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Objective:To investigate the protective effect and mechanism of isorhamnetin on myocardial injury in rats with acute myocardial infarction (AMI).Methods:The AMI rat model was established by coronary artery left anterior descending ligation. The SD rats were divided into sham-operated group, model group, Fasudil group (30 mg/kg) and low-, medium-, and high-dose (25, 50, 100 mg/kg) groups. Each group had 5 rats. They were administrated intragastric, once a day, and were treated for 14 d. Color Doppler ultrasonography was used to detect cardiac function by measuring left ventricular end-systolic diameter (LVESd), left ventricular end-diastolic diameter (LVEDd), and left ventricular long-axis shortening fraction (FS). Myocardial infarct size was detected by TCT staining. Superoxide dismutase (SOD), malondialdehyde (MDA), interleukin (IL)-6, IL-1β, and tumor necrosis factor-α (TNF-α) levels in rat serum were detected by ELISA and TUNEL assay for the cardiomyocyte apoptosis index. Cysteinyl aspartate specific proteinase-3 (Caspase-3) activity in myocardial tissue was detected by the Caspase-3 activity assay kit, and the expression of Bcl-2 associated X protein (Bax) and B-cell lymphoma-2 (Bcl-2) proteins in myocardial tissue was detected by Western Blot.Results:Compared with the sham operation group, the FS, SOD content, and expression levels of Bcl-2 protein in the model group were significantly decreased (all P<0.05), and LVEDd, LVESd, myocardial infarct size, MDA content, IL-6 content, TNF-α content, IL-1β content, apoptosis index, Caspase-3 activity, and Bax protein expression level were significantly increased (all P<0.05). There was no significant difference between the low concentration of isorhamnetin and the model group (all P>0.05). However, the above changes caused by the construction model after treatment with Fasudil and medium- and high-concentrations of isorhamnetin significantly reversed (all P<0.05). Conclusions:Isorhamnetin can improve cardiac function and protect myocardial injury in AMI rats by reducing oxidative stress and inflammatory damage to cardiomyocytes and inhibiting cardiomyocyte apoptosis.
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This study aims to explore underlying mechanism of Lonicerae Japonicae Flos(LJF) in protecting rats against acute alcoholic liver injury(ALI) based on mitogen-activated protein kinase(MAPK) pathway. First, the targets of LJF in preventing ALI were predicted by network pharmacology and the component-target-pathway network was constructed, so that the key targets of LJF components acting on MAPK pathway were screened. Second, male SD rats were randomized into the control(KB) group, model(MX) group, positive(YX) group, and LJF high-(GJ), medium-(ZJ), and low-(DJ) dose groups. Each administration group was given(ig) corresponding drugs for 7 days and KB group and MX group received(ig) equal volume of distilled water every day. Except for KB group, rats were given Chinese spirit(56%, 3 days) for ALI modeling. The levels of aspartate transaminase(AST), alanine transaminase(ALT), interleukin-6(IL6) and tumor necrosis factor-α(TNF-α) in serum and malondialdehyde(MDA), glutathione(GSH), superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in liver tissue of rats in each group were detected. Furthermore, we employed quantitative real-time PCR(qRT-PCR) to probe the effects of LJF on the key targets of MAPK pathway in ALI rats. A total of 28 active components of LJF were screened from TCMSP database, and 317 intersected with ALI-related targets. According to Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis, the 317 targets involved 226 pathways, which were mainly liver disease, inflammation, immunity, apoptosis and other related pathways. According to the MAPK pathway-target-active component network, the key active components of LJF, such as chlorogenic acid, hederagenol, and hyperoside, acted on 25 key targets of MAPK pathway. The results of in vivo experiments showed decreased levels of AST, ALT, and MDA in DJ, ZJ, and GJ groups(P<0.01 or P<0.05), reduced levels of IL6 in DJ and GJ groups(P<0.01 or P<0.05), and improved levels of SOD and GSH in ZJ and GJ groups(P<0.01 or P<0.05). The results of qRT-PCR demonstrated that the expression levels of mitogen-activated protein kinase kinase 4(MAPK2 K4) and mitogen-activated protein kinase 3(MAPK3) were decreased in DJ, ZJ, and GJ groups(P<0.01). The network pharmacology and experimental verification showed that the active components in LJF can reduce the inflammatory factor level and enhance the activities of SOD and GSH-Px by inhibiting the expression of key targets of MAPK pathway, thus alleviating and preventing liver damage caused by alcohol.
