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Objective@#To investigate and analyze the clinical features, epidemiologic information and pathogenic characteristics of a rabies patient.@*Methods@#Clinical data of the patient(boy) was collected and epidemiological survey was conducted, fluorescence quantitative reverse transcription-polymerase chain reaction (FQRT-PCR) and reverse transcription-polymerase chain reaction (RT-PCR) were used to detect the samples of saliva, cerebrospinal fluid (CSF), skin tissue with hair follicle at the back of the neck for rabies laboratory diagnosis.@*Results@#Early symptoms of the boy were vomiting, diarrhea, fever and irritability, followed by coma and death. The boy had nasal trauma one month ago and the domestic dog died of illness during the same period. He did not accept the rabies post-exposure prophylaxis (PEP). The result of the saliva sample was positive by FQRT-PCR. The predicted segments of the glycoprotein(G), nucleoprotein (N) genes of rabies virus were amplified from the positive saliva sample of the patient by RT-PCR. Compared with rabies virus strains in Henan province, the nucleotide homology and amino acid homology in G gene segment were 96.5%-98.8% and 96.5%-99.2% respectively.@*Conclusions@#The case was diagnosed in laboratory as rabies case. The pathogenic rabies virus strain was endemic in Henan province. The nasal trauma, the dead domestic dog were probably related to the infection of the boy.
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Objective To observe the dynamic changes of high mobility group protein B1 ( HMGB1 )expression in the lung of rats with Vibrio vulnificus sepsis so as to unravel the role of HMGB1 in lung injury.Methods Sixty rats of clean grade were randomly divided into normal control group ( A group, n = 10) and Vibrio vulnificus sepsis group (B group, n =50). Sepsis model was made in rats with subcutaneous injection of Vibrio vulnificus with concentration of 6 × 108 cfu/ml in dose of 0. 1 ml/100 g into left lower limb.The rats of group B were sacrificed 1 h, 6 h, 12 h, 24 h and 48 h after infection for taking lung tissues to detect the water content of lung and to observe the histopathological changes in lung under light microscope.The expression of HMGB1 mRNA and the level of HMGB1 protein in the lungs were detected by RT-PCR and Western blot, respectively. Data were analysed with ANOVA and LSD method for comparison between groups, and P ＜0.05 was considered statistically significant. Results Compared with the group A (0.652±0. 177), the expressions of HMGB1 mRNA in lung of rats of group B were significantly higher in 12 hours (1. 161 ±0.358, P=0.013), 24 hours (1.679 ±0.235, P =0.000) and 48 hours (1.258 ±0.274, P=0.004) and reached the peak in 24 h. Compared with group A (0.594 ±0. 190), the level of HMGB1 protein in rats of group B 6 h after infection ( 1. 408 ± 0. 567, P = 0. 026) was significantly increased (P＜0.05), and it reached peak in 24 h (2.415 ± 1.064, P =0.000) after infection. Compared with group A (0.699 ± 0.054), the lung water contents in rats of group B were significantly increased in 6 h (0.759±0.030, P=0.001), in 12 h (0.767 ±0.023, P =0.000), in 24 h (0.771 ±0.043, P=0.000) and in 48 h (0.789 ±0.137, P=0.000) after infection. Compared with group A, the pathological changes in the lung of rats in group B showed clearly marked pulmonary vascular congestion, interstitial edema and inflammatory cell infiltration, and those changes became more and more serious until alveolar sacs entirely collapsed and the boundaries of the alveolar septa could not be clearly identified in 48 h. Conclusions Vibrio vulnificus sepsis leads to the lung injury of infected rats, and the increase in the expression of HMGB1 mRNA in lung might be one of the mechanisms of lung injury in rats with Vibrio vulnificus sepsis.
