.05], Bax [Bax mRNA expression: (0.711 ± 0.200) vs. (1.000 ± 0.000), t = 2.507, P > 0.05; BAX protein expression: (0.089 ± 0.005) vs. (0.097 ± 0.003), t = 2.232, P > 0.05] and Bak [Bak mRNA expression: (1.255 ± 0.049) vs. (1.00 ± 0.00), t = 0.897, P > 0.05; BAK protein expression: (0.439 ± 0.048) vs. (0.571 ± 0.091), t = 2.231, P > 0.05] between in SEA- and PBS-treated macrophages. S. japonicum SEA induced macrophage apoptosis in the presence of a caspase inhibitor (F = 0.411, P > 0.05); however, SEA failed to induce macrophage apoptosis in the presence of H2O2 or NAC (F = 11.880 and 9.897, both P values < 0.05).@*CONCLUSIONS@#S. japonicum SEA may induce macrophage apoptosis through promoting reactive oxygen species expression during S. japonicum infections in mice.

Study on immunological functions of heat shock protein 40 kDa of Schistosoma japonicum / 中国血吸虫病防治杂志

Objective To investigate the immunological functions of heat shock protein 40 kDa of Schistosoma japonicum (SjHSP40). Methods The homology of the SjHSP40 protein sequence was analyzed and the B and T cell epitopes of SjHSP40 were predicted using bioinformatics tools. The full-length SjHSP40 gene was amplified using a PCR assay, and cloned into the prokaryotic expression vector pGEX-6P-1, which was transformed into Escherichia coli BL-21. The protein expression was induced with isopropyl β-D-thiogalactoside (IPDG), and then, the recombinant protein was purified with glutathione-sepharose 4B resin to yield the fusion protein GST-SjHSP40, which was checked with SDS-PAGE and Western blotting. Following immunization with GST-SjHSP40, the serum levels of anti-SjHSP40 IgG antibody and IgG1 and IgG2a subtypes were detected in BALB/c mice using ELISA. In addition, the effect of SjHSP40 on CD4+ T-cell subset differentiation was examined using flow cytometry. Results SjHSP40 contained 7 potential B cell epitopes and multiple T cell epitopes (CTL epitopes and Th epitopes). The prokaryotic expression plasmid pGEX-6p-1-SjSHP40 was successfully constructed, and the fusion protein GST-SjHSP40 was obtained following IPDG induction and protein purification. Significantly higher serum levels of anti-SjHSP40 IgG, IgG1 and IgG2a antibodies were detected in mice immunized with GST-SjHSP40 than in other groups; however, SjHSP40 showed no remarkable effects on CD4+ T-cell subset differentiation. Conclusions SjHSP40 may induce specific humoral immune responses in mice; however, it does not affect the balance of Th immune responses. It is suggested that SjHSP40 may be a potential vaccine candidate.

Impact of gender on hepatic pathology and antibody - mediated immunity caused by Schistosoma japonicum infection in C57BL/6 mice / 中国血吸虫病防治杂志