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Animals , Male , Rats , Chlorogenic Acid , Drugs, Chinese Herbal , Liver , Liver Diseases , Rats, Sprague-DawleyABSTRACT
BACKGROUND@#Generic drugs are bioequivalent to their brand-name counterparts; however, concerns still exist regarding the effectiveness and safety of generic drugs because of small sample sizes and short follow-up time in most studies. The purpose of this study was to evaluate the long-term antihypertensive efficacy, cost-effectiveness and cardiovascular outcomes of generic drugs compared with brand-name drugs.@*METHODS@#In a multicenter, community-based study including 7955 hypertensive patients who were prospectively followed up for an average of 2.5 years, we used the propensity-score-matching method to match the patients using brand-name drugs to those using generic drugs in a ratio of 1:2, 2176 patients using brand-name drugs and 4352 patients using generic drugs.@*RESULTS@#There were no significant differences between generic drugs and brand-name drugs in blood pressure (BP)-lowering efficacy, BP control rate, and cardiovascular outcomes including coronary heart disease and stroke. The adjusted mean (95% confidence interval [CI]) of systolic BP (SBP)-lowering was -7.9 mmHg (95% CI, -9.9 to -5.9) in the brand-name drug group and -7.1 mmHg (95% CI, -9.1 to -5.1) in the generic drug group after adjusting for age, sex, body mass index, number of antihypertensive drugs and traditionally cardiovascular risk factors. Among patients aged <60 years, brand-name drugs had a higher BP control rate (47% vs. 41%; P = 0.02) and a greater effect in lowering SBP compared with generic drugs, with the between-group difference of 1.5 mmHg (95% CI, 0.2-2.8; P = 0.03). BP control rate was higher in male patients using brand-name drugs compared with those using generic drugs (46% vs. 40%; P = 0.01). Generic drugs treatment yielded an average annual incremental cost-effectiveness ratio of $315.4 per patient per mmHg decrease in SBP compared with brand-name drugs treatment.@*CONCLUSIONS@#Our data suggested that generic drugs are suitable and cost-effective in improving hypertension management and facilitating public health benefits, especially in low- and middle-income areas.
Subject(s)
Aged , Humans , Male , Antihypertensive Agents/therapeutic use , Blood Pressure , China , Drugs, Generic/therapeutic use , Prospective StudiesABSTRACT
Objective CARD9 can activate several pathways involved in immunity, such as NF-ΚB, MAPK, etc. However the mechanism of this process has not yet been elucidated. For conducting relevant experiments in vitro, a prokaryotic expression vector of CARD9-MBP fusion protein has to been construct, and the fusion protein need to be expressed and purified. Methods The coding sequence of CARD9 and MBP genes were amplified by PCR and the recombinant plasmid was correctly inserted into the pET-30a(+) vector. The recombinant plasmid was transformed into E.coli DH5α competent cells and proceeded PCR identification, restriction analysis and gene sequencing. The correct recombinant plasmid was transformed into E.coli BL21(DE3) competent cells. The target protein was induced to express by IPTG under different conditions. Relative molecular weight of the target protein was detected by SDS-PAGE electrophoresis. The CARD9-MBP fusion protein was purified by MBP maltose chromatography column and gel filtration chromatography column, and identificated by MALDI-TOF mass spectrometry after MBP-tag to be removed by HRV3C enzyme. Results The CARD9-MBP fusion protein was successfully constructed and confirmed by PCR and restriction analysis. The result of gene sequencing was consistent with the target sequence. The SDS-PAGE electrophoresis showed that the target protein with molecular mass (MR) about 105 000 was successfully induced to express in E.coli BL21 (DE3). A quite pure CARD9-MBP fusion protein was obtained by purification of MBP maltose chromatography column. Identification by MALDI-TOF mass spectrometry demonstrated that the target protein after MBP-tag to be removed by HRV3C enzyme is CARD9 protein. In the later stage, gel filtration chromatography column was used to obtain further pure CARD9-MBP fusion protein. Conclusion The prokaryotic expression vector of CARD9-MBP fusion protein was successfully constructed and a large number of soluble protein expressed. The purified target protein can be obtained by purification with MBP maltose chromatography column and gel filtration chromatography column.
