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Background: Detecting epistatic interactions (EIs) involves the exploration of associations among single nucleotide polymorphisms (SNPs) and complex diseases, which is an important task in genome-wide association studies. The EI detection problem is dependent on epistasis models and corresponding optimization methods. Although various models and methods have been proposed to detect EIs, identifying EIs efficiently and accurately is still a challenge., Results: Here, we propose a linear mixed statistical epistasis model (LMSE) and a spherical evolution approach with a feedback mechanism (named SEEI). The LMSE model expands the existing single epistasis models such as LR-Score, K2-Score, Mutual information, and Gini index. The SEEI includes an adaptive spherical search strategy and population updating strategy, which ensures that the algorithm is not easily trapped in local optima. We analyzed the performances of 8 random disease models, 12 disease models with marginal effects, 30 disease models without marginal effects, and 10 high-order disease models. The 60 simulated disease models and a real breast cancer dataset were used to evaluate eight algorithms (SEEI, EACO, EpiACO, FDHEIW, MP-HS-DHSI, NHSA-DHSC, SNPHarvester, CSE). Three evaluation criteria (pow1, pow2, pow3), a T-test, and a Friedman test were used to compare the performances of these algorithms. The results show that the SEEI algorithm (order 1, averages ranks = 13.125) outperformed the other algorithms in detecting EIs., Conclusions: Here, we propose an LMSE model and an evolutionary computing method (SEEI) to solve the optimization problem of the LMSE model. The proposed method performed better than the other seven algorithms tested in its ability to identify EIs in genome-wide association datasets. We identified new SNP-SNP combinations in the real breast cancer dataset and verified the results. Our findings provide new insights for the diagnosis and treatment of breast cancer., Availability and Implementation: https://github.com/scutdy/SSO/blob/master/SEEI.zip ., (© 2024. The Author(s).)

The genus Neoseiulus in Shanxi Province is reviewed and seven species are recorded from the province. Four of these are redescribed and detailed taxonomic information are provided. Neoseiulus paraki (Ehara) is recorded for the first time in China and Neoseiulus neoreticuloides (Liang and Hu) is considered a new junior synonym of Neoseiulus bicaudus (Wainstein). Additionally, a diagnostic key to the known species of Neoseiulus in Shanxi is provided.

Background: Succinate dehydrogenase deficient gastrointestinal stromal tumors (SDH-deficient GISTs), which lack KIT or PDGFRA mutations demonstrate unique clinical and pathological features, and they respond poorly to standard targeted therapy. We herein present a novel case of SDH-deficient GIST in a three-month-old infant's colon mesentery, and he is the youngest patientto date., Case Presentation: The infantpresented with complaints of blood in the stool. CT showed a 6.3 × 4.6 cm mass in the left lower retroperitoneal. Complete resection of tumor and segmental bowel resection was performed without regional lymphadenectomy. Histologically, tumor cells were distinctive in their multinodular colon wall involvement with interspersed tracts of colon wall smooth muscle. The tumor was composed mainly of epithelioid cells. Immunohistochemically, the tumor cells were positive for Vim, CD117, PDGFR, while negative for SDHB. Mutational analysis showed a synonymous mutation for SDHB and wild-type for KIT and PDGFRA. Two months after surgery, metastases were found and Imatinib was administered. Unfortunately, the disease continued to progress, and the infant died 5 months after surgery., Conclusions: SDH-deficient GISTs comprise a subgroup of a relatively rare tumor type and show a number of clinically and biologically unique features, especially for infants. It is of great importance to developing new therapeutic targets and novel specific drugs.

Background: Hepatocellular carcinoma (HCC) is the sixth most common type of cancer and the fourth leading cause of cancer-related death worldwide. Sarcomatoid HCC, which contains poorly differentiated carcinomatous and sarcomatous components, is a rare histological subtype of HCC that differs from conventional HCC. It is highly aggressive and has a poor prognosis. Its clinicopathological characteristics, surgical outcomes and underlying mechanisms of its highly aggressive nature have not been fully elucidated., Aim: To examine the clinicopathological characteristics and surgical outcomes of sarcomatoid HCC and explore the histogenesis of sarcomatoid HCC., Methods: In total, 196 patients [41 sarcomatoid HCC and 155 high-grade (Edmondson-Steiner grade III or IV) HCC] who underwent surgical resection between 2007 and 2017 were retrospectively reviewed. The characteristics and surgical outcomes of sarcomatoid HCC were compared with those of patients with high-grade HCC. The histological composition of invasive and metastatic sarcomatoid HCCs was evaluated., Results: Sarcomatoid HCC was more frequently diagnosed at an advanced stage with a larger tumor and higher rates of nonspecific symptom, adjacent organ invasion and lymph node metastasis than high-grade HCC (all P < 0.05). Compared with high-grade HCC patients, sarcomatoid HCC patients are less likely to have typical dynamic imaging features of HCC (44.4% vs 72.7%, P = 0.001) and elevated serum alpha-fetoprotein levels (> 20 ng/mL; 36.6% vs 78.7%, P < 0.001). The sarcomatoid group had a significantly shorter median recurrence-free survival (5.6 mo vs 16.4 mo, log-rank P < 0.0001) and overall survival (10.5 mo vs 48.1 mo, log-rank P < 0.0001) than the high-grade group. After controlling for confounding factors, the sarcomatoid subtype was identified as an independent predictor of poor prognosis. Pathological analyses indicated that invasive and metastatic lesions were mainly composed of carcinomatous components., Conclusion: Sarcomatoid HCC was associated with a more advanced stage, atypical dynamic imaging, lower serum alpha-fetoprotein levels and a worse prognosis. The highly aggressive nature of sarcomatoid HCC is perhaps mediated by carcinomatous components., Competing Interests: Conflict-of-interest statement: The authors have no conflict-of interest to disclose., (©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.)