Objective To investigate the effect of gender on hepatic pathology and antibody-mediated immunity in Schistosoma japonicum-infected C57BL/6 mice. Methods Female and male C57BL/6 mice were infected with S. japonicum, and the hepatic pathological changes were observed using HE and picrosirius red staining in mice 8 weeks post-infection. The serum specific IgG antibody levels against the soluble adult worm antigen (SWA) and soluble egg antigen (SEA) were measured in mice using enzyme-linked immunosorbent assay (ELISA), and the percentages of follicular helper T (Tfh) cells and regulatory T (Treg) cells were detected in mouse spleen and lymph nodes using flow cytometry. Results HE staining showed no significant difference in the mean area of a single hepatic egg granuloma between female and male mice 8 weeks post-infection with S. japonicum [(28.050 ± 3.576) × 104 μm2 vs. (26.740 ± 4.093) × 104 μm2; t = 0.241, P = 0.821], and picrosirius red staining revealed no statistical differences between female and male mice in terms of the mean proportion of picrosirius red stained hepatic tissues [(7.667 ± 1.856)% vs. (7.667 ± 1.764)%; t = 0, P = 1] or the mean optical density [(0.023 ± 0.003) vs. (0.027 ± 0.007); t = 0.447, P = 0.678]. ELISA detected no significant differences in the serum IgG antibody levels against SWA [(2.098 ± 0.037) vs. (1.970 ± 0.071); t = 1.595, P = 0.162] or SEA [(3.738 ± 0.039) vs. (3.708 ± 0.043); t = 0.512, P = 0.623] between female and male mice 8 weeks post-infection with S. japonicum. Flow cytometry detected significantly greater percentages of Tfh cells in the spleen [female mice, (8.645 ± 1.356)% vs. (1.730 ± 0.181)%, t = 5.055, P = 0.002; male mice, (8.470 ± 1.161)% vs. (1.583 ± 0.218)%, t = 5.829, P = 0.001] and lymph nodes [female mice, (3.218 ± 0.153)% vs. (1.095 ± 0.116)%, t = 11.040, P < 0.001; male mice, (3.673 ± 0.347)% vs. (0.935 ± 0.075)%, t = 8.994, P = 0.001) of both female and male mice 8 weeks post-infection with S. japonicum than in uninfected mice; however, no significant differences were seen between female and male mice 8 weeks post-infection with S. japonicum in terms of the percentages of Tfh cells in the spleen [(8.645 ± 1.356)% vs. (8.470 ± 1.161)%; t = 0.098, P = 0.925] or lymph nodes [(3.218 ± 0.153)% vs. (3.673 ± 0.347)%; t = 1.332, P = 0.241]. There was no significant difference in the proportion of Treg cells in the spleen of male mice between infected and uninfected mice [(10.060 ± 0.361)% vs. (10.130 ± 0.142)%; t = 0.174, P = 0.867], while a higher proportion of Treg cells was seen in the spleen of female mice 8 weeks post-infection with S. japonicum than in uninfected mice [(10.530 ± 0.242)% vs. (9.450 ± 0.263)%; t = 3.021, P = 0.023]. There was no significant difference in the proportion of Treg cells in the spleen between female and male mice infected with S. japonicum [(10.530 ± 0.242)% vs. (10.060 ± 0.361)%; t =1.077, P = 0.323]. In addition, the proportions of Treg cells were significantly greater in the lymph node of S. japonicum -infected female [(17.150 ± 0.805)% vs. (13.100 ± 0.265)%; t = 4.781, P = 0.003] and male mice [(18.550 ± 0.732)% vs. (12.630 ± 0.566)%; t = 6.402, P = 0.001] than in uninfected mice; however, no significant difference was seen between female and male mice 8 weeks post-infection [(17.150 ± 0.805)% vs. (18.550 ± 0.732)%; t = 1.287, P = 0.246]. Conclusion There are no gender-specific hepatic pathological changes or antibody-mediated immunity in C57BL/6 mice post-infection with S. japonicum.

Alteration in serum concentrations of FGF19, FGF21, and FGF23 in patients with urothelial carcinoma.

Fibroblast growth factors (FGF) 19, 21, and 23 have been reported as functional factors in human metabolic diseases and malignancies. We performed a prospective survey to compare circulating FGF levels in urothelial carcinoma (UC) patients and normal controls. Between 2016 and 2017, 39 patients with UC of the urinary bladder or upper urinary tract who received surgical intervention were included. All the serum samples were obtained before surgeries. The control group included 28 healthy volunteers. Analysis of the circulating FGF19, 21, and 23 levels among all 67 subjects, as well as a subgroup analysis of the 39 UC patients were performed. The median levels of serum FGF19, 21, and 23 in the UC patients were 84.2, 505.3, and 117.6 pg/mL, respectively, which were statistically different from levels found in the healthy controls (P = 0.015, <0.001 and < 0.001, respectively). In the subgroup analysis, the FGF19 and FGF21 levels were significantly higher in end-stage renal disease UC patients, while FGF21 was also higher in the UC patients with cardiovascular diseases and history of recurrent UC. In the receiver operating characteristic (ROC) curve analysis, FGF19, 21, and 23 were all significant predictors of UC [area under the curve (AUC)] 0.674, P = 0.015; AUC 0.918, P < 0.001; AUC 0.897, P < 0.001, respectively). In UC patients, serum FGF19 level was significantly lower, while FGF21 and 23 were significantly higher, than respective levels in healthy controls. All three markers may serve as good predictors of UC occurrence, and FGF21 level was associated with disease recurrence. © 2018 BioFactors, 45(1):62-68, 2019.

