BACKGROUND: Regulatory dendritic cell (DCreg) subset exhibits a unique capacity for inducing immune tolerance among the variety subsets of dendritic cells (DCs) within gut-associated lymphoid tissues (GALTs). Fms-like tyrosine kinase 3 ligand (FLT3L) is involved in the differentiation of DCregs and the subsequent expansion of regulatory T-cells (Tregs) mediated by DCregs, though the precise mechanism remains poorly understood. This study aimed to explore the expansion mechanism of Treg induced by DCreg and the role of FLT3L in this process. METHODS: DCregs were distinguished from other DC subsets isolated from GALTs of BALB/c mice through a mixed lymphocyte reaction assay. The functions and mechanisms by which FLT3L promoted Treg expansion via DCregs were investigated in vitro through co-culture experiments involving DCregs and either CD4 + CD25 - T-cells or CD4 + CD25 + T-cells. Additionally, an in vivo experiment was conducted using a dextran sulfate sodium (DSS)-induced colitis model in mice. RESULTS: CD103 + CD11b + DC exhibited DCreg-like functionality and was identified as DCreg for subsequent investigation. Analysis of Foxp3 + Treg percentages within a co-culture system of CD4 + CD25 - T-cells and DCregs, with or without FLT3L, demonstrated the involvement of the FLT3/FLT3L axis in driving the differentiation of precursor T-cells into Foxp3 + Tregs induced by DCregs. Cell migration and co-culture assays revealed that the FLT3/FLT3L axis enhanced DCreg migration toward Tregs via the Rho pathway. Additionally, it was observed that DCregs could promote Treg proliferation through the Notch pathway, as inhibition of Notch signaling by DAPT (N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester) suppressed Treg expansion within the co-culture system of DCregs and CD4 + T-cells or CD4 + CD25 + T-cells. Furthermore, the FLT3/FLT3L axis influenced JAG1 expression in DCregs, indirectly modulating Treg expansion. In vivo experiments further established that FLT3L promoted DCreg expansion and restored Treg balance in DSS-induced colitis models, thereby ameliorating colitis symptoms in mice. CONCLUSION The FLT3/FLT3L axis is integral to the maintenance of DCreg function in Treg expansion.
Animals ; T-Lymphocytes, Regulatory/immunology* ; Dendritic Cells/immunology* ; Mice ; Mice, Inbred BALB C ; Membrane Proteins/metabolism* ; Receptors, Notch/metabolism* ; Lymphoid Tissue/metabolism* ; Signal Transduction/physiology* ; Coculture Techniques ; Flow Cytometry

By consulting ancient materia medica， medical books， prescription books and modern literature， this paper systematically combed and reviewed the name， origin， scientific name evolution， producting area， quality evaluation， medicinal parts， harvesting and processing and traditional efficacy of Lasiosphaera Calvatia. The results show that Mabo was first recorded in Mingyi Bielu. Since then， all dynasties have taken Mabo as a legitimate name. Before the Song dynasty， only Calvatia lilacina was used as the original plant of Lasiosphaera Calvatia， which was expanded after the Song dynasty with the appearance of C. gigantea， Lasiosphaera fenzlii， Bovistella radicata and other varieties. Until modern times， there was an addition of Lycoperdon perlatum， L. pyriforme and other original plants of Lasiosphaera Calvatia. Since 1975， the original plant of Lasiosphaera Calvatia in various regulations and academic monographs has been basically uniform for C. lilacina， Lasiosphaera fenzlii and C. gigantea. Resource of the medicinal fungus was widely distributed in China and was mainly wild. From ancient times to the present， the medicinal parts of Lasiosphaera Calvatia are all fruiting body， which is harvested in summer and autumn， and its processing method was to take powder in ancient times， but to cut blocks in modern times. In recent times， its quality has been summarized as large， thin-skinned， intact， full， loose-bubbled and elastic. The medicinal efficacy has been developed from very good for all scores， and after the Ming and Qing dynasties， it is consistent with the 2020 edition of Chinese Pharmacopoeia， with the efficacy of clearing the lung， promoting pharynx， relieving fever and hemostasis， mainly treating cough aphonia， throat obstruction and pharyngeal pain， vomiting blood， epistaxis， hemoptysis， and external treating sores and bleeding from cuts and wounds. Based on the results of herbal textual research， it is suggested that C. lilacina is the first choice for the origin of Lasiosphaera Calvatia involved in famous classical formulas， and it is processed into block or powder for medicine.