Background: Lymphoma is a common hematological malignancy with many subtypes and considerable heterogeneity. Traditional treatments include chemotherapy, radiotherapy, and surgery. Patients with relapsed, refractory or advanced stage lymphoma have a dismal prognosis. In recent years, chimeric antigen receptors (CARs) have been recognized as powerful tools that redirect antigen-specific T cells independent of human lymphocyte antigen (HLA) restriction and specifically kill tumor cells. Satisfactory results with CAR-based treatments have been achieved in relapsed/refractory B cell leukemia/lymphoma. Our center explored the strategy of subcutaneous injections combined with intravenous drip to overcome certain issues., Case Presentation: A patient with stage IV refractory and relapsed diffuse large B cell lymphoma was treated with regional and intravenous CAR-T cells. During the observation period, the temperature of the skin at the abdominal wall mass was slightly elevated, and tolerable pain in the injection area was reported. Imaging showed regional liquefactive necrosis. After the sequential administration of ibrutinib and venetoclax, the abdominal wall mass significantly decreased in size., Conclusion: The regional injection of CAR-T cells might be safe and feasible for the treatment of regional lesions in patients with refractory and relapsed advanced lymphoma., (Copyright © 2020 Wei, He, Lin, Ren, Zhu, Cheng, Nan, Liu, Yu and Guo.)

Background: Acute coronary syndrome (ACS) is closely related to unstable plaques and secondary thrombosis. The inflammatory cells in plaques and their inflammatory products may be the cause for plaque instability and ruptures. The study aimed to disclose the changes of inflammatory factors including serum intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40), and lipoprotein-associated phospholipase A2 (Lp-PLA2) in patients with ACS and its clinical significance., Methods: A total of 120 patients with coronary heart disease (CHD) were categorized into 2 groups: 69 with ACS and 51 with stable angina pectoris (SAP); 20 patients with chest pain and normal angiography served as a control group. The 120 patients with CHD were categorized into single-vessel disease group, double-vessel disease group, and three-vessel disease group based on the number of coronary artery stenosis. The severity of coronary artery stenosis was quantified based on coronary angiography using Gensini score. They were further divided into mild CHD group with its Gensini score <26 (n = 36), moderate CHD group with its Gensini score being 26-54 (n = 48) and severe CHD group with its Gensini score >54 (n = 36). Serum levels of ICAM-1, YKL-40, and Lp-PLA2 of different groups were determined by enzyme-linked immunosorbent assay. Correlation between ICAM-1, YKL-40, Lp-PLA2, and Gensini score was analyzed., Results: The levels of serum inflammatory factors ICAM-1, YKL-40, and Lp-PLA2 were significantly higher in the ACS group than those in control group and SAP group (all P < 0.05); and compared with control group, no significant difference was observed in terms of the serum ICAM-1, YKL-40, and Lp-PLA2 levels in the SAP group (P > 0.05).The levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not significantly different among control group, single-vessel disease group, double-vessel disease group, and three-vessel disease group (all P > 0.05). The levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not significantly different among control group, mild CHD group (Gensini score <26), moderate CHD group (Gensini score 26-54), and severe CHD group (Gensini score >54) (all P > 0.05). Nonparametric Spearman correlation analysis showed that the levels of serum ICAM-1, YKL-40, and Lp-PLA2 were not correlated with the Gensini score in CHD patients (r = 0.093, r = -0.149, and r = -0.085, all P > 0.05; respectively)., Conclusions: The serum levels of ICAM-1, YKL-40, and Lp-PLA2 were correlated with different clinical types of CHD, but not well correlated the severity and extent of artery stenosis, suggesting that ICAM-1, YKL-40, and Lp-PLA2 might be involved in occurrence of instability of atherosclerotic plaque, and might reflect the severity of CHD mostly through reflecting the plaque stability., Competing Interests: There are no conflicts of interest

