Animals ; Endothelium, Vascular ; Humans ; Lipid Metabolism ; Metabolic Diseases

Objective:To explore bromodomain and extra-terminal (BET) inhibition in the regulation of vascular endothelial cells activation and early atherosclerosis formation and its potential molecular mechanisms.Methods:1.Human umbilical vein endothelial cells (HUVEC) and mouse heart endothelial cells (MHEC) were isolated,and tumor necrosis factor α (TNFα) was used to activate in flammatory genes transcription in the presence or absence of JQ1,a specific BET inhibitor.The groups are as follows:(1)Normal control group;(2) TNFα(25 ng/mL)group;(3) TNFα+JQ 1 group.The gene mRNA and protein expression of inflammatory cytokines were measured by both real-time PCR and flow cytometry (FCM).2.LDL receptor-deficient (LDLR-/-) mice were randomly divided into 2 groups:JQ1 group (n=8,JQlintraperitoneal,50 mg/kg,daily) and control group (n=8,DMSO,daily).After 8 weeks feeding with high cholesterol diet,vascular cell adhesion molecule-1 (VCAM-1) expression in aortic arch was measured by immunohistochemistry.The activity of nuclear factor kappa B (NF-κB) signaling was monitored by 5XκB luciferase reporter assay in HEK293.Results:TNFα dramatically induced the mRNA and protein expression of inflammatory genes and JQ 1 significantly downregulated the induction of them (E-selectin,P-selectin,VCAM-1,IL-8)(P＜0.01).Immunohistochemistry detection indicated that JQ1 significantly downregulated the expression of VCAM-1 in aortic arch induced by 8 weeks high cholesterol diet feeding comparing to control group.In addition,BET bromodomain inhibition downregulated TNFα upregulated NF-κB transcriptional activity (P＜0.01).Conclusions:Our study demonstrated that BET bromodomain was involved in NF-κB mediated inflammatory genes expression;inhibition of BET bromodomain suppressed vascular endothelial activation in vitro,and attenuated early atherogenesis in vivo.

Tablet 912 - II. active ingredients from Fructus Tribuli, was applied for treating 40 cases of cerebral vascular disorders. After a course of 3 months, it was found that the tablet markedly improved the clincal symptoms, lowered the blood viscosity, regulated platelet functions, ameliorated the microcirculation, increased the cerebral blood flow. When compared with Carlan tablet, the action of improving clinical manifestations was basically the same, while for that of correlated clinical indices, 912-II is better than the latter tablet.

Objective To investigate the regularity and significance of serum interleukin-6(IL-6) levels in the patients with sepsis.Methods Forty sepsis patients with APACHEⅡ scores over 12 and 30 normal controls were involved in the study.The IL-6 levels in serum were detected with radio-immunity analysis assay and the correlations among the IL-6 levels,the prognosis and APACHEⅡ score were investigated.Results In the patients with sepsis,the average level of IL-6 in serum was(152.02?55.77) pg/ml,which was significantly higher than that of the control group [(85.79?30.96) pg/ml](P

Abstract: Objective To investigate the prevalence and pathogenic characteristics of Yersinia enterocolitica infection in children with diarrhea under 5 years of age in western Yunnan, and to provide a basis for the prevention and treatment of infectious diarrhea in children. Methods Feces were collected from under five-year-old children with diarrhea in the First Affiliated Hospital of Dali University from 2020 to 2021. Clinical information of the cases was also collected. Yersinia enterocolitica was isolated from the samples after cold enrichment on selective culture plates, and the pathogenic characteristics of Yersinia enterocolitica were analyzed by biological type and serotype and virulence gene detection. Results A total of 397 feces were collected. Seven strains of Yersinia enterocolitica were isolated in three samples, and the prevalence of Yersinia enterocolitica infection was 0.76% (3/397). Among the three positive samples, two Yersinia frederiksenii or Yersinia intermedia were isolated in specimen No. 212 , and five Yersinia enterocolitica were detected in specimens No. 24 and 226. Two Yersinia enterocolitica isolated from one sample were biological type 1A, and the virulence gene test results were ail-/ystA-/ ystB+ /yadA-/virF-, which were non-pathogenic Yersinia enterocolitica. Three Yersinia enterocolitica isolated from the other sample were biological type 3, serotype O∶3 (rfbc+), and virulence gene detection results were ail+/ystA+/ystB-/yadA+ /virF+, which were pathogenic Yersinia enterocolitica. While pathogenic Yersinia enterocolitica was detected from feces of children with diarrhea at 11 months of age with a infection rate of 0.50%(2/397). Conclusion Sporadic infection of pathogenic Yersinia enterocolitica was found in under five-year-old children in western Yunnan Province. It is necessary to strengthen the monitoring and research of Yersinia enterocolitica.

