Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 29
Filtrar
1.
Chinese Journal of Microbiology and Immunology ; (12): 182-190, 2023.
Artigo em Chinês | WPRIM | ID: wpr-995272

RESUMO

Objective:To isolate and culture WU polyomavirus (WUPyV), and to analyze the genome-wide evolutionary patterns, homology and population dynamics.Methods:Real-time quantitative PCR was used to detect the nasopharyngeal aspirate samples of hospitalized children with respiratory tract infection in Beijing Friendship Hospital during 2020 to 2022. Primary human airway epithelial cells cultured at the air-liquid interface were used to isolate and culture WUPyV. Whole genome sequence of the isolated strain was obtained by Sanger sequencing. For phylogenetic and evolutionary dynamics analysis, the whole genome was compared with the published whole genome sequences in GenBank database.Results:The detection rate of WUPyV was 4.7% (31/659) during 2020 to 2022, and a clinical strain BJ0593 of WUPyV type Ⅲc was successfully isolated. The homology of the whole genome and gene fragments of WUPyV was high. The average evolutionary rate of VP2 gene was about 1.256×10 -4 substitution/site every year, and the population dynamics of WUPyV tended to be flat in the last decade. Conclusions:This study successfully isolated a clinical WUPyV type Ⅲ strain for the first time, which provided the basis for further investigation on the molecular evolution and pathogenicity of WUPyV.

2.
Journal of Breast Cancer ; : 330-343, 2021.
Artigo em Inglês | WPRIM | ID: wpr-899003

RESUMO

Purpose@#Non-coding RNA activated by DNA damage (NORAD) has been reported to be a cancer-related long non-coding RNA (lncRNA) implicated in the progression of several cancers; however, its role in breast cancer (BC) has not yet been clarified. @*Methods@#Quantitative real-time polymerase chain reaction was used to examine NORAD, microRNA (miR)-155-5p, and suppressor of cytokine signaling 1 (SOCS1) mRNA expression levels. Western blotting was used to analyze SOCS1 protein expression. The malignancy of BC cells was assessed using the cell counting kit-8 (CCK-8), BrdU, and Transwell assays.Bioinformatics analysis, RNA immunoprecipitation assay, and dual-luciferase reporter gene assays were used to verify the targeted relationship between NORAD and miR-155-5p.Additionally, the regulatory effects of NORAD and miR-155-5p on SOCS1 expression were determined by western blotting. @*Results@#NORAD expression was significantly reduced in BC cell lines and tissues, and its low expression was associated with poor tumor tissue differentiation. NORAD overexpression repressed BC cell proliferation, migration, and invasion, whereas its knockdown produced the opposite effects. Additionally, miR-155-5p was found to be a target of NORAD, and the biological functions of miR-155-5p and NORAD were counteractive. MiR-155-5p was confirmed to target SOCS1, and SOCS1 was found to be positively regulated by NORAD. @*Conclusion@#NORAD suppresses miR-155-5p to upregulate SOCS1, thereby repressing the proliferation, migration, and invasion of BC cells.

3.
Chinese Journal of Medical Education Research ; (12): 1295-1298, 2021.
Artigo em Chinês | WPRIM | ID: wpr-909010

RESUMO

Problem-based learning (PBL) has been applied in many teaching fields and achieved satisfying effect. Dermatovenology is a special secondary discipline of clinical medicine which includes dermatosurgery, dermatopathology, laser cosmetology and other different sub-professional subjects, and it covers a wide variety of diseases and it is difficult to identify skin lesions. However, the teaching time for graduate students is limited and the traditional teaching mode can't meet the needs of learning for them. In this paper, the application of PBL method in the teaching of medical graduate students and the application of PBL combined with other teaching methods have been expounded through the analysis of domestic and foreign literature, and its role in the teaching of dermatology graduate students will be discussed.

