Early childhood caries (ECC) is a term used to describe dental caries in children aged 6 years or younger. Oral streptococci, such as Streptococcus mutans and Streptococcus sorbrinus, are considered to be the main e...Early childhood caries (ECC) is a term used to describe dental caries in children aged 6 years or younger. Oral streptococci, such as Streptococcus mutans and Streptococcus sorbrinus, are considered to be the main etiological agents of tooth decay in children. Other bacteria, such as Prevotella spp. and Lactobacillus spp., and fungus, that is, Candida albicans, are related to the development and progression of ECC. Biomolecules in saliva, mainly proteins, affect the survival of oral microorganisms by multiple innate defensive mechanisms, thus modulating the oral microflora. Therefore, the protein composition of saliva can be a sensitive indicator for dental health. Resistance or susceptibility to caries may be significantly correlated with alterations in salivary protein components. Some oral microorganisms and saliva proteins may serve as useful biomarkers in predicting the risk and prognosis of caries. Current research has generated abundant information that contributes to a better understanding of the roles of microorganisms and salivary proteins in ECC occurrence and prevention. This review summarizes the microorganisms that cause caries and tooth-protective salivary proteins with their potential as functional biomarkers for ECC risk assessment. The identification of biomarkers for children at high risk of ECC is not only critical for early diagnosis but also important for preventing and treating the disease.展开更多
Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury(EBI).In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,p...Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury(EBI).In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,poor initial clinical grades,and some radiographic findings are used,but these markers are somewhat subjective.Thus,it is imperative to find biomarkers of EBI that have beneficial prognostic and therapeutic implications.In our opinion,an ideal biomarker is a molecule that is implicated in the pathogenesis of both EBI and subsequently developing delayed cerebral ischemia(DCI),being a therapeutic target,and can be measured easily in the peripheral blood in an acute stage.A good candidate of such a biomarker is a matricellular protein,which is a secreted,inducible and multifunctional extracellular matrix protein.There are many kinds of matricellular proteins reported,but only tenascin-C,osteopontin,galectin-3 and periostin are reported relevant to EBI and DCI.Reliable biomarkers of EBI may stratify aneurysmal subarachnoid hemorrhage patients into categories of risk to develop DCI,and allow objective monitoring of the response to treatment for EBI and earlier diagnosis of DCI.This review emphasizes that further investigation of matricellular proteins as an avenue for biomarker discovery is warranted.展开更多
Early brain injury(EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage(SAH). This study investigated the role of glucose-regulated protein 78(GRP78) in EBI after SAH. Male Sprague-Dawley rats(n...Early brain injury(EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage(SAH). This study investigated the role of glucose-regulated protein 78(GRP78) in EBI after SAH. Male Sprague-Dawley rats(n=108) weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated d UTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH.展开更多
Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from ...Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from cerebral ischemia/reperfusion and to determine whether these effects are associated with the expression level of HSPA5 in the ischemic penumbra.Methods A total of 72 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group[middle cerebral artery occlusion(MCAO)-Ex,n=18],ischemia and sedentary group(MCAO-St,n=18),sham-surgery and exercise group(Sham-Ex,n=18),or sham-surgery and sedentary group(Sham-St,n=18).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 min,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 min once a day for 5 consecutive days.After reperfusion,the motor function of the rats was scored by the Bederson neurological function test,balance beam test,and screen test.Nissl staining was conducted to assess morphological and structural change of nerve cells in the ischemic penumbra.The reverse transcription-quantitative polymerase chain reaction was applied to detect the mRNA expression of HSPA5.Western blot analysis was conducted to determine the protein expression of HSPA5.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was carried out in the ischemic penumbra after MCAO.Results Rats receiving early treadmill exercise had lower Bederson neurological function,balance beam,and screen test scores on the 3rd,7th,and 14th days after MCAO;in addition,more neurons survived in the ischemic penumbra after MCAO,and higher mRNA and protein expression of HSPA5 and fewer TUNEL-positive stained cells were observed.Conclusion Our study demonstrated that early aerobic exercise can improve neurological function recovery after ischemia/reperfusion.Furthermore,the increased level of HSPA5 in the ischemic penumbra might be one of the mechanisms of enhanced neurological function recovery.展开更多
Dynamic protein-protein interactions are essential for proper cell functioning.Homointeraction events—physical interactions between the same type of proteins—represent a pivotal subset of protein-protein interaction...Dynamic protein-protein interactions are essential for proper cell functioning.Homointeraction events—physical interactions between the same type of proteins—represent a pivotal subset of protein-protein interactions that are widely exploited in activating intracellular signaling pathways.Capacities of modulating protein-protein interactions with spatial and temporal resolution are greatly desired to decipher the dynamic nature of signal transduction mechanisms.The emerging optogenetic technology,based on genetically encoded light-sensitive proteins,provides promising opportunities to dissect the highly complex signaling networks with unmatched specificity and spatiotemporal precision.Here we review recent achievements in the development of optogenetic tools enabling light-inducible protein-protein homo-interactions and their applications in optical activation of signaling pathways.展开更多
One hundred and three Hereford × Angus crossbred, spring-calving cows were used to determine the influence of early weaning performance of their male progeny and the need for cowherd winter supplementation. Treat...One hundred and three Hereford × Angus crossbred, spring-calving cows were used to determine the influence of early weaning performance of their male progeny and the need for cowherd winter supplementation. Treatments were early weaned (EW;115-d of age) and normal weaned calves (NW;220-d of age). Both intact and castrated male progeny were randomly allotted to early and normal weaning calf management treatments. Following summer grazing and weaning protocols, cows within each weaning group were randomly assigned to receive winter supplementation treatments of LOW (1.26 kg/hd/d) or HIGH (1.81 kg/hd/d) amounts of a protein supplement containing 450 g/kg CP (DM basis). Cows whose calves were weaned early gained more BW and BCS (P P P P = 0.04) BCS but similar (P = 0.74) BW when compared with cows with normally weaned calves that received 100% of their supplement (NWHIGH). Early weaning has the potential to reduce the amount of supplement required during the winter grazing period.展开更多
Background: Caries detection in school oral examinations insufficient accuracy. Objective: To evaluate advantages of introducing quantitative light-induced fluorescence-digital (QLF-D) in school oral examinations. Met...Background: Caries detection in school oral examinations insufficient accuracy. Objective: To evaluate advantages of introducing quantitative light-induced fluorescence-digital (QLF-D) in school oral examinations. Methods: Experiment No. 1. Early demineralized lesions in the upper and lower incisors and canines were visually inspected by three dentists and by QLF-D. The numbers of tooth planes with early demineralized lesions were compared between the methods. Experiment No. 2. Approximal demineralized lesions in molars were assessed by visual inspection, x-ray imaging, and QLF-D. The numbers of tooth planes with demineralized lesions were compared among the methods. Experiment No. 3. Plaque distribution was evaluated by QLF-D and a traditional staining method. The ratio of the diameter of plaque to tooth crown in the tooth axis direction in each method was calculated. The results were evaluated by Pearson’s