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Objective:To explore the effect of scenario simulation teaching combined with flipped classroom teaching for nursing interns in the department of gastroenterology.Methods:A total of 40 undergraduate nurses of 2019 from the Department of Gastroenterology of Renji Hospital, Shanghai Jiao Tong University School of Medicine were selected as the control group, and 40 undergraduate nurses of 2020 were selected as the research group. The control group adopted the traditional clinical teaching mode, and the research group adopted the scenario simulation combined with flipped classroom teaching mode. After 3 months of clinical teaching, theoretical and nursing operational assessment were performed on the nurses, and questionnaire survey was used to evaluate the teaching effect of clinical nursing teaching and the satisfaction of teaching quality. SPSS 20.0 was used for t test and chi-square test. Results:The theoretical and nursing operation skills assessment scores of the research group were higher than those of the control group, and the difference was statistically significant ( P<0.001). The research group was significantly better than the control group in improving nursing operation skills, improving autonomous learning ability, improving nurse-patient communication ability, improving response ability, improving overall analysis ability, enhancing humanistic care awareness, cultivating team spirit, and cultivating clinical thinking ability( P<0.001). The satisfaction rate of the practice nurses in the study group with the scenario simulation combined with the flipped classroom teaching was 100.00%, and that in the control group with the traditional clinical teaching mode was 70.00%, with statistical significance ( P<0.001). Conclusion:Scenario simulation teaching combined with flipped classroom can improve the theoretical foundation and operational skills of practical nurses in the department of gastroenterology, and improve the clinical nursing teaching effect and satisfaction of practical nurses, which is worthy of application and promotion.
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N-glycosylation modification, one of the most common protein post-translational modifications, occurs in heat shock protein gp96. The purpose of this study is to investigate the effect of N-glycosylation modification on immunologic function of the recombinant gp96 using the mutant gp96 in N-glycosylation sites. Firstly, wild-type and mutant gp96 proteins were expressed by insect expression system and their glycosylation levels were detected. To determine the effect of N-glycosylation on gp96 antigen presentation function, the IFN-γ+ CD8+ T cells in gp96-immunized mice and secretion level of IFN-γ were examined by flow cytometry and ELISA. The ATPase activity of gp96 was further detected by the ATPase kit. Finally, the effect of N-glycosylation on adjuvant function of gp96 for influenza vaccine was investigated in immunized mice. It was found that total sugar content of mutant recombinant gp96 was reduced by 27.8%. Compared to the wild type recombinant gp96, mutations in N-glycosylation sites resulted in decreased antigen presentation ability and ATPase activity of gp96. Furthermore, influenza vaccine-specific T cell levels induced by mutant gp96 as adjuvant were dramatically reduced compared to those by wild type recombinant gp96. These results demonstrate that N-glycosylation modification is involved in regulation of ATPase activity and antigen presentation function of gp96, thereby affecting its adjuvant function. The results provide the technical bases for development of gp96- adjuvanted vaccines.
Subject(s)
Animals , Mice , Adjuvants, Immunologic , CD8-Positive T-Lymphocytes/metabolism , Glycosylation , Heat-Shock Proteins , Influenza VaccinesABSTRACT
Objective The ADVIA Centaur XP automatic chemiluminescence immunoassay system detec-tion performance of the thyroid hormones and antibodies for validation and evaluation.Methods With refer-ence to the American association of clinical laboratory standardization guide and other related documents,the ADVIA Centaur XP automatic chemiluminescence immunoassay system test 7 items thyroid hormones and an-tibodies(T3,T4,FT3,FT4,TSH,anti-TG,anti-TPO)of precision,accuracy,linear range and carry pollution rate for validation.Results Within the seven thyroid hormones and antibodies batch testing precision of CV and CV between batch precision between 1.51% -6.17% and 1.86% -6.17%.Is the average accuracy of bias of the largest testing project FT3(8.47%),but in the acceptable range,good correlation,correlation coefficient R of 0.994 or higher,Average bias <1/2 CLIA′88 TEa(12.5%).T3,T4,FT3,FT4,TSH,anti-TG and anti-T PO in 1.24-5.59 nmol/L respectively,and 60.07 -195.10 nmol/L,3.40 -22.87 pmol/L,14.59 -40.54 pmol/L,1.63-74.13 μIU/mL,60.10 -381.30 μIU/mL and 180.10 -531.50 μIU/mL range is linear,Carry pollution rate is between 0.04% -0.97%.Conclusion ADVIA Centaur XP automatic chemiluminescence im-munoassay system detecting thyroid hormones and antibodies results consistent with the data provided by the manufacturer,the test results are accurate and reliable,and can be used for the detection of clinical samples.