Objective: To evaluate whether baseline interleukin-6 (IL-6), interleukin-10 (IL-10) as well as their ratio was associated with overall mortality risk over 7 years of follow-up in 11 communities of Beijing. Methods: Data from a prospective cohort study conducted between 2005 and 2012 in 11 communities of Beijing was analyzed to examine the above associations. Serum IL-6 and IL-10 were analyzed using enzyme-linked immunosorbent assay (ELISA) kits. Follow-up surveys were conducted in 2007, 2010 and 2012 to collect data about participant's survival. Cox regression model was used to estimate the impact of IL-6, IL-10 and their ratio on overall mortality risk. Results: Among 1 539 eligible participants (10 263 total person-years), 77 deaths occurred in 7 years of follow-up. The rates of all-cause death were 4.86, 7.24, and 10.56 per 1 000 person-years ( P =0.009) in the first, second, and third tertile of IL-6, respectively. The corresponding age-sex-adjusted hazard ratios ( HR ) were 1.00, 1.18 (95% CI: 0.64-2.19), and 1.80 (95% CI: 1.01-3.23) and full-adjusted HR were 1.00, 1.17 (95% CI: 0.63-2.19) and 1.87 (95% CI: 1.04-3.36). The corresponding rates of all-cause deaths were not significantly different among three tertiles of IL-10. The age-sex and full-adjusted HR were not significantly different in Cox model. The rates of all-cause death were 4.63, 8.99, and 8.93 per 1 000 person-years ( P =0.043) in the first, second, and third tertile of IL-6/IL-10 ratio, respectively. The corresponding age-sex-adjusted HR were 1.00, 1.67 (95% CI: 0.91-3.06), and 1.98 (95% CI: 1.08-3.64) and full-adjusted HR were 1.00, 1.66 (95% CI: 0.90-3.06), and 2.09 (95% CI: 1.13-3.87). Conclusion: High IL-6 and IL-6/IL-10 ratio may be new risk factors to all-cause death. However, IL-10 is not significantly associated with death.

Objective: To investigate the relationship between inflammatory factors and two Chinese medicine (CM) syndrome types of qi stagnation and blood stasis (QSBS) and qi deficiency and blood stasis (QDBS) in patients with acute coronary syndrome (ACS)., Methods: Sixty subjects with ACS, whose pathogenesis changes belongs to qi disturbance blood stasis syndrome, were divided into 2 groups: 30 in the QSBS group and 30 in the QDBS group. The comparative analysis on them was carried out through comparing general information, coronary angiography and inflammatory factors including intracellular adhesion molecule-1 (ICAM-1), chitinase-3-like protein 1 (YKL-40) and lipoprotein-associated phospholipase A2 (Lp-PLA2)., Results: Compared with the QSBS group, Lp-PLA2 and YKL-40 levels in the QDBS group showed no-significant difference (P>0.05); ICAM-1 was significantly higher in the QDBS group than in the QSBS group in the pathological processes of qi disturbance and blood stasis syndrome of ACS (P<0.05)., Conclusions: Inflammatory factor ICAM-1 may be an objective basis for syndrome typing of QSBS and QDBS, which provides a research direction for standardization research of CM syndrome types.

Recently, studies on the pathogenesis of dilated cardiomyopathy (DCM) have focused on the underlying molecular biology and the association between single nucleotide polymorphisms (SNPs) and disease. This study was designed to explore the association between the rs4641 SNP of the LMNA gene and DCM in order to identify a new gene locus related to DCM. Polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing were employed to detect and genotype rs4641 in 198 patients with DCM and 160 healthy controls. Genotype and allele frequencies were compared to discover their relationship and logistic regression was used to assess the risk of DCM associated with the polymorphic variants. In the DCM group, the frequencies of the TC and TT genotypes and the T allele of rs4641 were remarkably higher than those in the control group (P < 0.01). According to risk analysis, taking the CC genotype as a reference, both the TC and TT genotypes increased the risk of DCM pathogenesis, with OR (95%CI) values of 5.957 (2.903- 12.222) and 6.424 (2.156-19.141), respectively. Taking the C allele as the reference, presence of the T allele was found to increase DCM risk, with OR (95%CI) of 5.295 (3.121-8.983). These results suggested that the C to T mutation at the rs4641 locus of LMNA could enhance the risk of DCM, and that rs4641 represented a genetic susceptibility locus. Therefore, it was concluded that the LMNA rs4641 SNP was associated with DCM risk, which indicated that LMNA is a susceptibility gene for DCM.

This article marks the first report on high-performance liquid chromatography (HPLC) coupled with diode-array detection (DAD) and quadruple time-of-flight mass spectrometry (Q-TOF/MS) for the identification and quantification of main bioactive constituents in Baeckea frutescens. In total, 24 compounds were identified or tentatively characterised based on their retention behaviours, UV profiles and MS fragment information. Furthermore, a validated method with good linearity, sensitivity, precision, stability, repeatability and accuracy was successfully applied for simultaneous determination of five flavonoids and one chromone in different plant parts of B. frutescens collected at different harvest times, and their dynamic contents revealed the appropriate harvest times. The established HPLC-DAD-Q-TOF/MS using multi-bioactive markers was proved to be a validated strategy for the quality evaluation on both raw materials and related products of B. frutescens.

