The stable isotope labeling by amino acids in culture (SILAC) based quantitative proteomics serves as a gold standard because of the high accuracy and throughput for protein identifications and quantification. In this study, we discussed the application of SILAC technology in mammal model, and developed quantitative internal standard for comparative proteomics of disease model. The C57BL/6J mice fed by special diet containing the 13C6-Lysine and bred F2 generation. We identified and analyzed total proteins of 9 mice tissues of F2 generation, including brain, lung, heart, stomach, intestine, liver, spleen, kidney, and muscle. Quantitative analysis information could evaluate the mice and different tissues' labeling efficiency. Liver was the most efficient, brain the least, and the labeling efficiency were 96.34%±0.90% and 92.62%±1.98% respectively. The average of the labeling efficiency of F2 generation was 95.80%±0.64%, which met the international standard (≥ 95%) for SILAC quantitative proteomics effective study. SILAC technology was successfully extended to mammalian model system, which will provide powerful tools for the mechanism study of the pathophysiology process with mouse model.
Amino Acids ; chemistry ; Animals ; Diet ; veterinary ; Isotope Labeling ; Lysine ; chemistry ; Mice ; Mice, Inbred C57BL ; Proteins ; chemistry ; Proteomics ; methods

BACKGROUNDHypertensive target organ damage (TOD) is the main reason for mortality or disability in elderly hypertensive patients. The studies on TOD of hypertension in Asia, especially in Chinese elderly hypertensive patients, are very limited. The aim of this study was to evaluate the prevalence and correlative factors of TOD in older Chinese hypertensive inpatients.

METHODSThis is a retrospective survey and data were collected from the computerized medical files of hypertensive inpatients from January of 1993 to December of 2008. The analysis was done on 17 682 inpatients, aged 60 years or older, with a diagnosis of essential hypertension (EH). The evidence of hypertensive TOD and associated factors with TOD were collected.

RESULTSThe prevalence of any hypertensive target organ involvement among these subjects was high. In multivariable Logistic regressions adjusted for potentially confounding factors, older age, male gender, diabetes, EH grade 3, systolic blood pressure (SBP), and low-density lipoprotein cholesterol (LDL-C), were independently associated with coronary artery disease. Age, duration of EH, EH grade 3, SBP, pulse pressure (PP), and homocysteine (Hcy) were independently associated with cerebrovascular disease. Age, diabetes, duration of EH, EH grade 3, SBP, PP and estimated glomerular filtration rate (eGFR) were independently associated with chronic kidney disease. Male gender, EH grade 3 and SBP were independently associated with aortic dissection.

CONCLUSIONSThe prevalence of hypertensive TOD is high in older Chinese hypertensive inpatients. Various cardiovascular risk factors are associated with hypertensive TOD. The level of SBP and severe hypertension (grade 3 hypertension) are common independent risk factors of TOD.

Aged ; Aged, 80 and over ; Aortic Diseases ; blood ; Cerebrovascular Disorders ; blood ; China ; Cholesterol, LDL ; blood ; Coronary Artery Disease ; blood ; Female ; Glomerular Filtration Rate ; physiology ; Homocysteine ; blood ; Humans ; Hypertension ; physiopathology ; Kidney Failure, Chronic ; blood ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo investigate the effects of different anesthetic and analgesic protocols on the cellular immune function and stress hormone in patients undergoing lobectomy for esophagus cancer.

METHODSSixty ASA I or II patients undergoing lobectomy for esophagus cancer were randomly divided into two groups to receive postoperative general anesthesia and intravenous analgesia (group A, n=30) or intraoperative general anesthesia combined with thoracic epidural anesthesia with postoperative epidural analgesia (group B, n=30). The cervical venous blood samples were obtained from the patients at 30 min before anesthesia induction (T(0)), 2 h after skin incision (T(1)), and at 4 h (T(2)), 24 h (T(3)) and 48 h (T(4)) after the end of operation. The T-lymphocyte subsets (CD4(+) and CD8(+)) were analyzed by flow cytometry, serum concentrations of sIL-2R and IL-2 determined by ELISA, and the levels of growth hormone (GR), prolactin (PRL), IL-8 and cortisol (Cor) measured by radioimmunoassay. Visual analogue scale (VAS) was used for assessment of the postoperative analgesic effects.

RESULTSThe VAS scores were significantly lower in group B than in group A at T(2) and T(3) (P<0.05). The percentage of CD4(+) cells and the CD4(+)/CD8(+) ratio in the two groups began to decrease significantly at T(1) (P<0.05), reducing to the lowest level at T(2) in group B and at T(3) in group A. From T(1) to T(4), the percentage of CD4(+) in group B remained significantly higher than those in group A (P<0.05), and from T(3) to T(4), the CD4(+)/CD8(+) ratio in group B were significantly higher than those in group A (P<0.05). The IL-2 level in the two groups began to decrease significantly at T(1) (P<0.05), reaching the lowest level at T(2) in group A and at T(3) in group A. IL-2 level was significantly higher in group B than in group A from T(3) to T(4) (P<0.05). sIL-2R level in group A began to increase at T(1) and peaked at T(3), showing significant differences from the T(0) level, but the level showed no significant variations in group B compared with the T(0) level. From T(2) to T(4), sIL-2R level was significantly higher in group A than in group B (P<0.05). The levels of GH, PRL and Cor increased significantly, while IL-8 decreased in the two groups from T(1) to T(4) (P<0.05), but remained stable in group B.

