Fertility-preserving treatment outcomes in endometrial cancer and atypical hyperplasia patients with different molecular profiles / 中华妇产科杂志

Objective: To investigate the impact of molecular classification and key oncogenes on the oncologic outcomes in patients with endometrial carcinoma (EC) and atypical endometrial hyperplasia (AEH) receiving fertility-preserving treatment. Methods: Patients with EC and AEH undergoing progestin-based fertility-preserving treatment and receiving molecular classification as well as key oncogenes test at Obstetrics and Gynecology Hospital, Fudan University from January 2021 to March 2023 were reviewed. Hysteroscopic lesion resection and endometrial biopsy were performed before initiating hormone therapy and every 3 months during the treatment to evaluate the efficacy. The risk factors which had impact on the treatment outcomes in EC and AEH patients were further analyzed. Results: Of the 171 patients analyzed, the median age was 32 years, including 86 patients with EC and 85 patients with AEH. The distribution of molecular classification was as follows: 157 cases (91.8%) were classified as having no specific molecular profile (NSMP); 9 cases (5.3%), mismatch repair deficient (MMR-d); 3 cases (1.8%), POLE-mutated; 2 cases (1.2%), p53 abnormal. No difference was found in the cumulative 40-week complete response (CR) rate between the patients having NSMP or MMR-d (61.6% vs 60.0%; P=0.593), while the patients having MMR-d had increased risk than those having NSMP to have recurrence after CR (50.0% vs 14.4%; P=0.005). Multi-variant analysis showed PTEN gene multi-loci mutation (HR=0.413, 95%CI: 0.259-0.658; P<0.001) and PIK3CA gene mutation (HR=0.499, 95%CI: 0.310-0.804; P=0.004) were associated with a lower cumulative 40-week CR rate, and progestin-insensitivity (HR=3.825, 95%CI: 1.570-9.317; P=0.003) and MMR-d (HR=9.014, 95%CI: 1.734-46.873; P=0.009) were independent risk factors of recurrence in EC and AEH patients. Conclusions: No difference in cumulative 40-week CR rate is found in the patients having NSMP or MMR-d who received progestin-based fertility-preserving treatment, where the use of hysteroscopy during the treatment might be the reason, while those having MMR-d have a higher risk of recurrence after CR. Oncogene mutation of PTEN or PIK3CA gene might be associated with a lower response to progestin treatment. The molecular profiles help predict the fertility-preserving treatment outcomes in EC and AEH patients.

Comparison of the effects and safety of dydrogesterone and medroxyprogesterone acetate on endometrial hyperplasia without atypia: a randomized controlled non-inferior phase Ⅲ clinical study / 中华妇产科杂志