Attitudes of prostate cancer patients towards postoperative penile rehabilitation and their influencing factors / 中华男科学杂志

Objective@#To investigate the attitudes of prostate cancer (PCa) patients towards postoperative penile rehabilitation and their influencing factors.@*METHODS@#Seventy-nine PCa patients underwent radical prostatectomy from January through June 2017 and all received a questionnaire investigation before surgery on IIEF-5 and their attitudes towards postoperative penile rehabilitation. We analyzed the reasons for the patients' rejection of postoperative penile rehabilitation.@*RESULTS@#Totally 56 (71%) of the patients accepted and the other 23 (29%) refused postoperative penile rehabilitation. The factors influencing their attitudes towards penile rehabilitation mainly included age (P = 0.023), income (P = 0.040), tumor stage (P = 0.044), and preoperative sexual activity (P = 0.004). The patients who accepted penile rehabilitation had significantly higher IIEF-5 scores than those who refused it (14.75 ± 0.88 vs 8.48 ± 1.16, P = 0.000 2). During the follow-up period, only 29 (36.7%) of the patients bought the vacuum erection device but not the other 50 (63.3%). The tumor stage (P = 0.004), income (P < 0.01) and preoperative androgen-deprivation therapy (P = 0.039) significantly influenced the patients' decision on the purchase of the device. Relevant admission education achieved a 45% decrease in the number of the patients unwilling to accept penile rehabilitation for worrying about its negative effect on cancer treatment, a 25% decrease in those rejecting penile rehabilitation because of age, and a 20% decrease in those refusing it due to the tumor stage. The cost of treatment was an important reason for the patients' rejection of postoperative penile rehabilitation.@*CONCLUSIONS@#The tumor stage and income are the main factors influencing PCa patients' decision on postoperative penile rehabilitation. Relevant admission education and reduced cost of rehabilitation are important for popularization of postoperative penile rehabilitation in PCa patients.

The Impact of Local Intervention Combined with Targeted Therapy on Metastatic Renal Cell Carcinoma.

BACKGROUND/AIM: We performed a retrospective survey on our metastatic renal cell carcinoma (MRCC) patients who had received targeted therapies, and afterwards evaluated the clinical impacts of local interventions on the patient outcomes. MATERIALS AND METHODS: Between 2006 and 2016, 124 patients with MRCC who had received at least one line of tyrosine kinase inhibitors or mammalian target of rapamycin were included in the study. Seventy-five patients (60.5%) received targeted therapies only, twenty-six patients received complete resection and the remaining 23 received incomplete local interventions for their metastatic lesions. Analysis of the basic characteristics, overall survival and multi-variant regression amongst the three groups was performed. RESULTS: The age, gender distribution, tumor cell type, targeted therapy selection, line of therapies and sites of metastases were not different amongst the three groups. The targeted therapy-only group had a significantly higher percentage of Memorial Sloan Kettering Cancer Center (MSKCC) poor-risk patients compared with the other two groups (22.7% vs. 3.8% and 0%, p=0.006 respectively). The targeted treatment duration and follow-up duration was significantly shorted in the targeted therapy-only group. Of the twelve variables analyzed, complete resection and MSKCC poor-risk group showed a significant impact on the overall survival rate (HR=0.5, 95%CI=0.25-0.98, p=0.045; HR=2.97, 95%CI=1.05-8.4, p=0.04 respectively). CONCLUSION: Complete resection of metastatic sites for MRCC patients, combined with targeted therapy, could provide better overall survival rates than targeted therapy alone. Poor MSKCC risk is still correlated to a poor outcome in the current targeted therapy era.

[Effects of glycolytic pathway on generation of regulatory T cells during Schistosoma japonicum infection].

OBJECTIVE: To assess the influence of glycolytic pathway on the proportion and numbers of regulatory T cells during Schistosoma japonicum infection. METHODS: A S. japonicum -infected mouse model was established, and C57/BL6 male mice infected with S. japonicum were subjected to intraperitoneal injections of with the glycolytic inhibitor 2-Deoxy-D-glucose (2DG) or PBS for 6 times, and then the cells from spleen or mesenteric lymph nodes (LNs) were isolated and analyzed by flow cytometry (FCM) to detect the percentage of Glut1+CD4+ T cells and Treg cells. RESULTS: The proportions of Glut1+CD4+ T cells in the spleen (43.58%±2.50% vs. 21.15%±0.96%; t = 8.834, P < 0.01) and mesenteric LNs (38.97%±1.97% vs. 28.40%±2.11%; t = 3.662, P < 0.05) were higher in the normal mice than those in the infected mice, and the percentages of Treg cells in the spleen (6.83%±0.21% vs. 13.30%±0.35%; t = 15.65, P < 0.01) and LNs (8.26%±0.15% vs. 14.37%±0.44%; t = 13.14, P < 0.01) were lower in the normal mice than those in the infected mice. In addition, the proportions of Treg cells in the spleen (15.50%±0.76% vs. 13.07%±0.15%; t = 3.130, P < 0.05) and LNs (17.00% ±0.41% vs. 13.83%±0.18%; t = 6.947, P < 0.01) were higher in the infected mice injected intraperitoneally with 2DG than those in the infected mice injected intraperitoneally with PBS. CONCLUSIONS: Glycolytic pathway inhibits Treg differentiation in the spleen and mesenteric LNs of S. japonicum-infected mice.

[Study on role of TIGIT signal in Th1/Th2 balance in Schistosoma japonicum-infected mice].

OBJECTIVE: To investigate the role of TIGIT signal in Th1/Th2 balance in the process of Schistosoma japonicum infection. METHODS: Male C57BL/6 mice were infected with cercariae of S. japonicum, and normal uninfected mice served as the controls. The percentages of TIGIT+ cells, Ki67+CD3+CD4+TIGIT+ cells, Ki67+CD3+CD4+TIGIT- cells, IFN-γ+CD3+CD4+TIGIT+ cells, IFN-γ+CD3+CD4+TIGIT- cells, IL-4+CD3+CD4+TIGIT+ cells and IL-4+CD3+CD4+TIGIT- cells were evaluated in mouse spleen by flow cytometry. RESULTS: The proportion of TIGIT+CD4+ T cells was higher in the spleen of S. japonicum-infected mice than in the normal uninfected mice (29.30%±0.70% vs. 3.09%±0.50%; t = 8.834, P < 0.01) . However, no significant difference in the percentages of TIGIT+ CD8+ T cells between the infection group and normal controls (3.61%±0.26% vs. 3.58%±0.16%; t = 0.108, P > 0.05), and no significant difference was detected in the percentages of TIGIT+ cells in non-T cells between the infection group and controls (1.86%±0.19% vs.1.37%±0.17%; t = 1.931, P > 0.05) . In addition, the proportion of Ki67 in the TIGIT+ cells was higher than that in the TIGIT- cells (17.03%±0.64% vs. 6.59%±0.37%; t = 14.09, P < 0.01) . The Th2/Th1 ratio was higher in the TIGIT+CD4+ T cells than in the TIGIT-CD4+ T cells (39.28%±3.75% vs. 11.79%±1.64%; t = 6.721, P < 0.01) . CONCLUSIONS: The TIGIT signaling may be involved in the development of Th2 responses in the mice infected with S. japonicum.

