RESUMO
SummaryHaplotype network is becoming popular due to its increasing use in analyzing genealogical relationships of closely related genomes. We newly proposed McAN, a minimum-cost arborescence based haplotype network construction algorithm, by considering mutation spectrum history (mutations in ancestry haplotype should be contained in descendant haplotype), node size (corresponding to sample count for a given node) and sampling time. McAN is two orders of magnitude faster than the state-of-the-art algorithms, making it suitable for analyzation of massive sequences. AvailabilitySource code is written in C/C++ and available at https://github.com/Theory-Lun/McAN and https://ngdc.cncb.ac.cn/biocode/tools/BT007301 under the MIT license. The online web service of McAN is available at https://ngdc.cncb.ac.cn/ncov/online/tool/haplotype. SARS-CoV-2 dataset are available at https://ngdc.cncb.ac.cn/ncov/.
RESUMO
Genomic epidemiology is important to study the COVID-19 pandemic and more than two million SARS-CoV-2 genomic sequences were deposited into public databases. However, the exponential increase of sequences invokes unprecedented bioinformatic challenges. Here, we present the Coronavirus GenBrowser (CGB) based on a highly efficient analysis framework and a movie maker strategy. In total, 1,002,739 high quality genomic sequences with the transmission-related metadata were analyzed and visualized. The size of the core data file is only 12.20 MB, efficient for clean data sharing. Quick visualization modules and rich interactive operations are provided to explore the annotated SARS-CoV-2 evolutionary tree. CGB binary nomenclature is proposed to name each internal lineage. The pre-analyzed data can be filtered out according to the user-defined criteria to explore the transmission of SARS-CoV-2. Different evolutionary analyses can also be easily performed, such as the detection of accelerated evolution and on-going positive selection. Moreover, the 75 genomic spots conserved in SARS-CoV-2 but non-conserved in other coronaviruses were identified, which may indicate the functional elements specifically important for SARS-CoV-2. The CGB not only enables users who have no programming skills to analyze millions of genomic sequences, but also offers a panoramic vision of the transmission and evolution of SARS-CoV-2.
RESUMO
On 22 January 2020, the National Genomics Data Center (NGDC), part of the China National Center for Bioinformation (CNCB), created the 2019 Novel Coronavirus Resource (2019nCoVR), an open-access SARS-CoV-2 information resource. 2019nCoVR features a comprehensive integration of sequence and clinical information for all publicly available SARS-CoV-2 isolates, which are manually curated with value-added annotations and quality evaluated by our in-house automated pipeline. Of particular note, 2019nCoVR performs systematic analyses to generate a dynamic landscape of SARS-CoV-2 genomic variations at a global scale. It provides all identified variants and detailed statistics for each virus isolate, and congregates the quality score, functional annotation, and population frequency for each variant. It also generates visualization of the spatiotemporal change for each variant and yields historical viral haplotype network maps for the course of the outbreak from all complete and high-quality genomes. Moreover, 2019nCoVR provides a full collection of SARS-CoV-2 relevant literature on COVID-19 (Coronavirus Disease 2019), including published papers from PubMed as well as preprints from services such as bioRxiv and medRxiv through Europe PMC. Furthermore, by linking with relevant databases in CNCB-NGDC, 2019nCoVR offers data submission services for raw sequence reads and assembled genomes, and data sharing with National Center for Biotechnology Information. Collectively, all SARS-CoV-2 genome sequences, variants, haplotypes and literature are updated daily to provide timely information, making 2019nCoVR a valuable resource for the global research community. 2019nCoVR is accessible at https://bigd.big.ac.cn/ncov/.