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Cancer stem cells are currently under intensive investigation due to their capabilities for tumor initiation, self-renewal, and resistance to chemotherapy. CD133 is implicated in stemness and the malignancy of tumor cells. Here, we explored heat shock protein gp96 adjuvanted CD133 epitope vaccine against leukemia. We screened and identified three H2-Kd-restricted cytotoxic T lymphocyte (CTL) epitopes derived from CD133, CD133₄₁₉₋₄₂₈, CD133₇₀₂₋₇₁₀ and CD133₇₆₀₋₇₆₉. The immunogenicity and antitumor activity of the epitope vaccine using heat shock protein gp96 as adjuvant were further determined in CD133⁺ leukemia xenograft mice. Finally, we demonstrate that adoptive transfer of epitope-specific CTLs led to suppression of leukemia growth. Our data therefore provide the basis for designing a CD133 epitope vaccine to activate specific CTLs against CD133⁺ leukemia and other cancers.
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Objective To verify and valuate the performance of small dense low-density lipoprotein cholesterol (sdLDL-C) detection by the direct clearance method and evaluate its preliminary clinical application in acute coronary syndrome (ACS).Methods Case control study:The performance (accuracy,precision,linearity) of sdLDL-C was assessed by direct clearance method.In 143 cases of ACS patients selected from Cardiology Department and Emergency Department of Shangdong Provincial Hospital from April to October in 2016,with 100 cases male,female 43 cases,including acute myocardial infarction (AMI)group of 59 cases,unstable angina pectoris (UAP) group of 84 cases;83 cases of healthy volunteers as a control group selected from health physical examination center of Shandong Provincial Hospital,with 59cases male,female 24 cases.Levels of sdLDL-C,total cholesterol (TCH),triglyceride (TG),low-density lipoprotein cholesterol (LDL-C),high-density lipoprotein cholesterol (HDL-C),apolipoprotein A (ApoA I),apolipoprotein B (ApoB),lipoprotein (a) (Lpa) and high sensitive C-reactive protein (Hs-CRP) were detected separately by automatic biochemical analyzer.Non high density lipoprotein cholesterol (non-HDL-C) equals TCH minus HDL-C.x2 test,t test,one-way ANOVA,Pearson correlation and multiple linear regression analysis were used as statistical methods.Results The within-lot or between-lot variation was 2.85％ and 3.36％.Methodological comparison:regression equation Y =0.984X + 0.018,r2 =0.966,t =-0.191,P =0.850.There was a good linear correlation (Y =1.026X + 0.007,r2 =0.999) between theoretical values and actual detection results in range of 0.15-2.65 mmol/L.SdLDL-C concentrations were positive correlated with TCH,non-HDL,LDL-C,TG,ApoB (r =0.758,0.848,0.839,0.514,0.885,respectively,P ＜0.01),and negative correlated with HDL-C (r =-0.224,P =0.001),but no correlation with APOA I,Lpa and Hs-CRP(r =-0.021,0.050,0.003,respectively,P ＞ 0.05).Multiple linear regression analysis showed that the factors influencing sdLDL-C level were HDL-C,ApoB,LDL-C and TG.The levels of sdLDL-C,TG in the ACS group were significantly higher than those in the control group (t =3.415,4.660,respectively,P ＜ 0.01),but no difference between the two groups in the levels of TCH,non-HDL-C and LDL-C (t=-1.831,-0.452,-1.398,respectively,P ＞0.05).Comparing AMI group with control group,sdLDL-C,TG and Hs-CRP were significantly higher than the control group (P =0.000,0.000,0.000,respectively),but TCH,LDL-C and non-HDL were similar between the two groups (P =0.800,0.320,0.120,respectively);Comparing UAP group with control group,TG and Hs-CRP were higher than control group (P =0.001,0.047,respectively),TCH and LDL-C were significantly lower than the control group (P =0.003,0.008,respectively),but sdLDL-C had no difference (P =0.305);Comparing AMI group with UAP group,sdLDL-C,TCH,LDL-C and Hs-CRP were significantly higher than UAP group (P =0.000,0.003,0.001,0.000,respectively),and TG were no statistical significance (P =0.473).Conclusions Direct clearance method can meet the requirement of sdLDL-C detection.sdLDL-C level can assess the metabolism of blood lipids and be used as an independent risk factor and predictive index of ACS,superior to LDL-C.