Objective: To examine the relationship of interleukin (IL)-6 and IL-10 genetic variants and cardiovascular factors [oxygenized low density lipoprotein (ox-LDL), lower physical activity, overweight, etc.] with IL-6 and IL-10 secreted by monocytes., Methods: In the study, 40 health persons, aged from 51 to 80 years, without stroke and myocardial infarction, were randomly sampled from a community-based population in Beijing in 2010. Their data on smoking, drinking, blood pressure, fasting glucose, and lipid were collected. The single nucleotide polymorphisms (SNPs) of IL-6 (rs1800796, rs1524107, rs2066992) and IL-10 (rs1800872, rs1554286, rs3021094) were genotyped. The human monocytes were cultivated in RPMI 1640 medium for 24 h; then divided into two equal parts, in which ox-LDL (50 mg/L) and phosphate buffer solution (PBS) were added for another 48 h. Finally, the secretions of IL-6 and IL-10 in the culture supernatants were measured with ELISA., Results: Paired Wilcoxon tests showed that the IL-6, IL-10, and IL-6/IL-10 were significantly higher in ox-LDL medium than in PBS one (all P < 0.01). The concentrations in PBS/ox-LDL taken as repeated measurements, and adjusted for age and gender, the repeated general linear models showed: IL-10 was significantly lower for those overweight (BMI ≥ 26 kg/m(2)) than for those normal weight (P = 0.007), and IL-6/IL-10 was significantly higher in those overweight (P = 0.003). The IL-6/IL-10 was significantly higher in those with lower physical activity [metabolic equivalent of energy, METS < 166 kJ/(kg.d)] than those with higher physical activities (P = 0.046). IL-6 and IL-10 were significantly higher in alcohol drinkers (P = 0.049 and P = 0.006). IL-6 was significantly higher in those with higher high-density lipoprotein-cholesterol (HDL-c, ≥ 56.4 mg/dL, P = 0.027). There were significant interactions between IL-10 SNPs and ox-LDL on IL-10 (all P < 0.05), but no significant interactions between IL-6 gene SNPs and ox-LDL on IL-6., Conclusion: The ox-LDL together with lower physical activity and overweight shifts the balance of pro-inflammatory and anti-inflammatory in the direction of pro-inflammatory. The interaction between IL-10 gene and ox-LDL is intensively correlated with the secretion of the anti-inflammatory cytokine IL-10.

Baeckea frutescens is an aromatic shrub used in South China as an ornamental and as a spice. Four unusual C-methylated biflavonoids named baeckeins F-I (1-4) were isolated from the roots of B. frutescens. The baeckeins F-I possessed a unique carbon skeleton, a flavonol conjugated with a coumaronochromone molecule via the unusual linkages of C-2-C-8 and C-3-O-C-7. Their structures were elucidated by analysis of the 1D ((1)H/(13)C) and 2D NMR (HSQC/HMBC/NOESY) and HR-ESI-MS spectroscopic data, and the absolute stereochemistry for chiral carbons of C-2 and C-3 was established by CD spectrometry combined with quantum chemical calculations. Baeckeins F-I (1-4) were also evaluated for their anti-inflammatory activities by detecting the NO production of LPS-induced RAW264.7 murine macrophage cell line; baeckein I (4) with the β-d-glucose unit and configuration of (2R,3R) exhibited the highest NO inhibitory activity (IC50=15.2 μM), which was similar to that of the positive control indomethacin., (Crown Copyright © 2014. Published by Elsevier Ltd. All rights reserved.)

Angelica polymorpha Maxim. is a plant of the Angelica genus (Umbelliferae). The root and stem of this plant is a folk medicine known to have the actions of relieving rheumatism and cold and subsiding swelling and pains. To investigate the chemical constituents in the root of A. polymorpha Maxim., seven compounds were isolated from an 80% ethanol extract by column chromatography. Their structures were elucidated according to the spectroscopic analysis. Compound 1 is a new sesquiterpene, named as bisabolactone. Its absolute configuration was determined by 1D NOESY and CD analysis. The others were identified as 5-hydroxymethylfurfural (2), hycandinic acid ester 1 (3), ferulic acid (4), isooxypeucedanin (5), noreugenin (6) and cimifugin (7). Compound 2 and 3 were isolated from this genus for the first time and compound 4 was isolated from this plant for the first time.

A phytochemical study of Baeckea frutescens afforded a new biflavonoid named as baeckein E (1), along with four known C-methylated flavonols, baeckeins A (2) and B (3), 6-methylquercetin (6) and 6-methylquercetin-4'-O-β-D-glucopyranoside (7), and two known biflavonoids baeckeins C (4) and D (5). The structure of compound 1 was elucidated on the basis of spectroscopic analysis, including HR-ESI-MS, 1D and 2D NMR spectra, and the absolute configurations of C-2 and C-3 for compound 1 were assigned by circular dichroism spectrometry combined with quantum chemical calculations. The antioxidant and cytoprotective activities of compounds 1-7 were also investigated.