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Genetic as well as genomic study has advanced the development of precision medicine. We are marching on the road for right patients who are receiving more and more right treatment at right time. In hypertension field, precision medicine is available, actionable and affordable. First and the most practical advancement is monogenic hypertension, the disease-genes have been found for at least 17 types of monogenic hypertension. These patients can be precisely treated according to their carried gene mutation. Secondly, pharmacogenetic and pharmacogenomic guided anti-hypertensive drug selection, very promising but lack of clinic outcome data to support widely clinical application. Majority of hypertension are due to multiple genetic and environmental factors. GWAS fund some genetic variants related to primary hypertension, but these variants can only be responsible for 1-10% of blood pressure variation. We have a long way to go in exploring the real cause of primary hypertension.
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Objective Erythroderma is a very serious disease that affects nearly the entire cutaneous surface and are highly subjected to secondary hypoalbuminemia, infection, cardiovascular diseases, complex causes and high death rates. The article aimed to explore the etiology, comorbidities and complicated infection of erythroderma.Methods Retrospective analysis was conducted on clinical data of 95 cases of erythroderma in our department from January 2009 to August 2016. Observations were made on the patients' clinical characteristics, etiology and inducement, lab examination, complications and complicated infection.Results There were 73 first-episode and 22 recurrent patients, among which 14 cases are psoriasis as the basic disease. As to etiological factors, there were 57 cases secondary to other skin diseases (60%) and 25 cases by drug reactions (26%). As to inducing factors, there were 6 cases by upper respiratory tract infection, 38 cases by irrational application of glucocorticoids, and 7 cases by external stimulants (traditional Chinese medicine scrubbing and external medicinal liquor). The main complications were 38 cases of cardiovascular diseases (40%). The complicated infection rates of plasma albumin in patients <35g/L and ≥35g/L were 65.78% and 12.28%(P<0.01). The complicated infection rates of the patients with hypoalbuminemia and electrolyte disturbance were 44.2% and 25% respectively (P<0.05).Conclusion The erythroderma is mainly secondary to previous skin diseases, mostly psoriasis, with cardiovascular diseases as the main comorbidities. In clinical practice, importance should be attached to monitoring decreased plasma albumin level and electrolyte disturbances in order to reduce the risk of infection.
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Objective To observe optic nerve axons degenerative disorder and microglial responses by establishing unilateral optic nerve crush model.Methods YFP mouse group with axonal markers and GFP mouse group with microglia markers were divided into surgery and control group,the optic nerve were dissected at 4 hours,1 day,3 days,5 days,10 days after optic nerve crush,and the neuronal degenerative disorder and microglial responses were observed by confocal laser scanning microscope.Rrsults Compared with control group,the optic nerve axons in YFP mouse group were fractured in injury region at postoperative 4 hours;The partial axon became beadlike change at postoperative 1 day;Most of the axons turned into the process of beadlike change at postoperative 3 days;The axons became to debris from beadlike at postoperative 5 days;The axons changed into many debris at postoperative 10 days.Compared with control group,the formation of glial scar and resting microglia in GFP mouse group began to emerge at postoperative 4 hours;The microglia gradually activated and began to cover the injury region at postoperative 1 day;The activated miacroglia basically covered the injury region at postoperative 3 days;The number of microglia roughly remained stable,although the axons continued to deteriorate at postoperative 5 days and 10 days.Conclusion The optic nerve occur irreversible degenerative disorder after being injured,meanwhile with the microglial increase and activation.This phenomenon suggests that microglia is closely associated with optic nerve degeneration.