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ST-segment elevation myocardial infarction (STEMI) is the most serious type of coronary heart disease, accounting for 25%to 40%of acute myocardial infarction (AMI). The key to treat STEMI is to restore myocardial perfusion in the infarct area, to rescue the ischemic myocardium, and to reduce the size of infarction. About 41%to 67%of patients with STEMI have multiple vascular disease (MVD). Compared with single vessel disease, the clinical outcome of MVD is worse. In these patients, it still remains a controversial topic in emergency interventions for STEMI patients, the infarct-related artery only revascularization or multi-vessel revascularization, and the timing of revascularization. The clinical studies of revascularization strategy for MVD in STEMI patients have been ongoing, and the results have also led to the continuous updating of guidelines and treatment strategies.
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It is an important influencing factor that the generated wear particles lead to periprosthetic osteolysis after artificial joint replacement. Current research suggests that the primary cause of wear particles results in periprosthetic osteolysis is relate to the prosthetic materials and the stimulations because of these materials generated wear particles to relevant cells such as macrophage, osteoblast, osteoclast. Induced a variety of inflammatory cytokines, activating and openning the cell signal and channels, producing the long term chronic inflammation leads to periprosthetic osteolysis. Therefor, the paper mainly studies the different types of wear particles influence on periprosthetic osteolysis, and the wear particles around the periprosthetic osteolysis mechanism in the process of progress, moreover, to explore how to reduce the occurrence of wear particles and blocking its role in the periprosthetic osteolysis, in order to achieve the purpose of prevention and treatment of periprosthetic osteolysis.
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Gatifloxacin (GFX) is a kind of chiral fluoroquinolones compound due to the methyl group at the C-3 position of the piperazine ring[1]. Although the enantiomers of GFX show similar levels of antimicrobial activity and pharmacokinetics[2], the other biological activities (i.e., toxicity or enantioselective recognition to various receptors in vivo) of GFX enantiomers have not yet been studied. With this in mind, we developed a rapid and cost-effective high performance liquid chromatographic (HPLC) separation procedure for GFX enantiomers with a pre-column esterification strategy. With significant enhancement of drug solubility and optimization for chromatographic conditions, the proposed method was scaled up to preparative HPLC to obtain optical active S-(-)- and R-(+)-GFX. The antibacterial activities of GFX enantiomers after preparative separation were further verified by measuring the Minimum Inhibitory Concentration (MIC) values against Escherichia coli ATCC 25922. In addition, the binding selectivity of GFX enantiomers to protein receptor were evaluated by antibody using enzyme-linked immunosorbent assay (ELISA) for the first time.