CONCLUSIONGeneral anesthesia combined with thoracic epidural anesthesia may reduce the perioperative stress reaction and adverse effect on cellular immune function in patients undergoing lobectomy for esophagus cancer.

Adult ; Aged ; Analgesia, Epidural ; methods ; Anesthesia, Epidural ; methods ; Anesthesia, Intravenous ; methods ; Esophageal Neoplasms ; immunology ; surgery ; Female ; Humans ; Immunity, Cellular ; drug effects ; Male ; Middle Aged ; Pain, Postoperative ; drug therapy ; Postoperative Period ; Stress, Physiological ; drug effects ; T-Lymphocytes ; immunology

Actins ; analysis ; Animals ; Cell Division ; drug effects ; Cells, Cultured ; Collagen Type I ; analysis ; Estradiol ; pharmacology ; Liver ; cytology ; drug effects ; Liver Cirrhosis ; etiology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo study the prevalence of hypertension and glucose/lipid metabolism induced by chronic obstructive pulmonary disease (COPD) in hospitalized patients and their correlations to age and gender.

METHODSA retrospectively prevalence survey was conducted among the in-patients with COPD treated in General Hospital of PLA, Peking Union Medical College Hospital and Beijing Hospital between January 1st, 2000 and March 20th, 2010.

RESULTSA total of 4960 COPD patients were selected, including 3570 male and 1390 female patients with a mean age of 72.2∓10.4 years. Among these patients, hypertension had the highest prevalence (40.3%), followed by diabetes/impaired glucose tolerance (18.8%) and then by hyperlipidemia/fat protein metabolism (6.9%). The prevalence of hyperlipidemia/lipoprotein metabolism in the COPD patients over 80 years old decreased significantly with age. The prevalence of hypertension increased with age. In patients over 50 years old, the prevalence of hypertension was significantly higher in female than in male patients (P<0.05). The prevalence of diabetes/impaired glucose tolerance increased with age in male patients but decreased in female patients aged :80 years (P<0.05). Female patients in 60-69.9 and 70-79.9 years groups had significantly higher incidence of diabetes/impaired glucose tolerance than male patients (P<0.05), but not at older ages. The incidence of hyperlipidemia/lipoprotein metabolism disorder increased with age in both male and female patients aged below 80 years (P<0.05), but in 60-69.9 and 70-79.9 year groups, female patients had a significantly higher incidence than male patients (P<0.05); at the ages :80 years, the incidences was lowered in both women and men without a gender-specific differences (P>0.05).

CONCLUSIONSThe data we obtained concerning the prevalence of hypertension and glucose/lipid metabolism disorder, age distribution, and gender characteristics of the COPD patients provide a clear target for secondary prevention of COPD.

Aged ; Aged, 80 and over ; Female ; Glucose Metabolism Disorders ; complications ; Humans ; Hypertension ; complications ; Lipid Metabolism Disorders ; complications ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; complications ; Retrospective Studies

OBJECTIVETo screen the MYBPC3 gene mutations in Han Chinese patients with hypertrophic cardiomyopathy (HCM).

METHODSSixty-six patients with HCM were enrolled for the study. The exons in the functional regions of MYBPC3 were amplified with PCR and the products were sequenced.

RESULTSFour novel mutations and four common polymorphisms were identified in this patient cohort. A Lys301fs mutation in exon10 was evidenced in a H30, and when he was 47 years old, he had the chest tightness, shortness of breath with septal hypertrophy of 18.7mm; a Asp463stop mutation in exon17 was detected in a H48, he was 24 years old 24-year-old when a medical examination showed ventricular septal hypertrophy of 15.4 mm; both Gly523Arg mutation in exon18 and Tyr847His mutation in exon26 were found in a H53 with onset age 36 years old, feeling chest tightness after excise and his ventricular septal hypertrophy was 27 mm that time. MYBPC3 mutations occurred in 4.5% patients in this cohort. These mutations were not found in 100 non-HCM control patients.

CONCLUSIONMYBPC3 mutation is presented in a small portion of Han Chinese patients with HCM.

Adult ; Asian Continental Ancestry Group ; genetics ; Cardiomyopathy, Hypertrophic ; genetics ; Carrier Proteins ; genetics ; DNA Mutational Analysis ; Exons ; Female ; Genotype ; Humans ; Male ; Middle Aged ; Mutation ; Phenotype ; RNA, Messenger ; genetics

OBJECTIVEThe aim of this study was to screen the disease-causing gene mutations and investigate the genotype-phenotype correlation in 10 Chinese pedigrees with familial hypertrophic cardiomyopathy (HCM).