Objective: To evaluate the prediction value of blood lipid levels on the newly-identified carotid plaque in middle-aged and elderly Chinese population., Methods: All study subjects were recruited from two cohorts from 2002 to 2007 [the People's Republic of China/United States of America collaborative study (USA-PRC study) and the Chinese multi-provincial cohort study (CMCS)]. The baseline examination including cardiovascular disease risk factors and B-mode ultrasound of carotid artery was performed in 2002 and the second examination was made in September to October, 2007. The relationship between baseline lipids and carotid plaque incidence was analyzed in a total of 2000 subjects aged 47 to 79 years (mean 63 years)., Results: (1) During these 5 years, the prevalence of carotid plaque increased from 30.3% to 62.2% and from 21.5% to 51.5% for men and women, respectively. The newly-identified carotid plaque incidence was 41.8% for men and 34.1% for women. (2) The incidence of artery plaque significantly increased in both sexes in proportion to increase of baseline total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), non-high-density lipoprotein cholesterol (non-HDL-C), and total to high-density cholesterol ratio (TC/HDL-C) levels (P < 0.05 or P < 0.01). (3) Cross-stratification analysis of LDL-C, triglyceride (TG) and HDL-C for carotid plaque incidence indicated the existence of conjoint effects between LDL-C and HDL-C, LDL and TG, as well as between TG and HDL-C, on the increased incidence of carotid plaque. (4) Multi-factorial analysis showed that higher LDL-C, non-HDL-C and TC/HDL-C were independent risk factors for development of new carotid plaque [OR = 1.44 (95%CI = 1.07 - 1.94), OR = 1.45 (95% CI = 1.08 - 1.96), OR = 1.59(95% CI = 1.14 - 2.23) in men;OR = 1.47 (95% CI = 1.13 - 1.92), OR = 1.35 (95% CI = 1.04 - 1.75), OR = 1.64 (95% CI = 1.20 - 2.23) in women]., Conclusions: The prevalence of carotid plaque increased rapidly in this cohort between 2002 and 2007. Elevated LDL-C, non-HDL-C and TC/HDL-C levels were independent predictors of newly developed carotid plaque.

The objective of this study was to investigate the molecular basis of the inferior nodal extension (INE) in the atrioventricular junctional area that accounts for arrhythmias. The INE was separated from the adult rat heart by laser capture microdissection. The mRNA expression of ion channels was detected by quantitative real-time PCR. Hierarchical clustering was used to demonstrate clustering of expression of genes in sections. The mRNA expression of HCN4, Ca(v)3.1 and Ca(v)3.2 was high in the INE, atrioventricular node and sino-atrial node, and that of Ca(v)3.2 high in Purkinje fibres. Although the expression of HCN1 and Ca(v)1.3 was low in the rat heart, it was relatively higher in the INE, atrioventricular node and sino-atrial node than in right atrial and right ventricular (working) myocytes. Both HCN2 and Ca(v)1.2 were expressed at higher levels in working myocytes than in nodal tissues and in the INE. Hierarchical clustering analysis demonstrated that the expression of the HCN and calcium channels in INE was similar to that in the slow-response automatic cells and different from that in working myocytes and Purkinje fibres. The expression of HCN and calcium channels in the INE of the adult rat heart is similar to that of slow-response automatic cells and provides a substrate for automatic phase 4 depolarization in cells.

To investigate the chemical constituents of hemostatic extract of Pyrola calliantha H. Andres, the extract was subjected to chromatographic separation and purification. Along with some known compounds, a new naphthaquinone derivative was isolated and identified as 2-(1, 4-dihydro-2, 6-dimethyl-1, 4-dioxo-3-naphthalenyl)-3, 4, 5-trihydroxylbenzoic acid by physicochemical and spectroscopic analysis.

Objective: To describe the distribution characteristics of carotid atherosclerosis and to analyze its association with cardiovascular risk factors in middle-aged and elderly Chinese., Methods: In 2007, the second cross-sectional examination for cardiovascular risk factors and B-mode ultrasound imaging on carotid arteries were performed in a Chinese population consisting of 2681 subjects aged 43-81 years old from two cohorts participants of the People's Republic of China/United States of America collaborative study (USA-PRC study) and the Chinese multi-provincial cohort study (CMCS). The association of cardiovascular risk factors with intima-media thickening (ITM) and plaque of carotid arteries was analyzed., Results: (1) The mean intima-media thickening (Mean-IMT) of carotid artery was 0.68 mm, the maximal intima-media thickening (Max-IMT) was 1.07 mm. (2) The prevalence rate of carotid plaque was 60.3% in all subjects. As for the different sites of carotid arteries, the plaque prevalence rate at carotid bulb was 61.2% for male and 51.6% for female (chi2 = 23.44, P < 0.01), and at the internal carotid artery was 24.7% for male and 12.2% for female (chi2 = 69.57, P < 0.01), at the common carotid artery was 20.9% for male and 13.8% for female (chi2 = 23.18, P < 0.01). (3) After adjusting for age and other cardiovascular risk factors, the Mean-IMT and Max-IMT increased with elevated levels of systolic blood pressure, plasma glucose and low density lipoproteins cholesterol (LDL-C) (all P < 0.05). Hypertension, diabetes, smoking and high LDL-C (P < 0.05) were independent predict factors for the risk of carotid plaque., Conclusion: The carotid atherosclerosis was common in middle and older age Chinese and was positively associated with classical cardiovascular disease risk factors.

A simple, four-step synthesis of D-lividosamine starting from N-acetyl-D-glucosamine via a furanosyl oxazoline intermediate is described., (2009 Elsevier Ltd. All rights reserved.)

