Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis an...Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis and experimental autoimmune myocarditis and MIP-2γ mRNA expression in mouse was studied by TaqMan real-time fluorescent quantitative RT-PCR. Results: MIP-2γ mRNA expression rose on 3 to 5 d after CVB3 infection, reached peak on 7 d, and returned to normal level until 14 d, which corresponded well with the disease course. The MIP-2γ mRNA expression level rose significantly on the day 18 d after immunization with porcine cardiac myosin, which was consistent with pathological examination. Conclusion: MIP-2γ may be involved in the pathogenesis of myocarditis.展开更多
Rapid and accurate laboratory diagnosis of SARS-CoV-2 infection is crucial for the management of COVID-19 patients and control of the spread of the virus. At the start of the COVID-19 pandemic, Bangladesh had only one...Rapid and accurate laboratory diagnosis of SARS-CoV-2 infection is crucial for the management of COVID-19 patients and control of the spread of the virus. At the start of the COVID-19 pandemic, Bangladesh had only one government molecular laboratory where real-time RT-PCR would be performed to diagnose SARS-CoV-2 infection. With the increasing number of suspected cases requiring confirmation diagnostic testing, there is a requirement to expand capacity for large-scale testing quickly. The government of Bangladesh established over 100 molecular laboratories within one year to test COVID-19. To expand the testing capacity, the government was compelled to recruit laboratory staff with limited experience and technical expertise, especially in molecular assays, to process specimens, interpret results, troubleshoot. As a result, the risk of diagnostic errors, such as cross-contamination, increased, potentially undermining the efficacy of public health policies, public health response, surveillance programs, and restrictive measures aimed toward containing the outbreak. In this piece, we discuss the different sources of cross-contamination in the COVID-19 RT-PCR laboratories and proffer practical preventive measures to avoid them.展开更多
<strong>Background: </strong>The prevalence of anxiety disorders is increasing in the world. Studies revealed that generalized anxiety disorder may lead to change in platelet size, volume and functions. Th...<strong>Background: </strong>The prevalence of anxiety disorders is increasing in the world. Studies revealed that generalized anxiety disorder may lead to change in platelet size, volume and functions. Thus, the changes in platelet indices may increase the future risk of thrombotic diseases in patients with Generalized Anxiety Disorder (GAD). <strong>Aim: </strong>To evaluate platelet indices (total count of platelet, mean platelet volume, platelet distribution width, plateletcrit) levels in patients with generalized anxiety disorder. <strong>Material and Methods: </strong>A cross-sectional study was performed in the Department of Physiology, Dhaka Medical College, Dhaka from July 2019 to June 2020. After obtaining ethical clearance, a total of 144 individuals were selected based on inclusion and exclusion criteria with ages ranging from 18 - 50 years. Group A was the study group selected from Out Patient Department of Psychiatry of Dhaka Medical College Hospital, Dhaka, diagnosed by an experienced psychiatrist. Group B was the control group who were apparently healthy adults selected from different areas of Dhaka city. The subjects were interviewed and detailed history regarding personal, family, medical and drug history were taken. Prior to sample collection, informed written consent was taken from the participants. Platelet indices (total count of platelet, mean platelet volume, platelet distribution width, plateletcrit) were measured in the Department of Laboratory Medicine, Dhaka Medical College Hospital, Dhaka. <strong>Statistical Analysis:</strong> For statistical analysis, the Unpaired Student’s “t” test was considered using SPSS 25.0 version. <strong>Results: </strong>Mean platelet volume of generalized anxiety disorder patients was significantly higher (p < 0.001) than the control group. Platelet distribution width and plateletcrit were higher in the study group than the control group. <strong>Conclusion:</strong> It can be concluded that generalized anxiety disorder patients may have more chance of thrombotic diseases due to significantly higher mean platelet volume and higher platelet distribution width, plateletcrit than healthy adults.展开更多