METHODSThere are 91 family members from these 10 pedigrees and 5 members were normal mutated carriers, 23 members were HCM patients (14 male) aged from 1.5 to 73 years old. The functional regions of myosin heavy chain gene (MYH7), cardiac myosin-binding protein C (MYBPC3) and cardiac troponin T gene (TNNT2) were screened with PCR and direct sequencing technique. Clinical information from all patients was also evaluated in regard to the genotype.

RESULTSMutations were found in 5 out of 10 pedigrees. Mutations in MYH7 (Arg663His, Glu924Lys and Ile736Thr) were found in 3 pedigrees and 3 patients from these pedigrees suffered sudden death at age 20-48 years old during sport. Mutations in MYBPC3 were found in 2 pedigrees, 1 with complex mutation (Arg502Trp and splicing mutation IVS27 + 12C > T) and 1 with novel frame shift mutation (Gly347fs) and the latter pedigree has sudden death history. No mutation was identified in TNNT2.

CONCLUSIONSAlthough the Han Chinese is a relatively homogeneous ethnic group, different HCM gene mutations were responsible for familiar HCM suggesting the heterogeneity nature of the disease-causing genes and HCM MYH7 mutations are associated with a higher risk of sudden death in this cohort. Furthermore, identical mutation might result in different phenotypes suggesting that multiple factors might be involved in the pathogenesis of familiar HCM.

Adolescent ; Adult ; Aged ; Asian Continental Ancestry Group ; genetics ; Cardiac Myosins ; genetics ; Cardiomyopathy, Hypertrophic, Familial ; ethnology ; genetics ; Carrier Proteins ; genetics ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Middle Aged ; Mutation ; Myosin Heavy Chains ; genetics ; Pedigree ; Phenotype ; Troponin T ; genetics ; Young Adult

OBJECTIVEProstate cancer (PCa) has the highest incidence among male malignancies in Western industrialized countries and, as a most common malignant disease in urology, its incidence has been increasing in recent years in Chinese men. This study was to investigate the risk loci associated with PCa susceptibility in Han Chinese by analyzing single nucleotide polymorphisms (SNP).

METHODSWe collected peripheral blood samples from 1 667 PCa patients and 1 525 healthy men, and detected 40 loci associated with PCa susceptibility by analyzing SNPs using Sequenom technology.

RESULTSOf the 40 known loci, 16 were confirmed to be significantly associated with PCa susceptibility (P < 0.05). The loci 1, 2 and 5 at 8q24, 10q11 and 22q13.2 also contributed to PCa susceptibility in different ethnic groups.

CONCLUSIONPCa susceptibility is obviously associated with the risk loci rs1465618, rs721048, rs12621278, rs7679673, rs12653946, rs339331, rs1512268, rs10086908, rs16901979, rs1447295, rs10993994, rs10896449, rs902774, rs9600079, rs11649743 and rs5759167 in Chinese Han population.

Aged ; Asian Continental Ancestry Group ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Prostatic Neoplasms ; epidemiology ; genetics ; Risk Factors

Aged ; Blood Glucose ; metabolism ; Diabetes Mellitus ; diagnosis ; Female ; Glucose Tolerance Test ; Humans ; Male ; Middle Aged ; ROC Curve

BACKGROUNDBoth ischemic preconditioning (IPC) and limb remote ischemic postconditioning (LRIPOC) have been shown to possess significantly different cardioprotective effects against the myocardial ischemia reperfusion injury (IRI), but no study has compared the anti-inflammatory effects of IPC and LRIPOC during myocardial IRI process. We hypothesized that IPC and LRIPOC would produce different anti-inflammatory effects in an in vivo rat model with myocardial IRI.

METHODSEighty rats were randomly allocated into four equal groups: sham group, IRI group, IPC group and LRIPOC group. In 10 rats randomly selected from each group, serum levels of TNF-α, HMGB1, ICAM1, IL-1, IL-6 and IL-10 were assessed, and infarct size was determined. In another 10 rats of each group, myocardial levels of TNF-α, HMGB1, ICAM1, IL-1, IL-6 and IL-10 in both ischemic and non-ischemic regions were measured.

RESULTSThe infarct size was significantly lower in IPC and LRIPOC groups than in IRI group. The serum and myocardial levels of pro-inflammatory cytokines including TNF-α, HMGB1, ICAM1, IL-1 and IL-6 during reperfusion were significantly reduced in IPC and LRIPOC groups compared to IRI group. As compared to the IPC group, infarct size, serum level of TNF-α at 60 minutes of reperfusion, serum levels of HMGB1 and ICAM1 at 120 minutes of reperfusion, myocardial levels of TNF-α, ICAM1, IL-1 and IL-6 in the ischemic region, myocardial levels of ICAM1, IL-1 and IL-6 in the non-ischemic region were significantly increased in the LRIPOC group.

CONCLUSIONSIn the rats with myocardial IRI, IPC produces more powerful inhibitory effects on local myocardial and systemic inflammatory responses than LRIPOC. This may be partly attributed to more potent cardioprotection produced by IPC.

Animals ; HMGB1 Protein ; metabolism ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1 ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-6 ; metabolism ; Ischemic Postconditioning ; methods ; Ischemic Preconditioning ; methods ; Male ; Myocardial Reperfusion Injury ; immunology ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism