Objective To develop an UPLC-MS/MS method for simultaneously determination of five components (adenosine,cytidine,guanosine,mannitol and adenine) in Qiangshenpaidu capsules. Methods The UPLC separation was performed on an Agilent ZOBAX SB-C18(2.1 mm×150 mm,5 μm) column.Isocratic elution was carried out with mobile phase consisting of methanol-0.1%formic acid (5∶95) at a flow rate of 0.2 mL.min-1.The mass spectrometer was operated in the positive ionization electrospray ( ESI) mode using multiple monitoring ( MRM) for analysis of five components. Results Adenosine,cytidine,guanosine,mannitol and adenine were all analyzed with good precision and accuracy. The linear ranges were 35-1 120 ng.mL-1( r=0.998 1) ,10-320 ng.mL-1( r=0.996 4) , 30-980 ng.mL-1( r=0.999 3) , 40-1 280 ng.mL-1( r=0.993 4), 25-800 ng.mL-1(r=0.996 5),respectively.The recoveries of six analytes ranged from 97.4% to 103.6% and the relative standard deviations were all below 4.7%. Conclusion A sensitive,accurate and suitable UPLC-MS/MS method has been developed,and the method could be applied for the determination of adenosine,cytidine,guanosine,mannitol,and adenine in Qiangshenpaidu Capsules.

Objective To observe the effects of iguratimod ( IT) combined with methotrexate ( MTX) in patients with refractory rheumatoid arthritis ( rRA) . Methods Sixty patients with rRA were randomly divided into 2 groups ( n=30 each group) . The cases in treatment group received 50 mg.d-1 of iguratimod and 10 mg of MTX for 16 weeks. The cases in control group were treated by 10-15 mg of MTX. DAS28 was analyzed. Levels of VEGF and endostatin ( ES) were quantified. Results In the treatment group,after 16-week treatment,DAS28,levels of VEGF and ES were (3.0±1.2),(818.9±178.8) pg.mL-1, (337.8±132.6) ng.mL-1,and those in the control group were (5.7±1.9),(1000.2±245.9) pg.mL-1,(253.8±77.8) ng.mL-1,respectively. In the treatment group,DAS28 and VEGF after the treatment were significantly decreased as compared with those before the treatment ( P<0.01) . The decrement was more significant in the treatment group than in the control group ( P<0.01) . At the 16th week of treatment,ES was significantly increased as compared with that before the treatment ( P<0.01) , and there was a significant difference between the treatment group and the control group (P<0.01). Conclusion Iguratimod combined with MTX have a prominent effect on rRA with high safety. The efficacy of IT on RA might be related with decreasing VEGF release,increasing ES production and alleviating synovium angiogenesis.

Objective To analyse the high-dose dexamethasone suppression test(HDDST)-related differences in the clinical and biochemical features of the patients with Cushing's disease Methods Cases were drawn from 60 consecutive patients with Cushing's disease,who were then divided into two groups according to the response to the HDDST.The clinical and biochemical features between two groups were compared.Results(1) Of the 60 patients with Cushing's disease,23.3%(14/60)of patients(group A)did not yield results of suppression with the HDDST,and the others(group B)did.No difference was found in the age[(33.8?10.4 vs 36.2?11.2)years]and duration of illness[(2.1?1.6 vs 3.9?3.1)years]between two groups.(2)In clinical features,the patients in group A were more likely to have edema of lower limbs(64.3% vs 32.6%),hypokalemia (71.4% vs 28.3%),secondary diabetes(57.1% vs 26.1%)and purple striae(85.7% vs 54.3%,all P

Objective To study the relationship between intake of nutrients and bone density in the middle-aged and the old people. Methods Dietary investigation was conducted and bone densities of lumbar 2-4, tuberositas of femur and the neck of femur were measured in 200 healthy subjects who received physical check-up in our hospital. Subjects were distributed into 6 groups including young or middle-aged,pre-old and old groups of men and women.Comparison was conducted within those groups.Results The average intake of protein, carbohydrate and energy per day in old men-group were (71.4±11.7)g,(294.2±54.7)g and (2196.9±311.3) kcal, respectively, which were significantly lower than those in young or middle-aged groups,and the average amounts of protein, carbohydrate and energy intake per day in old men-group were (91.7±19.5)g, (81.8±85.2)g and (2716.0±451.7)kcal, respectively. The average intake of calcium in women-old group was (362.0±167.1)mg, the lowest in the groups. The incidence of abnormal bone densities of lumbar 2-4, the tuberositas of femur and the neck of femur in old men group was significantly higher than those in young or middle-aged men groups (P＜0.05 ) and the preold men group. There were significantly different incidence rates of abnormal bone densities between the 3 women groups (P＜0.05), especially between young or middle-aged groups and in the old women-group(P＜0. 0001 ). The incidences of abnormal bone density in pre-old men group and old men group were significantly lower than those in-pre-old and old women groups. Conclusion The decrease of calcium intake seemed to be one of the factors leading to abnormity of bone density in old women. While the increase of calcium intake served as one of the nutritional factors affecting the bones.