RESUMO
Purpose To study the expression of HIF-1αand Glut-1 in the molecular subtypes of breast carcinoma and their correlation with basal-like breast carcinoma. Methods 803 cases of invasive breast carcinoma from our database were identified. The clinicopath-ologic findings and the biologic markers including estrogen receptor ( ER) , progesterone receptor ( PR) , and human epidermal growth factor receptor-2 (HER-2) status were reviewed. Immunohistochemical MaxVision stains for cytokeratin 5/6 (CK5/6) and epidermal growth factor receptor ( EGFR) were performed. All breast carcinomas were subclassified into Luminal A, Lumincal B, HER-2 over-expression, normal-like, and basal-like subtypes according to Nielsen criteria. Immunohistochemical stain was also used to detect the expression of HIF-1αand Glut-1. Results Positive expression rates of HIF-1αprotein in basal-like, HER-2 over-expression, normal-like, Luminal A and Luminal B substypes were 77. 89% (74/95), 56. 06% (37/66), 55. 76% (92/165), 31. 97% (141/441), 25. 00% (9/36), respectively. The positive expression rates of Glut-1 protein were 80. 00% (76/95), 57. 58% (38/66), 58. 18%(96/165), 34. 01% (150/441), 25. 00% (9/36), respectively. The positive expression rates of HIF-1α and Glut-1 in the basal-like, HER-2 over-expressing and normal-like subtypes were remarkably higher than that in Luminal A and Luminal B subtypes ( P<0. 004 5) and the expression of HIF-1a and Glut-1 was negatively correlated with the expression of ER (P<0. 01). In the ER-negative breast cancers, the positive expression rates of HIF-1a and Glut-1 in basal-like substype were much higher than that in the other sub-types (P<0. 004 5), and the expression of HIF-1α was positively correlated with expression of Glut-1 in basal-like breast carcinoma (P<0. 01). Conclusion The overexpression of HIF-1αand Glut-1 may be closely related to the ER-negative breast cancer and HIF-1α and Glut-1 might play an important role in the development of basal-like breast carcinoma.
RESUMO
Objective To explore the application value of detecting HPV E6/E7 mRNA in cervical cancer screening and prognosis.Methods bDNA technology was used to detect the HPV E6/E7 mRNA levels in 301 cases of paraffin-embedded cervical samples that were grouped based on pathological diagnosis.Statistic differece between multiple groups were assessed by chi-square segmentation method.Results The positive detection rate of HPV E6/E7 mRNA is higher in cervical cancer group than that in CIN1-3 groups and cervicitis group,which were 94.29 ％ (66/70),30.30 ％ (20/66),50.00 ％ (19/38),59.46 ％ (44/74),5.66 ％ (3/50),respectively (all P ＜ 0.004 5).The HPV E6/E7-positive rates in all three CIN groups were higher than that in cervicitis group (P ＜ 0.004 5).The positive rates in CIN2 and CIN3 group were higher than that in CIN1 group (P ＜ 0.004 5).HPV E6/E7 mRNA levels in cervical cancer group were higher than those in CIN groups and cervicitis group (16 508.730±8 741.402,929:337±460.880,3 239.681±1 447.399,5 286.224±2 933.445,4.941±2.263,respectively,all P ＜ 0.05).CIN groups had higher copy numbers of HPV E6/E7 mRNA than cervicitis group (P ＜ 0.05).More copies of HPV E6/E7 mRNA were detected in CIN3 group than those in CIN1 group (P ＜ 0.05).Conclusions The positive rate and expression levels of HPV E6/ E7 mRNA was related to the severity of cervical diseases.HPV E6/E7 mRNA detection in paraffin-embedded samples have a high clinical application value in predicting the risk of cervical cancer.It' s an effective measure for cervical cancer screening and follow-up.
RESUMO
Objective To detect the protein expressions of S100A7 and CD34 in basal-like breast carcinoma and analyze the clinical relationship between S100A7 and basal-like breast cancer and the connection with CD34 markers microvessel denisity (MVD).To study the function of S100A7 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expressions of S100A7 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.Results The expressions of S100A7 in basal-like breast cancer,non basallike breast cancer and normal breast tissues were 83.33％,62.50％ and 35.00％ respectively.There were significant differences among them (x2 =5.556,P ＜ 0.051; x2 =17.107,P ＜ 0.05; x2 =4.051,P ＜ 0.05).There was positive correlation between S100A7 and lymph node metastasis,TNM staging and MVD of basal-like breast cancer patients.But there was no significant correlation between S100A7 and patient's age and tumor sizes.Conclusion The up-regulation of S100A7 expressions may be associated with the occurrence and development,invasion and metastasis,angiogenesis of basal-like breast cancer.The effects of S100A7 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basallike breast cancer.
