Sennoside A (SA), a typical prodrug, exerts its laxative effect only after its transformation into rheinanthrone catalyzed by gut microbial hydrolases and reductases. Hydrolases have been identified, but reductases remain unknown. By linking a photoreactive group to the SA scaffold, we synthesized a photoaffinity probe to covalently label SA reductases and identified SA reductases using activity-based protein profiling (ABPP). From lysates of an active strain, Bifidobacterium pseudocatenulatum (B. pseudocatenulatum), 397 proteins were enriched and subsequently identified using mass spectrometry (MS). Among these proteins, chromate reductase/nicotinamide adenine dinucleotide (NADH) phosphate (NADPH)-dependent flavin mononucleotide (FMN) reductase/oxygen-insensitive NADPH nitroreductase (nfrA) was identified as a potent SA reductase through further bioinformatic analysis and The Universal Protein Resource (UniProt) database screening. We also determined that recombinant nfrA could reduce SA. Our study contributes to further illuminating mechanisms of SA transformation to rheinanthrone and simultaneously offers an effective method to identify gut bacterial reductases.

Objective:To explore disorienting dilemmas during postgraduate study period experienced by postgraduates with master of nursing specialist who participated in the national postgraduate entrance examination and provide a basis for teaching reform based on transformative learning theory.Methods:Using the purposive sampling method and the maximum difference method, 23 full-time masters of nursing specialist who participated in the national postgraduate entrance examination from 3 universities of Jiangsu Province were selected as the research objects from March to June 2019. The descriptive qualitative research method was used to conduct semi-structured interviews, and the traditional content analysis method was used to analyze the interview data.Results:Four themes of disorienting dilemmas of masters of nursing specialist were extracted: the expected hypothesis of challenges of new environment in course learning, a gap between scientific research ability and training target, adjustment of self-positioning in clinical practice and insufficient preparations for graduation of master of nursing specialist.Conclusions:Master of nursing specialist have disorienting dilemmas in course study, scientific research, clinical practice and graduation preparation, and nursing educators can use this as a trigger point to carry out teaching reform based on transformative learning theory.

Objective:To evaluate the effects of different growth conditions on E.faecalis growth in the microfluid chip and the penetration of E.faecalis into microtubes.Methods:Four units on the chip were randomly selected as control,BHI,nutrient-Tdeprived (PBS) and pH 10 groups.The growth of E.faecalis was monitored by microscope for a period of 72 h after the suspension of E.faecalis had been added into the chip.Results:The microscopic analysis showed a distinct variation in the growth rate and morphological feature under different experimental conditions.he depth of bacterial penetration was significantly greater in BHI group.Conclusion:This study demonstrated that environmental changes can significantly influence the growth and penetration of E.faecalis into the micro tubes.

Objective To evaluate the value of the neutrophil-lymphocyte ratio (NLR) in elderly type 2 diabetic patients (T2DM) with coronary heart disease (CHD).Methods We performed a retrospective observational study on 228 patients undergoing coronary angiography in Guizhou Provincial People's Hospital from April 2014 to July 2015.Patients were divided into three groups:the simple T2DM group (n=77),simple CHD group (n=72),and T2DM complicated with CHD group (n=79).Meanwhile,70 healthy elderly subjects served as the control group.The white blood cell count,high-sensitivity C-reactive protein (hs-CRP) and other clinical and laboratory parameters were collected,and NLR was calculated.Risk factors for CHD in T2DM patients were determined by logistic regression analysis.Multiple stepwise regression analysis was adopted to identify factors influencing NLR.Results The white blood cell count,neutrophil count,NLR and hs-CRP level in the simple T2DM,CHD,and T2DM+CHD groups were higher than in the control group [(7.48 1.81) 109/L,(7.72± 1.89) 109/L,(7.98±2.12) 109/L vs.(6.22± 1.61) 109/L;(4.49±1.38) 109/L,(4.88±1.56) 109/L,(5.35±1.40) 109/L vs.(3.52±0.84) 109/L;(2.84± 0.77),(3.07±0.79),(3.34±0.83) vs.(1.58±0.42);(2.92±0.65) mg/L,(3.20±0.86) mg/ L,(4.98±1.10) mg/L vs.(1.105±0.23) mg/L;respectively,P＜0.05 or P＜0.01].The lymphocyte count in the simple T2DM,CHD,and T2DM+CHD groups were lower than in the control group [(1.57±0.41) × 109/L,(1.58±0.40) × 109/L,(1.61±0.48) × 109/L vs.(2.22± 0.51) × 109/L,P＜0.05].NLR and hs-CRP levels in the T2DM+CHD group were higher than in the former two groups (all P＜0.05).Pearson correlation analysis showed that NLR was positively correlated with the Gensini score and hs-CRP level (r=0.7455 and 0.7792,both P＜0.01).Logistic regression analysis showed that NLR,hs-CRP levels and glycosylated hemoglobin A1c (HbA1c) were the risk factors for CHD in T2DM patients (OR=4.331,3.997 and 2.928,all P＜0.05).Multiple stepwise regression analysis showed that NLR was positively correlated with fasting plasma glucose,HbA1 c levels and systolic blood pressure (β' =0.3133,0.4720 and 0.3069,all P＜0.05).Conclusions NLR may be a valuable predictive factor for CHD in elderly T2DM patients.