Objective: To understand the infection status, characteristics and drug resistance of non-O157 Shiga toxin-producing E. coli (STEC) in animal feces in Shandong Province. Methods: From 2015 to 2016, convient sampling method was used to collect 1 022 fresh feces of animals in Weishan county and Laizhou city, and 24 non-O157 STEC were isolated. The serotypes of non-O157 STEC strains were confirmed through serum agglutination test. The susceptibility was explored through the antimicrobial sensitivity experiments. ESBLs activity was confirmed by double-disc diffusion. PCR method was used to detect the resistance genes. PFGE typing was operated to assess the relatedness and variability of the strains. The multi-locus sequence typing (MLST) was adopted to get the allelic profile and ST sequence of strains. Analysis was made on the evolutionary relationship between different ST groups was made through CLC Sequence Viewer and Counting Express. Results: A total of 24 non-O157 STEC were isolated from animal feces. 23 strains were from pig feces, and 1 strain was from cow feces, and the serotypes were more dispersed. All of the 24 strains carried stx2 genes. The highest resistance rate was sulfamethoxazole(22 strains), the mount of cotrimoxazole and nalidixic acid was 18 strains, chloramphenicol was 13 strains, tetracycline was 19, and there was a phenomenon of multiple drug resistance. The drug resistance spectrum was sulfamethoxazole tetracycline-compound novammin-naphthidine-chloramphenicol. All strains were sensitive to cefepime and imipenem. The ESBLs confirmatory test showed that 4 strains of non O157 STEC produced beta lactamase. PCR detected 7 resistance genes, and 4 tetracycline resistance genes (Tet A, Tet B, tetC and tetD) were detected. The beta lactamase resistance genes (blaSHV-1, bla CTX-M, bla TEM) were all negative. 24 strains were divided into 15 PFGE types, and their clustering results were more dispersed and no dominant PFGE type. There were 11 kinds of MLST types, most of them are ST540 and ST5133 types, each of which was 4 strains, and clustered into 1 MLST genomes. Conclusion The serotypes of non-O157 STEC in animal feces O157 STEC were dispersed, and the resistant rate to common antibiotic was high. MLST typing results presents obvious polymorphism. Surveillance and manage ment of these strains should be strengthened.

Objective To investigate the risk factors of hemorrhagic transformation(HT) in patients with acute cerebral infarction(ACI).Methods A total of 335 patients with ACI from June 2013 to September 2016 was enrolled in this study,including 47 patients in hemorrhagic transform group(HT group) and 288 patients in non-hemorrhagic transformation group(NHT group).The general clinical situation,laboratory indexes,imaging features and treatment measures of the two groups were collected and compared,and then the risk factors of HT in the patients with ACI were analyzed by Logistic regression analysis.Results Univariate analysisshowed that there were significant differences between the two groups in the history of diabetes mellitus,hìstory of atrial fibrillation,NHISS score,systolic blood pressure at admission,fasting blood glucose,glycosylated hemoglobin,low density lipoprotein cholesterol,fibrinogen,infarct location,large area infarction and thrombolytic therapy(P＜0.05).Logistic regression analysis showed that history of atrial fibrillation(OR =2.703,95 ％ CI 1.169-6.250),high fasting blood glucose(OR =2.098,95 ％ CI 1.532-2.875),large area infarction(OR=9.999,95％CI 4.648-21.510) and thrombolytic therapy(OR=6.557,95％CI 1.954-22.003) were independent risk factors for HT.Conclusion The history of atrial fibrillation,high fasting blood glucose,large area infarction and thrombolytic therapy are the risk factors for HT in patients with ACI.Corresponding nursing measures should be arranged to facilitate the disease treatment.

OBJECTIVETo investigate the impact of NU7026 and Wortmannin, inhibitors of DNA-dependent protein kinase (DNA-PK), in HL60 cells apoptosis induced by 1, 4-benzoquinone (1, 4-BQ).