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BACKGROUND/AIMS: To evaluate the enhancement patterns of liver metastases and their influencing factors using dynamic contrast-enhanced ultrasound (CEUS). METHODS: A total of 240 patients (139 male and 101 female; 58.5±11.2 years of age) diagnosed with liver metastases in our hospital were enrolled in this study to evaluate tumor characteristics using CEUS. A comparison of enhancement patterns with tumor size and primary tumor type was performed using the chi-square test. The differences between quantitative variables were evaluated with the independent-sample t-test and one-way analysis of variance. RESULTS: The enhancement patterns of liver metastases on CEUS were categorized as diffuse homogeneous hyperenhancement (133/240, 55.4%), rim-like hyperenhancement (80/240, 33.3%), heterogeneous hyperenhancement (10/240, 4.2%), and isoenhancement (17/240, 7.1%). There were significant differences in the enhancement patterns during the arterial phase based on the nodule size (p=0.001). A total of 231 of the nodules showed complete washout during the portal phase, and 237 nodules were hypoenhanced during the delayed phase. The washout time was correlated with tumor vascularity, with a longer washout time observed in hypervascular metastases compared to hypovascular metastases (p=0.033). CONCLUSIONS: Diffuse homogeneous hyperenhancement followed by rapid washout was the most common enhancement pattern of liver metastases on CEUS and was affected by the nodule size and tumor vascularity. Small metastases were prone to show diffuse homogeneous hyperenhancement. Hyper-vascular metastases showed a significantly longer washout time compared to hypovascular metastases.
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Contrast Media/therapeutic use , Liver/diagnostic imaging , Liver Neoplasms/blood supply , Neovascularization, Pathologic/diagnostic imaging , Ultrasonography/methodsABSTRACT
New enrofloxacin microspheres were formulated, and their physical properties, lung-targeting ability, and tissue distribution in rats were examined. The microspheres had a regular and round shape. The mean diameter was 10.06 microm, and the diameter of 89.93% of all microspheres ranged from 7.0 microm to 30.0 microm. Tissue distribution of the microspheres was evaluated along with a conventional enrofloxacin preparation after a single intravenous injection (7.5 mg of enrofloxacin/kg bw). The results showed that the elimination half-life (t(1/2beta)) of enrofloxacin from lung was prolonged from 7.94 h for the conventional enrofloxacin to 13.28 h for the microspheres. Area under the lung concentration versus time curve from 0 h to infinity (AUC(0-infinity)) was increased from 11.66 h.microg/g to 508.00 h.microg/g. The peak concentration (Cmax) in lung was increased from 5.95 microg/g to 93.36 microg/g. Three lung-targeting parameters were further assessed and showed that the microspheres had remarkable lung-targeting capabilities.
Subject(s)
Animals , Female , Humans , Male , Rats , Anti-Bacterial Agents/adverse effects , Drug Delivery Systems/instrumentation , Fluoroquinolones/adverse effects , Half-Life , Injections, Intravenous , Lung/drug effects , Microspheres , Rats, Sprague-Dawley , Tissue DistributionABSTRACT
Residue after evaporation [RAE] from industrial vitamin C fermentation is emitted as a waste product at an amount of 60,000 tons per year in China. The disposal of RAE is difficult because of its high chemical oxygen demand [1.17[x]10[6] mg/1] and low pH [0.27]
We hypothesized that RAE could be used as an ameliorant for alkali-saline soils, and tried to verify it by carrying out a pot experiment of pakchoi cultivation and to explore its effect on soil chemical and microbial properties
The results showed that pakchoi yield was increased by 28.13% and pakchoi quality was also enhanced under RAE treatment. The improved chemical and microbial properties of treated soil were also observed: soil pH was decreased from 9.19 to 9.03; total organic carbon, available phosphorus and available potassium were increased by 49.15%, 34.91% and 42.02%, respectively; number of culturable bacteria, actinomycetes and fungi, microbial biomass carbon and enzyme activity number were improved by 52.97%, 104.05%, 79.09%, 57.82% and 31.16%, respectively
These results suggested the residue application led to an improved soil quality and subsequently a higher yield and quality of pakchoi. This study provided a strong evidence for the feasibility of RAE as an ameliorant for alkali-saline soil
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<p><b>OBJECTIVES</b>To establish a primary biliary cirrhosis (PBC) model by AMAM2 autoantigen injection into C57BL/6 mice.</p><p><b>METHODS</b>Mice of the model group were immunized intraperitonealy with 200 microl of purified recombinant AMAM2 autoantigen in complete Freund's adjuvant (CFA). Mice immunized with bovine serum albumin and CFA in the same way were used as negative controls. Sixty-six weeks later, mice were sacrificed and their sera were collected. Sera samples were assayed for AMAM2 autoantibody, alkaline phosphatase (ALP), ALT and total bilirubin (TBil). Their liver, stomach, muscle and kidney tissues were sectioned and stained using HE to observe the pathological changes.</p><p><b>RESULTS</b>Antibodies to AMAM2 autoantigen were readily induced in the model group. The mice in the model group had no significant changes in the level of serum ALT and TBil but had an obvious increase of ALP (P<0.05). The stomach, muscle and kidney tissues showed no evident damage while the livers had obvious pathological changes, including bile duct degeneration or proliferation, and mononuclear cell infiltration.</p><p><b>CONCLUSION</b>The AMAM2 autoantigen-induced PBC animal model was successfully established in C57BL/6 mice in our experiment and its characteristic biochemical and pathology are quite similar to that in the early stage of human PBC. This model may provide a useful experimental approach for further study of the pathogenesis and clinical treatment of human PBC.</p>
Subject(s)
Animals , Mice , Autoantigens , Allergy and Immunology , Disease Models, Animal , Liver Cirrhosis, Biliary , Mice, Inbred C57BL , Mitochondria , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the association between Chinese patients with autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and the polymorphisms of cytotoxic T lymphocyte -associated antigen-4 (CTLA-4) gene promoter (-318) and exon 1 (+49).</p><p><b>METHODS</b>The CTLA-4 promoter (-318 T/C) and exon 1 (+49A/G) polymorphisms were genotyped via restriction fragment length polymorphism methods in 62 Chinese AIH patients, 77 Chinese PBC patients and 160 healthy controls.</p><p><b>RESULTS</b>There was no difference in the distribution of CTLA-4 promoter -318 T/C polymorphisms between AIH patients and controls, but the C allele frequency was significantly increased in patients with AIH, compared to controls (P=0.02, OR=2.43). The distribution of CTLA-4 gene exon 1 49 A/G genotypes exhibited significant difference between PBC patients and controls (P=0.006), and the frequency of G allele showed a significant increase in PBC group as compared with controls (P=0.0046, OR=1.8). Although the genotype distribution of the CTLA-4 exon 1-promoter gene displayed no significant difference between AIH and PBC patients and controls, the occurrence of GG-CC was increased in the patients of the two groups (AIH: 32.3%, PBC: 37.7%; control: 22.5%).</p><p><b>CONCLUSION</b>The above findings suggest that the polymorphisms of CTLA-4 gene probably confer susceptibility to AIH and PBC in the Chinese population.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, CD , Genetics , Asian People , Genetics , CTLA-4 Antigen , China , Exons , Genetics , Genetic Predisposition to Disease , Genetics , Genotype , Hepatitis, Autoimmune , Ethnology , Genetics , Liver Cirrhosis, Biliary , Ethnology , Genetics , Polymerase Chain Reaction , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length , Promoter Regions, Genetic , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To determine the relationship between polymorphisms in the genes encoding IL-1, IL-6, and IL-10 with primary biliary cirrhosis (PBC) in Chinese population.