correlation coefficient analysis and Bland-Altman plot. Results: Experiment No. 1. The three dentists found 0.67 tooth planes on average. QLF-D found 22 tooth planes with early demineralized lesions in the same samples. Experiment No. 2. Fourteen approximal tooth planes of molars were found to have demineralized lesions by x-ray imaging. QLF-D detected 71.4% of the tooth planes out of the 14, whereas visual inspection found 7.1%. Experiment No. 3. The Pearson’s correlation coefficient for the evaluations of plaque distribution between the QLF-D and traditional staining methods was 0.77 (P Conclusion: The results support introduction of QLF-D for use in school dental examinations.展开更多
BACKGROUND: Insulin-like growth factor-I(IGF-1), as one of the important members of growth factor family, participants in the regulation of many physiological functions and behaviors, having very strong neuroprotec...BACKGROUND: Insulin-like growth factor-I(IGF-1), as one of the important members of growth factor family, participants in the regulation of many physiological functions and behaviors, having very strong neuroprotective effect. However, the expression of IGF-1 following cerebral ischemia/reperfusion is still disputed. OBJECTIVE: To observe the expression of IGF-1 and protein of corpora striata in ischemic side at the early stage of middle cerebral artery ischemia/reperfusion in rhesus monkey. DESIGN : A completely randomized grouping design, controlled animal experiment SETTING : Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University MATERIALS: ① Totally 17 rhesus monkeys , of either gender, aged 4 to 5 years, were enrolled . Seven rhesus monkeys observed with gene chip were randomly divided into 2 groups: sham operation group (n=3) and ischemia/reperfusion group 〈n=4〉. Ten rhesus monkeys observed with in situ hybridization and immunohistochemistry method were randomly divided into 2 groups: sham operation group 〈n=3 〉and ischemia/reperfusion group (n=7). Rhesus monkeys observed under microscope were divided into 2 groups: sham operation group (n=6) and ischamia/reperfusion group (n=-11).②Materials used in the experiment: cresyl violet (Sigma Company, America); immunohistochemical reagent kit ( Huamei Bio-engineering Company); In situ hybridization reagent kit (Boshide Bio-engineering Co.Ltd, Wuhan); 12 800 dots chip (Boxing Company, Shanghai). METHODS : This experiment was carried out at the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from January 2001 to December 2003.① The onset area of middle cerebral artery was blocked for 2 hours, middle cerebral artery ischemia/reperfusion models were created.② After ischemia/reperfusion for 24 hours, cerebral tissue sections of rhesus monkeys were prepared and stained with cresyl violet. Image analysis was performed with 5001W image analysis software. Morphological change of corpora striata of operative side was observed in the rhesus monkeys between two groups. Total RNA was extracted from cerebral tissue. ③ Detection of gene chip: Cy3-duTP and Cy5-duTP were used to respectively perform reverse transcription labeling. The sample was reversely transcribed into cDNA, then hybridized with cDNA of cerebral tissue. Genes with the separate absolute value of cy3 and cy5〉800, cY3/cy5 〉 2(high expression) or 〈 0.5 (low expression) were found out. Those were genes with differential expression. ④ The expressions of IGF-1 mRNA and protein level of corpora striata in ischemic side of rhe- sus monkeys were detected between sham operation group and ischemia/reperfusion group at 9 and 24 hours after ischemia/reperfusion with in situ hybridization method and immunohistochemical method. Brown granules were IGF-1 protein positive cells. ⑤ Analysis of variance was used in the difference comparison of measurement data among groups. MAIN OUTCOME MEASURES : ① Change of morphological structure of corpora striata at ischemic side in rhesus monkeys. ② Change of cerebral gene expression profiles at ischemia/reperfusion in rhesus monkeys between two groups.③ Expression of IGF-1 mRNA and protein level of corpora striata at ischemia/reperfu- sion in rhesus monkeys between two groups. RESULTS : ① Pathological change : Obvious pathological change of cerebral infarction appeared in the ischemia and reperfusion group, while there was no such pathological change in the sham operation group.② Change of gene expression profile : There were 4480 genes with difference expression in the ischemia/reperfusion group and sham-operation group, in which, 260 genes had high expression and their absolute value was over 800, and 63 genes had low expression, cy3/cy5 of IGF-1 was 0.379, being relative low ex- pression. ③ IGF-1 mRNA and protein positive cell counts in corpora striata at cerebral ischemic side[IGF-1 mRNA: 〈9.72±1.18),(9.11 ±0.76),(14.77±0.60) counts/field:lGF-1 protein: (15.11 ±1.83),(15.39±0.78), (34.62±0.97)counts/field, P 〈 0.05-0.01]. CONCLUSION: IGF-1 mRNA and protein are lowly expressed in middle cerebral artery of rhesus monkeys at ischemia/reperfusion.展开更多