<span style="font-family:""><span style="font-family:Verdana;">For just about 30 years, researchers have considered the likelihood to utilize </span><span style="font...<span style="font-family:""><span style="font-family:Verdana;">For just about 30 years, researchers have considered the likelihood to utilize </span><span style="font-family:Verdana;">nucleic acids as antiviral therapeutics. In principle, small single-stranded</span><span style="font-family:Verdana;"> nuc</span><span style="font-family:Verdana;">leotide sequence (oligonucleotide) could hybridize to a particular gene or</span><span style="font-family:Verdana;"> mes</span><span style="font-family:Verdana;">senger RNA and diminish transcription or translation, respectively, in this</span><span style="font-family:Verdana;"> manner decreasing the amount of protein that is synthesized. Until now, an incredible number of antisense oligonucleotides, double-stranded oligonucleotides, aptamers, ribozymes, deoxyribozymes, interfering RNAs, chimeric RNA</span></span><span style="font-family:Verdana;">-</span><span style="font-family:""><span style="font-family:Verdana;">DNA molecules, antibody genes has been created artificially and ap</span><span style="font-family:Verdana;">plied effectively for comprehension and manipulating biological processe</span><span style="font-family:Verdana;">s and in clinical preliminaries to treat a variety of diseases. Their versatility and potency make them similarly fit candidates for fighting viral infections. However, troubles with their efficiency, off-target effects, toxicity, delivery, and stability halted the development of nucleic acid-based therapeutics that can be utilized in the clinic. The potential for nucleic acid therapeutic agents is significant and is quite recently beginning to be realized. In this review, we have summarized some of the recent advancements made in the area of nucleic acid based therapeutics and focused on the methods of their delivery and associated challenges.展开更多
Objective: To explore the molecular regulation mechanism of carvedilol in attenuating the reversion back towards fetal energy metabolism during the development of cardiac hypertrophy induced by coarctation of abdomina...Objective: To explore the molecular regulation mechanism of carvedilol in attenuating the reversion back towards fetal energy metabolism during the development of cardiac hypertrophy induced by coarctation of abdominal aorta (CAA) in male Wistar rats. Methods: Hemodynamic and ventricular remodeling parameters, free fatty acid content in the serum were measured in the experimental animals at 16 weeks after the surgical CAA, the rats receiving carvedilol intervention (CAR) after CAA, and those with sham operation (SH). The expressions of muscle carnitine palmitoyltransferaseⅠ (M-CPTⅠ) and medium chain acyl-CoA dehydrogenase (MCAD) mRNA in the cardiac myocytes from every group were studied with RT-PCR. Results: Significant left ventricular hypertrophy were observed in the rats 16 weeks after coarctation operation (P<0.05), together with significant free fatty acids accumulation and downregulation of M-CPTⅠ and MCAD mRNA (P<0.05) in CAA group. Carvedilol at a dose of 30 mg/kg/d for 12 weeks inhibited the left ventricular hypertrophy induced by pressure overload and enhanced the gene expressions of rate-limiting enzyme (M-CPTⅠ) and key enzyme of fatty acid (MCAD) in the CAR group compared with CAA group (P<0.05). Conclusion: Pressure overload-induced hypertrophy in CAA rats causes the reversion back towards fetal enery metabolism, that is, downregulates the expressions of rate-limiting enzyme and key enzyme of fatty acid oxidation. The intervention therapy with carvedilol, a vasodilating alpha- and beta-adrenoreceptor antagonist, attenuates the reversion of the metabolic gene expression to fetal type through upregulating M-CPTⅠ and MCAD mRNA expressions. Thus, carvedilol may exert cardioprotective effects on heart failure by the mechanism of preserving the adult metabolic gene regulation.展开更多