RESUMO
Objective To detect the protein expression of Cripto-1 and CD34 in basal-like breast carcinoma and study the functions of Cripto-1 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expression of Cripto-1 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.The clinical relationship between Cripto-1 and basal-like breast cancer were analyzed as well as its connection with CD34 markers MVD.Results The expression of Cripto-1 protein in basal-like breast cancer (42/60,70.00 ％) was significantly higher than that of nonbasal-like breast cancer (19/40,47.50 ％) and normal breast tissue (4/20,20.00 ％) (both P ＜ 0.05).The expression of Cripto-1 protein was positively correlated with the tumor size,clinical stage,lymph nodes metastasis and MVD value in patients of basal-like breast cancer (all P ＜ 0.05),but there was no corelation with the age of the patients (P ＞ 0.05).Conclusions The up-regulation of Cripto-1 expression may be associated with angiogenesis and development in basal-like breast cancer.The effect of Cripto-1 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basal-like breast cancer.
RESUMO
Objective To study the expression and clinical significance of FoxM1 and Cep55 protein in basal-like breast carcinoma (BLBC).Methods The expression of FoxM1 and Cep55 protein in 66 cases of BLBC,70 cases of non-BLBC and 66 cases of normal adjacent breast tissue were detected by immunohistochemistry.Their relationship with the clinical pathological factors was analyzed.Results The positive rate of FoxM1 protein in BLBC,non-BLBC and normal adjacent breast tissue was 77.3％ (51/66),60.0％(42/70) and 13.6％(9/66),Cep55 protein was 74.2％(49/66),57.1％(40/70) and 16.7％(11/66),respectively.BLBC and non-BLBC tissue were higher than normal adjacent breast tissue,and BLBC tissue was higher than non-BLBC,there was significant difference (P ＜0.05).In BLBC tissue,the positive expression of FoxM1 and Cep55 protein correlated with lymph nodes metastasis and TNM staging (P ＜0.05),while did not correlate with age,menopause,tumor size (P ＞ 0.05).There was direct correlation between the positive expression of FoxM1 and Cep55 protein (r =0.259,P ＜ 0.05).Conclusion FoxM1 and Cep55 protein may play an important role in the carcinogenesis and development of BLBC,and both of them have the synergistic effect.
RESUMO
Objective To investigate the expressions and significances of SIRT1 and p53 in endometrial carcinoma with diabetes mellitus.Methods Immunohistochemistry was carried out to examine the expressions of SIRT1 and p53 in 39 cases of endometrial carcinoma with diabetes mellitus and 40 cases of endometrial carcinoma without diabetes mellitus and 25 cases of normal endometrium,and the correlations between these indexs were analysed.Results The positive rate of SIRT1 and p53 in experimental group was 64.1％,59.0％ respectively,and 85.0％,52.5％ in control group.The positive rate of SIRT1 in experimental group was lower than that of control group (x2 =4.561,P =0.033).The positive rates of SIRT1 and p53 in control group were higher than that of normal endometrium (x2 =21.462,P＜ 0.001; x2 =6.771,P =0.009).Expression of SIRT1 was positively correlated with p53 (r =0.360,P =0.024).Conclusions Hyperglycemia deregulates SIRT1 in endometrial carcinoma.SIRT1 may promote tumorigenesis by silence of p53.