A Modified Application of Peritoneal Dialysis Catheter Implantation: A Revolution from the Laparoscope- to the Nephroscope-Assisted Surgery.

INTRODUCTION: A laparoscope provides many advantages when establishing abdominal access for peritoneal dialysis (PD), particularly with direct observation and correction of the catheter's position. However, laparoscopic placement requires specialized equipment and usually requires using more than one working port, which may increase the potential for complications, including dialysis leakage. We modified the surgical technique by using a nephroscope, rather than a laparoscope. This study aimed to illustrate this modified technique step by step and compare the postoperative outcomes. MATERIALS AND METHODS: This study was based on a retrospective chart review of 397 consecutive patients who underwent either laparoscope- or nephroscope-assisted PD catheter insertion between September 2005 and December 2016 in our institute, as performed by a single surgeon. Data were collected and analyzed to compare the characteristics of the patients, including age and gender, along with surgical outcomes and complications between the two groups. RESULTS: Two-hundred fourteen patients underwent laparoscopy implantation, whereas 183 patients received the nephroscope-assisted method. More patients had previously undergone abdominal surgery in the nephroscopy group (29% vs 18.7%, p = 0.035) than those in the laparoscopy group. There was no significant difference in the 1-year catheter survival (82.5% vs 79.4%, p = 0.734) rate between the two groups. A total of five patients experienced dialysis leakage within the laparoscopy group, whereas none had dialysis leakage in the nephroscopy group. CONCLUSIONS: The surgical times were significantly shorter in the nephroscopy group. Although comparison of the complication rate between the two groups revealed no statistical significance, there were trends that showed there were less early surgical complications in the nephroscopy group.

[Schistosoma japonicum heat shock protein 60 enhances regulatory T cell immunosuppressive function by promoting the expressions of IL-10 and TGF-ß].

OBJECTIVE: To explore the possible mechanisms by which Schistosoma japonicum heat shock protein 60 (SjH-SP60) enhances CD4+CD25+ regulatory T cell (Treg) immunosuppressive function. METHODS: An in vitro method was used to investigate the effect of SjHSP60 on Treg immunosuppressive activity. Co-cultures in transwells and in vitro suppression assay were performed to investigate how SjHSP60 enhanced the immunosuppressive function of Tregs. Intracellular cytokine staining combined with flow cytometry was used to detect Treg-expressing IL-10 and TGF-ß, and flow cytometry was also used to analyze the expressions of Foxp3 and CTLA-4 in Tregs. RESULTS: SjHSP60 enhanced the immunosuppressive function of Tregs. Soluble cytokines IL-10 and TGF-ß mediated inhibitory activity of SjHSP60-triggered Tregs. SjHSP60 induced significant increases in both IL-10 and TGF-ß expressions of Tregs. Further investigation showed significant increased Foxp3 and CTLA-4 in SjHSP60-trggered Tregs. CONCLUSIONS: SjHSP60 enhances Treg immunosuppressive function by promoting the expressions of IL-10 and TGF-ß, possibly due to SjHSP60-mediated induction of Foxp3 and CTLA-4 in Tregs.

Study on role of TIGIT signal in Th1/Th2 balance in Schistosoma japonicum-infected mice / 中国血吸虫病防治杂志