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La Hepatitis C constituye un problema de salud pública y su transmisión está claramente asociada con la ruta parenteral. Sin embargo su agente causal, Virus de Hepatitis C (VHC), también ha sido aislado de otros fluidos incluyendo la saliva, aunque la relación existente entre VHC y la patología bucal no está completamente dilucidada. El objetivo del presente estudio fue determinar la presencia de ARN-VHC en la saliva de pacientes con Hepatitis C crónica. En la presente investigación se evaluaron 24 pacientes provenientes del Departamento de Hepatología del Hospital Clínico Universitario, Universidad Central de Venezuela, con infección por VHC. 5 ml de saliva no estimulada fue tomada de cada paciente. ARN-VHC fue detectada por la técnica de Transcriptasa Reversa- Reacción en cadena de la Polimerasa (TR-RCP). En 29 por ciento, (7/24) pacientes VHC+, se observó la presencia de ARN-VHC en saliva. En este estudio, observamos la presencia de ARN-VHC en la saliva de pacientes con infección crónica por VHC. Es necesario realizar estudios epidemiológicos a gran escala, para clarificar el significado biológico de la presencia de este agente viral en la saliva, incluyendo la potencial vía de transmisión por la exposición con este fluido
Hepatitis C is a worldwide public health problem and its transmisión is clearly associated with the parenteral route, however, the virus has also been isolated from other body fluids, including saliva, although the relationship between HCV and oral pathology is not clearly understood. The aim of this study was to determine the presence of HCV-RNA in saliva from patients with chronic C hepatitis. In the present investigation 24 patients, attended at the Hepatology Department, at the the Clinical Hospital University, Central University of Venezuela, with HCV infection were evaluated . 5ml of unstimulated saliva were taken of each patient. Saliva HCV-RNA was detected by Polymerase Chain Reaction (PCR). 29 percent (7/24) of HCV+ patients showed HCV-RNA in saliva. In this study, we observed the presence of HCV-RNA in saliva of patients infected with HCV. Further large-scale epidemiological studies are required to clarify the clinical significance of HCV in the saliva, including the potencial for viral transmisión through exposure to these fluids
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Female , Hepacivirus , Hepatitis C, Chronic , RNA , Polymerase Chain Reaction/methods , Saliva/cytology , Saliva/virologyABSTRACT
[Objective]To induce TGF-?1 gene which can increase ECM synthesis into adipose tissue derived stem cells(ADSCs) by employing gene transfer techniques and observe whether TGF-?1 gene could expresse continuously and whether type II collagen and aggrecan are synthesized in order to provide experimental data for constructing tissue-engineering cartilage. [Method]ADSCs were transferred with TGF-?1 gene ,TGF-?1,FN,ColⅡ and aggrecan were detected with RT-PCR and TGF-?1 protein was detected with Western-blot.[Result]RT-PCR demonstrated that the expression of TGF-?1,FN,ColⅡ and aggrecan in the TGF-?1 gene transferred groups increased more evidently than non-gene groups and control groups (P
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Objective To investigate the expression of protease activated receptors(PARs)in mast cells.Methods Reverse transcription polymerase chain reaction(RT-PCR),flowcytometry and immunofluorescent cell staining were used to detect the expression of PARs in mast cell lines P815 and MC/9 at the levels of protein and mRNA.Results Both the P815 and MC/9 of mast cell lines expressed PAR-1,PAR-2,PAR-3 and PAR-4 at either protein or mRNA level.Conclusion The expression of all the four PARs in mast cells were detectable,which may be of significance for the further study on the function of PARs in mast cells.
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@#ObjectiveTo detect the levels of bone morphogenetic protein-2(BMP-2) mRNA in gunshot fracture healing of the rabbits.MethodsA model of primary treating gunshot fracture of the rabbit with external fixator and the real time fluorescence quantitative reverse transcription polymerase chain reaction(FQ-RT-PCR) technology with SYBR Green I were established.The levels of BMP-2 mRNA 1,2,3 and 4 weeks after operation were detected with FQ-RT-PCR respectively.ResultsIn the control group,BMP-2 mRNA of the bone tissue began to rise at the first week after fracture and the peak of mRNA expression appeared at the second week.The expression returned to normal at the forth week.In the experimental group,the BMP-2 mRNA rose more slowly that it arrived peak stage at the third week and was significantly lower in experimental group than that in control group at the same phases.ConclusionReal time FQ-RT-PCR is a good method for measuring the levels of BMP-2 mRNA during gunshot fracture healing.The mRNA expression rose more slowly and was significantly lower in gunshot fracture than in common fracture at the same phases.
