目的探讨温肾宣痹汤联合碳酸钙D3咀嚼片治疗骨质疏松症的临床疗效。方法选取2018年1月—2020年1月期间濮阳市中医医院收治的骨质疏松症(Osteoporosis,OP)患者84例,按就诊顺序编号随机分为对照组42例和治疗组42例。对照组采用碳酸钙D3咀...目的探讨温肾宣痹汤联合碳酸钙D3咀嚼片治疗骨质疏松症的临床疗效。方法选取2018年1月—2020年1月期间濮阳市中医医院收治的骨质疏松症(Osteoporosis,OP)患者84例,按就诊顺序编号随机分为对照组42例和治疗组42例。对照组采用碳酸钙D3咀嚼片治疗,治疗组在此基础上加用温肾宣痹汤治疗,两组患者均以4周为1个疗程,治疗3个疗程。对两组患者临床疗效、中医症状积分、骨密度(L2~L4及双髋部)、骨代谢指标[血清骨钙素(Bone Glaprotein,BGP)、总Ⅰ型原胶原氨基端延长肽(Procollagen type I nterminal propeptide,PINP)、β胶原降解产物(Beta collagen degradation products,β-CTX)]、安全性进行分析及比较,采用视觉模拟疼痛评分(VAS)进行疼痛评估。结果治疗后治疗组总有效率95.24%(40/42)与对照组73.81%(31/42)比较,差异有统计学意义(P<0.05)。治疗后,两组患者中医症状积分及VAS评分与治疗前比较均降低,且组间比较,治疗组中医症状积分及VAS评分更低(P<0.05)。治疗后,两组患者L2~L4及双髋部骨密度与治疗前比较均增加,且组间比较,治疗组L2~L4及双髋部骨密度更高(P<0.05)。治疗后,两组患者BGP水平均高于治疗前,且组间比较,治疗组BGP水平更高(P<0.05);两组患者PINP及β-CTX水平均低于治疗前,且组间比较,治疗组PINP及β-CTX水平更低(P<0.05)。治疗组不良反应总发生率7.14%和对照组14.29%比较,差异无统计学意义(P>0.05)。结论温肾宣痹汤联合碳酸钙D3咀嚼片治疗OP肝肾不足证的疗效显著,能明显缓解患者疼痛,增加骨密度,改善骨代谢,且安全可靠。展开更多
Background Familial hypercholesterolemia （FH）, caused by low density lipoprotein （LDL） receptor （LDL-R） gene mutations, is associated with increased risk of premature coronary heart disease. Until now, limited m...Background Familial hypercholesterolemia （FH）, caused by low density lipoprotein （LDL） receptor （LDL-R） gene mutations, is associated with increased risk of premature coronary heart disease. Until now, limited molecular data concerning FH are available in China. The present study described the clinical profiles and cell biological defects of a Chinese FH kindred with novel LDL-R gene mutation. Methods The patient＇s LDL-R gene coding region was sequenced. The patient＇s lymphocytes were isolated and the LDL-R expression, binding and up-take functions were observed by immunohistochemistry staining and flow cytometry detection. The patient＇s heart and the major large vessels were detected by vessel ultrasound examination and myocardial perfusion imaging （MPI）. Results The patient＇s LDL-R expression, LDL binding and up-take functions were significantly lower than normal control （39%, 63% and 76% respectively）. A novel homozygous 1439 C→T mutation of the LDL-R gene was detected in the patient and his family. ECG showed atypical angina pectoris. Echocardiogram showed stenosis of the coronary artery and calcification of the aortic valve and its root. Blood vessel ultrasound examination showed the thickness of large vessel intima, and the vessel lumen was narrowed by 71%. MPI showed ischemic changes. Conclusions The LDL-R synthesis dysfunction of FH patients leads to arterial stenosis and calcification, which are the major phenotype of the clinical disorder. The mutation of the LDL-R gene is determined. These data increase the mutational spectrum of FH in China.展开更多
Background Familial hypercholesterolemia （FH） is an autosomal disorder associated with elevated plasma low density lipoprotein （LDL） levels leading to premature coronary heart disease （CHD）. As a result of long-...Background Familial hypercholesterolemia （FH） is an autosomal disorder associated with elevated plasma low density lipoprotein （LDL） levels leading to premature coronary heart disease （CHD）. As a result of long-term hyperlipemia, FH patients will present endarterium thickening and atherosclerosis. In the present study we scanned the related gene of a clinically diagnosed autosomal genetic hypercholesterolemia family for the possible mutations and established eukaryotic expression vector of mutation of proprotein convertase subtilisin/kexin type 9 （PCSK9） gene with gene recombination