Objective: To explore the electrophysiologic characteristics of Ito in myocytes from cardiac Koch triangle., Methods: Patch clamp technique was employed to investigate the I-V and D-V relation, steady-state activation and inactivation kinetics of Ito from myocytes in Koch triangle of rabbit hearts., Results: (1) The maximum peak current (pA) and peak current density (pA/pF) at +20 mV in PC (pacing cell), TC (transitonal cell) alpha,beta, AC (atrial cell) and PL (purkinje-like cell) cells were different from each other (P < 0. 05); The cells also had different current density (P < 0.05) except between TCalpha, TCbeta and PL cells (P > 0.05). (2) The half activation potential (V(mIto1/2), mV) of steady state activation among PC, TCalpha, TCbeta, AC and PL were significantly different (P < 0.05); The paired comparison between TCalpha and TCbeta, AC and PL, and TCbeta and PC showed significant difference (P < 0.05); but the differences between TCa and PC, TCbeta and AC, and TCbeta and PL were not significant (P > 0.05). (3) The half inactivation potential (V5(hIto1/2), mV) of steady state inactivation between TCalpha and PC, TCa and PL, TCbeta and PC, and TCbeta and PL demonstrated significant differences(P < 0.05). The differences of K(hIto) between TCalpha and TCbeta, PC and PL, TCbeta and PC, and AC and PL were significant (P < 0.05)., Conclusions: Ito of cardiac cells from Koch triangle in rabbit hearts are heterogeneous, but relatively specified and distributed in different groups.

Background: Few studies have explored the inward sodium current (INa) kinetics of transitional cardiomyocytes. This study aimed to explore the kinetics of transitional cardiomyocytes types alpha and beta., Methods: The whole-cell patch clamp technique was used to study the rapid INa of isolated transitional cardiomyocytes in the Koch triangle of rabbit hearts., Results: Maximal amplitude and density of INa in type alpha and type beta was (-1627 +/- 288) pA (alpha), (-35.17 +/- 6.56) pA/pF (beta) and (-3845 +/- 467) pA (alpha), (-65.64 +/- 10.23) pA/pF (beta) (P < 0.05). Steady state activation curves of INa, fitted to a Boltzmann distribution for both types, were sigmoid in shape. Half activation voltage and slope factors did not significantly differ between types at (-43.46 +/- 0.85) mV (alpha), (-41.39 +/- 0.47) mV (beta) or (9.04 +/- 0.66) mV (alpha), (11.08 +/- 0.89) mV (beta). Steady state inactivation curves of INa, fitted to a Boltzmann distribution in both types were inverse "S" shape. Half inactivation voltage and slope factors were (-109.9 +/- 0.62) mV (alpha), (-107.5 +/- 0.49) mV (beta) and (11.78 +/- 0.36) mV (alpha), (11.57 +/- 0.27) mV(beta), (P > 0.05), but time constants of inactivation were significantly different at (1.10 +/- 0.19) mV (alpha) and (2.37 +/- 0.33) ms (beta), (P < 0.05). Time constants of recovery from inactivation of INa for both types were (122.16 +/- 27.43) mV (alpha) and (103.84 +/- 28.97) ms (beta) (P < 0.05)., Conclusions: Transitional cardiomyocytes in rabbit hearts show a heterogeneous, voltage gated and time dependent fast inward sodium current. Types alpha and beta show the features of INa similar to those in slow- and fast-response myocytes, with probably better automaticity and conductivity, respectively.

Objective: To quantitatively detect the expression of connexins (Cx) mRNA in the posterior nodal extension (PNE) of adult rat heart and understand the relationship between Cx expression and atrial ventricular nodal reentrant tachycardia (AVNRT)., Methods: PNE was separated from adult rat heart by means of laser microdissection (LCM), and the cells were also isolated from the atrioventricular node (AVN), sinoatrial node (SAN), Purkinje fiber (PF), right atrium (RA) and right ventricle (RV), to serve as the controls. The Cx mRNA level was detected in these cells with quantitative real-time PCR (QRT-PCR)., Results: The cells were successfully isolated from the PNE and other regions of adult rat heart, where heterogeneous expression of the 3 Cx isoforms (Cx43, Cx45, and Cx40) were observed. Cx45 mRNA showed higher expression in the PNE than in the working myocardium, whereas Cx43 mRNA level was about 25 times higher (P<0.05) in the working myocardium and 18 times higher (P<0.05) in the PF than in the PNE. In the PF, Cx40 mRNA level was proximately 6.8 times (P<0.01) as much as that in the PNE. Cx expression in the PNE was, however, similar to that in the SAN and AVN., Conclusion: Cx mRNAs exhibit heterogeneous expression in the PNE to allow the formation of the slow pathway. In addition, Cx expression in the PNE is very different from that in the adjacent myocardium, resulting in conduction discontinuity at the cellular junction, where, on certain occasion, unidirectional block may occur to cause AVNRT.