Subject(s)
Anti-Bacterial Agents , Chemistry , Pharmacology , Escherichia coli , Esterification , Fluoroquinolones , Chemistry , Pharmacology , Microbial Sensitivity Tests , Molecular Structure , Structure-Activity RelationshipABSTRACT
Objective To investigate the distribution and drug resistance of pathogenic bacteria in hospital ICU patients with lower respiratory tract infection , and provide scientific reference for clinical rational drug use . Methods The strains were identified by VITEK -32 automatic bacterial identification instrument , the bacterial sensitivity was determined by K -B disk diffusion method , and the statistical analysis was performed by WHONET 5.4 software.Results The total isolated pathogenic bacteria of lower respiratory tract infection in ICU patients was 453 strains.332 strains of gram negative bacteria accounted for 73.3%,and the former three ones were Pseudomonas aeruginosa (115 strains,accounted for 25.4%),Klebsiella pneumonia (90 strains,accounted for 19.8%),Acineto-bacter baumannii (38 strains,accounted for 8.4%).102 strains of gram positive bacteria accounted for 22.5%,and the top three were Staphylococcus aureus (31 strains,accounted for 6.8%),coagulase negative Staphylococcus (22 strains,accounted for 4.9%),Enterococcus (18 strains,accounted for 4%).Meropenem,imipenem (Stenotroph-omonas maltophilia was excepted ) ,Cefoperazone /sulbactam and Amikacin were most sensitive against gram negative bacteria;Teicoplanin and vancomycin were highly sensitive against gram positive bacteria .Conclusion The main pathogenic bacteria of respiratory tract infection in ICU patients was gram negative bacilli ,which were seriously resist-ant to commonly used antimicrobial drugs .So strengthening the infection management of ICU and the control for risk factors,and rationally using of antimicrobial drugs has great significance in reducing the drug resistance of pathogenic bacteria rate .
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PURPOSE: The purpose of this study was to validate the Actiwatch with behaviorally determined sleepe-wake state in preterm infants and to explore the influence of postmenstrual age on the accuracy of Actiwatch. METHODS: A prospective and comparative research design was used. Twenty-four preterm infants with postmenstrual age ranging from 28-38 weeks were studied. The infants were studied for 2 hours between two feedings. Infant's sleep and wake state was measured every 30 seconds using Actiwatch and the Anderson Behavioral State Scale simultaneously. RESULTS: Actiwatch demonstrated high agreement, sensitivity, and predictivity of sleep state, when validated with the Anderson Behavioral State Scale at the setting of high and automatic activity thresholds, and was not influenced by the infant's postmenstrual age. However, lower specificity and predictivity were found in the wake state, and influenced by postmenstrual age. CONCLUSION: Results of this study suggest that high activity thresholds are the most accurate for determining sleep state in preterm infants, and health care professionals must take the limitations into consideration while using the Actiwatch to assess wake states.
Subject(s)
Humans , Infant , Infant, Newborn , Delivery of Health Care , Infant, Premature , Prospective Studies , Research Design , Sensitivity and SpecificityABSTRACT
BacKground:Helicobacter pylori ( Hp ) infection is an important etioIogicaI factor of gastric uIcer. AIthough eradicating Hp can heIp gastric uIcer heaIing,there is stiII a high recurrence rate of gastric uIcer. Therefore,other factors pIaying a roIe in the pathogenesis of gastric uIcer shouId be in concern. Aims:To investigate the non-reinfectious recurrence factors of gastric uIcer after Hp eradication. Methods:A totaI of 164 newIy diagnosed gastric uIcer patients with Hp infection from January 2011 to June 2012 at Ren Ji HospitaI,SchooI of Medicine,Shanghai Jiao Tong University were enroIIed. AII patients were given Hp eradication therapy and treatment of gastric uIcer. Patients with successfuI Hp eradication and heaIing of gastric uIcer were foIIowed up at 3,6,12 months after uIcer heaIing. Suspected risk factors of gastric uIcer recurrence were coIIected,and their reIationships with recurrence were anaIyzed. Results:A totaI of 123 patients positive for Hp infection compIeted the 12 months foIIow-up,gastric uIcer recurred in 18(14. 6%)of them. Chi-square test reveaIed that age,gender,smoking/drinking,non-steroidaI anti-infIammatory drugs( NSAIDs ),negative emotions and unheaIthy eating habits were significantIy associated with the recurrence of gastric uIcer(P<0. 05). Of the unheaIthy eating habits, irreguIar eating, and preference of indigestibIe food and fast food/carbonated drinks were significantIy associated with recurrence of gastric uIcer( P <0. 05 ). Conclusions:Patients with gastric uIcer shouId change their unheaIthy Iiving habits,foIIow rationaI use of drugs,and keep heaIthy mentaI status and eating habits for avoiding the recurrence of gastric uIcer.