technique to investigate the contributions of the variation on low density lipoprotein receptor （LDL-R） metabolism and function alternation.Methods Mutation detection was conducted for LDL-R, apolipoprotein B100 （apoB100） and PCSK9 gene with nucleotide sequencing in a Chinese FH family. The full-length cDNA of wild type PCSK9 gene （WT-PCSK9） was obtained from Bel-7402. Site mutagenesis was used to establish the recombinant eukaryotic expression vector carrying pathogenic type of PCSK9 gene and the inserted fragment was sequenced. With the blank vector as control, liposome transfection method was used to transfect the Bel-7402 cells with recombinant plasmid. The expression of LDL-R mRNA was examined by RT-PCR. PCSK9 and the expression of LDL-R protein were determined by Western blotting. Results The G→T mutation at the 918 nucleotide of PCSK9 gene resulted in the substitution of the arginine by a serine at the codon 306 of exon 6. After sequencing, it was confirmed that the inserted fragment of established expression vector had correct size and sequence and the mutant was highly expressed in Bel-7402 cells. There was no significant variation in the levels of LDL-R mRNA. LDL-R mature protein was decreased by 57% after the cells were transfected by WT-PCSK9 plasmid. Mature LDL-R was significantly decreased by 12% after the cells were transfected by R306S mutant as evidenced by gray scale scanning, suggesting that the new mutant R306S can significantly decrease the expression of mature LDL-R protein.Conclusions A novel missense mutation of PCSK9 gene, R306S, was found and the eukaryotic expression vectors of mutant and wild-type of PCSK9 gene were established. There was no significant variation in the levels of LDL-R mRNA. The R306S mutation could significantly lead to the decrease of LDL-R mature protein expression, which might be the pathogenic gene of the FH family.展开更多
Objective: This paper aims at measurement enhanced effect of oxidized lipoprotien(a) [ox Lp(a)] on permeability of monolayer endothelial cells and relationship with reactive oxygen species(ROS) generation and desmogle...Objective: This paper aims at measurement enhanced effect of oxidized lipoprotien(a) [ox Lp(a)] on permeability of monolayer endothelial cells and relationship with reactive oxygen species(ROS) generation and desmogleins(DSGs) expression.Methods and Results: Transendothelial permeability was assayed by transwell and reactive oxygen species(ROS) was determined by DCFH-DA staining. RT-PCR was carried out to determine DSGl and DSC2 expression in m RNA, respectively.Transendothelial permeability was enhanced by ox LP(a) dose and time dependently. The most marked effect appeared at a concentration of 100 mg/L, Transendothelial permeability reached the maximum value after 2 h of FITC-dextran addition, and then gradually decreased after 4 h. ox Lp(a) induces the generation of cellular reactive oxygen species(ROS), and this effect could be inhibited by superoxide dismutase(SOD).Incubation of HUVECs with ox Lp(a) resulted in a dose and time-dependent down-regulation of DSGl and DSC2 expression at transcriptional level. Conclusion:Permeability of monolayer endothelial cells was enhanced by ox Lp(a) which is related to up-regulating ROS formation and down-regulating desmogleins expression.展开更多