Background: The morphological and electrophysiological characteristics of cardiac cells in Koch triangle are still disputed. We studied the appearance and electrical properties of these diverse myocytes to elucidate their complex electrophysiological phenomena., Methods: Experiments were conducted using cooled charge coupling device (CCD) system and whole cell, patch clamp technique to determine the morphology, action potential and sodium current density of single viable myocytes enzymatically isolated from the Koch triangle of rabbit hearts., Results: Morphologically, cardiac cells in shape of spider, tiny spindle, slender spindle, rod and strip were observed in percentage of 3.0 +/- 0.3, 35.0 +/- 5.0, 15.0 +/- 2.0, 40.0 +/- 5.0 and 6.0 +/- 0.7 respectively. The cellular dimensions and capacitance gradually increased in the above order (all P < 0.05). Electrophysiologically, action potential configurations recorded from them were similar respectively to nodal (N), atrial nodal (AN), nodal Hisian (NH), atrial (A) and Hisian like potentials obtained from the intact atrioventricular nodal preparations. Diastolic depolarization appeared in all myocytes except for rod cells. Sodium current density increased in the order of tiny spindle, strip, rod, slender spindle cell (all P < 0.05), but could not be detected in spider-shaped cells. Linear regression analysis revealed that membrane capacitance was correlated negatively to the rate of diastolic depolarization r = -0.70, P < 0.001, but positively to maximum depolarization potential, amplitude of action potential, upstroke velocity and maximum peak value of sodium current density r = 0.84, 0.80, 0.87 and 0.75, respectively; all P < 0.001., Conclusions: The results demonstrated that spider-shaped, spindle, rod and strip cells in Koch triangle might correspond to pacemaking, transitional, atrial and Purkinje like cells, respectively. Furthermore, tiny spindle and slender spindle cells were referred to transitional cell alpha (TCalpha) and beta (TCbeta) accordingly considering their distinctive electrical properties. Different myocytes with diverse electrical properties constituted the infrastructure of sophisticated electrophysiological phenomena in Koch triangle. In view of the prominent percentage and electrical properties, tiny spindle and slender spindle cells were presumed to play important roles.

Objective: To use the laser capture microdissection (LCM) to isolate the pure atrial cells from heart. To extract tiny amount RNA and amplify the genes of beta-actin and GAPDH., Methods: First, the prepared sample RNA was extracted and verified. Second, the frozen sections were scraped after haematoxylin-eosin staining, and then the RNA of scraped tissues was extracted and verified. Last, the tiny amount RNA was extracted from atrial cells captured by LCM, and the genes of beta-actin and GAPDH were amplified by reverse transcriptase polymerase chain reaction (RT-PCR)., Results: The atrial cells isolated with LCM had clear morphology after quick staining. The high quality RNA was extracted from LCM sample and scraped tissues, of which A260/A280 was 1.9-2.0. And 18S rRNA and 28S rRNA were seen distinctly on the gel of RNA formaldehyde denaturing gel electrophoresis, which indicated that RNA didn't degrade before cells captured. Too small amount of RNA extracted from captured cells was below the detectable limit of gel electrophoresis or ultraviolet spectrophotometer. Bands of 300 bp and 357 bp could be amplified by RT-PCR from tiny amount RNA., Conclusion: Highly pure atrial cells could be obtained by LCM. Tiny amount RNA of captured atrial cells could be extracted and amplified with genes of beta-actin and GAPDH.

Objective: To investigate the expression and distribution of L-type calcium channel alpha1C subunits in adult rat heart., Methods: HE staining was applied on the frozen sections of adult rat heart to identify the sinoatrial node (SAN), atrioventricular node (AVN), and posterior nodal extension (PNE). The protein expression of L-type calcium channel alpha1C in adult rat heart and its cellular localization were examined by Western blotting and immunohistochemistry, respectively., Results: L-type calcium channel alpha1C subunit was immunolocalized on the membrane of the myocardial cells, and its expression increased gradually in the SAN, AVN, PNE, right atrium and right ventricle. The protein level of L-type calcium channel alpha1C in the AVN was similar to that in the PNE (P>0.05), and its level in the right atrium and ventricle were significantly higher than those in the SAN and AVN (P<0.01). The results of Western blotting were well consistent with the results of immunohistochemistry., Conclusion: The expression of L-type calcium channel alpha1C subunit may play a role in the electrophysiological functions of the heart.

We studied the effects of Chinese traditional medicine rhynchophylline (Rhy) on human ether-a-go-go related gene (HERG) channel and characterized the electrophysiological properties of Rhy's pharmacological effect on HERG channel using Xenopus oocytes. Xenopus oocytes were injected with either 23 nl (5.75 ng) HERG cRNA or 23 nl distilled water. Xenopus oocytes were randomly assigned to receive one of the following different concentrations of Rhy: (1) control, (2)10 mumol/L Rhy, (3)100 mumol/L Rhy, (4) 500 mumol/L Rhy, (5) 1 000 mumol/L Rhy, (6) 10 000 mumol/L Rhy. Cell currents were recorded in oocytes. The peak tail currents of HERG channel were inhibited by Rhy. The inhibition was in a dose-dependent manner [IC(50)=(773.4 +/- 42.5) mumol/L]. Experiment with 100 mumol/L Rhy indicated that the degree of HERG blockade showed some voltage dependence (within -40 mV to -20 mV ). Kinetic analyses revealed that Rhy decreased the rate of channel activation. The findings indicate that Rhy inhibits HERG encoded potassium channels. It may underline the molecular mechanism of myocardial electrophysiological characteristics associated with this drug.