METHODSHL60 cells were divided into three groups according to the exposures: the poisoned groups which were treated with 0, 5, 10, 25 and 50 µmol/L 1, 4-BQ for 24 h, respectively, the NU7026 groups which were preincubated with 10 µmol/L NU7026 for 1h prior to the 24h treatment of 0, 5, 10, 25 and 50 µmol/L 1, 4-BQ and the Wortmannin groups which were preincubated with 25 µmol/L Wortmannin for 1h prior to the 24 h treatment of 0, 5, 10, 25 and 50 µmol/L 1, 4-BQ. Then we detected the apoptosis via flowcytometry Annexin V-FITC/PI and the DNA Ladder, respectively. We also tested the expressions of Bax mRNA with Real-Time PCR in HL60 cells which were exposed to 10 µmol/L NU7026 for 24 h, 25 µmol/L Wortmannin 24 h, 10 µmol/L 1, 4-BQ 24 h, 10 µmol/L NU7026 1h+10 µmol/L 1, 4-BQ 24 h and 25 µmol/L Wortmannin 1 h+10 µmol/L 1, 4-BQ 24 h, as well as null (control). We also examed the expressions of p53 in HL60 cells with Western blot.

RESULTSAnnexin V-FITC/PI apoptosis tests suggested that apoptosis rates of NU7026+10 µmol/L 1, 4-BQ group and Wortmannin +10 µmol/L 1, 4-BQ were 17.6±1.19% and 15.2±1.22%, respectively. Both of results were higher than that of 10 µmol/L 1, 4-BQ group (6.3±1.04%); Apoptosis of NU7026+25 µmol/L 1, 4-BQ group was 46.2±3.55%, and Wortmannin +25 µmol/L 1, 4-BQ group 26.9±2.62%. Both of results were also higher than that of 25 µmol/L 1, 4-BQ group (14.1±1.54%); Apoptosis of NU7026+50 µmol/L 1, 4-BQ group (61.8±1.78%) was higher than that of 50 µmol/L 1, 4-BQ group (35.9±4.51%). The above results were all statistically significant (P < 0.05).

RESULTSof DNA-Ladder were basically consistent with those of Annexin V-FITC/PI apoptosis tests. In addition, both NU7026 and Wortmannin pretreatment elicited the higher expression of Bax mRNA in HL60 treated by 1, 4-benzoquinone with statistically significance (P < 0.05). However, p53 protein was not detected in HL60 cells as the western blot indicated.

CONCLUSIONInhibitors of DNA-PK, NU7026 and Wortmannin, promote p53-independent apoptosis induced by 1, 4-benzoquinone in HL60 cells.

Androstadienes ; pharmacology ; Apoptosis ; drug effects ; Benzoquinones ; toxicity ; Chromones ; pharmacology ; DNA-Activated Protein Kinase ; antagonists & inhibitors ; Flow Cytometry ; HL-60 Cells ; Humans ; Morpholines ; pharmacology ; RNA, Messenger ; Tumor Suppressor Protein p53

OBJECTIVETo investigate the association of human chorionic gonadotropin (HCG) level on day 3 of embryo culture with embryo development.

METHODSSpent culture media were collected from individually cultured embryos on day 3 of in vitro fertilization and embryo transfer (IVF-ET) cycles. HCG concentration in the culture media was measured using an ELISA kit and its association with embryo development was assessed.

RESULTSIn the 163 samples of embryo culture media from 60 patients, HCG was positive in 153 sample (93.8%) with a mean level of 0.85 ± 0.43 mIU/ml. The concentration of hCG in the culture media increased gradually as the number of blastomeres increased (F=2.273, P=0.03), and decreased as the morphological grade of the embryo was lowered (F=3.900, P=0.02).

CONCLUSIONELISA is capable of detecting HCG levels in spent culture media of embryos on day 3 of in vitro culture. The concentration of HCG in spent culture media is positively correlated with the status of early embryo development and implantation rate and thus serves as a useful marker for embryo selection in IVF-ET procedure.