RESUMO
Background: Isolated systolic hypertension (ISH) is a common disease in elderly people, threatening their health. Traditional Chinese medicine (TCM) treatment or integrative treatment had advantages in improving quality of life and protecting target organs, but need to be proved by large evidence-based researches. Objective: To observe the effects of TCM treatment (Jiangya Capsule) or integrative treatment (combination of Jiangya Capsule and nimodipine) on blood pressure and vasoactive agents, and their safety in elderly ISH patients. Design, setting, participants and interventions: A multicenter, randomized, double-blind controlled trial was adopted. A total of 270 elderly ISH patients recruited from Xiyuan Hospital, and TCM Hospital and Community Health Service Centers of Yanqing County of Beijing were randomly divided into 3 groups: TCM group (Jiangya Capsule plus nimodipine simulation, 90 cases), integrative group (Jiangya Capsule plus nimodipine, 90 cases) and Western medicine (WM) group (nimodipine plus Jiangya Capsule simulation, 90 cases). They were all treated for 4 weeks. Main outcome measures: Before and after 4-week treatment, office blood pressure, 24-hour ambulatory blood pressure, serum nitric oxide (NO), and plasma endothelin-1 (ET-1), thromboxane B2 (TXB2) and 6-keto-prostaglandin 1alpha (6-keto-PGF1alpha) were detected, and safety evaluation was conducted. Results: After 4-week treatment, 5 patients in TCM group were lost to follow-up and another 5 patients were excluded, and 80 patients finished the trial; 7 patients in integrative group were lost to follow-up and another 7 patients were excluded, and 76 patients finished the trial; 2 patients in WM group were lost to follow-up and another 3 patients were excluded, and 85 patients finished the trial. After treatment, systolic blood pressure (SBP) decreased in each group (P<0.05), and integrative treatment was superior to TCM or WM treatment in decreasing SBP (P<0.05). Twenty-four hour average SBP and day average SBP decreased significantly in each group, and night average SBP decreased in integrative group, and integrative treatment was superior to TCM or WM treatment in decreasing day average SBP. Serum NO and plasma 6-keto-PGF1alpha levels were elevated and plasma ET-1 and TXB(2) levels were reduced after treatment, and integrative treatment was superior to TCM or WM treatment in reducing plasma TXB(2) level. Conclusion: TCM treatment or integrative treatment has affirmative effects and safety in treating elderly ISH patients, and integrative treatment has superiority in improving some indexes, and deserves further study.
RESUMO
<p><b>OBJECTIVE</b>To observe the intervention effects of Huannao Yicong decoction on cognitive function and apoptosis and expression of relative regulative gene of Hippocampus in cognitive impairment rats induced by complex factors.</p><p><b>METHOD</b>60 SD rats were divided randomly into Huannao Yicong decoction high-dose group (HHG), Huannao Yicong decoction low-dose group (HLG), positive control group (PCG), model control group (MCG) and blank control group (BCG). Rats in the BCG were received daily hypodermic injection of tales doses of normal sodium for 10 weeks with normal feeder. Rats in other groups were received daily hypodermic injection of D-galactose with the concentration of 50 mg kg(-1) for 10 weeks, from the 5th week on, half fat feeder were fed until the end of the 10th week. From the 7th on, rats in HHG were administered with 0.01 mL g(-1) Huannao Yicong decoction suspension by gavage (crude drug 14 g kg(-1)). Rats in LHG were administered with 0.01 mL g(-1) Huannao Yicong decoction suspension by gavage (crude drug 7 g kg(-1)). Rats in PCG were administered with 0.01 mL g(-1) hydrochloricdonepezil suspension by gavage (0.4 mg kg(-1)). Rats in MCG and BCG were administered with 0.01 mL g(-1) distilled water by gavage, intragastric administration was given daily until the end of the 10th week. The behaviors of the rats were observed by morris water maze, the apoptosis and expression of relative regulative gene of hippocampus were measured.</p><p><b>RESULT</b>The morris water maze indicated that compared with the BCG, the platform locating latency of rats in the MCG was longer and the frequency of swimming through the platform was fewer(P <0.05, P < 0.01), compared with the MCG, there was significant difference on the frequency of swimming through the platform in the HHG and PCG (P <0.05, P <0.01). The number of apoptosis cells in the MCG was more than that in the BCG, the difference was significant (P <0.01), the number of apoptosis cells in the HHG, HLG and PCG was reduced and the ratio of Bcl-2 and Bax was increased (P < 0.01).</p><p><b>CONCLUSION</b>Huannao Yicong decoction could improve the learning and memory functions of cognitive impairment rats, inhibit the apoptosis of cells in hippocampus, regulate the expression of relative gene, accelerate the repairing of cells, protect the impaired brain tissue, and these may be part of the channels of clinical effects.</p>
Assuntos
Animais , Feminino , Humanos , Ratos , Apoptose , Transtornos Cognitivos , Tratamento Farmacológico , Genética , Metabolismo , Modelos Animais de Doenças , Medicamentos de Ervas Chinesas , Expressão Gênica , Hipocampo , Metabolismo , Proteínas Proto-Oncogênicas c-bcl-2 , Genética , Metabolismo , Distribuição Aleatória , Ratos Sprague-Dawley , Proteína X Associada a bcl-2 , Genética , MetabolismoRESUMO
OBJECTIVE: To observe the effect and explore the mechanism of Huannao Yicong capsule in treating senile patients with mild cognitive impairment (MCI). METHODS: The investigational drugs were packed by blind method. A randomized, double-blind and controlled trial was conducted on ninety senile patients with MCI. Other forty-five senile healthy persons were recruited to the healthy control group. The ninety senile patients were randomly divided into the Huannao Yicong capsule-treated group (45 patients administered with three Huannao Yicong capsules and two aniracetam capsule analogues) and aniracetam-treated group (45 patients treated with two aniracetam capsules and three Huannao Yicong capsule analogues). Patients in the two groups were treated three times daily for 16 weeks. Memory, traditional Chinese medicine syndrome, cerebral blood flow, free radicals and inflammatory mediators, such as superoxide dismutase (SOD), malondialdehyde (MDA), acetylcholinesterase (AchE), interleukin-1alpha (IL-1alpha) and interleukin-6 (IL-6) were determined before and after the treatment. Blood lipids, including triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), apolipoprotein A-1 (ApoA-1) and apolipoprotein B-100 (ApoB-100), were detected before and after the treatment. The safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and electrocardiogram (ECG) were taken before and after the treatment. RESULTS: Index score of clinical memory scale in senile healthy people was significantly higher than that in MCI patients before treatment (P<0.01), and the content of AchE, IL-1alpha and IL-6 was obviously lower (P<0.01, P<0.05), the activity of SOD was higher (P<0.05). No significant difference was found in direction memory of clinical memory scale between the two treatment groups. Other index scores of clinical memory scale and traditional Chinese medicine syndrome in patients of Huannao Yicong capsule-treated group were significantly improved as compared with those of the aniracetam-treated group (P<0.05, P<0.01). The blood flow parameters of anterior cerebral artery, posterior cerebral artery and resistant index in patients of Huannao Yicong capsule-treated group were increased significantly (P<0.01, P<0.05). Huannao Yicong capsule could significantly increase the activity of serum SOD and decrease the content of AchE, IL-1alpha and IL-6 (P<0.01, P<0.05), better than aniracetam. Furthermore, Huannao Yicong capsule could significantly improve the blood lipid, such as the level of TG, LDL-C, HDL-C, ApoA-1 and ApoB-100 (P<0.01, P<0.05), and better than aniracetam (P<0.01, P<0.05). No significant changes were found after treatment in safety indexes, such as routine tests of blood and urine, hepatic and renal function tests and ECG. CONCLUSION: Huannao Yicong capsule has better therapeutic effect than aniracetam capsule in treating senile mild cognitive impairment.
RESUMO
Objective To study the biological characteristics of dendritic cells (DCs) derived from peripheral blood mononuclear cells (PBMCs) of patients diagnosed with syphilis. Methods PBMCs were isolated from 16 patients clinically and serologically diagnosed with syphilis, and from 16 healthy human controls, then cultured with GM-CSF and IL-4. On day 10, the monocyte-derived dendritic cells (MoDCs)of the patients and controls were collected and subjected to the detection of surface molecules by flow cytometry; TpN17 was used to stimulate MoDCs from the controls, the expression of phosphorylated ERK was detected by Westem blotting 20 minutes following the stimulation. Results The positivity rate of CD80 was significantly increased in the patients with syphilis than that in the controls (51.90% vs 33.67,P ＜ 0.05), while no significant difference was observed in the expressions of CD83, CD86 or HLA-DR be tween the two groups (16.53% vs 15.99%, 66.13% vs 59.32%, 91.29% vs 90.51%, all P 0.05). The ex pressions of CD80 and CD83 on the surface of MoDCs were enhanced in a dose-dependent manner after ex-posure to TpN17. The expression of cytoplasmic phosphorylated ERK was observed in MoDC stimulated by TpN17, but not in those without the treatment. Conclusions Antigenic stimulation with Treponema pal-lidum may be a reason for phenotypic abnormality of MoDCs derived from patients with syphilis. TpN17 may stimulate the maturation of DCs through the ERK signal transduction pathway.
RESUMO
AIM: To solve the two difficulties of bone resorption and inflammation in rheumatoid arthritis, clone the recombinant human osteoprotegerin (OPG) and mycobacteria heat shock protein 70 (HSP70) functional gene,and study the expression and activity of OPG-HSP70 fusion protein in E.coli. METHODS: Experiments were performed at the Laboratory of Department of Immunology, Capital Medical University from May 2006 to September 2007. Complementary DNA encoding full length OPG protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR) from human osteosarcoma cell line MG63 and cloned into pGEMT-Easy vectors. Then, using the recombinant plasmid as the template,the DNA encoding the fusion protein OPG-HSP 70 was amplified by PCR,and was inserted into prokaryotic expression vector pET-28a. Construct pET-28a-OPG-HSP 70 was used to transform into competent E.coli. BL21(DE3) which were induced by isopropyl B-D-thiogalactopyranoside (IPTG),and the fusion protein from above E.coli was collected. Sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western-blotting were performed to identify OPG-HSP 70 fusion protein. The experiments of osteoclast inhibition and restraining inflammation were used to detect the bioactivity of fusion protein. RESULTS: ①The complementary DNA encoding full length OPG protein was obtained. OPG-HSP 70 fusion gene obtained in this experiment was successfully inserted into pET-28a vector. OPG-HSP70 fusion protein was expressed when transformed into E.coli.BL21(DE3). ②SDS-PAGE indicated that the fusion protein was large expressed at the molecular weight of Mr22 000, but there were no band in the total lysate of bacteria harboring pET-28a-OPG-HSP70 without IPTG induction group. ③Western-blotting indicated that the OPG-HSP70 fusion protein could specifically react with anti-human OPG monoclonal antibodies. ④The osteoclast inhibition test demonstrated that the fusion protein could reduce the number of osteoclast, and had the ability to inhibit bone absorptions in vitro. ⑤The experiment of restraining inflammation showed that the fusion protein could significantly reduce the inflammation of delayed type hypersensitivity (DTH) mice, which explained that HSP70 in fusion protein had the inflammation inhibitory bioactivity. CONCLUSION: OPG-HSP70 fusion protein is expressed in E.coli.BL21(DE3),and function study in vitro illustrates the bioactivity of fusion protein.
RESUMO
Clinical immunology should display the advanced and applied characteristics of immunology,and show the speciality of clinical practice.This course is a bridge between fundamental immunology and clinical teaching so that the medicos should get better foundation for the future learning.