Objective: To compare the effects and safety of dydrogesterone (DG) and medroxyprogesterone acetate (MPA) on the treatment in patients with endometrial hyperplasia without atypia (EH). Methods: This was a single-center, open-label, prospective non-inferior randomized controlled phase Ⅲ trial. From February 2019 to November 2021, patients with EH admitted to the Obstetrics and Gynecology Hospital of Fudan University were recruited. Enrolled patients were stratified according to the pathological types of simple hyperplasia (SH) or complex hyperplasia (CH), and were randomised to receive MPA or DG. Untill May 14, 2022, the median follow-up time after complete response (CR) was 9.3 months (1.1-17.2 months). The primary endpoint was the 6-month CR rate (6m-CR rate). The secondary endpoints included the 3-month CR rate (3m-CR rate), adverse events rate, recurrence rate, and pregnancy rate in one year after CR. Results: (1) A total of 292 patients with EH were enrolled in the study with the median age of 39 years (31-45 years). A total of 135 SH patients were randomly assigned to MPA group (n=67) and DG group (n=68), and 157 CH patients were randomly assigned to MPA group (n=79) and DG group (n=78). (2) Among 292 patients, 205 patients enrolled into the primary endpoint analysis, including 92 SH patients and 113 CH patients, with 100 patients in MPA group and 105 in DG group, respectively. The 6m-CR rate of MPA group and DG group were 90.0% (90/100) and 88.6% (93/105) respectively, and there were no statistical significance (χ2=0.11, P=0.741), with the rate difference (RD) was -1.4% (95%CI:-9.9%-7.0%). Stratified by the pathology types, the 6m-CR rate of SH patients was 93.5% (86/92), and MPA group and DG group were respectively 91.1% (41/45) and 95.7% (45/47); and the 6m-CR rate of CH patients was 85.8% (97/113), and MPA group and DG group were 89.1% (49/55) and 82.8% (48/58) respectively. The 6m-CR rates of the two treatments had no statistical significance either (all P>0.05). A total of 194 EH patients enrolled into the secondary endpoint analysis, including 88 SH patients and 106 CH patients, and 96 patients in MPA group and 98 in DG group, respectively. The 3m-CR rate of SH patients were 87.5% (77/88), while the 3m-CR rates of MPA group and DG group were 90.7% (39/43) and 84.4% (38/45), respectively; the 3m-CR rate of CH patients was 66.0% (70/106), and MPA group and DG group had the same 3m-CR rate of 66.0% (35/53). No statistical significance was found between the two treatments both in SH and CH patients (all P>0.05). (3) The incidence of adverse events between MPA group and DG group had no statistical significance (P>0.05). (4) A total of 93 SH patients achieved CR, and the cumulative recurrence rate in one year after CR were 5.9% and 0 in MPA group and DG group, respectively. While 112 CH patients achieved CR, and the cumulative recurrence rate in one year after CR were 8.8% and 6.5% in MPA group and DG group, respectively. There were no statistical significance between two treatment groups (all P>0.05). Among the 93 SH patients, 10 patients had family planning but no pregnancy happened during the follow-up period. Among the 112 CH patients, 21 were actively preparing for pregnancy, and the pregnancy rate and live-birth rate in one year after CR in MPA group were 7/9 and 2/7, while in DG group were respectively 4/12 and 2/4, and there were no statistical significance in pregnancy rate and live-birth rate between the two treatment groups (all P>0.05). Conclusions: Compared with MPA, DG is of good efficacy and safety in treating EH. DG is a favorable alternative treatment for EH patients.

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.

Clinical features related to lymphatic metastasis in grade 3 endometroid endometrial cancer: a retrospective cross-sectional study / 中华医学杂志(英文版)

BACKGROUND@#Endometrial cancer (EC) has been one of the most general cancers with respect to gynecological malignancies; however, there are debates on clinical strategies concerning treatments especially for patients with grade 3 (G3) endometroid endometrial cancer (EEC). Present study aimed to evaluate the lymphatic metastasis (LM) related factors and figure out the necessity of lymphadenectomy for G3 EEC patients.@*METHODS@#From January 2009 to April 2019, 3751 EC patients were admitted to Obstetrics and Gynecology Hospital of Fudan University. Clinical characteristics include age, grade, stage, and clinical pathological features. A total of 1235 EEC patients were involved in the multivariable analysis. Three hundred and eighty-one patients were involved in the survival analysis and the data attributed to sufficient follow-up information. Kaplan-Meier curve and log-rank test were utilized to analyze the survival rate.@*RESULTS@#Among the 1235 EEC patients, 181 (14.7%) were categorized as G3 and 1054 (85.3%) were grade 1 to grade 2 (G1-2). Multivariate analysis demonstrated that lymphovascular space invasion, adnexal involvement, and cervical stroma involvement were independent risk factors of LM in G3 cohort with odds ratio 3.4, 5.8, and 8.9; 95% confidence interval 1.1-10.6, 1.5-22.4, and 2.8-28.0, respectively. LM rates increased from 3.3% (3/92) to 75% (9/12) for G3 EEC cohort as related factor numbers increased from one to three. There were no differences between G3 and G1-2 EEC in overall survival and progression free survival. Additionally, no survival advantage was observed for G3 EEC patients at early stage with different plans of adjuvant treatment.@*CONCLUSIONS@#For G3 EEC patients without other pathological positive factor, the LM rate is lower than those with other pathological positive factor. Survival analysis showed no difference between G3 cohort and G1-2 cohort. Also, different adjuvant treatments had no impact on the overall survival for G3 EEC patients.

Sentinel lymph Node mapping versus systematic pelvic lymphadenectomy on the prognosis for patients with intermediate-high-risk Endometrial Cancer confined to the uterus before surgery: trial protocol for a non-inferiority randomized controlled trial (SNEC trial) / 부인종양

Background@#Sentinel lymph node (SLN) mapping has been recommended as an alternative staging approach to lymphadenectomy for apparent uterine-confined endometrial cancer (EC). However, the prognostic value of SLN mapping alone instead of systematic lymphadenectomy on EC patients remains unclear. @*Methods@#A multi-center, open label, non-inferiority randomized controlled trial has been designed to identify if SLN mapping alone is not inferior to pelvic lymphadenectomy on prognosis of patients with intermediate-high-risk EC clinically confined to uterus. Eligible patients will be 1:1 randomly assigned to accept SLN mapping or pelvic lymphadenectomy. The primary endpoint is the 2-year progression-free survival (PFS). The second points are the 5-year PFS, 5-year overall survival, surgery-related adverse events and life quality. A total of 780 patients will be enrolled from 6 hospitals in China within 3-year period and followed up for 5 years.