4.
Journal of Breast Cancer ; : 330-343, 2021.
Artigo em Inglês | WPRIM | ID: wpr-891299

RESUMO

Purpose@#Non-coding RNA activated by DNA damage (NORAD) has been reported to be a cancer-related long non-coding RNA (lncRNA) implicated in the progression of several cancers; however, its role in breast cancer (BC) has not yet been clarified. @*Methods@#Quantitative real-time polymerase chain reaction was used to examine NORAD, microRNA (miR)-155-5p, and suppressor of cytokine signaling 1 (SOCS1) mRNA expression levels. Western blotting was used to analyze SOCS1 protein expression. The malignancy of BC cells was assessed using the cell counting kit-8 (CCK-8), BrdU, and Transwell assays.Bioinformatics analysis, RNA immunoprecipitation assay, and dual-luciferase reporter gene assays were used to verify the targeted relationship between NORAD and miR-155-5p.Additionally, the regulatory effects of NORAD and miR-155-5p on SOCS1 expression were determined by western blotting. @*Results@#NORAD expression was significantly reduced in BC cell lines and tissues, and its low expression was associated with poor tumor tissue differentiation. NORAD overexpression repressed BC cell proliferation, migration, and invasion, whereas its knockdown produced the opposite effects. Additionally, miR-155-5p was found to be a target of NORAD, and the biological functions of miR-155-5p and NORAD were counteractive. MiR-155-5p was confirmed to target SOCS1, and SOCS1 was found to be positively regulated by NORAD. @*Conclusion@#NORAD suppresses miR-155-5p to upregulate SOCS1, thereby repressing the proliferation, migration, and invasion of BC cells.

5.
Journal of Zhejiang University. Medical sciences ; (6): 232-239, 2020.
Artigo em Chinês | WPRIM | ID: wpr-828563

RESUMO

Acute respiratory failure due to acute hypoxemia is the major manifestation in severe coronavirus disease 2019 (COVID-19) induced by severe acute respiratory syndrome coronavirus 2 infection. Rational and effective respiratory support is crucial in the management of COVID-19 patients. High-flow nasal cannula (HFNC) has been utilized widely due to its superiority over other non-invasive respiratory support techniques. To avoid HFNC failure and intubation delay, the key issues are proper patients, timely application and improving compliance. It should be noted that elder patients are vulnerable for failed HFNC. We applied HFNC for oxygen therapy in severe and critical COVID-19 patients and summarized the following experiences. Firstly, to select the proper size of nasal catheter, to locate it at suitable place, and to confirm the nose and the upper respiratory airway unobstructed. Secondly, an initial flow of 60 L/min and 37℃ should be given immediately for patients with obvious respiratory distress or weak cough ability; otherwise, low-level support should be given first and the level gradually increased. Thirdly, to avoid hypoxia or hypoxemia, the treatment goal of HFNC should be maintained the oxygen saturation (SpO) above 95% for patients without chronic pulmonary disease. Finally, patients should wear a surgical mask during HFNC treatment to reduce the risk of virus transmission through droplets or aerosols.


Assuntos
Idoso , Humanos , Betacoronavirus , Cânula , Infecções por Coronavirus , Terapêutica , Hipóxia , Terapêutica , Máscaras , Oxigênio , Oxigenoterapia , Padrões de Referência , Pandemias , Pneumonia Viral , Terapêutica
6.
Journal of Zhejiang University. Medical sciences ; (6): 232-239, 2020.
Artigo em Chinês | WPRIM | ID: wpr-828547

RESUMO

Acute respiratory failure due to acute hypoxemia is the major manifestation in severe coronavirus disease 2019 (COVID-19). Rational and effective respiratory support is crucial in the management of COVID-19 patients. High-flow nasal cannula (HFNC) has been utilized widely due to its superiority over other non-invasive respiratory support techniques. To avoid HFNC failure and intubation delay, the key issues are proper patients, timely application and improving compliance. It should be noted that elder patients are vulnerable for failed HFNC. We applied HFNC for oxygen therapy in severe and critical ill COVID-19 patients and summarized the following experiences. Firstly, to select the proper size of nasal catheter, to locate it at suitable place, and to confirm the nose and the upper respiratory airway unobstructed. Secondly, an initial ow of 60 L/min and 37℃ should be given immediately for patients with obvious respiratory distress or weak cough ability; otherwise, low-level support should be given first and the level gradually increased. Thirdly, to avoid hypoxia or hypoxemia, the treatment goal of HFNC should be maintained the oxygen saturation (SpO) above 95% for patients without chronic pulmonary disease. Finally, patients should wear a surgical mask during HFNC treatment to reduce the risk of virus transmission through droplets or aerosols.