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Objective To investigate the clinical and laboratory characteristics of acute myeloid leukemia (AML) with t (16;21)(p11;q22) translocation.Methods Two patients diagnosed by morphology,cytochemical stain,immunology,cytogenetics and genetic testing.Similarities and differences of clinical characteristics and laboratory examination were analysed,along with a review of the literatures.Results According to the FAB classification,one patient was M4 and the other one was M1.The cytogenetic aberrations were 46,XY,t(16;21)(p11;q22)[16]/47,XY,t(16;21)(p11;q22),+21[4] of ease 1 and 46,XX,t(16;21)(p11;q22)[20] of case 2.Cytophagocytosis and CD56 antigen expression were found in both cases.The prognosis was poor in both cases.Conclusions AML with t(16;21)(p11;q22) is a specific type,which has unusual characteristics of morphology,immunology,cytogenetics,clinical feature.The prognosis of the patients is poor,so stem-cell transplantation maybe the only and the first choice of treatment.
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To evaluate the characteristics of pituitary adenomas that produce both prolactin and adrenocorticotropin. Between 2002 and 2011, we reviewed the data of 336 patients undergoing transsphenoidal surgery at Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China. Patients were divided into 2 subgroups: patients with mixed prolactin and adrenocorticotropin secreting adenomas, and patients with merely prolactin-secreting adenomas. Clinical and endocrinological data, imaging, histopathological reports, and outcomes were reviewed. Differences between the 2 groups were statistically analyzed, and p<0.05 was considered statistically significant. Compared to patients with merely prolactin-secreting adenomas, patients with mixed prolactin and adrenocorticotropin secreting adenomas were younger [p<0.001], had higher incidences of headaches and dizziness [p=0.021], progressive obesity [p<0.001], menstrual disorders [p=0.006], polyuria and polydipsia [p<0.001], hypertension [p=0.001], diabetes mellitus [p=0.001], and had higher rates of postoperative hyponatremia [p<0.001]. Recurrence rates in patients with prolactin-secreting adenomas were 19.1% and patients with mixed prolactin and adrenocorticotropin secreting adenomas were 35.1% [p=0.023]. However, the endocrine normalization rate in mixed prolactin and adrenocorticotropin secreting adenomas was lower [p=0.004]. Careful long-term follow-up is needed for patients with mixed prolactin and adrenocorticotropin secreting adenomas
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Heat shock protein gp96 isolated from tumor tissues holds great promise for tumor immunotherapy. However, at present only very limited amount of gp96 protein can be isolated from tumor tissues. Here, we reconstituted the yeast-expressed gp96 (recombinant gp96, rgp96) with B16.F10 melanoma antigens in vitro to prepare new gp96 tumor vaccine on large-scale, and analyzed its induction of specific anti-tumor immunoresponses by ELISPOT, IFN-gamma intracellular staining and cytotoxicity assays. Immunization with rgp96-tumor antigen complexes significantly inhibited B16 tumor growth compared with either rgp96 or tumor antigens alone and led to enhancement of tumor-specific T-cell activities, which was found similar to that of tumor tissue derived gp96. Our results therefore may provide bases for large-scale preparation of the new generation of gp96 tumor vaccines.