Objective To investigate the role of TIGIT signal in Th1/Th2 balance in the process of Schistosoma japonicum in-fection.Methods Male C57BL/6 mice were infected with cercariae of S.japonicum,and normal uninfected mice served as the controls.The percentages of TIGIT+cells,Ki67+CD3+CD4+TIGIT+cells,Ki67+CD3+CD4+TIGIT-cells,IFN-γ+CD3+CD4+TIGIT+cells,IFN-γ+CD3+CD4+TIGIT- cells,IL-4+CD3+CD4+TIGIT+cells and IL-4+CD3+CD4+TIGIT- cells were evaluated in mouse spleen by flow cytometry.Results The proportion of TIGIT+CD4+T cells was higher in the spleen of S.japonicum-infected mice than in the normal uninfected mice(29.30%±0.70% vs.3.09%±0.50%;t=8.834,P<0.01).However,no significant differ-ence in the percentages of TIGIT+CD8+T cells between the infection group and normal controls(3.61% ± 0.26% vs. 3.58% ± 0.16%;t=0.108,P>0.05),and no significant difference was detected in the percentages of TIGIT+cells in non-T cells be-tween the infection group and controls(1.86%±0.19% vs.1.37%±0.17%;t=1.931,P>0.05).In addition,the proportion of Ki67 in the TIGIT+cells was higher than that in the TIGIT-cells(17.03%±0.64% vs.6.59%±0.37%;t=14.09,P<0.01).The Th2/Th1 ratio was higher in the TIGIT+CD4+T cells than in the TIGIT-CD4+T cells(39.28%±3.75% vs.11.79%±1.64%;t=6.721,P<0.01).Conclusion The TIGIT signaling may be involved in the development of Th2 responses in the mice infected with S.japonicum.

Effects of glycolytic pathway on generation of regulatory T cells during Schistosoma japonicum infection / 中国血吸虫病防治杂志

Objective To assess the influence of glycolytic pathway on the proportion and numbers of regulatory T cells dur-ing Schistosoma japonicum infection. Methods A S. japonicum-infected mouse model was established,and C57/BL6 male mice infected with S.japonicum were subjected to intraperitoneal injections of with the glycolytic inhibitor 2-Deoxy-D-glucose (2DG)or PBS for 6 times,and then the cells from spleen or mesenteric lymph nodes(LNs)were isolated and analyzed by flow cytometry(FCM)to detect the percentage of Glut1+CD4+T cells and Treg cells. Results The proportions of Glut1+CD4+T cells in the spleen(43.58%±2.50% vs.21.15%±0.96%;t=8.834,P<0.01)and mesenteric LNs(38.97%±1.97% vs.28.40%± 2.11%;t=3.662,P<0.05)were higher in the normal mice than those in the infected mice,and the percentages of Treg cells in the spleen(6.83% ± 0.21% vs. 13.30% ± 0.35%;t = 15.65,P < 0.01)and LNs(8.26% ± 0.15% vs. 14.37% ± 0.44%;t =13.14,P<0.01)were lower in the normal mice than those in the infected mice.In addition,the proportions of Treg cells in the spleen(15.50%±0.76% vs.13.07%±0.15%;t=3.130,P<0.05)and LNs(17.00% ± 0.41% vs.13.83% ± 0.18%;t=6.947, P<0.01)were higher in the infected mice injected intraperitoneally with 2DG than those in the infected mice injected intraperi-toneally with PBS. Conclusion Glycolytic pathway inhibits Treg differentiation in the spleen and mesenteric LNs of S.japoni-cum-infected mice.

Schistosoma japonicum heat shock protein 60 enhances regulatory T cell im-munosuppressive function by promoting the expressions of IL-10 and TGF-β / 中国血吸虫病防治杂志

Objective To explore the possible mechanisms by which Schistosoma japonicum heat shock protein 60(SjH-SP60)enhances CD4+CD25+regulatory T cell(Treg)immunosuppressive function.Methods An in vitro method was used to investigate the effect of SjHSP60 on Treg immunosuppressive activity.Co-cultures in transwells and in vitro suppression assay were performed to investigate how SjHSP60 enhanced the immunosuppressive function of Tregs.Intracellular cytokine staining combined with flow cytometry was used to detect Treg-expressing IL-10 and TGF-β,and flow cytometry was also used to analyze the expressions of Foxp3 and CTLA-4 in Tregs.Results SjHSP60 enhanced the immunosuppressive function of Tregs.Soluble cytokines IL-10 and TGF-β mediated inhibitory activity of SjHSP60-triggered Tregs.SjHSP60 induced significant increases in both IL-10 and TGF-β expressions of Tregs.Further investigation showed significant increased Foxp3 and CTLA-4 in SjHSP60-trggered Tregs.Conclusion SjHSP60 enhances Treg immunosuppressive function by promoting the expressions of IL-10 and TGF-β,possibly due to SjHSP60-mediated induction of Foxp3 and CTLA-4 in Tregs.