Assuntos
Idoso , Humanos , Betacoronavirus , Cânula , Infecções por Coronavirus , Terapêutica , Oxigênio , Pandemias , Pneumonia Viral , Terapêutica
7.
Chinese Journal of Medical Education Research ; (12): 593-595, 2019.
Artigo em Chinês | WPRIM | ID: wpr-753428

RESUMO

To explore the organ-system oriented integrated program, this project constructed a new clinical skill training program of sensory system for"5+3" medical students. According to the current trend and requirement of medical teaching reform, taking into consideration of the present status of clinical skills teaching, this project integrated the basic and clinical teaching contents of otolaryngology, ophthalmology and dermatology to design a new training program of clinical skills for"5+3"medical students. Specifically, the project group edited a new textbook and syllabus, and set up a specialized teaching group. In this system, the training models included simulation training and practical bedside training, the assessment included operation ability and analyzing ability for clinical cases, and the training contents involved all conventional skills of each specialty. It has been demonstrated that the new training program is capable of promoting the clinical skills of students quickly and effectively.

8.
The Korean Journal of Pain ; : 235-243, 2018.
Artigo em Inglês | WPRIM | ID: wpr-742202

RESUMO

Postherpetic neuralgia (PHN) is the most troublesome side effect of Herpes Zoster (HZ), which mainly affects the elderly and immunocompromised populations. Despite the current advancement of treatments, PHN persists in many individuals influencing their daily activities and reducing their quality of life. Anticonvulsants, antidepressants, topical therapies including lidocaine and capsaicin, and opioids, are the most widely used therapies for the treatment of PHN. These medications come with their adverse effects, so they should be used carefully with the elderly or with patients with significant comorbidities. Other measures like botulinum toxin, nerve blocks, spinal cord stimulation, and radiofrequency have also contributed significantly to the management of PHN. However, the efficacy, safety, and tolerability of these invasive methods need to be carefully monitored when administering them. Early diagnosis and early initiation of treatment can reduce the burden associated with PHN. The zoster vaccine has effectively reduced the incidence of HZ and PHN. In this article, we discuss the treatment options available for the management of PHN, mainly focusing on the efficacy and safety of different therapeutic modalities.


Assuntos
Idoso , Humanos , Analgésicos Opioides , Anticonvulsivantes , Antidepressivos , Toxinas Botulínicas , Capsaicina , Comorbidade , Diagnóstico Precoce , Herpes Zoster , Vacina contra Herpes Zoster , Incidência , Lidocaína , Bloqueio Nervoso , Neuralgia Pós-Herpética , Qualidade de Vida , Estimulação da Medula Espinal
9.
Chinese Journal of Experimental and Clinical Virology ; (6): 558-561, 2017.
Artigo em Chinês | WPRIM | ID: wpr-808833

RESUMO

Objective@#To establish mouse intestinal tissue and its application in the proliferation of Marmota Himalayana hepatovirus (MHHAV).@*Methods@#The intestinal crypts of Balb/C mice were digested with EDTA at a high concentration, cultured in Matrigel and passaged into frozen intestinal enteroids. The monolayers were inoculated on 96-well plates and infected with MHHAV. Real-time qPCR virus concentrations at different time points were compared with those of a number of cell lines routinely used to develop hepatitis A virus.@*Results@#The mouse intestinal tissue culture system was successfully established. MHHAV did not proliferate in the cell lines commonly used for the culture of hepatitis A virus, but proliferated well in the intestinal tissues of mice. The virus multiplied about 3 700 fold in 4 days.@*Conclusions@#Mice intestinal enteroids can be used for the isolation and culture of MHHAV.

10.
Chongqing Medicine ; (36): 2085-2088,2093, 2017.
Artigo em Chinês | WPRIM | ID: wpr-610032

RESUMO

Objective To evaluate the clinical efficacy and adverse reaction of acitretin combined with total glucosides of peony for patients with psoriasis.Methods Clinical controlled trials that compared acitretin with combination therapy of acitretin and total glucosides of peony for patients with psoriasis were searched in the Cochrane Library,PubMed,Embase and so on(from establishment to October 2016).All extractive data analysis was performed by Review Manage version 5.3 after the data was extracted and the quality was assessed according to the inclusion criteria.Results A total of 8 articles were included.The results of meta-analysis showed that the total effective rate[OR=3.15,95%CI(2.04,4.86),P0.05).Conclusion Acitretin combined with total glucosides of peony for patients with psoriasis can improve the clinical efficacy and reduce the incidence of adverse reaction.