Subject(s)
Animals , Female , Mice , Cancer Vaccines , Genetics , Allergy and Immunology , Therapeutic Uses , Heat-Shock Proteins , Genetics , Allergy and Immunology , Therapeutic Uses , Melanoma, Experimental , Therapeutics , Mice, Inbred C57BL , Neoplasm Transplantation , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Therapeutic Uses , Skin Neoplasms , Therapeutics , Yeasts , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To establish the stable inhibition of HER2/neu expression by vector-mediated small hairpin RNA in malignant transformed human bronchial epithelial cell line induced by anti-benzo(a)pyrene-trans-7, 8-dihydrodiol-9, 10-epoxide (anti-BPDE).</p><p><b>METHODS</b>The pSIREN-RetroQ-neu recombinant vector targeting HER2/neu was constructed and confirmed by restriction and sequencing analysis, then it was transfected into anti-BPDE malignant transformed 16HBE cells (16HBE-T) through lipofectamine 2000. The control groups included the 16HBE-T cells transfected with negative control vector (negative control) and 16HBE-T. The cells transfected with vectors were screened by puromycin. The HER2/neu mRNA and protein expressions in the vector-transfected 16HBE-T cells were detected by RT-PCR and Western blot method respectively.</p><p><b>RESULTS</b>The pSIREN-RetroQ-neu recombinant vector which inhibited HER2/neu mRNA and protein expressions in the 16HBE-T was constructed. The level of HER2/neu mRNA in the 16HBE-T cells transfected with pSIREN-RetroQ-neu was significantly reduced as compared to the negative control and blank control cells (0.114 +/- 0.003 vs.blank control 0.186 +/- 0.001, t = 39.154, P < 0.05; and negative control 0.182 +/- 0.015, t = 7.564, P < 0.05), while its level did not differ significantly between negative control cells and blank control of 16HBE-T (t = -0.409, P > 0.05). HER2/neu protein level in pSIREN-RetroQ-neu transfected cells was inhibited by 40% and 39% respectively.</p><p><b>CONCLUSION</b>Plasmid-based shRNA expression systems targeted against HER2/neu gene were generated successfully, which resulted in down-regulation of HER2/neu gene expression in the 16HBE-T efficiently.</p>
Subject(s)
Humans , 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide , Toxicity , Base Sequence , Cell Line, Transformed , Epithelial Cells , Metabolism , Gene Expression , Genes, erbB-2 , Molecular Sequence Data , RNA, Small Interfering , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
Elevated serum levels of interleukin-10 (IL-10) have been reported in patients with Kawasaki disease (KD). IL-10 reduces the inflammatory actions of macrophages and T cells and it may play a significant role in the regulation of inflammatory vascular damage associated with systemic vasculitis. The aim of this study was to examine whether -592 IL-10 promoter polymorphism is a susceptibility or severity marker of KD in Chinese patients in Taiwan. The study included 105 KD patients and 100 normal controls. Genotype and allelic frequencies for the IL-10 gene polymorphism in both groups were compared. There were no significant between-group differences in the genotype distribution of IL-10 A-592C gene polymorphism (P=0.08). However, the frequency of the -592*A allele was significantly increased in the patients with KD compared with controls (71.9% vs. 61.0%, P=0.019). The odds ratio for developing KD in individuals with IL-10-592*A allele was 1.64 (95% confidence interval, 1.06-2.52) compared to individuals with the IL-10-592*C allele. No significant difference was observed in the genotype and allelic frequencies for the IL-10 A-592C polymorphism between patients with and without coronary artery lesions. The IL-10-592*A allele may be involved in the development of KD in Taiwanese children.
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Alleles , Asian People/genetics , Gene Frequency , Genotype , Interleukin-10/blood , Mucocutaneous Lymph Node Syndrome/diagnosis , Polymorphism, Genetic , Promoter Regions, Genetic , TaiwanABSTRACT
Elevated serum levels of interleukin-10 (IL-10) have been reported in patients with Kawasaki disease (KD). IL-10 reduces the inflammatory actions of macrophages and T cells and it may play a significant role in the regulation of inflammatory vascular damage associated with systemic vasculitis. The aim of this study was to examine whether -592 IL-10 promoter polymorphism is a susceptibility or severity marker of KD in Chinese patients in Taiwan. The study included 105 KD patients and 100 normal controls. Genotype and allelic frequencies for the IL-10 gene polymorphism in both groups were compared. There were no significant between-group differences in the genotype distribution of IL-10 A-592C gene polymorphism (P=0.08). However, the frequency of the -592*A allele was significantly increased in the patients with KD compared with controls (71.9% vs. 61.0%, P=0.019). The odds ratio for developing KD in individuals with IL-10-592*A allele was 1.64 (95% confidence interval, 1.06-2.52) compared to individuals with the IL-10-592*C allele. No significant difference was observed in the genotype and allelic frequencies for the IL-10 A-592C polymorphism between patients with and without coronary artery lesions. The IL-10-592*A allele may be involved in the development of KD in Taiwanese children.