Objective: To screen the differentially expressed gene in liver regeneration with cDNA array and gain insight of the pathogenesis of the acute hepatic failure. Methods: Acute liver failure model in Sprague-Dawley (SD)...Objective: To screen the differentially expressed gene in liver regeneration with cDNA array and gain insight of the pathogenesis of the acute hepatic failure. Methods: Acute liver failure model in Sprague-Dawley (SD) rat was induced by 95% hepatectomy. The differential expression profiles in regenerating rat liver were studied by a cDNA array representing 1176 cDNA clusters. Some genes were randomly selected from a pool of differentially expressed genes and subjected to RT-PCR to further confirm the result from microarray hybridization. Results: The alterations of transcription levels were noted in the expressions of 188 genes. There were 138 genes with their expression up-regulated such as growth factors, PCNA, ribosomal proteins, IL6 and CDKs which were associated with cell cycle regulation, stress, metabolism and proliferation. Conclusion: cDNA array is a powerful tool to explore the gene expression of acute liver failure and is potential for the diagnosis and treatment of liver regeneration.展开更多
The effects of inhibitors of TNFα converting enzyme (TACE) on TNFα secretion were studied to develop an approach to interfere inflammation processes. The HL-60 cell lines were stimulated in vitro with LPS intravenou...The effects of inhibitors of TNFα converting enzyme (TACE) on TNFα secretion were studied to develop an approach to interfere inflammation processes. The HL-60 cell lines were stimulated in vitro with LPS intravenously for different time to establish the cellular model of inflammation and simultaneously induce in vivo inflammation animal model by LPS The cytotoxic effects of soluble TNFa were checked using MTT colorimetric method to determine the rate of cell proliferation. The level of expression of TACE was detected by using RT-PCR, FCM and immuno-histochemical technique respectively. It was found Chinese medicine Reduqing (RDQ) could inhibit the transcription of TNFa mRNA induced by LPS stimulation (P<0. 01, compared with the control). The anti-oligodeoxyribonucleotide (anti-ODN) of TNFα mRNA could inhibit 78. 9 % of TNFα secretion. The mimic peptides of TACE substrates with hydroxamine group showed potency in vivo and in vitro a-gainst converting of pro-TNFα. It was concluded that all the three types of TACE inhibitors can regulate the expression of TACE at different levels and inhibit sTNFα secretion, indicating TACE is a novel target for inflammation therapy.展开更多
Objective To characterize the cellular properties of ovarian cancer, we examined the correlation between the expression of apoptosis-related gene survivin and those of Bcl-2 and Bax proteins. Methods Expressions of su...Objective To characterize the cellular properties of ovarian cancer, we examined the correlation between the expression of apoptosis-related gene survivin and those of Bcl-2 and Bax proteins. Methods Expressions of survivin mRNA, and Bcl-2 and Bax proteins in 35 cases of ovarian carcinoma, 10 cases of borderline carcinoma, 10 cases of benign tumors and 10 cases of normal tissue were evaluated by reverse transcription polymer-ase chain reaction (RT-PCR) and immunohistochemistry SABC method, respectively. Results Expression of survivin gene was detected in a significantly greater proportion in ovarian carcinoma and borderline carcinoma than those in benign tumors and normal tissues. Although there was no relationship between expression of survivin gene and FIGO stage, histologic grade, pathological type and lymphatic metastasis, expressions of Bcl-2 and Bax proteins were positively and negatively correlated with that of survivin gene, respectively. Conclusion Survivin may play an important role in pathogenesis of ovarian carcinoma, with a synergistic role of apoptosis-related gene Bcl-2 protein and an antagonistic role of Bax protein in formation and progression of ovarian carcinoma.展开更多
Objective To investigate the possibility of the transfection of glial-cell line derived neurotro-phic factor (GDNF) gene into Schwann cells (SCs). Methods SCs cultures from sciatic nerves of neonatal rats were establi...Objective To investigate the possibility of the transfection of glial-cell line derived neurotro-phic factor (GDNF) gene into Schwann cells (SCs). Methods SCs cultures from sciatic nerves of neonatal rats were established. A recombinant retrovirus vector containing GDNF gene was constructed and transferred into SCs. Expression levels of GDNF mRNA and protein were respectively identified with reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry. Determination of GDNF synthesis rates from Retro. pLNCX2-GDNF-transduced SCs (GDNF-SCs) in vitro by enzyme-linked immunoassay sensitive assay (ELISA). Biololgical activity of conditioned medium from GENF-SCs was analysed by co-culture with rat motoneurons. Results Transfection of GDNF gene into SCs lead to significantly enhanced expression of GDNF mRNA and protein. The rate of GDNF secreted by GDNF-SCs was also enhanced(5. 1-fold) ,and more motoneurons survived co-cultured with conditioned medium of GNDF-SCs than with that of normal SCs. Conclusion GNDF gene transfection may be a better way to graft SCs promoting regeneration and repairing demyelination in PNS and CNS.展开更多