11.
Chinese Circulation Journal ; (12): 676-678, 2016.
Artigo em Chinês | WPRIM | ID: wpr-497301

RESUMO

Objective: To study the features of spontaneous isolated superior mesenteric artery dissection (SISMAD) by unexpected detection of renal artery dual-source CT (DSCT) imaging in hypertension patients. Methods: A total of 4107 patients with suspected secondary hypertension received renal artery DSCT examination in our hospital from 2010-03 to 2015-04 were studied and SISMAD was unexpectedly found in 12 patients. There were 3 patients with mild abdominal pain and the rest without obvious abdominal symptoms. The position and length, true and false lumens, detached tunica intimal flap and branch involvement of dissection, intestinal wall edema and ileus were recorded. Results: SISMAD in all 12 (0.3%) patients were found unexpectedly. Axial CT with post-processing technique clearly displayed the ruptured tunica intimal orifice, true and false lumens, detached intimal flap; the branches were all originated from true lumen. According to Sakamoto classification, all 12 patients were belong to Type I as the true and false lumens were with an entry and re-entry respectively, no filling defect in false lumen. The distance from orifice of dissection to root of abdominal aorta was (26.7 ± 11.3) mm and the length of dissection was (35.1 ± 11.7) mm.There were 10 patients with aneurysmal expansion with the diameter of (11.9 ± 2.5) mm. Conclusion: Unexpected detection of SISMAD by renal artery CT imaging was about 0.3%, radiologist should pay special attention to find superior mesenteric artery dissection in hypertension patients.

12.
Chinese Journal of cardiovascular Rehabilitation Medicine ; (6): 498-501, 2015.
Artigo em Chinês | WPRIM | ID: wpr-478243

RESUMO

Objective:To explore the relationship among cardioversion ,cerebral infarction (CI) and N terminal pro brain natriuretic peptide (NT‐proBNP) level in patients with heart failure (HF) complicated atrial fibrillation (AF) . Methods :A total of 150 HF + AF patients received intravenous drip of amiodarone for cardioversion therapy .Ac‐cording to cardioversion results ,they were divided into cardioversion group (n=100 ) and non‐cardioversion group (n=50) ,NT‐proBNP level change was observed in two groups before and after cardioversion .According to CI on‐set or not ,patients were divided into CI group (n=20) and non‐CI group (n= 130) ,NT‐proBNP level was com‐pared between two groups before and after onset .Results :Within 48h after administration ,a total of 100 patients (66.67% ) recovered to sinus rhythm .Compared with before cardioversion ,NT‐proBNP level significantly reduced [(967.04 ± 366.16) pg/ml vs .(496.21 ± 142.54) pg/ml] after cardioversion in cardioversion group ,and was signifi‐cantly lower than that of non‐cardioversion group (996.76 ± 351.28) pg/ml , P<0.01 all . In CI group ,compared with small size CI group ,there were significant rise in NT‐proBNP level [ (784.21 ± 231.26) pg/ml vs .(1983.24 ± 32.96) pg/ml ,(3562.19 ± 1468.32) pg/ml] in medium and large size CI group , P< 0.05 or <0.01 .Conclusion:NT‐proBNP level at hospitalization possesses predictive value for drug cardioversion effect in HF + AF patients . NT‐proBNP level is related with CI onset .After acute CI ,the higher NT‐proBNP level is ,the larger infarct size is , the poorer prognosis is .