Embolization, Therapeutic ; Humans ; Hypertension, Portal ; therapy ; Portasystemic Shunt, Surgical

Objective:To express and purify hemoglobin receptor(HgbA) and its partial fragment(HgbAF) from Haemophilus ducreyi and to develop a sandwich ELISA for the detection of H.ducreyi infection.Methods:The HgbA,a hemoglobin-binding outer membrane protein of H.ducreyi and its partial fragment(HgbAF) were expressed by cloning the genes of hgbA and its 705bp fragment into pET30a and pET28a respectively,and the expressing products were purified from E.coli BL21 with Ni-NTA-His affinity chromatography.The polyclonal antibodies were developed by immunizing rabbits with the rHgbA and rHgbAF.The anti-rHgbA IgG and anti-rHgbAF IgG were purified respectively by saturated amine sulphate precipitation,and their immunoactivity with rHgbA and rHgbAF was tested by Westen blot and ELISA analysis.A Sandwich ELISA was developed for the detection of chinical infection of H.ducreyi using the specific polyclonal antibodies.Results:The HgbA and its partial fragment,HgbAF of H.ducreyi,were expressed and purified successfully by cloning their genes respectively.The results obtained by Western blot analysis showed that each of the antibodies could react with both antigens,rHgbA and rHgbAF.The results of the ELISA analysis showed that H.ducreyi strain was strongly positive,and all other bacteria,including H.influenzae and the bacteria known to relate to genital ulcers were negative.The results of the ELISA analysis showed that the minimum amount of rHgbA detected was 1.56 ng/ml and the minimum number of CFU of H.ducreyi detected was 2?105 cfu/ml in buffer and 1?106 cfu/ml in pus.Conclusion:HgbA and its partial fragment,HgbAF of H.ducreyi are expressed and purified successfully.The polyclonal antibodies developed by immunizing rabbits using rHgbA and rHgbAF could react not only with rHgbA and rHgbAF,but also with H.ducreyi specifically.They do not react with other bacteria,including H.influenzae and the bacteria known to relate to genital ulcers.So the ELISA based on the polyclonal antibodies was specific for the detection of H.ducreyi.Although the level of sensitivity of the ELISA may not be sufficient to detect H.ducreyi in all clinical specimens,further work to increase the sensitivity could potentially make this assay a valuable tool in areas where chancroid is endemic.

Objective To explore the therapeutic mechanism of kidney-reinforcing,blood-activating and phlegmresolving therapy for left ventricular fibrosis of spontaneously hypertensive rats (SHR).Methods Twenty SHR were randomly assigned to Chinese medicine group and model group.Additionally,ten Wistar-Kyoto(WKY) rats served as normal control group.After 12-week prevention,Masson staining method was used to measure the degree of fibrosis of left ventricular myocardial tissues,reverse transcription-polymerase chain reaction(RT-PCR) was used to detect Smad3 mRNA expression,and Western blotting method was used for the detection of Smad3 protein expression.Results The degree of left ventricular fibrosis myocardial tissue in Chinese medicine group was milder than that in the model group,and Samd3 protein and mRNA expression levels in Chinese medicine group were lower than those in the model group (P ＜ 0.05).Conclusion Kidney-reinforcing,blood-activating and phlegmresolving therapy can improve left ventricular fibrosis in SHR by inhibiting Smad3 expression.

Objective To explore the significance of nuclear factor-?B(NF-?B)activation in peripheral blood mononuclear cells(PBMC)during acute Kawasaki disease(KD).Methods Peripheral blood was collected from children with acute KD(n=30)and healthy age-matched children(n=20).PBMC were cultured in vitro and divided into 3 groups:naturally cultured blank control group,protein kinase C(PKC)activator stimulated phorbol 12-myristate 13-acetate(PMA)group and PMA plus NF-?B inhibitor treated PMA plus pyrrolidine dithiocarbamate(PDTC)group.Percentages of NF-?B activation were detected by immunohistochemistry.Results Under natural culturing,the percentage of cells with activated NF-?B was significantly higher in acute KD blank control group than that in healthy blank control group.The percentage of cells with activated NF-?B was significantly higher in acute KD PMA group than that in acute KD blank group and that in normal control PMA group,respectively(Pa0.05).Conclusions NF-?B activation in PBMC during acute KD is markedly increased,which suggests that NF-?B activation plays an important role in the formation of vasulitis and CAL in this disease.NF-?B activation in PBMCs in children with KD is regulated by the PKC signaling pathway and PDTC obviously inhibits the activation of NF-?B.J Appl Clin Pediatr,2009,24(1):35-37

Objective To study the telomerase expression in peripheral blood mononuclear cells(PBMCs) in children with acute Kawasaki disease(KD) and its clinical significance.Methods The PBMCs of 64 children with acute KD [25 cases of them with coronary artery lesions(CAL),while the rest without] from 2 months to 6 years old admitted into Jiangxi Children's Hospital from Mar.2005 to Dec.2008 and those of 52 sex-age-matched healthy children (healthy control group) from 5 months to 7 years old were all assayed by Roche telomerase polymerase chain reaction enzymelinked immunosorbent assay(PCR ELISA).WBC,ESR and CRP were also detected.SPSS 11.0 software was used to analyze the data.Results The telomerase expression frequency of PBMCs in children with KD was 32.8%(21/64 cases),while that in healthy control group was only 15.4%(8/52 cases),the difference between the 2 groups was significant (?2= 4.65,P0.05).There were no significant difference of WBC,ESR and CRP between the telomerase of PBMCs positive group and negative group.Conclusions The higher frequency of telomerase expression in peripheral blood lymphocytes might be related to the development and progression of KD.