Objective: To develop a general quality of life (QOL) instrument for Chinese in accordance with the Chinese culture and to assess its reliability, validity and sensitivity., Methods: A 35-item QOL questionnaire(QOL-35) was developed with reference to the World Health Organization QOL questionnaire(WHO-100) and the 36-item medical outcomes study on short-form health status(SF-36). Thirty five items were divided into six domains (general, physical, independent, psychological, social, environment) and one item on QOL transition. The reliability of QOL-35 was assessed by a test-retest survey among 127 adults with an interval of 24-72 hours. The internal consistency and validity were evaluated by a survey on 135 adults from outpatients or general population, using QOL-35, WHO-100 and SF-36. The adaptability was assessed by application to 1356 community-based samples in Beijing., Results: (1)Test-retest reliability of QOL-35: weighted Kappa indexes for items were from 0.86 to 1.00. Intraclass correlation coefficients were from 0.68 to 0.94 for domains, and 0.94 for total score. (2) On internal consistency: Cronbach's Alphas were 0.93, 0.97 and 0.89 for QO1-35, WHO-100 and SF-36. (3)On construct validity. The accumulated proportions of variances of the preceding seven factors were 66.5%, 50.3% and 65.3% for QOL-35, WHO-100 and SF-36. (4) On criterion validity. Spearman correlation coefficients of total QOL score of QOL-35 with those of WHO-100 and SF-36 were 0.805 and 0.745. (5)The rates of chronic diseases were 53.1%, 33.1%, 26.4% and 25.1% from first to fourth quantile of the total QOL scores of QOL-35(P<0.05). (6)Cronbach's Alpha was from 0.68 to 0.93 in 135 subjects, and from 0.71 to 0.91 in 1356 individuals of natural population., Conclusion: The QOL-35 instrument satisfied test-retest reliability and was highly correlated with WHO-100 and SF-36, having fewer items but better construction validity, better internal consistency, and better discrimination ability. We suggested that QOL-35 be used as a replicable tool to assess quality of life in the Chinese general population.

Objective: To investigate the molecular pathology in families with long QT syndrome (LQTS) including Jervell-Longe-Nielsen syndrome (JLNS) and Romano-ward syndrome (RWS) and Brugada syndrome (BS) in Chinese population., Methods: Polymerase chain reaction and DNA sequencing were used to screen for KCNQ1, KCNH2, KCNE1, and SCN5A mutation., Results: We identified a novel mutation N1774S in the SCN5A gene of the BS family, a novel mutation G314S in a RWS family which had also been found in Europe, North America, and Japan, and a single nucleotide polymorphisms (SNPs) G643S in the KCNQ1 of the JLNS family. In this JLNS family, another heterozygous novel mutation in exon 2a was found in KCNQ1 of the patients., Conclusion: New mutations were found in our experiment, which expand the spectrum of KCNQ1 and SCN5A mutations that cause LQTS and BS.

Objective: To identify the mutation of a Chinese family with inherited long QT syndrome(LQTS)., Methods: The disease-causing gene was tentatively determined in light of the clinical manifestations and electrophysiological properties, and then polymerase chain reaction and DNA sequencing were used for screening and identifying mutation., Results: A missense mutation G940A(G314S) in the KCNQ1 gene was identified, which was the 'hot spot' of long QT syndrome mutation., Conclusion: The mutation that is involved with long QT syndrome in Chinese patients is the same as that in the European, American and Japanese patients.

To study PCR site-directed mutagenesis of long QT syndrome KCNQ1 gene in vitro. The site-directed mutagenesis of LQTS gene KCNQ1 was made by PCR. Two sets of primers were designed according to the sequence of KCNQ1 cDNA, and mismatch was introduced into primers. Mutagenesis was performed in a three-step PCR. The amplified fragments from the third PCR which contained the mutation site were subcloned into the T-vecor PCR2.1. Then the fragments containing the mutation site was obtained from PCR2.1 with restriction enzyme digestion and was inserted into the same restriction site of pIRES2-EGFP-KCNQ1. With Effectene Transfection Reagent, pIRES(2)-EGFP-KCNQ1 was transfected into HEK293 cell. The sequencing analysis showed that the mutation site was correct. Mutation from T to C in 934 site of KCNQ1 cDNA was found. Under the fluorescence microscope, the green fluorescence was spread in the transfected HEK293 cell, meaning the pIRES(2)-EGFP-KCNQ1 containing the mutation site was expressed correctly.

Objective: To establish a RP-HPLC method for determination of glycosides in Traditional Chinese Medicine Liu-wei Di-huang., Method: The samples were analyzed on an ODS column at 30 degrees C, with mobile phase of methanol/water (33:67) at flow rate 1.0 mL.min and detection at wavelength of 236 nm., Result: Three major components reached base-line separation and were identified to be mononiside, loganin, paeoniflorin. Respectively for the three components, linear correlations were found between peak areas and concentrations in the ranges of 7.4-60, 7.7-62 mg.L-1 and 8.5-68 mg.L-1, and the recoveries were 98.8%, 98.3%, 99.6%., Conclusion: The established method is proved to be suitable for simultaneous quantification of three major glycosidic components in Liuwei Dihuang decoction and can be used for evaluation of the quality of Liuwei Dihuang preparations.