We tested urine albumin excretion rate (UAER), urine transrerrin (TRF ), retinolulnding protein (RBP), N --acetyl--aD-gi ucosamlnldase (N AG ), P, - mi c rogl obu I in (P, -- M G ) and lgGIn 45 cases of NIDDM. Thirty ...We tested urine albumin excretion rate (UAER), urine transrerrin (TRF ), retinolulnding protein (RBP), N --acetyl--aD-gi ucosamlnldase (N AG ), P, - mi c rogl obu I in (P, -- M G ) and lgGIn 45 cases of NIDDM. Thirty cases with UAER<300 mg/d were divided into two groups. Data wereshown with urine protein index (urine protein/urine creatlne). ResultS showed that urine transferrinwas more seusltlve than albumlu, and the combined test or urine Protein is nontraumatic, which hadsteatncance to diagnose early diabetic nepkropatby.展开更多
基金supported by the National Natural Science Foundation of China81400502(Qiong Zhang)the Young Teacher Research Start Funding of Sichuan University 2016SCU11049(Yuan Zhou)
文摘Early childhood caries (ECC) is a term used to describe dental caries in children aged 6 years or younger. Oral streptococci, such as Streptococcus mutans and Streptococcus sorbrinus, are considered to be the main etiological agents of tooth decay in children. Other bacteria, such as Prevotella spp. and Lactobacillus spp., and fungus, that is, Candida albicans, are related to the development and progression of ECC. Biomolecules in saliva, mainly proteins, affect the survival of oral microorganisms by multiple innate defensive mechanisms, thus modulating the oral microflora. Therefore, the protein composition of saliva can be a sensitive indicator for dental health. Resistance or susceptibility to caries may be significantly correlated with alterations in salivary protein components. Some oral microorganisms and saliva proteins may serve as useful biomarkers in predicting the risk and prognosis of caries. Current research has generated abundant information that contributes to a better understanding of the roles of microorganisms and salivary proteins in ECC occurrence and prevention. This review summarizes the microorganisms that cause caries and tooth-protective salivary proteins with their potential as functional biomarkers for ECC risk assessment. The identification of biomarkers for children at high risk of ECC is not only critical for early diagnosis but also important for preventing and treating the disease.
基金supported by a Grant-in-Aid for Scientific Research from Novartis Pharmaceuticals to HS
文摘Aneurysmal subarachnoid hemorrhage remains devastating,and the most important determinant of poor outcome is early brain injury(EBI).In clinical settings,as a surrogate marker of EBI,loss of consciousness at ictus,poor initial clinical grades,and some radiographic findings are used,but these markers are somewhat subjective.Thus,it is imperative to find biomarkers of EBI that have beneficial prognostic and therapeutic implications.In our opinion,an ideal biomarker is a molecule that is implicated in the pathogenesis of both EBI and subsequently developing delayed cerebral ischemia(DCI),being a therapeutic target,and can be measured easily in the peripheral blood in an acute stage.A good candidate of such a biomarker is a matricellular protein,which is a secreted,inducible and multifunctional extracellular matrix protein.There are many kinds of matricellular proteins reported,but only tenascin-C,osteopontin,galectin-3 and periostin are reported relevant to EBI and DCI.Reliable biomarkers of EBI may stratify aneurysmal subarachnoid hemorrhage patients into categories of risk to develop DCI,and allow objective monitoring of the response to treatment for EBI and earlier diagnosis of DCI.This review emphasizes that further investigation of matricellular proteins as an avenue for biomarker discovery is warranted.