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Objective: To investigate the molecular mechanism of Pingchuanning in treating asthma.Methods: Using reverse transcription-polymerase chain reaction(RT-PCR),cavies' lung tissue Eotaxin mRNA expression were given semi-quantitative analysis.Results: The Eotaxin mRNA levels in asthma model group were increased than the control group,P
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Objective To explore the clinical significance of the differential expression of PTEN in glioma tissue.Methods The mRNA and protein expressions of PTEN were assayed by reverse transcription polymerase chain reaction(RT-PCR) and immunohistochemistry in 75 human brain glioma cases.Results The positive expression rate mRNA of PTEN differed significantly between brain glioma tissue and normal brain tissue(?2=22.66,P
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PURPOSE: The cause of aseptic meningitis remains mostly unknown because viral culture and identification is difficult. Thus, we report a study on 123 children with aseptic meningitis in Gyeongju in 2002 to identify the causing virus and the relationship with the clinical manifestation. METHODS: We prospectively investigated the patients, admitted to Dongguk University Hospital, into two groups between April and October 2002. Group 1 included 123 patients diagnosed as aseptic meningitis. Group 2, the adimssion control, included 120 patients, who suffered from none-enteroviral diseases. Specimens of CSF and stool were collected to perform reverse transcription polymerase chain reaction(RT-PCR), and enteroviral culuture was done in RT-PCR positive patients as well. RESULTS: The male to female sex ratio was 2.2:1 and the mean age was 6.2+/-3.7 years. The clinical manifestations were fever, headache and vomiting. The RT-PCR for enterovirus, performed in 58 cases of CSF in group 1, showed 5.2% positive results and negative result in viral culture. The RT-PCR for enterovirus used in stool specimens showed 89.3% and 41.1% of positive results in group 1 and group 2, respectively. Viral culture of stool specimens showed five cases of echovirus 13 and four cases of echovirus 6 in group 1, whereas three cases of echovirus 6 and one case of coxsackie B4 were detected in group 2. CONCLUSION: The etiologic viruses of the aseptic meningitis outbreak in Gyeongju in 2002 is presumed to be echovirus 13 and echovirus 6. Since echovirus 13 firstly appeared with various age distributions, the outbreak may have emerged due to a lack of acquisition of immunity to this virus.
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Child , Female , Humans , Male , Age Distribution , Echovirus 6, Human , Enterovirus , Enterovirus B, Human , Fever , Headache , Meningitis, Aseptic , Prospective Studies , Reverse Transcription , Sex Ratio , VomitingABSTRACT
OBJECTIVE: To investigate the influence of transforming growth factor-alpha (TGF-alpha) on the expression of Matrix metalloproteinase-2 (MMP-2) and Matrix metalloproteinase-9 (MMP-9) mRNA in mouse embryos. MATERIALS AND METHOD: Eight-cell stage mouse embryos were cultured for 48hours with TGF-alpha at concentrations of 1, 10 and 100 ng/ml. Embryos not treated with TGF-alpha served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2 and MMP-9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with analysis of variance (ANOVA) and statistical significance was defined as p<0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of 10 and 100 ng/m of TGF-alpha treatment groups were significantly increased than in the control and 1 ng/ml of TGF-alpha treatment group (67.2+/-7.5 and 77.4+/-11.6 vs. 38.6+/-4.5 and 43.4+/-6.1, p<0.001). The relative quantities of MMP-9 mRNA of 100 ng/ml TGF-alpha treatment group was significantly increased than in the control and 1 ng/ml TGF-alpha treatment groups (67.6+/-6.5 vs. 36.6+/-14.2 and 40.2+/-11.3, p<0.001, p<0.01, respectively). CONCLUSION: This study suggests that TGF-alpha itself may induce the expression of MMP-2 and 9 mRNA in mouse embryos.