The outcome and influencing factors in the reversion of impaired glucose regulation(IGR)to normal glycemia(NG)after health education for one year were analyzed by the criterion of American Diabetes Association 2003.The results showed that the improvement of glucose regulation well accorded with the improvement of insulin resistance and islet 13-cell function.Fasting plasma glucose,fasting insulin,triglycerides, insulin resistance and islet?-cell function were the influencing factors for the reversion of IGR to NG.

We report a case of 51-year-old woman with pure unroofed coronary sinus without persistent left superior vena cava and other cardiac anomaly. She presented with dyspnea on exertion during several years. Her chest film showed prominent cardiomegaly and dilated hilar vessels. Cardiac rhythm was atrial fibrillation. Transthoracic echocardiography demonstrated the enlarged coronary sinus with defect toward left atrium on parasternal long axis view and significant flow from coronary sinus into right atrium on subxyphoid view, and its other findings were dilated right ventricle and right atrium, paradoxical septal motion, moderate tricuspid regurgitation and mild mitral regurgitation, which were mimicking of large secundum atrial septal defect. Radionuclide cardioangiography and cardiac catheterization showed the existence of significant shunt. There was no evidence of persistent left superior vena cava on chest CT. Closure of Coronary sinus opening was done. Thereafter her symptoms of congestive heart failure were much improved. We think that careful examination of 2-D echocardiography can be valuable tool for diagnosis of unroofed coronary sinus in adult patient.
Adult ; Atrial Fibrillation ; Axis, Cervical Vertebra ; Cardiac Catheterization ; Cardiac Catheters ; Cardiomegaly ; Coronary Sinus* ; Diagnosis ; Dyspnea ; Echocardiography ; Female ; Heart Atria ; Heart Failure ; Heart Septal Defects, Atrial ; Heart Ventricles ; Humans ; Middle Aged ; Mitral Valve Insufficiency ; Thorax ; Tomography, X-Ray Computed ; Tricuspid Valve Insufficiency ; Vena Cava, Superior*

OBJECTIVETo clarify the differential expression of the genes related to the lipid metabolism in the early stage of atherosclerosis in the young LDLR-/- mice of different ages.

METHODSA RT-PCR assay was used to analyse the gene expression patterns in the livers of LDLR-/- mice and wild type (WT) mice from 14 to 90 days. The characteristics of early lipid deposition in intima were evaluated using biochemical and pathological techniques.

RESULTSIn LDLR-/- mice, when compared to WT mice, the mRNA level of the apolipoprotein A IV (apoA IV), fatty acid translocase (Fat/CD36) and carnitine palmitoyl transferase I (CPT I) changed prominently at the age of 14-days (P < 0.05). At 30 days, the mRNA level of apolipoprotein A I (apoA I) was up regulated, but apolipoprotein F (apoF), CD36 and CPT I were down regulated (P < 0.05). At 60 days, the mRNA levels of apoA I, CPT I and liver X receptor alpha (LXRalpha) were up regulated, but apoA IV was down regulated (P < 0.05). At 90 days, the level of the apoA I was higher, but the expression of the apoA IV, apoF and acyl-coenzymeA oxidase 1 (ACOX1) were down regulated (P < 0.05), whereas the expression of apolipoprotein A V (apoA V), apolipoprotein E (apoE), peroxidase proliferator-activated receptor alpha (PPARalpha) and angiopoietin-like protein 3 (angptl 3) had no significant changes (P > 0.05). The serum levels of TC (P < 0.05), TG (P < 0.05) and LDLC (P < 0.05) in LDLR-/- mice were significantly higher than those in wild type mice with the same age.

CONCLUSIONSThe mRNA levels of the apoA I, apoA IV, apoF, FAT/CD36, CPT I, ACOX1 and LXRalpha of the LDLR-/- mice were significantly changed compared to the WT mice. The genes may be of some relevance to the complicated lipid metabolism network, and have effect in the early stage of atherogenesis.

Animals ; Apolipoprotein A-I ; genetics ; metabolism ; Apolipoproteins A ; genetics ; metabolism ; Apolipoproteins E ; genetics ; metabolism ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Liver X Receptors ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Orphan Nuclear Receptors ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency

OBJECTIVETo explore the relationship between the expression characteristics of lipid metabolism-related genes in the liver and early atherosclerotic lesions in apolipoprotein E and low density lipoprotein receptor gene double knockout (apoE(-/-)/LDLR(-/-)) mice.

METHODSRT-PCR was used to detect the differential expression of lipid metabolism-related genes in the liver of apoE(-/-)/LDLR(-/-) and wild type (WT) mice. Serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) level as well as aortic morphology were also analyzed.