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.

Fertility-preserving treatment for early stage endometrial cancer / 中国实用妇科与产科杂志

The fertility preservation treatment has become one of the main issues with the increasing incidence of endometrial cancer.Strict selection of the young endometrial cancer patients who are eligible for fertility preservation treatment and individualizedtreatment plan are fundamental for improving treatmenteffect while decreasing the risk of treatment-relatedcomplications and treatment failure.This article willdiscuss the evaluation and treatment choice for fertilitypreservation in young endometrial cancer patients.

Anti-inflammatory effects of Eucommia ulmoides Oliv. male flower extract on lipopolysaccharide-induced inflammation / 中华医学杂志(英文版)

BACKGROUND@#Eucommia ulmoides Oliv. is a medicinal plant native to China, with its bark (Eucommiae Cortex) traditionally being used for medicinal purposes. Previous research has shown that Eucommia male flowers can exert anti-inflammatory, analgesic, antibacterial, and other pharmacological effects, including immune regulation. This study explored the anti-inflammatory effects of the 70% ethanol extract of male flowers (EF) of E. ulmoides in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and LPS-administered mice.@*METHODS@#Cytotoxicity of EF for RAW 264.7 cells was investigated using Cell Counting Kit-8. The production of proinflammatory mediators, nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 was determined using enzyme-linked immunosorbent assays. IL-17, IL-23, and IL-10 mRNA levels were determined using quantitative real-time polymerase chain reaction. Activation of the nuclear factor (NF)-κB pathway in RAW 264.7 cells was investigated via Western blotting. In vivo anti-inflammatory effects of EF were studied in an LPS-induced acute inflammation mouse model by analyzing lung tissue histopathology, serum TNF-α and IL-6 levels, and myeloperoxidase (MPO) activity in lung tissue.@*RESULTS@#EF showed no significant cytotoxicity at concentrations from 10 to 60 μg/mL (cell viability > 80%) in the CCK-8 cell viability assay. EF inhibited the RAW 264.7 cell proliferation (EF 60 μg/mL, 120 μg/mL, and 250 μg/mL vs. negative control: 87.31 ± 2.39% vs. 100.00 ± 2.50%, P = 0.001; 79.01 ± 2.56 vs. 100.00 ± 2.50%, P < 0.001; and 64.83 ± 2.50 vs. 100.00 ± 2.50%, P < 0.001), suppressed NO (EF 20 μg/mL and 30 μg/mL vs. LPS only, 288.81 ± 38.01 vs. 447.68 ± 19.07 μmol/L, P = 0.004; and 158.80 ± 45.14 vs. 447.68 ± 19.07 μmol/L, P < 0.001), TNF-α (LPS+EF vs. LPS only, 210.20 ± 13.85 vs. 577.70 ± 5.35 pg/mL, P < 0.001), IL-1β (LPS+EF vs. LPS only, 193.30 ± 10.80 vs. 411.03 ± 42.28 pg/mL, P < 0.001), and IL-6 (LPS+EF vs. LPS only, 149.67 ± 11.60 vs. 524.80 ± 6.24 pg/mL, P < 0.001) secretion, and downregulated the mRNA expression of IL-17 (LPS+EF vs. LPS only, 0.23 ± 0.02 vs. 0.43 ± 0.12, P < 0.001), IL-23 (LPS+EF vs. LPS only, 0.29 ± 0.01 vs. 0.42 ± 0.06, P=0.002), and IL-10 (LPS+EF vs. LPS only, 0.30 ± 0.01 vs. 0.47 ± 0.01, P=0.008) in LPS-stimulated RAW 264.7 cells. EF inhibited the LPS-induced NF-κB p65 (LPS+EF 20 μg/mL and 30 μg/mL vs. LPS only: 0.78 ± 0.06 vs. 1.17 ± 0.08, P < 0.001; and 0.90 ± 0.06 vs. 1.17 ± 0.08, P =0.002) and inhibitor of kappa B (IκBα) phosphorylation (LPS+EF 20 μg/mL and 30 μg/mL vs. LPS only: 0.25 ± 0.01 vs. 0.63 ± 0.03, P < 0.001; and 0.31 ± 0.01 vs. 0.63 ± 0.03, P < 0.001), LPS+EF 30 μg/mL inhibited IκB kinase (IKKα/β) phosphorylation (LPS+EF 30 μg/mL vs. LPS only, 1.12 ± 0.14 vs. 1.71 ± 0.25, P = 0.002) in RAW 264.7 cells. Furthermore, EF 10 mg/kg and EF 20 mg/kg inhibited lung tissue inflammation in vivo and suppressed the serum TNF-α (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 199.99 ± 186.49 vs. 527.90 ± 263.93 pg/mL, P=0.001; and 260.56 ± 175.83 vs. 527.90 ± 263.93 pg/mL, P = 0.005), and IL-6 (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 41.26 ± 30.42 vs. 79.45 ± 14.16 pg/ ml, P = 0.011; and 42.01 ± 26.26 vs. 79.45 ± 14.16 pg/mL, P = 0.012) levels and MPO (LPS+EF 10 mg/kg and 20 mg/kg vs. LPS only, 3.19 ± 1.78 vs. 5.39 ± 1.51 U/g, P = 0.004; and 3.32 ± 1.57 vs. 5.39 ± 1.51 U/g, P = 0.006) activity in lung tissue.@*CONCLUSIONS@#EF could effectively inhibit the expression of inflammatory factors and overactivation of neutrophils. Further investigation is needed to evaluate its potential for anti-inflammation therapy.