</p><p><b>METHODS</b>Whole-blood samples were taken from 77 patients with PBC and 160 healthy controls. DNA was extracted and the polymorphisms at positions IL-1 +3953, IL-1RN intron 2, IL-6 -174, and IL-10 -1082, -819, and -592 were determined by using sequence-specific polymerase chain reaction (SSP) or polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).</p><p><b>RESULTS</b>The frequency of IL-1RN1,1 allele in PBC group was significantly higher than in control group (90.9% vs 79.4%, P=0.026), and the frequency of IL-1RN1,2 in PBC group was significantly lower than in control group (6.5% vs 18.8%, P=0.013). There was no significant difference in the frequence of IL-1RN*2 allele between PBC group and control group (P=0.06). Of the 77 patients with PBC, 4 patients were IL-6 -174GC, 73 were IL-6 174GG. All the 160 health controls are IL-6 -174GG (P=0.0036). The frequence of IL-6 -174C allele in PBC group was significantly higher than that in control group (P=0.0038). No significant differences of polymorphisms for IL-1 +3953 and IL-10 (-1082, -819 and -592) were found between PBC group and control group.</p><p><b>CONCLUSION</b>The polymorphisms of IL-1RN and IL-6 -174G/C appear to be associated with PBC, and the polymorphisms of IL-1 +3953 and IL-10 promoter gene are not associated with PBC in a Chinese population.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Interleukin-1 , Genetics , Interleukin-10 , Genetics , Interleukin-6 , Genetics , Liver Cirrhosis, Biliary , Genetics , Polymerase Chain Reaction , Methods , Polymorphism, Restriction Fragment LengthABSTRACT
<p><b>OBJECTIVE</b>To study the effects of arsenic trioxide (As(2)O(3)) on cell cycle and expression of cyclin dependent kinase inhibitors (CDKIs) in multiple myeloma (MM) cells, and explore its pharmacological mechanism.</p><p><b>METHODS</b>The DNA content of MM cells line HS-Sultan was analyzed by flow cytometry after exposure to As(2)O(3), the effects on expression of CDKI P15, P16 AND P21 were studied by reverse transcriptase PCR.</p><p><b>RESULTS</b>DNA flow cytometric analysis showed that As(2)O(3) induced most of HS-Sultan cells, arrest at G(0)/G(1) phase and a small fraction at G(2)/M phase and apoptosis occurred mainly in S phase. There was no expression of P15 and P16 mRNA in untreated HS-Sultan cells and 1.0 micromol/L As(2)O(3) could make them expressed after exposed 24 or 48 hours respectively. Expression of P12 mRNA was obviously elevated by As(2)O(3) comparing with that of control.</p><p><b>CONCLUSION</b>One of the pharmacological mechanisms of As(2)O(3) is to activate the expression of CDKI P15, P16 and P21, and consequently affect cell proliferation cycle.</p>
Subject(s)
Humans , Antineoplastic Agents , Pharmacology , Arsenicals , Pharmacology , Cell Cycle , Physiology , Cyclin-Dependent Kinase Inhibitor p15 , Genetics , Cyclin-Dependent Kinase Inhibitor p16 , Genetics , Cyclin-Dependent Kinase Inhibitor p21 , Genetics , Multiple Myeloma , Drug Therapy , Metabolism , Pathology , Oxides , Pharmacology , RNA, Messenger , Genetics , Tumor Cells, CulturedABSTRACT
Objective To investigate the risk factors of intrahepatic recurrence of hepatocellular carcinoma(HCC) after radiofrequency ablation(RFA).Methods Forty-seven patients with total of 55 HCC mass were treated with RFA between March 2001 to August 2006.The patients were either Child- Pugh class A or B with total number of mass
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Objective: To study retrovirus (RV)-mediated transduction of gastric carcinoma cells with the herpes simplex virus thymidine kinase (HSV-tk) gene and the subsequent treatment with ganciclovir(GCV). Methods: The TK gene was transfected into human gastric carcinoma cell line MKN28 using HSV-TK that packed with PA317 cell, the sensitivity of MKN28TK cells to GCV was examined in vitro. Results: The retroviral-mediated HSV-TK gene can be transfected to MKN28 cells. The growth rate of MKN28 cells transfected with HSV-TK gene did not change. MKN28TK cells became significantly sensitive to GCV and had bystander effect. Conclusion: Transfection of gastric carcinoma with HSV-TK has higher transfection efficiency. MKN28TK cells are significantly sensitive to GCV.
