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Objective@#To construct lentiviral vector of late endosomal/lysosomal adaptor, mitogen-activated protein kinase and mammalian target of rapamycin activator 2 (lamtor2) gene, and to explore its regulatory role on inflammatory response of macrophages after Klebsiella pneumoniae infection.@*Methods@#Two pairs of mouse lamtor2 short hairpin RNA (shRNA) were designed and sub-cloned into PLKO.1-puro to construct lentiviral vector, and were transfected into the murine RAW264.7 macrophage. There were two experimental groups including pLKO.1-puro shlamtor 2-1(sh1 group) and pLKO.1-puro shlamtor 2-2 (sh2 group), and the RAW264.7 cells transfected with non-treated pLKO.1-puro was set as control. The expession of lamtor2 were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot. The levels of inflammatory factors including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α secreted by the cells were detected by RT-qPCR. T test was used for comparison between groups.@*Results@#The recombinant lentiviral vector PLKO.1-shlamtor 2 transfected RAW264.7 cells successfully. The relative expressions of lamtor2 mRNA in the control group, the sh1 group and the sh2 group were 1.000±0.000, 0.596±0.125 and 0.120±0.080, respectively. The expression of lamtor2 in the sh2 group was significantly lower than that in the sh1 group (t=3.399, P=0.015), and they were both significantly lower than the control group (t=3.333 and 9.734, respectively, both P< 0.05). After infection with Klebsiella pneumoniae, expression levels of IL-1β (t=15.20), IL-6 (t=43.30) and TNF-α (t=12.67) were significantly higher than those in the control group (all P<0.01).@*Conclusion@#The lentiviral vector of lamtor2 can stably down-regulate the expression of lamtor2 gene in macrophages through RNA interference mechanism, which has a significant effect on the secretion of inflammatory factors of macrophages that are infected with Klebsiella pneumoniae.
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Objective To construct lentiviral vector of late endosomal／lysosomal adaptor, mitogen-activated protein kinase and mammalian target of rapamycin activator 2 ( lamtor2) gene, and to explore its regulatory role on inflammatory response of macrophages after Klebsiella pneumoniae infection.Methods Two pairs of mouse lamtor2 short hairpin RNA (shRNA) were designed and sub-cloned into PLKO.1-puro to construct lentiviral vector, and were transfected into the murine RAW264.7 macrophage.There were two experimental groups including pLKO.1-puro shlamtor 2-1(sh1 group) and pLKO.1-puro shlamtor 2-2 (sh2 group), and the RAW264.7 cells transfected with non-treated pLKO.1-puro was set as control.The expession of lamtor2 were detected by real-time quantitative polymerase chain reaction ( RT-qPCR ) and Western blot.The levels of inflammatory factors including interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-αsecreted by the cells were detected by RT-qPCR.T test was used for comparison between groups.Results The recombinant lentiviral vector PLKO.1-shlamtor 2 transfected RAW264.7 cells successfully.The relative expressions of lamtor2 mRNA in the control group, the sh1 group and the sh2 group were 1.000 ±0.000, 0.596 ±0.125 and 0.120 ±0.080, respectively.The expression of lamtor2 in the sh2 group was significantly lower than that in the sh 1 group (t＝3.399, P＝0.015), and they were both significantly lower than the control group ( t ＝3.333 and 9.734, respectively, both P ＜0.05).After infection with Klebsiella pneumoniae, expression levels of IL-1β( t ＝15.20), IL-6 (t＝43.30) and TNF-α(t＝12.67) were significantly higher than those in the control group (all P＜0.01).Conclusion The lentiviral vector of lamtor2 can stably down-regulate the expression of lamtor2 gene in macrophages through RNA interference mechanism , which has a significant effect on the secretion of inflammatory factors of macrophages that are infected with Klebsiella pneumoniae.