First Line Androgen Deprivation Therapy Duration Is Associated with the Efficacy of Abiraterone Acetate Treated Metastatic Castration-Resistant Prostate Cancer after Docetaxel.

Introduction: We performed a chart review study in our castration-resistant prostate cancer (CRPC) patients who received Abiraterone acetate (AA) treatment after docetaxel and identified clinical markers which can predict treatment outcome. Materials and Methods: From 2012 to 2016, 64 patients who received docetaxel after CRPC followed by AA treatment were included. Clinical parameters were recorded and analysis was performed to identify associations between pre-treatment variables and treatment outcome. Results: Thirty three patients (51.6%) achieved a decrease in PSA of 50%. The median PSA progression-free survival and overall survival in the total cohort of 64 patients were 6.6 and 24 months, respectively. Adverse events (AEs) in all grades developed in 35.9% (23/64) patients and mostly were grade 1 or 2. The most common AEs were gastric upset, hypokalemia and elevated liver function tests. Of the eight variables analyzed, first line androgen deprivation therapy (ADT) duration showed positive association to progression free survival (HR 0.98, 95% CI [0.96-0.99], p = 0.012) and overall survival (HR 0.97, 95% CI [0.94-0.99], p = 0.019). Pre-AA PSA and PSA progression ratio showed negative association only to progression free survival (HR 1.0, 95% CI [1.000-1.002], p = 0.025, HR 1.01, 95% CI [1.00-1.01], p < 0.001, respectively). Conclusion: First line ADT duration was positively associated with AA treatment efficacy in progression free survival and overall survival. It can be used as a pre-treatment predictor.

Evaluation of 99mTc-labeled PSMA-SPECT/CT imaging in prostate cancer patients who have undergone biochemical relapse / 亚洲男科学杂志(英文版)

Using conventional imaging modalities, it is difficult to detect recurrent lesions in prostate cancer patients who have undergone biochemical relapse, especially in patients with low prostate-specific antigen (PSA) levels. We retrospectively reviewed the files of fifty patients with histopathologically confirmed prostate cancer who underwent 99mTc-labeled prostate-specific membrane antigen (PSMA) single-photon emission computed tomography (SPECT)/computed tomography (CT), magnetic resonance imaging (MRI), and bone scan within a 30-day period. PSMA-SPECT/CT indicated metastatic lesions in 39 patients and had a higher detection rate (78.0%) than bone scan (34.0%) or MRI (40.0%). The diagnostic efficiency of PSMA-SPECT/CT imaging for bone and lymph node metastases (50.0% and 42.0%) was better than bone scan (34.0% and 0.0%) or MRI (24.0% and 20.0%). PSMA-SPECT/CT provided a higher detection rate at serum PSA levels of ≤1 ng ml-1, 1-4 ng ml-1, 4-10 ng ml-1, and >10 ng ml-1. No correlation was found between Gleason score, PSA level, and the tracer tumor/background ratio of metastatic lesions. With the aid of PSMA-SPECT/CT imaging, the therapeutic strategy was changed for 31 patients, and this may have enhanced their clinical outcome. In conclusion, PSMA-SPECT/CT imaging could detect more metastatic lesions and achieve a higher detection rate than conventional imaging modalities at different serum PSA levels in prostate cancer patients who had undergone biochemical relapse.

[Sex bias in generation and functional phenotypes of peripheral T follicular helper cells in schistosomiasis japonica].