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Alleles , Asian People/genetics , Gene Frequency , Genotype , Interleukin-10/blood , Mucocutaneous Lymph Node Syndrome/diagnosis , Polymorphism, Genetic , Promoter Regions, Genetic , TaiwanABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of small hairpin interfering RNA (shRNA) in suppressing cytochrome P450 3A4 (CYP3A4) gene expression in CHL-3A4 cells.</p><p><b>METHODS</b>Three shRNA expression vectors targeting CYP3A4 gene (CYP3A4 I, C YP3A4 II, and CYP3A4 III, respectively) were designed, synthesized and transfected into CHL-3A4 cells via liposomes. The inhibitory effect of shRNA on CYP 3A4 gene expression was detected by Western blotting and RT-PCR, and the effect of shRNA transfection in suppressing cyclophosphamide-induced cytotoxicity was measured using MTT assay.</p><p><b>RESULTS</b>The vector carrying CYP3A4 III shRNA significantly reduced the expression of CYP3A4 gene at both the mRNA (75%) and protein levels (80%) in CHL3A4 cells. The cytotoxicity of cyclophosphamide was markedly inhibited by CYP3A4 III-mediated suppression of CYP3A4 gene expression by 75% in CHL-3A4 cells.</p><p><b>CONCLUSION</b>The vector-mediated RNA interference can suppress CYP3A4 gene expression in CHL-3A4 cells, and RNA interference technique provides a new means for studying cytochrome P450 gene function in mammalian cells.</p>
Subject(s)
Animals , Cricetinae , Humans , Cells, Cultured , Cytochrome P-450 CYP3A , Genetics , Fibroblasts , Cell Biology , Metabolism , Lung , Cell Biology , RNA Interference , RNA, Small Interfering , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To observe the effect of ketoconazole on the activity of cytochrome P450 (CYP) 1A2 and 3A4 in hepatic microsomes of healthy adults.</p><p><b>METHODS</b>Human hepatic microsomes obtained from healthy adults were randomly divided into control group and ketoconazole-treatment groups at different concentrations. After 15 min of culture, the substrates (testosterone for CYP3A4 and phenacetin for CYP1A2) were added and incubated for another 20 min. The metabolites (6-testosterone and acetaminophen) were then measured with high-performance liquid chromatography (HPLC) to assess the activities of CYP3A4 and 1A2.</p><p><b>RESULTS</b>Significant difference was found between the groups in the quantity of 6-testosterone and the relative activity of CYP3A4 (P<0.05). The IC(50) of ketoconazole for CYP3A4 was 0. 16 mg/L. Both the quantity of 6-testosterone and the relative activity of CYP3A4 were reduced gradually with the increment of ketoconazole concentration. Significant differences were found between the ketoconazole groups and the control group in both the quantity of acetaminophen and the relative activity of CYP1A2 (P<0.05). Ketoconazole at low doses reduced CYP1A2 activity and but increased the activities at high doses (P<0.05).</p><p><b>CONCLUSION</b>In the range of maximum clinical blood concentration, ketoconazole can inhibit the activity of CYP3A4, but not that of CYP1A2, in the hepatic microsomes in healthy adults.</p>
Subject(s)
Adult , Female , Humans , Male , Antifungal Agents , Pharmacology , Cytochrome P-450 CYP1A2 , Metabolism , Cytochrome P-450 CYP3A , Metabolism , Dose-Response Relationship, Drug , Ketoconazole , Pharmacology , Microsomes, LiverABSTRACT
OBJECTIVE: In order to study the mechanism of the aqueous humor circulation and its relationship to the glaucoma macroscopically with engineering methods. METHODS: A dynamistic model was presented, which can be used to simulate the situation and the treatment of the primary open angle glaucoma (POAG). The frame of the model was built based on the ophthalmically accepted feedback mechanism between the aqueous humor circulation and the intraocular pressure (IOP). The transfer functions and the parameters were educed from the analysis of physiological theories, the basic elements of hydrodynamics, and the clinical parameters. The relation between the parameters of the system and the episode mechanism of POAG was discussed. A digital method was used to simulate the Challenge test and some medicines' treatment of POAG, and the results were consistent with clinical observations. RESULTS: The results of simulation illuminate that the model can simulate the mechanism of the aqueous humor circulation and the curative mechanism of some medicines under the physiological condition and the pathological condition of the POAG. CONCLUSION: A few parameters which can hardly be captured with clinical method could be obtained from the model. These parameters can be helpful for the diagnosis and prediction of the curative effect.