Objective To investigate the expression of TLR9 mRNA of peripheral blood mononuclear cells (PBMCs) in patients with acute pancreatitis .Methods Fifty two AP patients with the disease duration in 24 h were collected ,peripheral EDTAK2 coag‐ulation vein blood were collected on the first ,third and fifth day ,then plasma were cryop reserved to detect pancreatic elastase , proinflammatory cytokines and anti‐inflammatory cytokines .Then the peripheral EDTAK2 coagulation vein blood two to three months after treatment were collected in the same method to undertake these tests ,and act as the reference level value .Peripheral blood was collected from 36 acute pancreatitis patients .Three months later ,peripheral blood was collected again from these 36 peo‐ple as controls .PBMCs were isolated by Ficoll‐Hypaque gradient centrifugation .RT‐PCR was adopted to determine the relative con‐tent of the expression of TLR9 mRNA of the PBMCs .Results The relative content of expression of TLR9 mRNA were significant‐ly up‐regulated in the patients with acute pancreatitis ,compared with that of controls (P<0 .05) .The up‐regulated expression of TLR9 mRNA was related to expression of TNF‐a and IL‐1 .Conclusion The up‐regulated expression of TLR9 mRNA in acute pan‐creatitis patients indicates that infective factors might be mediated by TLR 9 .

Objective To establish and assess the high-birth-weight offspring model of the diabetic rat induced by stueptozotocin,and the long-term metabolic impact of maternal hyperglycemia of those offsprings.Methods Streptozotocin (STZ,25 mg/kg) was given to Wistar rats (G group,n =14) once intraperitoneally to induce maternal hyperglycemia model (blood glucose between 10-20 mmol/L),and there still had a number of rats defined as severe hyperglycemia model group (SG group,n =5).The Control group (C group,n =7) were given the same volume citrate buffer solution.The body weight and blood glucose were recorded,and the lavaging glucose tolerance test (LGTT) was performed by a glucose meter in the gestation.The offsprings were corresponding allocated into 2 groups,and the birth weight were recorded.All the offsprings were observated body weight,blood glucose blood pressure (male rats only),and so on.Results (1) The blood glucose of G group (16.8 ±5.4 mmol/L) and SG group (20.5 ±5.6 mmol/L) were increased significantly as compared with C group (7.0 ± 1.4 mmol/L) 5 days after the model was established (P ＜ 0.01) ; and the average blood glucose of G group (16.6 ± 3.4 mmol/L) and SG group (23.8 ± 1.5 mmol/L) increased too as comparede with C group (5.8 ± 1.1 mmol/L),the difference was significance according to statistics (P ＜ 0.01).(2) According to the LGTT result,which operationed on generation day 4 and day 10,the blood glucose of every time point of G group were increased significantly as compared with C group (P ＜ 0.01).(3) The male and female birth weight of G group was remarkably higher than the C group and the SG group (P ＜ 0.05),and the blood glucose of SG /G/C group was (6.5 ±1.2) mmol/L,(4.1 ± 0.8) mmol/L,(4.1 ± 0.8) mmol/L respectively,according to the statistics results,the difference between SG group and G/C group respectively both remarkable (P ＜ 0.05).(4) The body weight,Lee's index,fat weight,and the fat weight of mass ratio in C group mother rats after lactation presented dressed compared with the SG group (P ＜ 0.05),and so as to the G group compared with the SG group (P ＜ 0.05).(5) In the female offsprings of G group,the birth weight was remarkably increased compared with the C group (P ＜ 0.05) ; the body weight of the female offsprings presented an increased trend compared with the C group since the 12 weeks,but had no statistical significance; there were significant differences of body weight between G group and C group since 15 weeks (P ＜ 0.05),and the trend kept up until 26 weeks; in the male offsprings of G group,the body weight on birth day and 4 weeks had a marked rise compared with the C group (P ＜ 0.05) ; and from then on,the body weight of the male offsprings presented an increased trend compared with the C group,but had no statistical significance until 26 weeks (P＞0.05).(6) In G group,the blood glucose on 30 min and 60 min of LGTT in female offsprings were increased than the C group since 20 weeks (P ＜ 0.05) ; the blood glucose of LGTT (30 min) still had a marked rise until 24 weeks (P ＜ 0.05) ; in G group,the blood glucose on 30 min of LGTT in male offsprings was remarkably incrcascd than the C group since 16 weeks (P ＜0.05) ; the blood glucose of LGTT (30 min) still had a marked rise until 24 weeks (P ＜ 0.05).(7) The blood pressure of male offsprings in G group had a marked rise on 12 weeks compared with the C group (P ＜ 0.05) ; from then on the blood pressure of G group kept up a rise trend until 26 weeks,but had no statistical significance (P ＞0.05).Conclusion The diabetic high-birth-weight rat model could be duplicated with STZ (25 mg/kg) once intrapertoneally on the first day of gestation,which were observed some evidently metabolic changes in weight,glucose tolerance and blood pressure.These results could represent an forward step in the clinical study of human gestational diabetes mellitus and their macrosomia babies,which may suffer some metabolic disease in their later life.