Objective To investigate the relationship between NF-κB activity and IFN-γ gene expression, as well as the histopathological changes following liver transplants, both with and without cyclosporin A (CsA) treatment.M...Objective To investigate the relationship between NF-κB activity and IFN-γ gene expression, as well as the histopathological changes following liver transplants, both with and without cyclosporin A (CsA) treatment.Methods Sixty male Wistar and Thirty male SD rats were subjected to orthotopic liver transplants. Fourty-five of the Wistar rats were used as recipients, and were divided into 3 groups: group Ⅰ , syngeneic control (Wistar-to-Wistar); group Ⅱ , acute rejection (SD-to-Wistar); and group Ⅲ: acute rejection treated with cyclosporin A by intramuscular injection (SD-to-Wistar + CSA). After the liver transplants, electrophoretic gel mobility shift assay was used to analyze NF-κB activity in the splenocytes of recipient rats, and RT-PCR was used to measure IFN-γ gene expression in grafted liver specimens. In addition, histopathological examinations were performed to assess the severity of acute liver rejection.Results In group Ⅰ , low levels of NF-KB activity were only detectable on day 5 and 7 post-transplant, and only weak IFN-γ mRNA expression was observed at all time points. By contrast, both high NF-κB activity and high expression levels of IFN-γ mRNA were detected at all time points in group Ⅱ. In group Ⅲ, NF-κB activity and IFN-γ mRNA expression were significantly inhibited, as compared to group Ⅱ (P < 0. 05). A good correlation was found between NF-κB activity and IFN-γ mRNA expression (r = 0.815) . In addition, NF-κB activity and IFN-γ mRNA expression mirrored histopathological changes in all three experimental groups.Conclusions Changes in IFN-γ mRNA expression levels after liver transplantation are at least partially due to changes in levels of NF-κB activity. CsA appears to downregulate NF-κB activity, thus inhibiting IFN-γ gene transcription. Blocking the NF-κB mediated transcription pathway may be of benefit in preventing liver transplant rejection.展开更多
文摘Objective: To study the role of macrophage inflammatory protein (MIP)-2γ in myocarditis pathogenesis in BALB/c mice. Methods: The relationship between the progression of Coxsarckie virus B3(CVB3) viral myocarditis and experimental autoimmune myocarditis and MIP-2γ mRNA expression in mouse was studied by TaqMan real-time fluorescent quantitative RT-PCR. Results: MIP-2γ mRNA expression rose on 3 to 5 d after CVB3 infection, reached peak on 7 d, and returned to normal level until 14 d, which corresponded well with the disease course. The MIP-2γ mRNA expression level rose significantly on the day 18 d after immunization with porcine cardiac myosin, which was consistent with pathological examination. Conclusion: MIP-2γ may be involved in the pathogenesis of myocarditis.
文摘Rapid and accurate laboratory diagnosis of SARS-CoV-2 infection is crucial for the management of COVID-19 patients and control of the spread of the virus. At the start of the COVID-19 pandemic, Bangladesh had only one government molecular laboratory where real-time RT-PCR would be performed to diagnose SARS-CoV-2 infection. With the increasing number of suspected cases requiring confirmation diagnostic testing, there is a requirement to expand capacity for large-scale testing quickly. The government of Bangladesh established over 100 molecular laboratories within one year to test COVID-19. To expand the testing capacity, the government was compelled to recruit laboratory staff with limited experience and technical expertise, especially in molecular assays, to process specimens, interpret results, troubleshoot. As a result, the risk of diagnostic errors, such as cross-contamination, increased, potentially undermining the efficacy of public health policies, public health response, surveillance programs, and restrictive measures aimed toward containing the outbreak. In this piece, we discuss the different sources of cross-contamination in the COVID-19 RT-PCR laboratories and proffer practical preventive measures to avoid them.
文摘<strong>Background: </strong>The prevalence of anxiety disorders is increasing in the world. Studies revealed that generalized anxiety disorder may lead to change in platelet size, volume and functions. Thus, the changes in platelet indices may increase the future risk of thrombotic diseases in patients with Generalized Anxiety Disorder (GAD). <strong>Aim: </strong>To evaluate platelet indices (total count of platelet, mean platelet volume, platelet distribution width, plateletcrit) levels in patients with generalized anxiety disorder. <strong>Material and Methods: </strong>A cross-sectional study was performed in the Department of Physiology, Dhaka Medical College, Dhaka from July 2019 to June 2020. After obtaining ethical clearance, a total of 144 individuals were selected based on inclusion and exclusion criteria with ages ranging from 18 - 50 years. Group A was the study group selected from Out Patient Department of Psychiatry of Dhaka Medical College Hospital, Dhaka, diagnosed by an experienced psychiatrist. Group B was the control group who were apparently healthy adults selected from different areas of Dhaka city. The subjects were interviewed and detailed history regarding personal, family, medical and drug history were taken. Prior to sample collection, informed written consent was taken from the participants. Platelet indices (total count of platelet, mean platelet volume, platelet distribution width, plateletcrit) were measured in the Department of Laboratory Medicine, Dhaka Medical College Hospital, Dhaka. <strong>Statistical Analysis:</strong> For statistical analysis, the Unpaired Student’s “t” test was considered using SPSS 25.0 version. <strong>Results: </strong>Mean platelet volume of generalized anxiety disorder patients was significantly higher (p < 0.001) than the control group. Platelet distribution width and plateletcrit were higher in the study group than the control group. <strong>Conclusion:</strong> It can be concluded that generalized anxiety disorder patients may have more chance of thrombotic diseases due to significantly higher mean platelet volume and higher platelet distribution width, plateletcrit than healthy adults.
文摘<span style="font-family:""><span style="font-family:Verdana;">For just about 30 years, researchers have considered the likelihood to utilize </span><span style="font-family:Verdana;">nucleic acids as antiviral therapeutics. In principle, small single-stranded</span><span style="font-family:Verdana;"> nuc</span><span style="font-family:Verdana;">leotide sequence (oligonucleotide) could hybridize to a particular gene or</span><span style="font-family:Verdana;"> mes</span><span style="font-family:Verdana;">senger RNA and diminish transcription or translation, respectively, in this</span><span style="font-family:Verdana;"> manner decreasing the amount of protein that is synthesized. Until now, an incredible number of antisense oligonucleotides, double-stranded oligonucleotides, aptamers, ribozymes, deoxyribozymes, interfering RNAs, chimeric RNA</span></span><span style="font-family:Verdana;">-</span><span style="font-family:""><span style="font-family:Verdana;">DNA molecules, antibody genes has been created artificially and ap</span><span style="font-family:Verdana;">plied effectively for comprehension and manipulating biological processe</span><span style="font-family:Verdana;">s and in clinical preliminaries to treat a variety of diseases. Their versatility and potency make them similarly fit candidates for fighting viral infections. However, troubles with their efficiency, off-target effects, toxicity, delivery, and stability halted the development of nucleic acid-based therapeutics that can be utilized in the clinic. The potential for nucleic acid therapeutic agents is significant and is quite recently beginning to be realized. In this review, we have summarized some of the recent advancements made in the area of nucleic acid based therapeutics and focused on the methods of their delivery and associated challenges.
