Objective To study the effects of estrogens ( 17α-ethynylestradiol and 17β-estradiol ) on the ATP-sensitive potassium channel ( KATP ) activities in the enzymatically ( collagenase and protease ) isolated ICR mouse ventricular cardiomyocytes using inside-out and cell-attached patch clamp techniques .Methods In the inside-out patch configurations, the two estrogens reduced the K ATP channel activities in a dose-dependent manner at -60 mV holding potential ( HP) .Results The half-maximal inhibitory concentration ( IC50 ) of the 17α-esthynylestradiol and the 17β-estradiol were 0.3μmol/L and 0.1 nmol/L, respectively.However, these agents had no effects on the pinacidil-and DNP-induced KATP channel activities in the cell-attached patch configuration.However, the inhibitory effect of KATP channel activities by the 17-estrogens or the anti-estrogens in the excised inside-out patch configuration were abolished by 1 pmol/L PDBu.Conclusions The results of this study demonstrate that some estrogens decrease the K ATP channel activity acting on the inside of the isolated cardiomyocytes , and play a protective role to cardiomyocytes .

Objective To study the effects of different transport protein on the transport of 7,4'-dihydroxyflavone (7,4'-DHF) and its metabolite (7,4'-DHF-S) in Caco-2 cell model.Methods Ultra performance liquid chromatography was employed to determinethe content of 7,4'-DHF and 7,4'-DHF-S incubation buffer,their structures were identified by LC-MS/MS.Bidirectional transport of Caco-2 cells model was used to investigate the influence of ko143 (the inhibitor of BCRP) and MK571 (the inhibitor of MRP2) on the transport of 7,4'-DHF and 7,4'-DHF-S,respectively.Results Metabolic product of 7,4'-DHF in Caco-2 monolayer cell was identified as one monosulfate;PDR of 7,4'-DHF was (1.43 ± 0.11),PDR of ko143 and MK571 on the apparent permeability of 7,4'-DHF was (1.59 ± 0.04) and (1.48 ± 0.07) (P ＞ 0.05);PDR of 7,4'-DHF-S was (1.60 ± 0.06);ko143 could significantly reduce the apparent permeability of 7,4'-DHF-S,and the PDR was (0.23 ±0.03) (P ＜ 0.01);MK571 had no significant effect on the apparent permeability of the 7,4'-DHF-S,and the PDR was (1.51±0.04) (P ＞ 0.05).Conclusion Caco-2 cells can mediate the suffonated reaction of 7,4'-DHF;7,4'-dihydroxyflavone sulfonated combination product may be a substrate for BCRP.

Objective To investigate the protective effect of nimodipine on neuron of the rats with focal cerebral ischemia-reperfusion injury and the expressions of Bax and Bcl-2,and to clarify their mechanisms.Methods The focal cerebral-ischemia reperfusion model was induced by the middle cerebral artery occlusion(MCAO)method. 30 male Wistar rats were randomly divided into sham operation,model,and nimodipine groups(n=10).The neurological deficit score was performed after 2 h ischemia following 2 h reperfusion.The infarction was observed by TUNEL staining and the expressions of Bax and Bcl-2 were detected by SP immunohistochemistry method. Results Compared with model group, the number of apoptotic cells of the rats in nimodipine group was significantly decreased(P<0.05),the expression of Bax was significantly decreased (P<0.05),and the Bcl-2 expression was increased significantly(P<0.05).The morphological examination showed that the neurons of the rats in model group had serious necrosis and edema while the number of dead cells in nimodipine treatment group was reduced and the edema was improved.Conclusion Nimodipine has a protective effect on brain tissue of the rats with focal cerebral ischemia-reperfusion inj ury, which is closely related to the down-regulation of Bax and up-regulation of Bcl-2 and inhibition of the apoptosis of neuron.

Objective To investigate the metabolic profile of chrysin in SULT1A3 and human small intestine S9.Methods After incubation of chrysin using in vitro SULT1A3 and human small intestine S9 system, high-performance liquid chromatography was utilized to determine the sulfates of chrysin.Mass spectrum(MS) were employed to elucidate the structures of metabolite.Results In the SULT1A3 with PAPS, Km were (3.06 ±1.04) and (0.41±0.06) μM, Vmax were (12.13 ±1.30) and (6.72 ±1.61) nmol/(min· mg), Vmax/Km were 3.96 and 16.39 mL/(min· mg), respectively.In the human small intestine S9 with PAPS, Km were (1.92 ±0.35) and (0.01 ±0.00) μM, Vmax were (0.52 ±0.02) and (0.08 ± 0.02) nmol/(min· mg), Vmax/Km were 0.27 and 8.00 mL/(min· mg).The metabolic behavior of chrysin in SULT1A3 and human small intestine S9 both were followed biphasic kinetics.The sulfation of chrysin in SULT1A3 showed a significant correlation with that in human small intestine S9(R2 =0.985).Conclusion The result indicates that SULT1A3 is the major enzyme to the metabolism of chrysin, human small intestine may be the main metabolic organs of chrysin.