[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.

[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.

Management of whole process of healthcare-associated infection based on new media / 中国感染控制杂志

Objective To promote the meticulous management on whole process of healthcare-associated infection (HAI)through the development of new media function.Methods In 2016,with the help of WeChat enterprise number of the new media network tool,existing procedures were deeply integrated and optimized by means of infor-mation push as well as image and questionnaire survey,HAI prevention and control information platform was con-structed,participation rate of the missing reporting and reporting of HAI during the process of implementation and operation of the platform in 2014－2016 were compared.Results In 2016,the participation rate of reporting of HAI cases was 90.43%,with an increase of 26.34% compared with 64.09% in 2014,there was significant difference in reporting rate of HAI in three years(χ2＝104.53,P＜0.001).From 2014 to 2016,missing reporting rate of HAI were 14.63%,10.81%,and 4.24% respectively,difference was statistically significant (χ2＝53.85,P＜0.001). From 2014 to 2016,the active reporting rate of HAI increased from 0 to 14.22%,the normal reporting rate of HAI increased from 12.88% to 44.17%,and the delayed reporting rate of HAI decreased from 87.12% to 41.61%. Conclusion The innovative management mode of HAI based on new media is helpful for conducting prospective pre-vention and control of HAI,achieve the active management and real-time monitoring on HAI,and promote the closed-loop management on the whole process of HAI prevention and control.

Antithrombotic effects of ethyl acetate fraction of Curcuma kwangsiensis / 中成药

AIM To study the antithrombotic effects and mechanism of ethyl acetate fraction of Curcuma kwangsiensis S.G.Lee et C.F.Liang (CKEAF).METHODS To observe the antithrombotic effects of CKEAF on tail thrombotic mouse models and arteriovenous bypass thrombotic rat models induced by carrageenan,and in vivo thrombosis rat models induced by FeCl3.The serum levels of ET-1,6-keto-PGF1α,TXB2 in rats were measured by enzyme linked immunosorbent assay (ELISA),and the level of NO was determined by nitrate reductase method.Rat models of acute blood stasis were further established for hemorheological study.RESULTS CKEAF significantly decreased the number and length of blackened thrombotic tail of carrageenan-treated mice.Rat models shared significant wet weight loss in arteriovenous bypass thrombosis and in vivo thrombosis,increased levels of NO,6-keto-PGF1α,decreased levels of ET-1,TXB2,and decreased whole blood and plasma viscosity (rats of acute blood stasis model).CONCLUSION Significant antithrombotic effects by CKEAF may contribute to elevated levels of NO,6-keto-PGF1α,and decreased levels of ET-1,TXB2,and lowered whole blood and plasma viscosity.

