OBJECTIVE: To investigate the characteristics of mutations in exon 3-20 of Wilson disease (WD) gene and their consequences in Chinese population. METHODS: Sixty unrelated normal Chinese and forty-four unrelated WD pat...OBJECTIVE: To investigate the characteristics of mutations in exon 3-20 of Wilson disease (WD) gene and their consequences in Chinese population. METHODS: Sixty unrelated normal Chinese and forty-four unrelated WD patients were studied. Genomic DNA was prepared from peripheral blood leukocytes by a salt-out method. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and subsequently direct sequencing were used to identify the mutations and polymorphisms of WD gene. RESULTS: Ten different mutations have been found, accounting for 52% of the mutant genes. Five of them are identified as novel missense mutations. Mutations Arg778Leu, Thr935Met and Ala874Val were represented respectively in 28.4%, 6.8% and 3.4% of WD chromosomes. The remaining mutations were found rare and limited to one or two patients. A total of 11 patients were homozygous for a single mutation, and 17 patients were in a compound heterozygous state with or without a known mutation. CONCLUSION: In Chinese, WD seems to result from two or three relatively common mutations and a large number of rare mutations. Arg778Leu and Thr935Met might be hotspots of mutation in Chinese population. The results indicated that the feature of mutations of WD gene is different between Chinese and the Western. Instead of exon 14 and exon 18, we had to select exon 8 and exon 12 first to detect mutations of WD gene in Chinese. It is of great importance to establish a direct diagnostic method for WD. This study improves our knowledge on functional domains of the WD gene, and helps elucidate the wide spectrum of manifestations of the disease as well.展开更多
Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute...Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury （RIRI）. Methods： Fifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/（kg.d）], SHT low-dose group [1.5 g/（kg,d）] and SHT high-dose group [3.0 g/（kg.d）], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope. Results： Compared with the sham surgery group, the serum creatinine （SCr） and blood urea nitrogen （BUN） levels were significantly increased, Na＋-K＋-ATPase protein level was decreased, and kidney injury molecule-1 （KIM-1） protein level was increased in the model group after the surgery （P〈0.01 or P〈0.05）. Compared with the model group, the SCr, BUN, pathological scores, Na＋-K＋-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery （P〈0.05 or P〈0.01）. And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group （P〈0.05）. The Iocalizations of Na＋-K＋-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group. Conclusions： The SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.展开更多
文摘OBJECTIVE: To investigate the characteristics of mutations in exon 3-20 of Wilson disease (WD) gene and their consequences in Chinese population. METHODS: Sixty unrelated normal Chinese and forty-four unrelated WD patients were studied. Genomic DNA was prepared from peripheral blood leukocytes by a salt-out method. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and subsequently direct sequencing were used to identify the mutations and polymorphisms of WD gene. RESULTS: Ten different mutations have been found, accounting for 52% of the mutant genes. Five of them are identified as novel missense mutations. Mutations Arg778Leu, Thr935Met and Ala874Val were represented respectively in 28.4%, 6.8% and 3.4% of WD chromosomes. The remaining mutations were found rare and limited to one or two patients. A total of 11 patients were homozygous for a single mutation, and 17 patients were in a compound heterozygous state with or without a known mutation. CONCLUSION: In Chinese, WD seems to result from two or three relatively common mutations and a large number of rare mutations. Arg778Leu and Thr935Met might be hotspots of mutation in Chinese population. The results indicated that the feature of mutations of WD gene is different between Chinese and the Western. Instead of exon 14 and exon 18, we had to select exon 8 and exon 12 first to detect mutations of WD gene in Chinese. It is of great importance to establish a direct diagnostic method for WD. This study improves our knowledge on functional domains of the WD gene, and helps elucidate the wide spectrum of manifestations of the disease as well.
基金Supported by National Science and Major Project(No.2010ZX09102-204)National Natural Sciences Foundation of China(No.81072914 and 81273968)the Medicine and Health Foundation of PLA(No.10ZYZ255)
文摘Objective： To observe the effect of Compound Shenhua Tablet （复方肾华片, SHT） on the sodium- potassium-exchanging adenosinetriphosphatase （Na＋-K＋-ATPase） in the renal tubular epithelial cells of rats with acute ischemic reperfusion and to investigate the mechanisms underlying the effects of SHT on renal ischemic reperfusion injury （RIRI）. Methods： Fifty male Wistar rats were randomly divided into the sham surgery group, model group, astragaloside group [150 mg/（kg.d）], SHT low-dose group [1.5 g/（kg,d）] and SHT high-dose group [3.0 g/（kg.d）], with 10 rats in each group. After 1 week of continuous intragastric drug administration, surgery was performed to establish the model. At either 24 or 72 h after the surgery, 5 rats in each group were sacrificed, blood biochemistry, renal pathology, immunoblot and immunohistochemical examinations were performed, and double immunofluorescence staining was observed under a laser confocal microscope. Results： Compared with the sham surgery group, the serum creatinine （SCr） and blood urea nitrogen （BUN） levels were significantly increased, Na＋-K＋-ATPase protein level was decreased, and kidney injury molecule-1 （KIM-1） protein level was increased in the model group after the surgery （P〈0.01 or P〈0.05）. Compared with the model group, the SCr, BUN, pathological scores, Na＋-K＋-ATPase, and the KIM-1 protein level of the three treatment groups were significantly improved at 72 h after the surgery （P〈0.05 or P〈0.01）. And the SCr, BUN of the SHT low- and high-dose groups, and the pathological scores of the SHT high-dose group were significantly lower than those of the astragaloside group （P〈0.05）. The Iocalizations of Na＋-K＋-ATPase and megalin of the model group were disrupted, with the distribution areas overlapping with each other and alternately arranged. The severity of the disruption was slightly milder in three treatment groups compared with that of the model group. The results of immunofluorescence staining showed that the SHT high-dose group had a superior effect as compared with the astragaloside group and the SHT low-dose group. Conclusions： The SHT effectively alleviated RIRI caused by ischemic reperfusion, promoted the recovery of the polarity of renal tubular epithelial cells, and protected the renal tubules. The therapeutic effects of SHT were superior to those of astragaloside as a single agent.