Background Aortic dissection(AD)is a life-threatening vascular disease caused by various etiologies including Marfan’s syndrome.Angiotensin receptor blocker(ARB)drugs have been confirmed beneficial for patients with ...Background Aortic dissection(AD)is a life-threatening vascular disease caused by various etiologies including Marfan’s syndrome.Angiotensin receptor blocker(ARB)drugs have been confirmed beneficial for patients with Marfanoid AD but remain unclear for non-Marfanoid case.Methods Patients with DeBakey typeⅢ,Standford A aortic dissection were consecutively recruited to Guangdong Provincial People’s Hospital and Jieyang People’s Hospital between January 2010 and January 2015,with 25 individuals in the ARB group and 32 individuals in the non-ARB group(control group),respectively.The follow-up was performed by database or telephone.The follow-up data consisted of patient’s symptoms and chief complaints,blood pressure,heart rate,and aortic computed tomography angiography(CTA)data.Endpoints and in-hospital events were recorded.Results Baseline data,including age,gender,heart rate,smoking status,and the incidence of diabetes,demonstrated no significant difference between two groups.There was neither death nor urgent surgical case in either group during the follow-up period.The aortic CTA follow-up demonstrated the absorption of the proximal inversely avulsed hematoma for both groups,whereas the reduction was more evident in the ARB group as compared with the non-ARB group[-(9.33±1.99)mm vs.-(4.08±2.35)mm,respectively,P<0.05],suggesting that hematoma absorption was more remarkable in the ARB group.The maximum vascular diameter at the dissection in both groups declined during the follow-up,whereas the reduction in the ARB group was significantly more striking relative to that in the non-ARB group[-(5.50±1.15)mm vs.-(0.31±0.69)mm,respectively,P<0.001],indicating a potential role of ARB drugs in curbing aortic enlargement and remodeling.Conclusions Independent of blood pressure(BP)reduction,ARB utilization in patients with non-Marfan’s aortic dissection may facilitate the absorption of reverse-torn hematoma at the proximal end of dissection,as well as delaying the aortic dilation and remodeling.展开更多
Background Aortic dissection(AD)is a lethal medical emergency,which lacks specific biomarkers and effective pharmaceutical therapies.Increasing evidences have shown beneficial effect of angiotensin receptor blocker(AR...Background Aortic dissection(AD)is a lethal medical emergency,which lacks specific biomarkers and effective pharmaceutical therapies.Increasing evidences have shown beneficial effect of angiotensin receptor blocker(ARB)drugs on downregulating transforming growth factor-β(TGF-β)pathway in Marfanoid AD.However,for non-Marfanoid AD,the effectiveness of ARB drugs,as well as the possible mechanisms,remains unclear.Methods Sprague Dawley(SD)rats were fed by gavage(i.g.)with either 150 mg/(kg·d)Hydroxyethyl diamine(AEEA)or isovolumic saline(normal saline group).AEEA-induced SD rats were further randomly divided into three groups,including the AEEA+Losartan group[AEEA induction+20 mg/(kg·d)i.g.Losartan],the AEEA+Amlodipine group[AEEA induction+6.5 mg/(kg·d)i.g.Amlodipine]and the AEEA+normal saline group(AEEA induction+isovolumic saline i.g.)group.Thus there were 4 groups with 12 mice in each.Tail blood pressure,aortic diameter and the number of aortic dissected lesions were measured in the above 4 groups 4 weeks thereafter.Western-blot was used to detect the expression of components of TGF-β/SMADs pathway,such as TGF-β1,drosophila mothers against decapentaplegic protein 2(Smad2),Smad3,Smad4,protein kinase B(AKT)and phosphorylated AKT(p-AKT).Results No significant difference of blood pressure was seen between the AEEA+Losartan group and the AEEA+Amlodipine group(P=0.81).Ultrasound data indicated a significant reduction in aortic dilation of ascending aorta,aortic arch and descending aorta in Losartan intervention group relative to the Amlodipine intervention group(P<0.001).Hematoxylin-eosin(HE)staining of aortic tissue demonstrated that under the setting of AEEA induction,AEEA+Losartan group had a lower incidence of aortic dissection than the AEEA+normal saline group and the AEEA+Amlodipine group(all P<0.01).Losartan significantly reduced the expression of TGF-β1,Smad2,Smad3,Smad4 in aortic tissues of AEEA-induced rats(all P<0.05).Conclusions Independent of BP reduction,Losartan,as an ARB drug,can prevent aortic dissection by inhibiting TGF-β/SMADs signaling pathway.展开更多
文摘目的探讨温肾宣痹汤联合碳酸钙D3咀嚼片治疗骨质疏松症的临床疗效。方法选取2018年1月—2020年1月期间濮阳市中医医院收治的骨质疏松症(Osteoporosis,OP)患者84例,按就诊顺序编号随机分为对照组42例和治疗组42例。对照组采用碳酸钙D3咀嚼片治疗,治疗组在此基础上加用温肾宣痹汤治疗,两组患者均以4周为1个疗程,治疗3个疗程。对两组患者临床疗效、中医症状积分、骨密度(L2~L4及双髋部)、骨代谢指标[血清骨钙素(Bone Glaprotein,BGP)、总Ⅰ型原胶原氨基端延长肽(Procollagen type I nterminal propeptide,PINP)、β胶原降解产物(Beta collagen degradation products,β-CTX)]、安全性进行分析及比较,采用视觉模拟疼痛评分(VAS)进行疼痛评估。结果治疗后治疗组总有效率95.24%(40/42)与对照组73.81%(31/42)比较,差异有统计学意义(P<0.05)。治疗后,两组患者中医症状积分及VAS评分与治疗前比较均降低,且组间比较,治疗组中医症状积分及VAS评分更低(P<0.05)。治疗后,两组患者L2~L4及双髋部骨密度与治疗前比较均增加,且组间比较,治疗组L2~L4及双髋部骨密度更高(P<0.05)。治疗后,两组患者BGP水平均高于治疗前,且组间比较,治疗组BGP水平更高(P<0.05);两组患者PINP及β-CTX水平均低于治疗前,且组间比较,治疗组PINP及β-CTX水平更低(P<0.05)。治疗组不良反应总发生率7.14%和对照组14.29%比较,差异无统计学意义(P>0.05)。结论温肾宣痹汤联合碳酸钙D3咀嚼片治疗OP肝肾不足证的疗效显著,能明显缓解患者疼痛,增加骨密度,改善骨代谢,且安全可靠。
基金This study was supported by grants from the National Natural Science Foundation of China （No, 30470722, 30771982, 30772356）, Beijing Natural Science Foundation （No. 7052021, 7062010）, and Science and Technology New Star Funds of Beijing （No. 2004B27, 2005A29）
文摘Background Familial hypercholesterolemia （FH）, caused by low density lipoprotein （LDL） receptor （LDL-R） gene mutations, is associated with increased risk of premature coronary heart disease. Until now, limited molecular data concerning FH are available in China. The present study described the clinical profiles and cell biological defects of a Chinese FH kindred with novel LDL-R gene mutation. Methods The patient＇s LDL-R gene coding region was sequenced. The patient＇s lymphocytes were isolated and the LDL-R expression, binding and up-take functions were observed by immunohistochemistry staining and flow cytometry detection. The patient＇s heart and the major large vessels were detected by vessel ultrasound examination and myocardial perfusion imaging （MPI）. Results The patient＇s LDL-R expression, LDL binding and up-take functions were significantly lower than normal control （39%, 63% and 76% respectively）. A novel homozygous 1439 C→T mutation of the LDL-R gene was detected in the patient and his family. ECG showed atypical angina pectoris. Echocardiogram showed stenosis of the coronary artery and calcification of the aortic valve and its root. Blood vessel ultrasound examination showed the thickness of large vessel intima, and the vessel lumen was narrowed by 71%. MPI showed ischemic changes. Conclusions The LDL-R synthesis dysfunction of FH patients leads to arterial stenosis and calcification, which are the major phenotype of the clinical disorder. The mutation of the LDL-R gene is determined. These data increase the mutational spectrum of FH in China.