Effects of Alcohol Extracts of Bark and Male Flower of Eucommia ulmoides Oliv. on Airway Allergic Inflammation of Model Mice / 中国中医药信息杂志

Objective To investigate the effects of alcohol extract of bark and male flower of Eucommia ulmoides Oliv. on airway allergic inflammation induced by chicken ovalbumin (OVA) in mice; To explore its mechanism of action. Methods On day 0, day 7, mice were intraperitoneally injected OVA for sensitization, followed by nasal stimulation for 21 days to establish airway allergic inflammation mice models. The mice were divided into normal group, model group, alcohol extract of bark of Eucommia ulmoides Oliv. group, alcohol extract of male flower of Eucommia ulmoides Oliv.group,and Dexamethasone group.Each medication group was given relevant medicine for gavage. The lung tissue was embedded in HE and PAS dyeing, to observe the pathological changes of bronchus and surrounding lung. The levels of serum OVA-IgE, IL-4, IFN-γ and IL-13 were measured by ELISA. The expression of ICAM-1, VEGF, MMP9 and TIMP1 were detected by immunohistochemistry. Flow cytometry was used to detect the expression of Th17 cells in peripheral blood. The expressions of TNF-α and IL-6 mRNA in lung tissue were detected by RT-PCR. Results The model group showed changes of airway allergic inflammatory such as eosinophils and other inflammatory cell infiltration, bronchial spasm, and mucus secretion. Lung histopathology in alcohol extract of bark and male flower of Eucommia ulmoides Oliv.groups was improved significantly(P<0.05).Compared with the normal group, the levels of serum OVA-IgE, IL-4 and IL-13 increased in model group, while the level of IFN-γ decreased (P<0.05, P<0.01). The expressions of ICAM-1, VEGF and MMP9 increased, while the expression of TIMP1 decreased (P<0.01); peripheral blood IL-17+cells increased (P<0.01); the expressions of TNF-α and IL-6 mRNA increased. Compared with the model group, the levels of serum OVA-IgE, IL-4 and IL-13 decreased in alcohol extract of bark and male flower of Eucommia ulmoides Oliv. groups (P<0.05, P<0.01); the expressions of ICAM-1 and VEGF decreased (P<0.05, P<0.01); the expression of TIMP1 increased. Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.could down-regulate IL-17+cells,reduce the expression of IL-6 mRNA(P<0.05,P<0.01). Alcohol extract of bark of Eucommia ulmoides Oliv. group could induce the secretion of IFN-γ (P<0.01), and down-regulate the expression of TNF-α mRNA(P<0.05).Alcohol extract of male flower of Eucommia ulmoides Oliv. group could significantly down-regulate the expression of MMP9 (P<0.05). Conclusion Alcohol extract of bark and male flower of Eucommia ulmoides Oliv.can induce the production of OVA-IgE,inhibit secretion of Th2 cytokines, inhibit the expression of adhesion molecules, depress Th17 cells, so as to inhibit the airway allergic inflammation.

Genetic diversity of 6 species in Polygonatum by SSR marker / 中国中药杂志

Polygonatum is a genus of the perennial herb family Liliaceae, with great potential in food, medicine and other field. In this study, genetic diversity and cluster structure analysis of 6 species in Polygonatum were investigated the molecular marker technique of simple sequence repeat (SSR). A total of 49 SSR makers were used to study genetic diversity and population structure within 60 germplasm resources which obtained from 38 counties and cities in 14 provinces of China. A total of 211 alleles were identified and the number of alleles ranged from 2 to 10, with an average of 4.306 1 alleles per SSR. The range of polymorphism information content (PIC) varied from 0.731 8 to 0.031 7, with the average of 0.309 6. The cluster analysis classified 60 germplasm resources into four defined groups at the genetic distance value of 0.26, among which most species with relatives were clustered into the same group. Extraordinarily, there were 6 germplasm resources clustered into other species, indicating that the classification of inter-genus and geographical distribution was crossed in Polygonatum. The genetic diversity index of the 4 geographical populations from high to low was: Western region, Central China, Southern China, Eastern China. The population structure analysis, also indicating divided the entire collection into four groups, which was similar to the assignment pattern of cluster structure analysis. These results suggested that the Polygonatum germplasm resources used in this study is rich in relatively high genetic diversity with large variation range, relatively fuzzy boundaries of species. It appeared the phenomenon that there is a difference decreases between the alternate leaf system and the rotation leaf system. The genetic diversity in the western region was higher than that in other regions, and the western region may be the origin center of the genus Polygonatum.