RESULTSAmong the 11 lipid metabolism-related genes, apolipoprotein B100 (apoB100) mRNA levels were significantly higher in apoE(-/-)/LDLR(-/-)mice compared with WT mice. At 14 days, 1, 2 and 3 months of age, the level of mRNA expression were 1.55, 1.47, 1.50 and 2.42 folds of those of the age matched WT mice respectively. The fatty acid transporter (FAT/CD36) mRNA expression levels were higher in 14-day and 3-month old mice at 1.30 and 1.35 folds of those of the age matched WT mice, respectively. Apolipoprotein A IV (apoA IV) and Apolipoprotein AV (apoAV) mRNA levels were significantly down-regulated (0.89 fold decrease in 14-day, and 0.90 folds decrease in 3-month, respectively). The mRNA expression levels of apolipoprotein AI (apo AI), apolipoprotein F (apo F), peroxidase proliferator-activated receptor alpha (PPAR-alpha), liver X receptor alpha (LXRalpha), angiopoietin-like protein 3 (ANGPTL3), acyl-coenzymeA oxidase 1 (ACOX1) and carnitine palmitoyl transferase 1 (CPT1) had no significant changes. Serum TC, TG and LDL-C were higher than those of age matched WT mice at 7, 2 and 30 folds, respectively. Furthermore, apoE(-/-)/LDLR(-/-) mice demonstrated typical early atherosclerotic lesions at sinus and root regions of aorta in an age dependent manner.

CONCLUSIONAlterations of the expression of lipid metabolism-related genes in liver play important roles in the development of AS in the apoE(-/-)/LDLR(-/-) mice at early ages.

Animals ; Aorta ; pathology ; Apolipoprotein A-V ; Apolipoprotein B-100 ; biosynthesis ; genetics ; Apolipoproteins ; biosynthesis ; genetics ; Apolipoproteins A ; biosynthesis ; genetics ; Apolipoproteins E ; deficiency ; Atherosclerosis ; etiology ; metabolism ; pathology ; CD36 Antigens ; biosynthesis ; genetics ; Gene Expression ; Lipid Metabolism ; Liver ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; RNA, Messenger ; metabolism ; Receptors, LDL ; deficiency

The work was aimed to investigate the differential expressions of lipid metabolism related genes in the early stage of atherosclerosis in the young apolipoprotein E deficient (apoE(-/-)) mice at different ages with normal chow diet. The genotypes of mice were identified by using multiplex polymerase chain reaction (multi-PCR) analysis. The semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and real-time quantitative RT-PCR were used to analyze the expressions of lipid metabolism related genes in the liver of apoE(-/-) and age-matched wild type (WT) mice of 14-day old, 1-month old, 2-month old, 3-month old. The serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) contents were assayed using COD-PAP and GPO-PAP methods. The serum apolipoprotein B100 (apoB100) content was quantitated by immune turbidimetry. The hearts were perfusion-fixed in 4% formaldehyde, infiltrated with 30% gum sucrose for 24 h at 4 °C, and embedded in OCT compound. The aortic sinus tissues were serially sectioned at -15 °C, stained with Sudan IV, and counterstained with light green. The results were shown as follows. Compared with that in WT mice, the mRNA levels of apoA I and apoA IV in apoE(-/-) mice aged from 14-day old to 3-month old changed prominently (P<0.05), with apoA I up-regulated and apoA IV down-regulated. At the age of 1 month, the expression of apoB100 in apoE(-/-) mice was higher than that in WT mice (P<0.05). The expression of apoA V was up-regulated (P<0.05) and there was obvious lipid deposition in the aortic intima in apoE(-/-) mice at the age of 2 months. The expressions of fatty acid translocase (Fat/CD36) and angiopoietin-like protein 3 (Angptl 3) in apoE(-/-) mice were higher than those in WT mice at the age of 3 months (P<0.05), while the expressions of peroxisome proliferator-activated receptor α (PPARα), liver X receptor α (LXRα), carnitine palmitoyl transferase I (CPT I) and acyl coenzyme A oxidase 1 (ACOX1) showed no significant changes. The serum TC, TG, LDL-C and HDL-C contents in apoE(-/-) mice aged from 14-day old to 3-month old were higher than those in age-matched WT mice. apoE(-/-) mice showed a marked increase in serum apoB100 content, consistent with the trend of serum LDL-C content and apoB100 mRNA content in the liver. The results suggest that the mRNA expressions of apoA I, apoA IV, apoA V, apoB100 and Angptl 3 in apoE(-/-) mice change significantly compared with those in WT mice, and these genes might be relevant to the complicated lipid metabolism network, and involved in the early stage of atherogenesis.
Animals ; Apolipoprotein A-I ; metabolism ; Apolipoprotein B-100 ; blood ; Apolipoproteins A ; metabolism ; Apolipoproteins E ; genetics ; Atherosclerosis ; genetics ; Gene Expression ; Lipid Metabolism ; genetics ; Lipoproteins, HDL ; blood ; Lipoproteins, LDL ; blood ; Liver ; metabolism ; Mice ; Mice, Knockout ; Triglycerides ; blood