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Objective To analyze the differences of clinical manifestations and organ damage between patients with severe fever with thrombocytopenia syndrome(SFTS)and patients with tsutsugamushi disease,and to investigate the prognostic factors of SFTS.Methods The research was performed on 49 patients with SFTS and 16 patients with tsutsugamushi disease who visited the First Affiliated Hospital of Anhui Medical University from October 2014 to June 2017.The general information of patients including region,age,gender and clinical manifestations were evaluated.Blood routine,liver and kidney function,myocardial enzyme levels,lipase,amylase,electrolytes,C-reactive protein,procalcitonin,prothrombin time(PT)and activated partial thromboplastin time(APTT)were continuously monitored during the course of disease.T test was used for continuous variables of normal distribution,and non-parametric test was used for variables of non-normal distribution.Chi-square test was used for categorical variables.Results The mean age of SFTS patients was 62.1±15.5(ranging from 17 to 87 years)and the mean age of tsutsugamushi patients was 56.1±9.2(ranging from 47 to 73 years).There was no significant difference between the two groups(t=1.47,P=0.147).There were 25 males(51%)in SFTS patients and 8 males(50%)in tsutsugamushi disease patients.There was no significant difference between the two groups(x2=0.005,P=0.943).The incidences of headache,vomiting,superficial lymphadenectasis,disturbance of consciousness,proteinuria,hematuria,pulmonary infection,multiple organ dysfunction and acute pancreatitis in SFTS patients were all significantly higher than those in tsutsugamushi disease patients(x2=8.82,4.38,8.71,11.17,7.88,5.56,4.35,9.43,and 8.13,respectively,P <0.05 or 0.01).The counts of leukocytes(Z=2.73),neutrophils(Z=2.46),lymphocytes(Z=3.15),platelets(Z=4.25),albumin(Z=2.65)and sodium ion(t=2.10)in SFTS patients were all significantly lower than those in patients with tsutsugamushi disease(P <0.05 or 0.01).The levels of aspartate aminotransferase(Z=2.94),lactate dehydrogenase(Z=3.42),creatine kinase(CK)(Z=2.88),amylase(Z=2.11),lipase(Z=2.82),creatinine(Z=2.07)and urea nitrogen(Z=2.50)in fatal SFTS patients were all significantly higher than those in patients with tsutsugamushi disease(P <0.05 or 0.01).Among 49 SFTS patients,16 patients died and 33 patients recovered finally.The age(t=3.33),platelet count(Z=2.55),alanine aminotransferase(ALT)(Z=2.10),aspartate aminotransferase(AST)(Z=2.22),lactate dehydrogenase(Z=2.26),CK(Z=3.50),CK-MB(Z=3.10),creatinine(Z=2.17),urea nitrogen(Z=2.36),and sodium(t=2.65)between the two subgroups had significant differences(P <0.05 or 0.01).Conclusions SFTS is more severe and has high mortality,while tsutsugamushi disease has a better prognosis.Early differential diagnosis and early rational treatment are important to reduce the mortality of patients with SFTS.
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The paper introduced the third-party satisfaction measurement and refined appraisal for quality continuous improvement programs launched by the hospital,and the PDCA management tools which optimized quality of care and patient satisfaction.The hospital is thus recommended to leverage databases for big data comparison,and carry out refined management on such basis,for the purposes of better performance management,higher doctor-patient friendship and patient trust.
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Objective To study the clinical efficacy of respiratory allergic diseases in children treated by nasal atomization of budesonide combined with 3% hypertonic saline. Methods Children diagnosed as upper air-way cough syndrome or mild to moderate asthma without being controlled by anti-asthma treatment were included in the study.They contracted with complications of nasal congestion and/or running noseand other symptoms,and na-sal CT confirmed the nasal lesions.Thirty children undergoing conventional treatment(conventional drug therapy+keeping away from allergen)were defined as the control group. 89 children treated with nasal spray therapy(con-ventional drug therapy + keeping away from allergen + nasal spray treatment)were defined as the therapy group. The treatment group was subdivided into IgE-mediated group and non-IgE-mediated group according to IgE level. The treatment course was 7 days.The clinical symptoms score,nasal symptom visual scale(VAS),peak expirato-ry flow(PEF)index were observed and analyzed. Results The clinical symptom score and nasal VAS showed a decreasing trend and the percentage of PEF showed an increasing trend in the two groups within 1 week after treat-ment.The clinical symptom score and nasal VAS score of the treatment group were significantly lower than those of the control group,while the PEF%value was significantly higher.The difference in treatment method was interact-ed with the treatment time(P<0.05).The percentage of PEF in the non-IgE-mediated group was significantly high-er than that of IgE-mediated group 1 week after treatment(P < 0.05). Conclusion The nasal atomization of budesonide combined with hypertonic saline in the treatment of children with respiratory allergic diseases is effec-tive and of a good clinical value.
