ObjectiveTo investigate the inhibitory effect of gingerol， an active component of ginger， on Staphylococcus aureus （S. aureus） infection， and to preliminarily explore its mechanism related to extracellular vesicles （EVs）. MethodsS.aureus infection models were established in human umbilical vein endothelial cell （HUVECs）， Vero E6 cells， and C57BL/6 mice. Experimental groups included control， infection， and gingerol-treated groups. Bacterial load and VE-cadherin protein expression were detected using immunofluorescence and Western blot. EVs were isolated by size exclusion chromatography and characterized by transmission electron microscopy and nanoparticle tracking analysis. miRNA sequencing was performed on EVs. ResultsGingerol treatment significantly reduced the bacterial load in both in vitro and in vivo infection models and upregulated VE-cadherin expression. miRNA sequencing of EVs revealed that S. aureus infection upregulated the expression of hsa-miR-320c， while gingerol treatment reversed this abnormal expression. Bioinformatic analysis further predicted that the target genes of hsa-miR-320c were significantly enriched in cell junction-related pathways. ConclusionGingerol exhibits clear antibacterial and host-protective effects， by regulating hsa-miR-320c in EVs to maintain endothelial barrier integrity.

Objective:To analyze the level of interleukin-36(IL-36) family cytokines in peripheral blood, and explore the regulatory role of recombinant human IL-36α in monocyte function in patients with diabetic kidney disease(DKD).Methods:A total of 41 type 2 diabetes mellitus(T2DM) patients, 36 DKD patients, and 20 controls were consecutively enrolled. Plasma and peripheral blood mononuclear cells(PBMCs) were isolated. Enzyme-linked immunosorbent assay(ELISA) was used to measure plasma levels of IL-36α, IL-36β, IL-36γ, and IL-36 receptor antagonist(IL-36Ra). PBMCs were sorted, and real-time quantitative PCR was performed to detect the mRNA expression of IL-36 receptor subunits in monocytes. Monocytes were stimulated with recombinant IL-36α, and levels of cytotoxic molecules and cytokines in the culture supernatant were measured. Flow cytometry was used to assess the expressions of programmed death receptor-1(PD-1) and cytotoxic T-lymphocyte-associated protein 4(CTLA-4). Co-culture of monocytes with Vero cells was performed to evaluate monocyte cytotoxicity.Results:Plasma levels of IL-36α and IL-36β in the T2DM and DKD groups were significantly higher than those in the control group. The DKD group also showed higher plasma levels of IL-36α compared to the T2DM group( P<0.05). There were no significant differences in IL-36γ and IL-36Ra levels among the three groups( P>0.05). The mRNA expression of IL-36 receptor subunits in monocytes was comparable among the three groups( P>0.05). The DKD group had higher level of tumor necrosis factor-alpha(TNF-α) compared to the control and T2DM groups( P<0.05). The levels of PD-1 and CTLA-4 were lower in the DKD group than those in the control and T2DM groups( P<0.05). The proportion of monocyte-induced Vero cell death was significantly higher in the DKD group compared to the control and T2DM groups( P<0.05). After stimulation with recombinant human IL-36α, monocytes from DKD patients showed a significant increase in the secretion of granzyme B and TNF-α( P<0.05), as well as an increased proportion of monocyte-induced Vero cell death( P=0.024). Conclusion:In DKD patients, elevated IL-36α and granzyme B levels in monocytes enhance monocyte function.

In recent years, adaptive laboratory evolution (ALE) has emerged as a powerful tool for basic research in microbiology (e.g., molecular mechanisms of microbial evolution) and efforts on evolutionary engineering of microbial strains (e.g., accelerated evolution of industrial strains by bringing beneficial mutations). The ongoing rapid development of next-generation sequencing platforms has provided novel insights into growth kinetics and metabolism of microbes, and thus led to great advances of this technique. In this review, we summarize recent advances in the applications of long-term and short-term ALE techniques mainly for microbial strain engineering, and different modes of ALE are also introduced. Furthermore, we discuss the current limitations of ALE and potential solutions. We believe that the information reviewed here will make a significant contribution to further advancement of ALE.
High-Throughput Nucleotide Sequencing ; Laboratories ; Mutation

Objective To explore the relationship between fibroblast growth factor 21(FGF21) and islet β cell function in pregnant women with different glucose tolerance status.Methods A total of 441 pregnant women were selected in this study from our hospital.Their 50 g GCT at 24~28 gestational weeks were all positive.One week later,all the subjects were treated with 75 g OGTT,and divided into three groups according to their test results:GDM group (n=228),GIGT group (n=112) and GNGT group (n=91).Serum FGF21 level was tested by ELISA.Islet β cell function was evaluated by HOMA-IR,ISI-Matsuda,HOMA-IS,Stumvoll first,second phase secretion and ISSI.The correlation between FGF21 and islet β cell function was evaluated by Pearson correlation analysis.Results (1) BMI,0 h,1 h,2 h,3 hPG and 1 h,2 h,3 hIns were higher in GDM group and GIGT group than in GNGT group,and highest in GDM group (P<0.05 or P<0.01).SBP,DBP,0 hIns and HbA1c were higher in GDM group than in GNGT group and GIGT group(P<0.05 or P<0.01);(2)From GNGT group,GIGT group to DGM group,the levels of FGF21[(101.74±20.40) vs (137.93±25.52) vs (185.69±31.61) ng/L]and HOMA-IR[1.74(0.91,2.85) vs 2.39(1.31,4.87) vs 3.38(2.19,6.75)]were increased,while ISI-Matsuda[(58.74±15.68) vs (41.62±15.65) vs (39.73±18.98)],HOMA-IS[(157.69±88.41) vs (144.35±78.98) vs (107.30±87.23)]and ISSI[(72253.55±15167.53) vs (42313.91±7112.47) vs (30032.50±11500.24)]were decreased (P<0.05 or P<0.01).The stumvoll first,second phase secretion were lower in GDM group than in GNGT group and GIGT group(P<0.01),but there was no statistical significance between GNGT and GIGT group(P>0.05).(3)Pearson correlation analysis showed that FGF21 was positively correlated with HOMA-IR(r=0.255,P=0.030) and was negatively correlated with ISI-Matsuda,HOMA-β,Stumvoll first,second phase secretion and ISSI(r=-0.289,-0.256,-0.224,-0.230,-0.277,P=0.019,0.037,0.045,0.040,0.023).Conclusion Along with the worsening of glucose metabolic damage,the FGF21 level is increased gradually.FGF21 is related to islet β cell function,and may enroll in the occurrence and development of GDM.

Objective:To investigate the efficacy and safety of insulin glarsine combined with sitagliptin in elderly type 2 diabetic patients whose blood glucose levels were inadequately controlled by oral anti-diabetic drugs ( OAD) . Methods: In the open-labeled, randomized and parallel study, 98patients (≥60 years) were randomly divided into two groups: insulin glargine/sitagliptin combina-tion group (n=52, the observation group ) and insulin Aspart 30 injection group (n=46, the control group). The dose was adjusted according to the blood glucose. The fasting blood glucose (FBG), 2h postprandial blood glucose (2hPBG), glycosylated hemoglobin (HbA1c), the incidence of hypoglycemia and body mass index (BMI) after the 12-week treatment were compared between the two groups. Results:The fasting glucose and the incidence of hypoglycemia in the observation group were lower than those in the control group (P<0. 05). There were no significant differences in 2h postprandial blood glucose, HbA1c and BMI between the two groups (P>0. 05). Conclusion:The treatment of insulin glargine combined with sitagliptin is safe, effective and convenient in elderly type 2 diabetes patients with poor glycemic control. By diabetic education, the lower incidence of hypoglycemia treatment will be a better choice for elderly type 2 diabetic patients.

Objective To observe the effects of different dosages of granulocyte-macrophage colony-stimulating factor (GM-CSF) on generating the routine bone marrow dendritic cells, and supply suitable dosage of GM-CSF on preparation of dendritic cell vaccines used for different purpose. Methods Using low (5 ng/ml) and conventional (20 ng/ml) and high dosage( 50 ng/ml ) of GM-CSF combined interleukin-4 ( IL-4 ) to induce murine bone marrow dendritic cells were performed, The phenotypes (CD_(11c), CD_(80), CD_(86)) and functional properties of the DC were compared by FACS analysis and MLR. Results The DC induced by low dosage of GM-CSF were immature DC, expressing low CD_(11c), CD_(80), and CD_(86). DC induced by conventional dosage were functional mature, expressing higher CD_(11c), CD_(80), CD_(86), which could induce allogeneic T lymphocyte responses. DC induced by the high dosage GM-CSF were the most phetotypicaUy and functional mature cells, expressing the highest CD_(11c), CD_(80) CD_(86), which could induce the strongest allogeneic T lymphocyte responses. Conclusion The dosages of GM-CSF affect the maturation stage of dendritic cells. Low dosage of GM-CSF generated immature dendritic cells, but conventional dosage and high dosage generated mature dendritic cells. DC generated through high dosage of GM-CSF were the most mature in phenotype and function.