Objective To study the roles of advanced glycation end products and its receptor on fetal brain injury of gestational diabetes mellitus (GDM) rats.Methods Twenty one adult pregnant Wistar rats were administered streptozotocin (STZ) intraperitoneally to induce GDM rats model.The fourteen pregnant rats were divided into two groups according to the fasting glucose on the 3rd day of pregnancy:severe GDM group with the fasting glucose ＞ 16.7 mmol/L and mild GDM group with the fasting glucose between 6.7 - 16.7 mmol/L Another seven pregnant rats were chosen as the severe GDM and intervention with micronutrient group,receiving gavage with micronutrient during the whole pregnancy.Five control rats received the same volume of citric acid buffer.All the pregnant rats were tested fasting glucose from the tailvein and their weight on the pregnant day 3,13 and 19.Maternal serum levels of AGE were measured by ELISA and RAGE levels in the embryonic brain tissues were tested by immunohistochemistry.Results ( 1 ) There was no statistically significant difference of pre-pregnancy fasting glucose level among all groups (P ＞ 0.05 ).The fasting glucose levels on the 3rd day and the mean fasting glucouse level of pregnancy in the severe GDM group and the severe GDM and intervention with micronutrient group were higher than those of the control group ( P ＜0.05 ).And there was no significant difference between the severe GDM group and the severe GDM and intervention with micronutrient group (P ＞0.05 ).(2)The serum AGE levels in the severe GDM group and the mild GDM group were( 1037 + 38) ng/L and( 880 ± 34) ng/L respectively,with no significant difference ( P ＞ 0.05 ).The serum AGE levels in the control group and the severe GDM and intervention with micronutrient group were (857 ± 32 ) ng/L and (988 ± 37 ) ng/L,and the difference was statistically significant ( P ＜ 0.05 ).The serum AGE levels in the severe GDM and intervention with micronutrient group and in the mild GDM group had no significant difference ( P ＞ 0.05 ).( 3 ) The serum AGE levels in the severe GDM group,mild GDM group and the control group were positively associated with the mean glucose level of pregnancy ( r =0.603,P ＜ 0.05 ) and the grlucose on the 3rd day of pregnancy (r =0.704,P ＜ 0.05 ).(4)The fetal brain nerve cell number and morphology in the control group were normal.While in the mild GDM group fetal brain nerve cells decreased,the proliferation and swelling of glial cells were seen.In the severe GDM group and the severe GDM and intervention with micronutrient group,the fetal brain cells furtherly reduced,and large vacuole around the cells,deformation and debris of the cells were seen. Glial scar formation was visible in some fetal brain tissues.There was a few RAGE expression in the control fetal brain tissues.In the mild GDM group and the severe GDM group,RAGE expression increased significantly.And the RAGE expression intensity in the severe GDM and intervention with micronutrient group was between the severe and the mild GDM groups.Conclusions( 1 ) Abnormal fetal brain development of GDM rats was associated with the increase of maternal serum AGE and the enhancement of RAGE expression in fetal brain tissues,which suggested that AGE/RAGE pathway may play an important role in the fetal brain injury of GDM rats.(2) Micronutrients can reduce the brain damage of GDM fetuses.

OBJECTIVETo investigate the apoptosis of NK cells induced by the erythroleukemia cell line K562 in vitro.

METHODSPrimary NK cells isolated from the peripheral blood of healthy donors by magnetic-activated cell sorting were cultured with stem cell medium containing recombinant human interleukin-2 (rhIL-2). The NK cells and K562 cells were mixed and co-cultured at different E:T ratios for different time lengths. The apoptosis of NK cells and K562 cells were detected using PE-AnnexinV/7-AAD labeling and flow cytometry.

RESULTSThe purity of isolated NK cells reached (93.99∓4.22)%. At the same E: T ratio, the apoptotic rate of NK cells induced by K562 cells increased significantly with time. As the E:T ratio reduced, the apoptotic rate of the NK cells increased and their cytotoxic activity against K562 cells was attenuated.

CONCLUSIONK562 cells can induce the apoptosis of activated NK cells, which is one of the probable mechanisms of immune escape of tumors.

Apoptosis ; Cytotoxicity, Immunologic ; Humans ; K562 Cells ; Killer Cells, Natural ; cytology ; immunology ; Tumor Escape