13.
Chongqing Medicine ; (36): 1851-1853,1856, 2014.
Artigo em Chinês | WPRIM | ID: wpr-572408

RESUMO

Objective To observe the clinical efficacy and safety of calcipotriol betamethasone ointment for the treatment of pso‐riasis vulgaris in the stable stage of patient .Methods This was a randomized ,parallel controlled clinical trials ,in which 90 patients with psoriasis vulgaris were randomly divided into experimental group and controlled group .All of them received 4 weeks of thera‐py .We compared the efficacy and safety 1 ,2 ,4 week after treatment by calcipotriol betamethasone ointment .Results About the clinical efficacy ,according to the decrease in percentage of psoriasis area and severity index (PASI) were:23 patients in the experi‐mental group were effective(efficacy rate is 76 .67% ) ,14 patients in control group Ⅰ was effective(efficacy rate is 46 .67% ) ,15 pa‐tients in control group Ⅱ was effective(efficacy rate is 50% ) .The differention between experimental group and the two control groups was significant(P< 0 .05 ,P< 0 .01) .After 1 ,2 ,4 weeks of therapy ,the decrease in percentage of PASI of experimental group is higher than that in the control groups .The scores of the observed erythema ,infiltration ,scale were decreased ,and the total scores of PASI are also decreased 4 weeks after therapy .The decrease score of experimental group is higher than that of control group Ⅰ(P<0 .01) and control group Ⅱ(P<0 .01) .The side effects of the experimental group are mild such as itching ,folliculi‐tis ,erythema ,and the laboratory examinations had no abnormal changes .Conclusion Calcipotriol betamethasone ointment was more effective to treat the stable stage psoriasis vulgaris patients compared with single use of halometasone or calcipotriol .It takes effect quickly ,and could be more feasible and safe .

14.
Chinese Journal of Ultrasonography ; (12): 511-515, 2014.
Artigo em Chinês | WPRIM | ID: wpr-450775

RESUMO

Objective To study the correlation between blood flow parameters by three-dimensional sonography and Nottingham prognostic index (NPI) of breast cancer.Methods A retrospective analysis of sonographic image data of 89 patients who were pathologically confirmed invasive ductal breast cancer was made.The technology of three-dimensional color power angiography (3D-CPA) was used to obtaining vascular index (VI),flow index (FI) and vascular-flow index (VFI) before the operation.NPI was obtained by a formula which was computed on a basis of postoperative pathological data.The case of VI,FI and VFI in different grades and NPI in various prognosis groups were counted,and the relationship between blood flow parameters and NPI was analyzed.Results The correlation coefficient between VI and NPI was 0.643 (P <0.0001) and there was no significant correlation between FI,VFI and NPI.Conclusions Among VI and NPI there was a considerable positive correlation,VI was helpful to evaluate the prognosis of patients before operation and to provide basis for clinical treatment programs.

15.
Modern Clinical Nursing ; (6): 24-26, 2014.
Artigo em Chinês | WPRIM | ID: wpr-452909

RESUMO

Objective To explore the effect of WeChat education and telephone follow-up on reducing depression incidence of primipara in stegmonth.Methods One hundred and six primiparas who have got labored were numbered by the order of discharge from hospital and ramdomized in equal number into observation group(n=53)and control group(n=53)by random digit table. Both groups were re-diagnosed on time and the observation group also received WeChat education and telephone follow-up in order to compare the depression incidence of primiparas in both groups.Result The depression incidence of the observation group was lower than that of the control group and before intervention with statistical difference(P<0.05).Conclusion A communication platform is established between nurses and primiparas by WeChat education and telephone follow-up with subsequent, dynamic and individualized management mode, which has improved self-nursing ability of primipara and reduced the depression incidence in stegmonth.

16.
Chinese Journal of Dermatology ; (12): 19-21, 2012.
Artigo em Chinês | WPRIM | ID: wpr-417696

RESUMO

Objective To study the expression of skin aspartic protease (SASPase) in lesions of cutaneous lupus erythematosus (CLE) and its role in the pathogenesis of CLE.Methods Skin samples were resected from the lesions and normal skin of 9 patients with CLE,including 3 cases of subacute cutaneous lupus erythematosus (SCLE),4 cases of discoid lupus erythematosus (DLE) and 2 cases of acute cutaneous lupus erythematosus (ACLE).Keratinocytes were isolated from the tissue samples and cultured in serum-free medium.Total proteins were extracted from the keratinocytes and separated by two-dimensional gel electrophoresis.ImageMaster 2D analysis software was used to assess differentially expressed proteins in keratinocytes between the lesional and normal skin,which were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS).The expression levels of SASPase were further determined by Western blot.The data were analyzed statistically by Student's t test.Results Keratinocytes were isolated from the tissue samples and successfully cultured in vitro.Two-dimensional electrophoresis profiles of proteins from the keratinocytes were obtained with high resolution and reproducibility,and the average matching protein spots were about 1200 with the matching rate higher than 80%.As Western blot showed,the relative expression level of SASPase was 0.463 ± 0.018 in keratinocytes from the lesional skin,and 0.145 ± 0.011 in those from the normal skin (P < 0.05).The Western blot results were consistent with those of two-dimensional electrophoresis.Conclusion The initiation and progression of CLE seem to be associated with the abnormal activation and overexpression of SASPase.