基金This study was supported by grants from the National Natural Science Foundation of China （No. 30470722, 30771986, 30772356）, Beijing Natural Science Foundation （No. 7092016, 7062010, 7052021）, and Key Project for Applicable Basic Research of Hunan Province （No. 2008FJ2006）.
文摘Background Familial hypercholesterolemia （FH） is an autosomal disorder associated with elevated plasma low density lipoprotein （LDL） levels leading to premature coronary heart disease （CHD）. As a result of long-term hyperlipemia, FH patients will present endarterium thickening and atherosclerosis. In the present study we scanned the related gene of a clinically diagnosed autosomal genetic hypercholesterolemia family for the possible mutations and established eukaryotic expression vector of mutation of proprotein convertase subtilisin/kexin type 9 （PCSK9） gene with gene recombination technique to investigate the contributions of the variation on low density lipoprotein receptor （LDL-R） metabolism and function alternation.Methods Mutation detection was conducted for LDL-R, apolipoprotein B100 （apoB100） and PCSK9 gene with nucleotide sequencing in a Chinese FH family. The full-length cDNA of wild type PCSK9 gene （WT-PCSK9） was obtained from Bel-7402. Site mutagenesis was used to establish the recombinant eukaryotic expression vector carrying pathogenic type of PCSK9 gene and the inserted fragment was sequenced. With the blank vector as control, liposome transfection method was used to transfect the Bel-7402 cells with recombinant plasmid. The expression of LDL-R mRNA was examined by RT-PCR. PCSK9 and the expression of LDL-R protein were determined by Western blotting. Results The G→T mutation at the 918 nucleotide of PCSK9 gene resulted in the substitution of the arginine by a serine at the codon 306 of exon 6. After sequencing, it was confirmed that the inserted fragment of established expression vector had correct size and sequence and the mutant was highly expressed in Bel-7402 cells. There was no significant variation in the levels of LDL-R mRNA. LDL-R mature protein was decreased by 57% after the cells were transfected by WT-PCSK9 plasmid. Mature LDL-R was significantly decreased by 12% after the cells were transfected by R306S mutant as evidenced by gray scale scanning, suggesting that the new mutant R306S can significantly decrease the expression of mature LDL-R protein.Conclusions A novel missense mutation of PCSK9 gene, R306S, was found and the eukaryotic expression vectors of mutant and wild-type of PCSK9 gene were established. There was no significant variation in the levels of LDL-R mRNA. The R306S mutation could significantly lead to the decrease of LDL-R mature protein expression, which might be the pathogenic gene of the FH family.
基金Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province,National and Natural Science Foundation of Chinagrant number:81070221Visiting Scholar Foundation of Key Laboratory of Biorheological Science and Technology (Chongqing University) of Ministry of Education(2010)
文摘Objective: This paper aims at measurement enhanced effect of oxidized lipoprotien(a) [ox Lp(a)] on permeability of monolayer endothelial cells and relationship with reactive oxygen species(ROS) generation and desmogleins(DSGs) expression.Methods and Results: Transendothelial permeability was assayed by transwell and reactive oxygen species(ROS) was determined by DCFH-DA staining. RT-PCR was carried out to determine DSGl and DSC2 expression in m RNA, respectively.Transendothelial permeability was enhanced by ox LP(a) dose and time dependently. The most marked effect appeared at a concentration of 100 mg/L, Transendothelial permeability reached the maximum value after 2 h of FITC-dextran addition, and then gradually decreased after 4 h. ox Lp(a) induces the generation of cellular reactive oxygen species(ROS), and this effect could be inhibited by superoxide dismutase(SOD).Incubation of HUVECs with ox Lp(a) resulted in a dose and time-dependent down-regulation of DSGl and DSC2 expression at transcriptional level. Conclusion:Permeability of monolayer endothelial cells was enhanced by ox Lp(a) which is related to up-regulating ROS formation and down-regulating desmogleins expression.