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Animals , Mice , Blastocyst , Digestion , DNA, Complementary , Embryonic Structures , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Reverse Transcription , RNA, Messenger , Sensitivity and Specificity , Sequence Analysis , Transforming Growth Factor alphaABSTRACT
OBJECTIVES: To investigate the effect of granulocyte colony stimulating factor (G-CSF) and granulocyte macrophage colony stimulating factor (GM-CSF) on expression of matrix metalloproteinase-2, 9 (MMP-2, 9) mRNA in mouse embryos. Materials and METHOD: From October 1997 to December 1998, morula stage mouse embryos were cultured for 48 hours with G-CSF and GM-CSF at concentrations of 0.1 pg/ml, 1 pg/ml, 10 pg/ml, 100 pg/ml, 1 ng/ml and 10 ng/ml, respectively. Embryos not treated with G-CSF or GM-CSF were served as control. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of MMP-2, 9 mRNA in developed blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with Kolmogorov-Smirnov test and analysis of variance (ANOVA). The statistical significance was defined as p< 0.05. RESULTS: The relative quantities (relative volume x intensity) of MMP-2 mRNA expressed in embryos of all G-CSF treatment groups were significantly increased than in the control, especially in 10, 100 pg/ml and 1 ng/ml treatment groups. The relative quantities of MMP-2 mRNA in all GM-CSF treatment groups were also significantly increased than in the control, especially in 100 pg/ml treatment group. The relative quantities of MMP-9 mRNA of all GM-CSF treatment groups except 10 ng/ml group were significantly increased than in the control, especially 10, 100 pg/ml and 1 ng/ml treatment group. However, the relative quantity of MMP-9 mRNA was significantly increased in only 10 ng/ml G-CSF treatment group than in the control and other treatment groups. CONCLUSION: This study suggests that G-CSF and GM-CSF may increase the m-RNA expression of MMP-2 or 9 in mouse blastocysts with the concentration-specific manner.
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Animals , Mice , Blastocyst , Colony-Stimulating Factors , Digestion , DNA, Complementary , Embryonic Structures , Granulocyte Colony-Stimulating Factor , Granulocyte-Macrophage Colony-Stimulating Factor , Granulocytes , Matrix Metalloproteinase 2 , Morula , Reverse Transcription , RNA, Messenger , Sensitivity and Specificity , Sequence AnalysisABSTRACT
This study was performed to investigate the influence of epidermal growth factor (EGF) on preimplantation development, implantation, and expression of epidermal growth factor receptor (EGFR) in mouse embryos. Riverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the presence of transcripts. Following reverse transcription, strategically designed nested primers, optimised for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Eight-cell stage mouse embryos were cultured for 48hrs with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/ml. Embryos not treated with EGF were served as control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 48hrs were determined. The percentages of fully expanded murine blastocysts at 48hrs in all EGF treated group were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in EGF treatment group at 0.1ng/ml (90.7%), 10 ng/ml (89.3%) compared to the control (82.1%; p < 0.05, p < 0.05). The percentages of implanted blastocyst in vitro were significantly higher following incubation with EGF at concentrations of O.lng/ml (38.1%; p < 0.05), 1.0ng/ml (33.3%; p < 0.05), 10ng/ml (22.2%; p < 0.05) compared to the control (10.7%). Embryo development and implantation in vitro were not significantly inhibited or enhanced in cultures supplemented with 100ng/ml EGF compared to the control. The mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF were significantly higher than those of the control and other EGF treatment groups. The implantation rate and mRNA concentration of EGFR in embryos treated with 0.1ng/ml of EGF group were significantly higher than those of other treatd groups. In conclusion, EGF may have a stimulatory role in embryonic development, implantation and expression of EGFR in embryo itself with concentration-specific manner. These results suggest that EGF may act directly on the mouse embryo and favor its implantaion, irtespective of the presence ar absence of the endometrium.