文摘Objective: To explore the molecular regulation mechanism of carvedilol in attenuating the reversion back towards fetal energy metabolism during the development of cardiac hypertrophy induced by coarctation of abdominal aorta (CAA) in male Wistar rats. Methods: Hemodynamic and ventricular remodeling parameters, free fatty acid content in the serum were measured in the experimental animals at 16 weeks after the surgical CAA, the rats receiving carvedilol intervention (CAR) after CAA, and those with sham operation (SH). The expressions of muscle carnitine palmitoyltransferaseⅠ (M-CPTⅠ) and medium chain acyl-CoA dehydrogenase (MCAD) mRNA in the cardiac myocytes from every group were studied with RT-PCR. Results: Significant left ventricular hypertrophy were observed in the rats 16 weeks after coarctation operation (P<0.05), together with significant free fatty acids accumulation and downregulation of M-CPTⅠ and MCAD mRNA (P<0.05) in CAA group. Carvedilol at a dose of 30 mg/kg/d for 12 weeks inhibited the left ventricular hypertrophy induced by pressure overload and enhanced the gene expressions of rate-limiting enzyme (M-CPTⅠ) and key enzyme of fatty acid (MCAD) in the CAR group compared with CAA group (P<0.05). Conclusion: Pressure overload-induced hypertrophy in CAA rats causes the reversion back towards fetal enery metabolism, that is, downregulates the expressions of rate-limiting enzyme and key enzyme of fatty acid oxidation. The intervention therapy with carvedilol, a vasodilating alpha- and beta-adrenoreceptor antagonist, attenuates the reversion of the metabolic gene expression to fetal type through upregulating M-CPTⅠ and MCAD mRNA expressions. Thus, carvedilol may exert cardioprotective effects on heart failure by the mechanism of preserving the adult metabolic gene regulation.
文摘Objective: To screen the differentially expressed gene in liver regeneration with cDNA array and gain insight of the pathogenesis of the acute hepatic failure. Methods: Acute liver failure model in Sprague-Dawley (SD) rat was induced by 95% hepatectomy. The differential expression profiles in regenerating rat liver were studied by a cDNA array representing 1176 cDNA clusters. Some genes were randomly selected from a pool of differentially expressed genes and subjected to RT-PCR to further confirm the result from microarray hybridization. Results: The alterations of transcription levels were noted in the expressions of 188 genes. There were 138 genes with their expression up-regulated such as growth factors, PCNA, ribosomal proteins, IL6 and CDKs which were associated with cell cycle regulation, stress, metabolism and proliferation. Conclusion: cDNA array is a powerful tool to explore the gene expression of acute liver failure and is potential for the diagnosis and treatment of liver regeneration.
基金This project was supported by a grant from National Natural Sciences Foundation of China(No.30070722).
文摘The effects of inhibitors of TNFα converting enzyme (TACE) on TNFα secretion were studied to develop an approach to interfere inflammation processes. The HL-60 cell lines were stimulated in vitro with LPS intravenously for different time to establish the cellular model of inflammation and simultaneously induce in vivo inflammation animal model by LPS The cytotoxic effects of soluble TNFa were checked using MTT colorimetric method to determine the rate of cell proliferation. The level of expression of TACE was detected by using RT-PCR, FCM and immuno-histochemical technique respectively. It was found Chinese medicine Reduqing (RDQ) could inhibit the transcription of TNFa mRNA induced by LPS stimulation (P<0. 01, compared with the control). The anti-oligodeoxyribonucleotide (anti-ODN) of TNFα mRNA could inhibit 78. 9 % of TNFα secretion. The mimic peptides of TACE substrates with hydroxamine group showed potency in vivo and in vitro a-gainst converting of pro-TNFα. It was concluded that all the three types of TACE inhibitors can regulate the expression of TACE at different levels and inhibit sTNFα secretion, indicating TACE is a novel target for inflammation therapy.