Objective To investigate the effects of tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis (RA) curative effect and peripheral blood of patients with IL-6, effects of IL-10 and TNF- alpha. Methods 80 patients with RA from February 2014 to August 2016 in our hospital were retrospectively analyzed. According to the different treatment methods by for the observation group, the control group (40 cases). To compare and analyze the curative effect before and after short-term treatment and treatment of patients in the 2 groups were the main clinical symptoms and signs of change and ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index. Results The total effective rate of observation group (92.50％) was significantly higher than the control group (77.50％), the difference was statistically significant (P<0.05), 2 groups of patients before treatment of main clinical symptoms and signs, there was no significant difference, after treatment, clinical symptoms and signs of 2 groups of patients were compared with those before treatment was statistically significant to alleviate the differences (P<0.05), and the observation group of main clinical symptoms and signs were lower than the control group and the remission (down), the difference was statistically significant (P<0.05).Before ESR, the treatment of 2 patients with RF, CRP, TNF- IL-6, alpha, IL-10 index level, no statistical significance, the 2 groups after treatment in patients with ESR, RF, CRP, IL-6, TNF- alpha, IL-10 index level than before treatment with significant difference (P<0.05), and the observation group ESR, RF, CRP, TNF- alpha, IL-6 average index of water compared with the control group decreased significantly, IL-10 index increased significantly than the control group. The difference was statistically significant (P<0.05). Conclusion Tripterygium glycosides combined with methotrexate in treatment of rheumatoid arthritis And it can effectively inhibit the expression of peripheral serum L-6 and TNF- alpha, promote the expression of IL-10 and alleviate the inflammatory reaction

Objective To investigate the kinectics characteristics of sulfation of apigenin mediated by SULTIA3.Methods After incubation of apigenin using in vitro SULT1A3 system, high-performance liquid chromatography was utilized to determine the sulfates of apigenin.Mass spectrum(MS) were employed to elucidate the structure of metabolite.The program GraphPad Prism 5 was used to perform the kinetic characterization of SULT1A3 catalyzed metabolism of apigenin.Results A liner calibration curve for the assay of apigenin was validated in the range of 0.15625 ~30 μM with the recoveries of at least 80% and intra-day and inter-day RSD of less than 15%.Metabolic product of apigenin and SULT1A3 in the incubated system was identified one monosulfate.The metabolic behavior of apigenin in SULT1A3 was followed substrate inhibition kinetics.Apparent kinetic parameters of metabolism of apigenin by SULT1A3, Kmwas(0.355 ±1.04) μM and Ksi was(23.62 ±0.06) μM,Vmax was(65.71 ±1.30) nmol/(min? mg),Vmax/Km was 185.10 mL/(min? mg).Conclusion SULT1A3 can mediate the binding of apigenin sulfonated reaction, and the character of enzymatic kinetics shows substrate inhibition.Sulfation of apigenin mediated by SULTIA3 may play an important role in phaseⅡmetabolic in vivo.

Objective To establish a high-throughput evaluation model for anaphylactic reactions; To screen and identify potential anaphylactogens from TCM monomeric compounds.MethodsCell model of stably expressed MrgX2 was established. Recombinate plasmid pmCherry-C1-MrgX2 was transfected to HEK293 to establish cell line for screening model. MrgX2 agonist and antagonist were used to identify the validation and stability of the cell line. A small library consisting of 180 compounds was profiled by using a cell-based calcium mobilization assay to find novel compounds targeting the MrgX2 receptor. EC50 test, IC50 test, specificity validation and cytotoxicity evaluation were carried out to detect the function of the positive agonist.ResultsThe EC50 of C48/80 to MrgX2 model was 2.7 μg/mL and the IC50 of 2-APB (evoked by 10 μg/mL C48/80) was 46.29 μmol/L. The first generation cell model of MrgX2 was similar to the 20th generation, and the Z factor of MrgX2 cell model was 0.78. In the primary screening for agonist, isoliensinine was identified as a novel agonist targeting receptor MrgX2 with an EC50 of 4.5 μmol/L and IC50 of39.47 μmol/L. Moreover, isoliensinine was validated to activate MrgX2 receptor specifically without cytotoxicity. Conclusion A high-throughput evaluation method for anaphylactic reactions can be established in vitro through calcium mobilization assay. A potential anaphylactogen isoliensinine is identified and validated.