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Animals , Female , Mice , Pregnancy , Blastocyst , Digestion , DNA, Complementary , Embryonic Development , Embryonic Structures , Endometrium , Epidermal Growth Factor , ErbB Receptors , Reverse Transcription , Rivers , RNA, Messenger , Sensitivity and Specificity , Sequence AnalysisABSTRACT
Otitis media with effusion(OME) is the most common cause of acquired hearing loss in children. Some children suffer from a chronic form of this disease known as chronic otitis media with effusion(COME), which is manifested by the retention of fluid and inflammatory products in the middle ear cleft and by the eustachian tube dysfunction. The etiology and pathogenesis of COME, however, have not been fully elucidated. Middle ear effusion(MEE) is a complex mixture of transudate, secretory products from glands of middle ear mucosa and products from inflammatory cells and infecting organisms. Recently, there has been a great interest in the pathogenetic roles of cytokines, a group of low molecular weight glycoproteins produced by macrophages, lymphocytes and other cells. Activities of cytokines include fever production, activation of osteoclasts, fibroblasts, phagocytes and cytotoxic cells, regulation of antibody formation and inhibition of growth of cartilage, bone and endothelial cells. In this study, we have utilized the reverse transcription polymerase chain reaction(RT-PCR) technique to determine accurately the existence of mRNAs for five cytokines in MEEs collected from 22 children with COME. Messenger RNAs for TNF-alpha, IL-1beta, IL-2 and IL-8 were detected in 68%, 86%, 59% and 95% of specimens, respectively, Interleukin-4 mRNA was absent in all the specimens. The persistent production of cytokines by the inflammatory cells in MEE of COME due to sustained presence of antigens or most-recent antigenic stimuli may play the central role in prolonged OME and responsible for the mucosal damage, bone erosion, fibrosis and resulting hearing loss seen in some cases of COME.
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Child , Humans , Antibody Formation , Cartilage , Cytokines , Ear, Middle , Endothelial Cells , Eustachian Tube , Exudates and Transudates , Fever , Fibroblasts , Fibrosis , Glycoproteins , Hearing Loss , Interleukin-2 , Interleukin-4 , Interleukin-8 , Lymphocytes , Macrophages , Molecular Weight , Mucous Membrane , Osteoclasts , Otitis Media with Effusion , Otitis Media , Otitis , Phagocytes , Reverse Transcription , RNA, Messenger , Tumor Necrosis Factor-alphaABSTRACT
Objective To investigate the effects of Buyang Huanwu Decoction(BHD) and two kinds of fractions extracted from BHD on expression of caspase-1,caspase-3,and caspase-8 mRNA after cerebral ischemia-reperfusion in rats.Methods The model of cerebral ischemia-reperfusion was induced by the middle cerebral artery occlusion(MCAO) in rats,via string ligation of arteria carotis interna.The expression of caspase-1,caspase-3,and caspase-8 mRNA in cerebral ischemic tissues was assessed by reverse transcription-polymerase chain reaction(RT-PCR).Results The expression of caspase-1,caspase-3,and caspase8 mRNA in cerebral tissues was significantly increased after 22 h reperfusion following 2 h cerebral ischemia in model group(P
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Objective:To explore the mRNA expression of macrophage migration inhibitory factor(MIF)in esophageal squamous cell carcinoma and its relationship with the clinical pathologic characteristics of that carcinoma.Methods:The expressions of MIF mRNA in esophageal carcinoma tissues and normal esophageal tissues were detected by RT-PCR in eighty patients.Results:MIF mRNA expression rate in esophageal carcinoma was 85%(68/80),while in the normal esophageal tissues it was 15%(15/80).the difference was significant(P0.05).Conclusion:Increased MIFmRNA expression contributed to carcinogenesis of esophageal squamous cell carcinoma,which might provide a new way for early diagnosis and therapy of esophageal carcinoma.