基金supported by grants from the National Natural Science Foundation of China(No.81360185)the Foundation of the First Affiliated Hospital of Medical College of Shihezi University of China(No.SS2011-095)
文摘Early brain injury(EBI) plays a key role in the pathogenesis of subarachnoid hemorrhage(SAH). This study investigated the role of glucose-regulated protein 78(GRP78) in EBI after SAH. Male Sprague-Dawley rats(n=108) weighing 260±40 g were divided into control, sham-operated, and operated groups. Blood was injected into the prechiasmatic cistern of rats in the operated group. Neurological scores, ultrastructures of neurons, apoptosis, and GRP78 expression in the hippocampus were examined using Garcia scoring system, transmission electron microscopy, terminal deoxynucleotidyl transferase-mediated d UTP nick-end labelling, and Western blotting at 1, 6, 12, 24, 48, and 72 h after SAH, respectively. The results showed that neurological scores were significantly decreased in the operated group as compared with those in control and sham-operated groups at 12, 24, 48, and 72 h. Metachromatin, chromatin pyknosis at the edge, endoplasmic reticulum swelling, and invagination of nuclear membrane were observed at 24 h in the operated group, indicating the early morphological changes of apoptosis. The number of apoptotic cells was significantly increased in the operated group as compared with that in control and sham-operated groups at 6, 12, 24, 48, and 72 h. The GRP78 protein expression levels in the operated group were significantly elevated at all time points and reached the peak at 12 h. GRP78 expression was positively associated with apoptosis cells and negatively with neurological scores. In conclusion, EBI was demonstrated to occur after SAH and GRP78 was involved in the development of EBI after SAH.
基金This study was funded by Basic Research Projects in Shanxi Province(No.2015021178)Start-up Funds for Doctoral Research at Shanxi Datong University(No.2014-B-01).
文摘Objective The neuroprotective function of heat shock protein A5(HSPA5)in ischemic stroke has been confirmed.This study aimed to investigate the effects of early aerobic exercise on neurological function recovery from cerebral ischemia/reperfusion and to determine whether these effects are associated with the expression level of HSPA5 in the ischemic penumbra.Methods A total of 72 male Sprague-Dawley rats were randomly assigned to the ischemia and exercise group[middle cerebral artery occlusion(MCAO)-Ex,n=18],ischemia and sedentary group(MCAO-St,n=18),sham-surgery and exercise group(Sham-Ex,n=18),or sham-surgery and sedentary group(Sham-St,n=18).The MCAO-Ex and MCAO-St groups were subjected to MCAO for 60 min,whereas the Sham-Ex and Sham-St groups were subjected to an identical operation without MCAO.Rats in the MCAO-Ex and Sham-Ex groups then ran on a treadmill for 30 min once a day for 5 consecutive days.After reperfusion,the motor function of the rats was scored by the Bederson neurological function test,balance beam test,and screen test.Nissl staining was conducted to assess morphological and structural change of nerve cells in the ischemic penumbra.The reverse transcription-quantitative polymerase chain reaction was applied to detect the mRNA expression of HSPA5.Western blot analysis was conducted to determine the protein expression of HSPA5.Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling(TUNEL)staining was carried out in the ischemic penumbra after MCAO.Results Rats receiving early treadmill exercise had lower Bederson neurological function,balance beam,and screen test scores on the 3rd,7th,and 14th days after MCAO;in addition,more neurons survived in the ischemic penumbra after MCAO,and higher mRNA and protein expression of HSPA5 and fewer TUNEL-positive stained cells were observed.Conclusion Our study demonstrated that early aerobic exercise can improve neurological function recovery after ischemia/reperfusion.Furthermore,the increased level of HSPA5 in the ischemic penumbra might be one of the mechanisms of enhanced neurological function recovery.