Objective To investigate risk factors of the prognosis of patients with severe fever with thrombocytopenia syndrome (SFTS).Methods From May 2012 to July 2014,17 cases of severe fever with thrombocytopenia syndrome in Renmin Hospital of Wuhan University were treated.Clinical data including history of epidemiology,clinical manifestations,complications,physical examination and laboratory test results on admission and the third day after admission were retrospectively analyzed and compared with the death group and recovery group by application of Spearman correlation analysis.Results Elderly male patients with neuropsychiatric symptoms,or abnormal liver function,or abnormal blood clotting function had higher risk of the poor prognosis.In SFTS patients,AST,ALT was significantly increased,AST 539 U/L (229.73,545.4) U/L (r =0.597,P =0.015) was a risk factor affecting prognosis.Elevated blood ammonia indicated serious liver dysfunction and neurological dysfunction which were manifested as irritability,delirium,and trembling limbs.In SFTS patients,platelets were significantly decreased accompanied with mouth ulcers / bleeding gums,gastrointestinal bleeding.PLT 24.88 × 10 9/L-1 (12.75,35.00) ×10 9/L-1 (r=0.557,P=0.005) or APTT 86.06 s (66.88,114.18) (r=0.798,P=0.001) or D-dimmer 9.79 mg / L (4.09,16.51) mg/L (r =0.597,P =0.015) are risk factors affecting poor prognosis.Conclusions On the third days after admission,AST,WBC,PLT,APTT,Ddimmer are risk factors for prognosis of patients with severe fever with thrombocytopenia syndrome infected by a novel bunyavirus.

Objective To investigate the prevalence of healthcare-associated infection (HAI)in a hospital in 2013-2015,and provide reference for the improvement of HAI management.Methods Prevalence rates of HAI among patients in the First Affiliated Hospital of Zhengzhou University in 2013 -2015 were investigated with cross-sec-tional survey method,prevalence rates of HAI and constituent ratios of HAI sites in patients of different genders and different age groups were analyzed.Results In 2013 -2015,29 605 patients should be investigated,29 581 (99.92%)were actually investigated.Prevalence rates of HAI in 2013-2015 were 2.83%,2.14%,and 1 .73%,re-spectively,difference was significant(χ2 =27.521 ,P <0.01),which showed a downward trend year by year.Preva-lence rates of HAI in male was significantly higher than that of female (2.47% vs 1 .98%,χ2 =7.954,P <0.01). The prevalence rates of HAI in the age dimension was roughly U-shaped distribution in 2013 -2015.Prevalence rates of HAI in ≥70 age group was the highest(4.15%),in 0 -30 age group decreased with age,in > 30 age group increased with age.The top three sites for prevalence of HAI were lower respiratory tract (45.67%),upper respiratory tract (9.92%),and urinary tract (8.52%).Conclusion The prevalence rates of HAI decreased year by year in 2013-2015,which suggests that HAI prevention and control measures are effectively implemented.

Objective:To analyze the variation and evolution characteristics of hemagglutinin (HA) and neuraminidase (NA) genes of influenza B virus circulating in Jining from 2017 to 2020.Methods:Throat swab specimens were collected from patients with influenza-like symptoms in sentinel hospitals and influenza outbreaks in Jining from 2017 to 2020 and tested for influenza B virus nucleic acid. After virus isolation, 20 representative strains of influenza B virus were selected to sequence the full length of HA and NA genes. Phylogenetic trees were constructed and the molecular characteristics were analyzed using bioinformatics software. Results:A total of 4 575 specimens were collected and 842 of them were positive for influenza virus, including 398 (8.7%, 398/4 575) influenza B virus-positive specimens. The positive rate of influenza B virus was 47.27% (398/842). The isolated influenza B virus strains of Victoria (BV) and Yamagata (BY) lineages from 2017 to 2020 shared 98.7%-98.8% and 98.5%-99.1% homology in HA gene with vaccine strains, respectively. The BV lineage strains isolated from 2018 to 2020 belonged to Victoria clade 1A branch and the BY lineage strains isolated from 2017 to 2018 belonged to Yamagata clade 3 branch. Mutations were detected in several antigenic sites, but not in the sites related to NA inhibitor resistance. Conclusions:Mutations in several antigenic sites caused antigenic changes in influenza B virus of BV and BY lineages, which might be related to the outbreaks of influenza B virus infection in Jining during 2017 to 2020.