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Objective: To investigate the effect of protein ki nase C on signal transduction such as tyrosine phosphorylation, c-fos and c-ju n mRNA expression in antigen activated mast cells. Methods: RBL-2H3 cells either untreated or treated with phorbol 12-myristate 13 -acetate (PMA) were sensitized with anti-DNP IgE, and activated with DNP-BSA, histamine release and tyrosine phosphorylation were quantitatively measured by ELISA and flow cytometry, respectively. The effect of PKC on the ex pression of c-fos and c-jun in serum-deprived RBL-2H3 cells activated by DNP-BSA detected by ethidium staining of PCR-amplified cDNA, the amplified cDNA products were subjected to Southern blot hybridization using specific prob es to determine the veracity of amplification. Results: Tyr osine phosphorylation and histamine release were significantly reduced from (4.4 7±0.03)% to (2.79±0.07)% and (104.47±1.31) nmol/L to (60.75±1.38) nm ol/L, respectively, 45 min after DNP-BSA stimulation in sensitized cells pre treated with PMA for 48 h. Bands of the size predicted for the amplified cDNA we re obtained: 299 bp for c-fos, and 651 bp for c-jun, a decrease of 91% and 82% , respectively, for c-fos and c-jun mRNAs was observed in antigen stimulated c ells pretreated with PMA for 48 h. Conclusion: PKC plays an impo rtant role in modulating the tyrosine phosphorylation and histamine release resp onses and may upregulate the expression of c-fos and c-jun in antigen activate d mast cell.
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Objective: To study retrovirus (RV)-mediated transduction of gastric carcinoma cells with the herpes simplex virus thymidine kinase (HSV-tk) gene and the subsequent treatment with ganciclovir(GCV). Methods: The TK gene was transfected into human gastric carcinoma cell line MKN28 using HSV-TK that packed with PA317 cell, the sensitivity of MKN28TK cells to GCV was examined in vitro. Results: The retroviral-mediated HSV-TK gene can be transfected to MKN28 cells. The growth rate of MKN28 cells transfected with HSV-TK gene did not change. MKN28TK cells became significantly sensitive to GCV and had bystander effect. Conclusion: Transfection of gastric carcinoma with HSV-TK has higher transfection efficiency. MKN28TK cells are significantly sensitive to GCV.
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Objective: To investigate the effect of protein ki nase C on signal transduction such as tyrosine phosphorylation, c-fos and c-ju n mRNA expression in antigen activated mast cells. Methods: RBL-2H3 cells either untreated or treated with phorbol 12-myristate 13 -acetate (PMA) were sensitized with anti-DNP IgE, and activated with DNP-BSA, histamine release and tyrosine phosphorylation were quantitatively measured by ELISA and flow cytometry, respectively. The effect of PKC on the ex pression of c-fos and c-jun in serum-deprived RBL-2H3 cells activated by DNP-BSA detected by ethidium staining of PCR-amplified cDNA, the amplified cDNA products were subjected to Southern blot hybridization using specific prob es to determine the veracity of amplification. Results: Tyr osine phosphorylation and histamine release were significantly reduced from (4.4 7±0.03)% to (2.79±0.07)% and (104.47±1.31) nmol/L to (60.75±1.38) nm ol/L, respectively, 45 min after DNP-BSA stimulation in sensitized cells pre treated with PMA for 48 h. Bands of the size predicted for the amplified cDNA we re obtained: 299 bp for c-fos, and 651 bp for c-jun, a decrease of 91% and 82% , respectively, for c-fos and c-jun mRNAs was observed in antigen stimulated c ells pretreated with PMA for 48 h. Conclusion: PKC plays an impo rtant role in modulating the tyrosine phosphorylation and histamine release resp onses and may upregulate the expression of c-fos and c-jun in antigen activate d mast cell.
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For purifying recombinant human IL—2 (rhIL—2),the columns of immunoabsorptionwere prepared with 4 anti—IL—2 McAb (9B12,9F5,9B2 and 8H7) purified by caprylic acid.Although 4 McAbs differ as regards their antigen—antibody binding characteristics,all they canserve as effective immnoabsorbents,provided optimum condition was adopted.The recoveryrate of 9B12,9F5,8H7 and 9B2 columns were 49.2%,37.5%,31.5% and 18.8% respec-tively.The purity of rhIL—2 obtained was more than 95% and biological activity remainedhigher.