基金supported by a Shun Hing Institute of Advanced Engineering Grant(No.4720247)a General Research Fund/Early Career Scheme(No.24201919)from the Research Grants Council of Hong Kong Special Administrative Region(to LD)。
文摘Dynamic protein-protein interactions are essential for proper cell functioning.Homointeraction events—physical interactions between the same type of proteins—represent a pivotal subset of protein-protein interactions that are widely exploited in activating intracellular signaling pathways.Capacities of modulating protein-protein interactions with spatial and temporal resolution are greatly desired to decipher the dynamic nature of signal transduction mechanisms.The emerging optogenetic technology,based on genetically encoded light-sensitive proteins,provides promising opportunities to dissect the highly complex signaling networks with unmatched specificity and spatiotemporal precision.Here we review recent achievements in the development of optogenetic tools enabling light-inducible protein-protein homo-interactions and their applications in optical activation of signaling pathways.
文摘One hundred and three Hereford × Angus crossbred, spring-calving cows were used to determine the influence of early weaning performance of their male progeny and the need for cowherd winter supplementation. Treatments were early weaned (EW;115-d of age) and normal weaned calves (NW;220-d of age). Both intact and castrated male progeny were randomly allotted to early and normal weaning calf management treatments. Following summer grazing and weaning protocols, cows within each weaning group were randomly assigned to receive winter supplementation treatments of LOW (1.26 kg/hd/d) or HIGH (1.81 kg/hd/d) amounts of a protein supplement containing 450 g/kg CP (DM basis). Cows whose calves were weaned early gained more BW and BCS (P P P P = 0.04) BCS but similar (P = 0.74) BW when compared with cows with normally weaned calves that received 100% of their supplement (NWHIGH). Early weaning has the potential to reduce the amount of supplement required during the winter grazing period.
文摘Background: Caries detection in school oral examinations insufficient accuracy. Objective: To evaluate advantages of introducing quantitative light-induced fluorescence-digital (QLF-D) in school oral examinations. Methods: Experiment No. 1. Early demineralized lesions in the upper and lower incisors and canines were visually inspected by three dentists and by QLF-D. The numbers of tooth planes with early demineralized lesions were compared between the methods. Experiment No. 2. Approximal demineralized lesions in molars were assessed by visual inspection, x-ray imaging, and QLF-D. The numbers of tooth planes with demineralized lesions were compared among the methods. Experiment No. 3. Plaque distribution was evaluated by QLF-D and a traditional staining method. The ratio of the diameter of plaque to tooth crown in the tooth axis direction in each method was calculated. The results were evaluated by Pearson’s correlation coefficient analysis and Bland-Altman plot. Results: Experiment No. 1. The three dentists found 0.67 tooth planes on average. QLF-D found 22 tooth planes with early demineralized lesions in the same samples. Experiment No. 2. Fourteen approximal tooth planes of molars were found to have demineralized lesions by x-ray imaging. QLF-D detected 71.4% of the tooth planes out of the 14, whereas visual inspection found 7.1%. Experiment No. 3. The Pearson’s correlation coefficient for the evaluations of plaque distribution between the QLF-D and traditional staining methods was 0.77 (P Conclusion: The results support introduction of QLF-D for use in school dental examinations.