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Prior to the formation of new metastases,a large number of cancer cells released from the primary tumor enter the dormancy period.Once the dorruancy is broken,tumor cells will recover the capability of proliferation.In recent years,various hypotheses and mechanisms of tumor metastasis have been studied.Primary tumor resection is considered to be an important factor to break tumor dormancy state.It is recognized as an important reason to promote tumor metastasis.The improvement of surgical technique and further study on the mechanism of dormancy may provide new ideas for the treatment of metastatic tumor.
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ObjectiveTo investigate the protective effects and mechanism of ursodeoxycholic acid (UDCA) on α-naphthylisothi (ANIT)-induced cholestatic liver injury in rats.MethodsA total of 48 Sprague-Dawley (SD) rats were selected.Fouty-two rats were gavaged with ANIT (100 mg/kg) to induce acute liver injury,six rats were sacrificed 24 hours after the liver injury and the rats left were evenly divided into control group which were gavaged with saline and UDCA group which were gavaged with UDCA (20 mg/kg).Six rats were sacrificed at 48 hours,72 hours and 96 hours after modeling.The six untreated rats were set as blank control group.Serum and liver tissues of all rats were kept after sacrificed.Serum levels of alanine transaminase (ALT),aspartate transaminase (AST),total bilirubin (TBil) and total bile acid (TBA) were tested,interleukin-10 (IL-10),interleukin-6 (IL-6),and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).The expression of multidrug resistance associated protein2 (Mrp2) at mRNA level in liver tissue was measured by quantitative real-time polymerase chain reaction (qRT-PCR) and the inflammatory reaction activity of liver tissues was inspected with Haematoidin-Eosin (HE)staining under microscope.ResultsAt 48 hours after liver injury modeling,serum TBil (143.80± 12.08) μmol/L vs.(178.50±15.19) μmol/L,TBA (13.15±3.81) μmol/L vs.(21.68±7.93)mol/L,IL-10 (44.13±3.68,37.15±6.25 ng/L),IL-6(50.80±2.09,57.32±4.63 ng/L) and TNF-α (17.53±0.84) ng/L vs,(19.10±1.64) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P ＜ 0.01 or P＜ 0.05).At 72 hours after liver injury modeling,serum ALT (721.67±97.54) U/L vs.(929.50±148.29) U/L and IL-10 (54.68±6.79)ng/L vs.(43.85±4.08) ng/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).At 96 hours after liver injury modeling,serum ALT (156.83±14.99) U/L vs.(250.67±42.29) U/L,AST (143.67±27.45) U/L vs.(206.00±63.94) U/L and TBil (23.53±5.08) μmol/L vs.(34.02±9.98) μmol/L of UDCA group and control group were compared,and the differences were statistically significant (P＜0.01 or P＜0.05).The differences of Mrp2 expression at mRNA level in liver tissues between UDCA group and control group at 48 hours (0.77 ± 0.21,0.46 ± 0.25),72 hours (2.27 ±0.84,1.10 ±0.38) and 96 hours (3.64±0.54,2.75±0.69) after liver injury modeling were statistically significant (P＜0.01 or P＜0.05).ConclusionThe mechanism of the protective effects of UDCA on ANIT-induced liver injury may be related with the regulation of serum cytokines and liver Mrp2 expression.