17.
Chinese Journal of Biotechnology ; (12): 747-754, 2011.
Artigo em Chinês | WPRIM | ID: wpr-324542

RESUMO

Matrix protein 2(M2) is an integral tetrameric membrane protein of influenza A virus, which functions as ion channel. M2 sequence has shown remarkable conservation, so there has been growing interest in it as "universal" vaccine. In order to establish a stable 293 cell line that express M2 protein under the control of the tetracycline operator, M2 gene was obtained by PCR amplification from the plasmid containing the segment 7 of influenza A virus strain A/PR/8/34 firstly. The PCR product was cloned into BamH I/Not I restriction site of pcDNA5/FRT/TO vector, and cotransfected with pOG44 which express Flp recombinase into Flp-In T-REx-293 cell. Integration of pcDNA5/FRT/TO-M2 into the cell genome at the Flp Recombination Target (FRT) site brought the SV40 promoter and the initiation codon in frame with the hygromycin resistance gene. Thus, stable cell lines were selected for hygromycin resistance. The expression of M2 protein from hygromycin-resistant cell was induced by addition of tetracycline into the cell culture media, and then tested by indirect immunofluorescence assay (IFA). 16 strains with high expression of M2 were selected. After subculturing for more than ten passages, the cell lines still stably expressed M2 protein. No M2 protein could be detected without tetracycline induction, suggesting that the expression was strictly controlled by tetracycline operator. The cell lines expressing M2 will be useful for further functional studies of M2 protein, detection of immune response against natural structure M2 protein and development of live attenuated influenza virus vaccine with reverse genetics technique.


Assuntos
Animais , Humanos , Clonagem Molecular , Expressão Gênica , Vetores Genéticos , Genética , Células HEK293 , Vírus da Influenza A , Genética , Metabolismo , Vacinas contra Influenza , Genética , Regiões Operadoras Genéticas , Proteínas Recombinantes , Genética , Tetraciclina , Farmacologia , Transfecção , Proteínas da Matriz Viral , Genética
18.
Chinese Journal of Biotechnology ; (12): 876-883, 2011.
Artigo em Inglês | WPRIM | ID: wpr-324491

RESUMO

In order to evaluate the response to vector-expressed M1 and HA genes of influenza virus in mice, we prepared recombinant plasmid pStar-M1/HA and recombinant adenovirus Ad-M1/HA containing both the full-length matrix protein 1(M1) and hemagglutinin (HA) genes of human H5N1 influenza virus strain A/Anhui/1/2005. We then combined the DNA vaccine and adenoviral vaccine in immunization of BALB/c mice with a prime-boost regime. We immunized the mice with DNA vaccine at day 0 and 28 and with recombinant adenoviral vaccines at day 14 and 42. We took blood samples before each injection and 14 days after the final injection for detection of humoral immune responses. At day 56, we sacrificed the mice and collected splenocytes for detection of cellular immune responses. ELISA and hemagglutination inhibition (HI) assay showed that specific IgG Abs against H5N1 influenza virus was induced in serum of the immunized mice. ELISPOT results confirmed that the specific cellular immune responses were successfully induced against the M1 and HA proteins of H5N1 influenza virus. This study provides new strategy for development of novel influenza vaccines.


Assuntos
Animais , Camundongos , Adenoviridae , Genética , Metabolismo , Anticorpos Antivirais , Sangue , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Genética , Alergia e Imunologia , Imunização , Virus da Influenza A Subtipo H5N1 , Alergia e Imunologia , Vacinas contra Influenza , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão , Genética , Alergia e Imunologia , Vacinas de DNA , Alergia e Imunologia , Proteínas da Matriz Viral , Genética , Alergia e Imunologia
19.
Chinese Journal of Biotechnology ; (12): 649-656, 2010.
Artigo em Chinês | WPRIM | ID: wpr-292224