RESUMO
<p><b>OBJECTIVE</b>To investigate the effects of oral administration of type II collagen (CII) on adjuvant arthritis (AA) in rats and its mechanisms, and to compare the effects of CII with those of the Chinese traditional medicine Tripterygium Polyglycoside administered similarly.</p><p><b>METHODS</b>Arthritis was induced in rats by immunization using Freund's complete adjuvant (FCA). After feeding rats either soluble CII or Tripterygium Polyglycoside, changes in degree of articular swelling and articular histological findings were observed in AA rats. Some correlative immunological indexes were measured, including delayed type hypersensitivity (DTH) reaction, anti-collagen and anti-Mycobacterium tuberculosis (MT) antibody in serum, and levels of IFN-gamma and TNF-alpha in articular steep in rats.</p><p><b>RESULTS</b>Oral administration of CII was able to alleviate both distinctly articular and general symptoms in AA rats, suppress synovium hyperplasia and inflammatory cells infiltration in arthrosis capsule. The effects brought about by CII were stronger than those by Tripterygium Polyglycoside. Oral administration of CII inhibited antigen-specific immune response, such as DTH and antibody reaction to CII. In addition, the expression of IFN-gamma and TNF-alpha in joints were locally downregulated.</p><p><b>CONCLUSIONS</b>The therapeutic effect of oral administration of CII is obvious on adjuvant arthritis in rats. Its remedial mechanisms are likely related to the downregulation of both IFN-gamma and TNF-alpha, and the suppression of cell immunity.</p>
Assuntos
Animais , Masculino , Ratos , Administração Oral , Anticorpos , Sangue , Artrite Experimental , Tratamento Farmacológico , Alergia e Imunologia , Colágeno Tipo II , Usos Terapêuticos , Hipersensibilidade Tardia , Tolerância Imunológica , Interferon gama , Mycobacterium tuberculosis , Alergia e Imunologia , Fitoterapia , Ratos Sprague-Dawley , Membrana Sinovial , Patologia , Tripterygium , Fator de Necrose Tumoral alfaRESUMO
Aim To investigate the effects of monocrotaline pyrrole on production of nitric oxide and expression of eNOS protein of cultured pulmonary artery endothelial cells and on contractility of cultured pulmonary artery smooth muscle cells.Methods DAF-2 fluorescence technique was used to determine NO level,Western blot analysis was performed to determine the level of eNOS protein,and collagen gel contraction system was adopted to analyze muscle contractility.Results NO production induced by ACh and expression of eNOS protein were obviously inhibited by monocrotaline pyrrole compared with those of control group and gel contraction area in MCTP-treated cells induced by Thapsigargin obviously decreased.Conclusions monocrotaline pyrrole could inhibit the level of the ACh-induced production of NO and expression of eNOS protein,and enhance the contractility of pulmonary artery smooth muscle cells,which may be one of the possible mechanisms of MCTP-induced pulmonary artery hypertension.
RESUMO
ObjectiveTo study Decorin biological effects,Decorin gene cDNA was cloned and expressed in E. coli. MethodsThe Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19. The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method, and then it was subcloned into expression plasmid vector pGEX-4T-1, the recombinant vector was indentified with BamHI and EcoRI in 1.2 ％ agarose gel electrophoresis. The expression of Decorin fusion protein with GST in E. coli JM109 was induced with IPTG and identified by SDS-PAGE. ResultsThe PCR amplified DNA fragment shared identical sequence with known Decorin gene reported in GenBank (Accession number: AF138300). The SDS-PAGE analysis revealed that the expressed fusion protein MW was approximatelly 65.6KD and in soluble format. ConclusionThe cloned Decorin gene is correct and it was expressed in fusion protein with GST.
RESUMO
AIM: To apply the solid phase extraction (SPE) on salvianolic acids in Radix Salvia Miltiorrhizae by means of high performance liquid chromatography with mass spectroscopic detector (HPLC MSn). METHODS: A C 18 solid phase column and negative ion electrospray ionization (ESI) mode were used by orthogonal design to optimize four factors which might affect recoveries of samples, which included vacuum tightness, sample size, amount of eluting agent and eluting power. RESULTS: The optimized results were as follows: vacuum tightness was 0.002?106 Pa, sample size was 0.75 mL, amount of eluting agent was 15mL and eluting power was 1.0mL?s -1 . CONCLUSION: The established method is simple, reliable and suitable for the usual quality control method of salviamolic acids in Radix Salvia Miltiorrhizae.
RESUMO
Objective:To study the relationship between the T cell clonotypes and RA,carried out the analysis of infiltrating T cell clonotypes in foot of a murine model of spontaneous RA Methods:RT PCR and SSCP were used for this study Results:The identical rate of V?2 and V?8 2 clonotypes of T cells in different foot joints is 68 1 percent and 59 2 percent separately The assay of RT PCR/SSCP showed there were identical V?2 and V?8 2 clonotypes in four foot joints of all SKG mice(100%) DNA sequencing showed that the same DNA bands in samples from different joints by SSCP were the same T cell clonotypes The assay of T cell transformation suggested that clonotypes of T cells showed up in joints is related to morbid formation of RA Conclusion:This investigation indicated that T cells of V?2 and V?8 2 TCR types probably play an important role in SKG murine RA