文摘Objective To characterize the cellular properties of ovarian cancer, we examined the correlation between the expression of apoptosis-related gene survivin and those of Bcl-2 and Bax proteins. Methods Expressions of survivin mRNA, and Bcl-2 and Bax proteins in 35 cases of ovarian carcinoma, 10 cases of borderline carcinoma, 10 cases of benign tumors and 10 cases of normal tissue were evaluated by reverse transcription polymer-ase chain reaction (RT-PCR) and immunohistochemistry SABC method, respectively. Results Expression of survivin gene was detected in a significantly greater proportion in ovarian carcinoma and borderline carcinoma than those in benign tumors and normal tissues. Although there was no relationship between expression of survivin gene and FIGO stage, histologic grade, pathological type and lymphatic metastasis, expressions of Bcl-2 and Bax proteins were positively and negatively correlated with that of survivin gene, respectively. Conclusion Survivin may play an important role in pathogenesis of ovarian carcinoma, with a synergistic role of apoptosis-related gene Bcl-2 protein and an antagonistic role of Bax protein in formation and progression of ovarian carcinoma.
文摘Objective To investigate the possibility of the transfection of glial-cell line derived neurotro-phic factor (GDNF) gene into Schwann cells (SCs). Methods SCs cultures from sciatic nerves of neonatal rats were established. A recombinant retrovirus vector containing GDNF gene was constructed and transferred into SCs. Expression levels of GDNF mRNA and protein were respectively identified with reverse transcriptase-polymerase chain reaction (RT-PCR) and immunocytochemistry. Determination of GDNF synthesis rates from Retro. pLNCX2-GDNF-transduced SCs (GDNF-SCs) in vitro by enzyme-linked immunoassay sensitive assay (ELISA). Biololgical activity of conditioned medium from GENF-SCs was analysed by co-culture with rat motoneurons. Results Transfection of GDNF gene into SCs lead to significantly enhanced expression of GDNF mRNA and protein. The rate of GDNF secreted by GDNF-SCs was also enhanced(5. 1-fold) ,and more motoneurons survived co-cultured with conditioned medium of GNDF-SCs than with that of normal SCs. Conclusion GNDF gene transfection may be a better way to graft SCs promoting regeneration and repairing demyelination in PNS and CNS.
基金a grant from the Bureau of Science & Technology of Zhejiang Province (No. 011106206).
文摘Objective To investigate the relationship between NF-κB activity and IFN-γ gene expression, as well as the histopathological changes following liver transplants, both with and without cyclosporin A (CsA) treatment.Methods Sixty male Wistar and Thirty male SD rats were subjected to orthotopic liver transplants. Fourty-five of the Wistar rats were used as recipients, and were divided into 3 groups: group Ⅰ , syngeneic control (Wistar-to-Wistar); group Ⅱ , acute rejection (SD-to-Wistar); and group Ⅲ: acute rejection treated with cyclosporin A by intramuscular injection (SD-to-Wistar + CSA). After the liver transplants, electrophoretic gel mobility shift assay was used to analyze NF-κB activity in the splenocytes of recipient rats, and RT-PCR was used to measure IFN-γ gene expression in grafted liver specimens. In addition, histopathological examinations were performed to assess the severity of acute liver rejection.Results In group Ⅰ , low levels of NF-KB activity were only detectable on day 5 and 7 post-transplant, and only weak IFN-γ mRNA expression was observed at all time points. By contrast, both high NF-κB activity and high expression levels of IFN-γ mRNA were detected at all time points in group Ⅱ. In group Ⅲ, NF-κB activity and IFN-γ mRNA expression were significantly inhibited, as compared to group Ⅱ (P < 0. 05). A good correlation was found between NF-κB activity and IFN-γ mRNA expression (r = 0.815) . In addition, NF-κB activity and IFN-γ mRNA expression mirrored histopathological changes in all three experimental groups.Conclusions Changes in IFN-γ mRNA expression levels after liver transplantation are at least partially due to changes in levels of NF-κB activity. CsA appears to downregulate NF-κB activity, thus inhibiting IFN-γ gene transcription. Blocking the NF-κB mediated transcription pathway may be of benefit in preventing liver transplant rejection.