基金the Natural Science Foundation of Shandong Province, No. Y2004C04
文摘BACKGROUND: Insulin-like growth factor-I(IGF-1), as one of the important members of growth factor family, participants in the regulation of many physiological functions and behaviors, having very strong neuroprotective effect. However, the expression of IGF-1 following cerebral ischemia/reperfusion is still disputed. OBJECTIVE: To observe the expression of IGF-1 and protein of corpora striata in ischemic side at the early stage of middle cerebral artery ischemia/reperfusion in rhesus monkey. DESIGN : A completely randomized grouping design, controlled animal experiment SETTING : Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University MATERIALS: ① Totally 17 rhesus monkeys , of either gender, aged 4 to 5 years, were enrolled . Seven rhesus monkeys observed with gene chip were randomly divided into 2 groups: sham operation group (n=3) and ischemia/reperfusion group 〈n=4〉. Ten rhesus monkeys observed with in situ hybridization and immunohistochemistry method were randomly divided into 2 groups: sham operation group 〈n=3 〉and ischemia/reperfusion group (n=7). Rhesus monkeys observed under microscope were divided into 2 groups: sham operation group (n=6) and ischamia/reperfusion group (n=-11).②Materials used in the experiment: cresyl violet (Sigma Company, America); immunohistochemical reagent kit ( Huamei Bio-engineering Company); In situ hybridization reagent kit (Boshide Bio-engineering Co.Ltd, Wuhan); 12 800 dots chip (Boxing Company, Shanghai). METHODS : This experiment was carried out at the Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University from January 2001 to December 2003.① The onset area of middle cerebral artery was blocked for 2 hours, middle cerebral artery ischemia/reperfusion models were created.② After ischemia/reperfusion for 24 hours, cerebral tissue sections of rhesus monkeys were prepared and stained with cresyl violet. Image analysis was performed with 5001W image analysis software. Morphological change of corpora striata of operative side was observed in the rhesus monkeys between two groups. Total RNA was extracted from cerebral tissue. ③ Detection of gene chip: Cy3-duTP and Cy5-duTP were used to respectively perform reverse transcription labeling. The sample was reversely transcribed into cDNA, then hybridized with cDNA of cerebral tissue. Genes with the separate absolute value of cy3 and cy5〉800, cY3/cy5 〉 2(high expression) or 〈 0.5 (low expression) were found out. Those were genes with differential expression. ④ The expressions of IGF-1 mRNA and protein level of corpora striata in ischemic side of rhe- sus monkeys were detected between sham operation group and ischemia/reperfusion group at 9 and 24 hours after ischemia/reperfusion with in situ hybridization method and immunohistochemical method. Brown granules were IGF-1 protein positive cells. ⑤ Analysis of variance was used in the difference comparison of measurement data among groups. MAIN OUTCOME MEASURES : ① Change of morphological structure of corpora striata at ischemic side in rhesus monkeys. ② Change of cerebral gene expression profiles at ischemia/reperfusion in rhesus monkeys between two groups.③ Expression of IGF-1 mRNA and protein level of corpora striata at ischemia/reperfu- sion in rhesus monkeys between two groups. RESULTS : ① Pathological change : Obvious pathological change of cerebral infarction appeared in the ischemia and reperfusion group, while there was no such pathological change in the sham operation group.② Change of gene expression profile : There were 4480 genes with difference expression in the ischemia/reperfusion group and sham-operation group, in which, 260 genes had high expression and their absolute value was over 800, and 63 genes had low expression, cy3/cy5 of IGF-1 was 0.379, being relative low ex- pression. ③ IGF-1 mRNA and protein positive cell counts in corpora striata at cerebral ischemic side[IGF-1 mRNA: 〈9.72±1.18),(9.11 ±0.76),(14.77±0.60) counts/field:lGF-1 protein: (15.11 ±1.83),(15.39±0.78), (34.62±0.97)counts/field, P 〈 0.05-0.01]. CONCLUSION: IGF-1 mRNA and protein are lowly expressed in middle cerebral artery of rhesus monkeys at ischemia/reperfusion.
文摘We tested urine albumin excretion rate (UAER), urine transrerrin (TRF ), retinolulnding protein (RBP), N --acetyl--aD-gi ucosamlnldase (N AG ), P, - mi c rogl obu I in (P, -- M G ) and lgGIn 45 cases of NIDDM. Thirty cases with UAER<300 mg/d were divided into two groups. Data wereshown with urine protein index (urine protein/urine creatlne). ResultS showed that urine transferrinwas more seusltlve than albumlu, and the combined test or urine Protein is nontraumatic, which hadsteatncance to diagnose early diabetic nepkropatby.