Biomarkers ; chemistry ; Chorionic Gonadotropin ; chemistry ; Culture Media ; chemistry ; Embryo Transfer ; Embryonic Development ; Fertilization in Vitro ; Humans

Objective The aim of this prospective study was to evaluate the changes in the ovarian reverse after myomectomy based on serum anti-Mullerian Hormone (AMH) levels. Methods This is a prospective longitudinal observational study. Serum AMH levels were measured at the baseline and 2 day , 3months after myomectomy in 35 women aging from 36 to 45years.Follicle stimulate hormone(FSH) and luteal hormone(LH) were measured at the same time. 35 women of the same age with fibroid who did not undergo operation were selected as control group. Result (1)AMH level is (1.54 ± 0.95)ng/mL,(1.18 ± 0.77)ng/mL,(1.50 ± 0.58 )ng/mL at 0 day, 2 days and 3 months after operation. AMH level decreased significantly at 2 days after operation (P < 0.05) and increased gradually 3 months after operation, but showed no significant change (P > 0.05).(2) Significant differences in the serial change of AMH levels existed at each time point between myomectomy group and control group (P <0.05). No significant differences in FSH or LH levels existed at each time point. Conclusion AMH is may be superior to FSH or LH in evaluating the changing of ovarian reverse. The study suggests that myomectomy affect the ovarian function for up to 3 months post-operatively , and hemorrhage during and after operation may decrease serum AMH levels.

Objective To evaluate the value of the new ovarian function marker serum anti-Mullerian hormone (AMH)and to clarify the effect of hysterectomy on the ovarian function of the younger women.Methods The serum AMH,follicle stimulating hormone (FSH)and luteinizing hormone (LH)levels in 35 women suffered uterus benign lesion aged 36-45 years and 35 women suffered hysteromyoma without operation were measured at different time.And the ovarian arterial blood flow resistance index (RI ) was measured by Doppler ultrasound. Results Compred with before operation, the serum AMH levels of the patients in hysterectomy group 2 d and 3 months after operation were significantly decreased (P<0.05 or P<0.01).Compared with control group,the serum AMH levels of the patients in hysterectomy group 2 d and 3 months after operation were also decreased (P<0.05).The ovarian arterial blood flow RI of the patients in hysterectomy group 1 and 3 months after operation were increased compared with control group (P<0.05).There were no significant differences in the serum FSH and LH levels in each group between different time points (P>0.05).Conclusion Hysterectomy can affect the ovarian function,and the serum AMH level 3 months after operation is decreased and the ovarian arterial blood flow RI is increased.AMH is superior to FSH or LH in evaluating the changes of ovarian function.

Objective To investigate the association of human chorionic gonadotropin (HCG) level on day 3 of embryo culture with embryo development. Methods Spent culture media were collected from individually cultured embryos on day 3 of in vitro fertilization and embryo transfer (IVF-ET) cycles. HCG concentration in the culture media was measured using an ELISA kit and its association with embryo development was assessed. Results In the 163 samples of embryo culture media from 60 patients, HCG was positive in 153 sample (93.8%) with a mean level of 0.85 ± 0.43 mIU/ml. The concentration of hCG in the culture media increased gradually as the number of blastomeres increased (F=2.273, P=0.03), and decreased as the morphological grade of the embryo was lowered (F=3.900, P=0.02). Conclusion ELISA is capable of detecting HCG levels in spent culture media of embryos on day 3 of in vitro culture. The concentration of HCG in spent culture media is positively correlated with the status of early embryo development and implantation rate and thus serves as a useful marker for embryo selection in IVF-ET procedure.

Objective To investigate the association of human chorionic gonadotropin (HCG) level on day 3 of embryo culture with embryo development. Methods Spent culture media were collected from individually cultured embryos on day 3 of in vitro fertilization and embryo transfer (IVF-ET) cycles. HCG concentration in the culture media was measured using an ELISA kit and its association with embryo development was assessed. Results In the 163 samples of embryo culture media from 60 patients, HCG was positive in 153 sample (93.8%) with a mean level of 0.85 ± 0.43 mIU/ml. The concentration of hCG in the culture media increased gradually as the number of blastomeres increased (F=2.273, P=0.03), and decreased as the morphological grade of the embryo was lowered (F=3.900, P=0.02). Conclusion ELISA is capable of detecting HCG levels in spent culture media of embryos on day 3 of in vitro culture. The concentration of HCG in spent culture media is positively correlated with the status of early embryo development and implantation rate and thus serves as a useful marker for embryo selection in IVF-ET procedure.