文摘Background Aortic dissection(AD)is a life-threatening vascular disease caused by various etiologies including Marfan’s syndrome.Angiotensin receptor blocker(ARB)drugs have been confirmed beneficial for patients with Marfanoid AD but remain unclear for non-Marfanoid case.Methods Patients with DeBakey typeⅢ,Standford A aortic dissection were consecutively recruited to Guangdong Provincial People’s Hospital and Jieyang People’s Hospital between January 2010 and January 2015,with 25 individuals in the ARB group and 32 individuals in the non-ARB group(control group),respectively.The follow-up was performed by database or telephone.The follow-up data consisted of patient’s symptoms and chief complaints,blood pressure,heart rate,and aortic computed tomography angiography(CTA)data.Endpoints and in-hospital events were recorded.Results Baseline data,including age,gender,heart rate,smoking status,and the incidence of diabetes,demonstrated no significant difference between two groups.There was neither death nor urgent surgical case in either group during the follow-up period.The aortic CTA follow-up demonstrated the absorption of the proximal inversely avulsed hematoma for both groups,whereas the reduction was more evident in the ARB group as compared with the non-ARB group[-(9.33±1.99)mm vs.-(4.08±2.35)mm,respectively,P<0.05],suggesting that hematoma absorption was more remarkable in the ARB group.The maximum vascular diameter at the dissection in both groups declined during the follow-up,whereas the reduction in the ARB group was significantly more striking relative to that in the non-ARB group[-(5.50±1.15)mm vs.-(0.31±0.69)mm,respectively,P<0.001],indicating a potential role of ARB drugs in curbing aortic enlargement and remodeling.Conclusions Independent of blood pressure(BP)reduction,ARB utilization in patients with non-Marfan’s aortic dissection may facilitate the absorption of reverse-torn hematoma at the proximal end of dissection,as well as delaying the aortic dilation and remodeling.
文摘Background Aortic dissection(AD)is a lethal medical emergency,which lacks specific biomarkers and effective pharmaceutical therapies.Increasing evidences have shown beneficial effect of angiotensin receptor blocker(ARB)drugs on downregulating transforming growth factor-β(TGF-β)pathway in Marfanoid AD.However,for non-Marfanoid AD,the effectiveness of ARB drugs,as well as the possible mechanisms,remains unclear.Methods Sprague Dawley(SD)rats were fed by gavage(i.g.)with either 150 mg/(kg·d)Hydroxyethyl diamine(AEEA)or isovolumic saline(normal saline group).AEEA-induced SD rats were further randomly divided into three groups,including the AEEA+Losartan group[AEEA induction+20 mg/(kg·d)i.g.Losartan],the AEEA+Amlodipine group[AEEA induction+6.5 mg/(kg·d)i.g.Amlodipine]and the AEEA+normal saline group(AEEA induction+isovolumic saline i.g.)group.Thus there were 4 groups with 12 mice in each.Tail blood pressure,aortic diameter and the number of aortic dissected lesions were measured in the above 4 groups 4 weeks thereafter.Western-blot was used to detect the expression of components of TGF-β/SMADs pathway,such as TGF-β1,drosophila mothers against decapentaplegic protein 2(Smad2),Smad3,Smad4,protein kinase B(AKT)and phosphorylated AKT(p-AKT).Results No significant difference of blood pressure was seen between the AEEA+Losartan group and the AEEA+Amlodipine group(P=0.81).Ultrasound data indicated a significant reduction in aortic dilation of ascending aorta,aortic arch and descending aorta in Losartan intervention group relative to the Amlodipine intervention group(P<0.001).Hematoxylin-eosin(HE)staining of aortic tissue demonstrated that under the setting of AEEA induction,AEEA+Losartan group had a lower incidence of aortic dissection than the AEEA+normal saline group and the AEEA+Amlodipine group(all P<0.01).Losartan significantly reduced the expression of TGF-β1,Smad2,Smad3,Smad4 in aortic tissues of AEEA-induced rats(all P<0.05).Conclusions Independent of BP reduction,Losartan,as an ARB drug,can prevent aortic dissection by inhibiting TGF-β/SMADs signaling pathway.