OBJECTIVE: To study the influence of sex on the generation and function of T follicular helper (Tfh) cells in the process of Schistosoma japonicum infection. METHODS: Totally 50 schistosomiasis patients were selected in the endemic areas of Chizhou City, Anhui Province, and totally 50 healthy people were selected in the non-endemic areas of Chizhou City as the controls. The peripheral bloods were collected from the above study subjects, and the human peripheral blood mononuclear cells from male/female schistosomiasis japonica patients or healthy controls were collected into the sodium heparin tubes and purified by Ficoll-paque plus density gradient centrifugation. Then, the percentages of total Tfh cell, ICOS+ Tfh cells, and PD-1+ Tfh cells in the schistosomiasis male/female patients or healthy controls were evaluated by flow cytometry. RESULTS: Compared with the male and female healthy controls, the frequency of the total Tfh cells (Umale = 149.0, Ufemale = 39.5, both P < 0.01), the percentages of PD-1+ Tfh cells (tmale = 5.9, tfemale = 7.7, both P < 0.01) and ICOS+ Tfh cells (tmale = 3.2, tfemale = 5.9, both P < 0.01) of the male and female patients with schistosomiasis increased, all the differences were statistically significant. The further analysis showed that the percentage of total Tfh cells (U = 187.5, P < 0.05), ICOS+ Tfh cells (t = 3.2, P < 0.05), and PD-1+ Tfh cells (t = 3.0, P < 0.05) of the female patients were markedly higher than those of the male patients, all the differences were also statistically significant. CONCLUSIONS: Sex may be a crucial role in the generation and function of Tfh cells in the process of Schistosoma japonicum infection.

Visual Detection of Human Coronavirus NL63 by Reverse Transcription Loop-Mediated Isothermal Amplification / 病毒学报

A simple and sensitive assay for rapid detection of human coronavirus NL63 (HCoV-NL63) was developed by colorimetic reverse transcription loop-mediated isothermal amplification (RT-LAMP). The method employed six specially designed primers that recognized eight distinct regions of the HCoV-NL63 nucleocapsid protein gene for amplification of target sequences under isothermal conditions at 63 degrees C for 1 h Amplification of RT-LAMP was monitored by addition of calcein before amplification. A positive reaction was confirmed by change from light-brown to yellow-green under visual detection. Specificity of the RT-LAMP assay was validated by cross-reaction with different human coronaviruses, norovirus, influenza A virus, and influenza B virus. Sensitivity was evaluated by serial dilution of HCoV-NL63 RNA from 1.6 x 10(9) to 1.6 x 10(1) per reaction. The RT-LAMP assay could achieve 1,600 RNA copies per reaction with high specificity. Hence, our colorimetric RT-LAMP assay could be used for rapid detection of human coronavirus NL63.

Studies on immunogenicity and immunoprotection induced by heat shock protein 60 kDa of Schistosoma japonicum in mice / 中国血吸虫病防治杂志

Objective To evaluate the immunogenicity and immunoprotective effect of heat shock protein 60 kDa (SjHSP60) of Schistosoma japonicum in mice after immunization and challenge infection, and explore the mechanism. Methods B cell/an?tibody?related databases and analysis tools were used to predict B?cell epitopes of SjHSP60. The mice were immunized with the recombinant SjHSP60 and challenged with S. japonicum cercariae. SjHSP60?specific antibodies in serum were detected by ELI?SA. The level of splenocyte proliferation was determined by 3H?TdR incorporation. Ex vivo suppression assay was performed to in?vestigate the effects of CD4 +CD25 + regulatory T cells (Tregs) induced by SjHSP60. Results SjHSP60 possessed multiple pre?dominant regions of B?cell epitopes. SjHSP60 induced a significant increase in both SjHSP60?specific IgG levels (P 0.05) and liver?accumulated eggs (P > 0.05) in S. japonicum?infected mice. Ex vivo assay showed that CD4+CD25+ Tregs from SjHSP60?immunized mice enhanced immunosuppressive activity. Conclusion SjH?SP60 has a dual role in host immune system, being involved in the induction of dominant humoral and cellular immune responses as well as in the enhancement of immunosuppression.