RESUMO

We developed vectors expressing two antigen of H5N1 influenza virus. Based on the human H5N1 avian influenza virus strain A/Anhui/1/2005 isolated in China, we amplified the matrix protein 2 (M2) and Hemagglutinin (HA) genes by PCR and subcloned them into pStar vector to construct two genes co-expressing recombinant DNA vaccine pStar-M2/HA. After transfection of 293 cells with the plasmid, we confirmed with indirect immunofluorescence assay (IFA) that M2 and HA genes cloned on plasmid pStar co-expressed successfully. Using Ad-Easy adenovirus vector system, by homologous recombination in bacteria and packaging in 293 cells, we constructed two recombinant adenoviruses, namely Ad-M2 and Ad-HA. After infection of 293 cells with the recombinant adenoviruses, we confirmed with IFA that M2 and HA genes cloned into adenoviruses expressed successfully. We then combined the recombinant DNA vaccine and adenoviral vector vaccines in immunization of BALB/c mice with a prime-boost regime. On day 0 and day 28, we immunized the mice with DNA vaccine and on day 14 and day 42, with recombinant adenovirus vaccines. We took blood samples before each injection and 14 days after the final injection. On day 56, we collected splenocytes from the mice. ELISA and hemagglutination inhibition (HI) assay showed that the vaccines successfully induced specific IgG antibodies against HA protein in serum of the immunized mice. ELISPOT confirmed that the vaccines successfully induced the special cellular immune response to M2 and HA protein of H5N1 influenza virus. The study on combined immunization with M2 and HA genes provided basis for development of novel influenza vaccine.


Assuntos
Animais , Feminino , Camundongos , Adenoviridae , Genética , Metabolismo , Vetores Genéticos , Genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Genética , Virus da Influenza A Subtipo H5N1 , Genética , Alergia e Imunologia , Vacinas contra Influenza , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Proteínas Recombinantes , Genética , Alergia e Imunologia , Vacinação , Vacinas de DNA , Alergia e Imunologia , Proteínas da Matriz Viral , Genética
20.
Chinese Journal of Dermatology ; (12): 605-608, 2008.
Artigo em Chinês | WPRIM | ID: wpr-398746

RESUMO

Objective To observe the expressions of two inhibitors of apoptosis proteins (LAPs), survivin and livin, as well as their related factors, Bci-xl and Caspase-3 in mycosis fungoides (MF), along with the effects of NB-UVB irradiation on them. Methods Totally, 30 patients with MF (5 at erythema stage, 16 at plaque stage and 9 at tumor stage) collected from 1995 to 2007 were included into this study. Of the patients, 11 received the treatment with NB-UVB irradiation. Tissue samples were resected from 30 untreated patients, 11 irradiated patients and 10 normal human controls. SABC immunohistochemistry (IHC) stain was used to evaluate the protein expression of survivin, livin, Bel-xi and caspase-3 in these samples. Also, the mRNA expression of these four factors and cell apoptosis were detected by hybridization in situ (ISH) and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP-fluorescence nick end labeling (TUNEL stain) respectively in the samples from 11 patients before and after NB-UVB irradiation. Results In samples of erythema-stage MF, plaque-stage MF and tumor-stage MF, the positivity rate was 40.00%, 75.00%, 77.78% for survivin respectively, 60.00%, 68.75%, 88.89% for Bcl-xl respectively, 40%, 25%, 44.44% for livin respectively, and 60.00%, 68.75%, 88.89% for caspase-3 respectively. No expression of survivin or Bcl-xl was observed in normal controls, while the expression of livin and caspase-3 was similar between MF and control samples. After NB-UVB irradiation, an increase was noticed in the count of apoptosis cells (t=6.49, P<0.001) and mRNA expression of caspase-3 (P<0.10), while a decrease in the mRNA expression of survivin and Bcl-xl in MF tissues, and no changes occurred to the mRNA expression of livin (P>0.10). Conehsions Survivin, Bcl-xl and caspase-3 may be associated with the pathogenesis of MF by regulating the cell apoptosis of T lymphocytes. NB-UVB could suppress the mRNA expression of survivin and Bcl-xl, lower the levels of LAP, enhance the transcription of caspase-3, and accelerate the apoptosis of tumor cells, which may partly explain the mechanism of therapeutic effect of NB-UVB in MF.

SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA