It was estimated that about 428 species of genus Corydalis are distributed all worldwide, with about 298, especially 10 groups and 219 species being uniquely spread in China. The genus Corydalis have been widely employed as folk medicines in China, especially as traditional Tibetan medicines, for treatment of fever, hepatitis, edema, gastritis, cholecystitis, hypertension and other diseases. The phytochemical studies revealed that isoquinoline alkaloids are its major bioactive ingredients. The extensive biological researches suggested its pharmacological activities and clinic applications against cardiovascular diseases and central nervous system, antibacterial activities, analgesic effects, anti-inflammatory, anti-oxidation and anti-injury for hepatocyte, and so on. As an effort in promoting the research of pharmacodynamic ingredients, this article presents an overview focusing on the distribution, phytochemical and pharmacological results of Corydalis species that have been applied in traditional Tibetan medicinal, hopefully to provide a reference for the new Tibetan medicine development from Corydalis plant resource.
Alkaloids ; chemistry ; pharmacology ; Animals ; Anti-Infective Agents ; chemistry ; pharmacology ; Corydalis ; chemistry ; classification ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Mice ; Molecular Structure ; Phytotherapy

OBJECTIVETo investigate the expression of E-cadherin in nasopharyngeal carcinoma (NPC) and its relationship with cervical lymph node metastasis.

METHODSThe expression of E-cadherin in 80 patients with NPC was detected by immunohistochemistry.

RESULTSLower expression of E-cadherin was associated with advanced N-stage of the tumor (P = 0.018). There was no significant correlation between the expression of E-cadherin and lymph node size (P = 0.435). The expression of E-cadherin was higher in patients with cervical lymph node metastasis limited to a single area than that distributing in some scattered areas (P = 0.000). There was a trend that the expression of E-cadherin in the cases with the tumor and lymph nodes in the same side was higher (56.5%) than that in the patients with bilateral lymph node metastases (32.6%), however, the difference was not significant (P = 0.059). The expression rates of E-cadherin in patients with lymph node metastasis in levels II, III and Va were higher than that in levels I, IV, Vb and VI, but with a non-significant difference (P = 0.059).

CONCLUSIONThe expression of E-cadherin has influence on the lymph node metastasis in nasopharyngeal carcinoma. E-cadherin expression is negatively correlated with the numbers of the lymph node metastases and the metastasis distance, i.e. a lower expression of E-cadherin leads to an advanced N-stage. The lymph node metastasis of nasopharyngeal cancer from above to below is more considerably influenced by E-cadherin expression than the metastasis towards contralateral lymph nodes.

Adolescent ; Adult ; Aged ; Cadherins ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplasm Staging ; Young Adult

Objective To explore the value of single source dual energy CT for quantitative measurement of liver fat fraction in the rabbit model of nonalcoholic fatty liver disease(NAFLD).Methods Thirty male New Zealand rabbits were randomly divided into five groups.Six rabbits were fed with standard chow as a control group for 3 weeks.TwentyGfour rabbits were divided into four groups and fed with highGfat, highGcholesterol diet to reach different stage of NAFLD model for 1 ,3 ,4 and 8 weeks respectively before dualGenergy CT scanning.1 40 keV polychromatic CT values (QC),70 keV monochromatic CT values (Mono 70 keV),slope,effective atomic number (EffectiveGZ)and fat concentration based on dualGenergy CT fat decomposition (Fat/Water)were measured.Liver samples were obtained to measure the fat fraction and staged according to Burnt staging system.Correlations between different CT indexes and fat fraction were analyzed.ROC was used to evaluate the diagnosis efficacy of different parameters.Results Correlation between fat concentration based on dualGenergy CT fat decomposition and fat fraction (r＝0.936)was better than that between 140 keV polychromatic CT values (r＝－0.838)and 70 keV monochromatic CT values (r＝－0.906),as well as effective atomic number (r＝－0.858)and slope (r＝0.863).In terms of diagnostic performance of material decomposition fat imaging,the values of area under the curve were 0.944 (stage 0 vs.stage 1 or more severe),0.995 (stage 1 or less severe vs.stage 2 or more severe)and 1 (stage 2 or less severe vs.stage 3)with optimal cutoff values of 59.310,99.5 17 and 22 3.02 3 mg/cm3 ,respectively.Conclusion The dualGenergy CT can quantitatively measure liver fat concentration as a noninvasive surrogate bioGmarker in the rabbit model of nonalcoholic fatty liver disease.DualGenergy CT derived material decomposition fat images can provide more diagnostic information at the early stage of NAFLD.

Congenital long QT syndrome (LQTS) is a genetically heterogeneous disease in which six ion-channel genes have been identified. The phenotype-genotype relationships of the HERG (human ether-a-go-go-related gene) mutations are not fully understood. The objective of this study is to identify the underlying genetic basis of a Chinese family with LQTS and to characterize the clinical manifestations properties of the mutation. Single strand conformation polymorphism (SSCP) analyses were conducted on DNA fragments amplified by polymerase chain reaction from five LQT-related genes. Aberrant conformers were analyzed by DNA sequencing. A novel splice mutation in C-terminus of HERG was identified in this Chinese LQTS family, leading to the deletion of 11-bp at the acceptor splice site of Exon9 [Exon9 IVS del (-12-->-2)]. The mutation might affect, through deficient splicing, the putative cyclic nucleotide binding domain (CNBD) of the HERG K(+) channel. This mutation resulted in a mildly affected phenotype. Only the proband had a history of syncopes, while the other three individuals with long QT interval had no symptoms. Two other mutation carriers displayed normal phenotype. No sudden death occurred in the family. The 4 affected individuals and the two silent mutation carriers were all heterozygous for the mutation. It is the first splice mutation of HERG reported in Chinese LQTS families. Clinical data suggest that the CNBD mutation may be less malignant than mutations occurring in the pore region and be partially dominant over wild-type function.
Asian Continental Ancestry Group ; DNA Mutational Analysis ; methods ; DNA, Recombinant ; genetics ; ERG1 Potassium Channel ; Ether-A-Go-Go Potassium Channels ; genetics ; Family ; Genetic Predisposition to Disease ; genetics ; Genetic Testing ; methods ; Humans ; Incidence ; Long QT Syndrome ; genetics ; metabolism ; Mutation ; genetics ; Pedigree ; Polymorphism, Genetic ; Risk Assessment ; methods ; Risk Factors

OBJECTIVETo investigate the effect of interleukin-1beta (IL-1beta) on expression and activity of matrix metalloproteinase-2 (MMP-2) of cultured human cardiac fibroblasts and related signaling pathway.

METHODSPrimary human cardiac fibroblasts seeded in 6-well tissue culture plates and cultured to 80% to 90% confluence were harvested at passage 3 to 6 and exposed to IL-1beta at various concentrations for 24 h, culture supernatant and cell protein were obtained. MMP-2 mRNA was determined by RT-PCR. The activity of MMP-2 was analyzed by zymography and the expression of inducible nitric oxide synthase (iNOS) protein level was detected by Western blot analysis. Assessment of NO production in the culture supernatant was performed using the Griess method.

RESULTSIL-1beta (4 ng/ml) significantly increased MMP-2 activity of cultured fibroblasts in a time-dependent manner. MMP-2 mRNA expression was significantly upregulated by IL-1beta (4 ng/ml and 10 ng/ml, all P<0.01). Moreover, IL-1beta also significantly increased NO production in supernatant (P<0.01) and these effects could be significantly blocked by cotreatment with L-NMMA (10(-3) mol/L, all P<0.01). Western blot analysis showed that iNOS could not be detected in unstimulated human cardiac fibroblasts but could be detected in cardiac fibroblasts exposed to IL-1beta.

CONCLUSIONIL-1beta increased MMP-2 activity and transcription of human cardiac fibroblasts via iNOS-NO pathway.

Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Gene Expression Regulation ; Humans ; Interleukin-1beta ; pharmacology ; Matrix Metalloproteinase 2 ; metabolism ; Myocytes, Cardiac ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; RNA, Messenger ; metabolism

The aim of the present study was to investigate whether fluvastatin augments the number of endothelial progenitor cells (EPCs), and promotes EPCs proliferation, migration and adhesion. Total mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation. The cells were then plated on fibronectin-coated culture dishes. After being cultured for 7 d, the attached cells were stimulated with fluvastatin (final concentrations: 0.01, 0.1, 1, 10 micromol/L), simvastatin (1 micromol/L) or a vehicle for the respective time points (6, 12, 24 and 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs were further documented by demonstrating the expression of KDR, VEGFR-2 and AC133 with flow cytometry. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed by MTT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating it on fibronectin-coated dishes, and the adherent cells were then counted. In addition, we also studied EPCs culture assay of peripheral blood from fluvastatin-treated animals in vivo. Incubation of isolated human MNCs with fluvastatin dose- and time-dependently increased the number of EPCs, while reached the maximum 24 h after the administration at 1 micromol/L, (2.5-fold increase, P<0.05). Moreover, treatment of rats with fluvastatins elevated the number of EPCs (3-fold increase, P<0.05), thus extending the in vitro data. In addition, fluvastatin also promoted EPC proliferation, migration, adhesion and in vitro vasculogenesis in a concentration-dependent manner. The effects of fluvastatin on EPCs were compared with those of simvastatin at the same concentration (1 micromol/L), with a result of no statistical difference. The results of the present study define a novel mechanism of the action of statins: the augmentation of EPCs with enhanced functional activity.
Cell Adhesion ; drug effects ; Cell Count ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; Fatty Acids, Monounsaturated ; pharmacology ; Humans ; Indoles ; pharmacology ; Leukocytes, Mononuclear ; cytology ; Simvastatin ; pharmacology ; Stem Cells ; cytology

OBJECTIVEo clone angiotensin-converting enzyme 2(ACE2) gene, to analyze its amino acids and nucleotides sequence and to investigate tissue distribution of ACE2 in adult mice.

METHODSThe full-length ACE2 encoding sequence was amplified from the RNA of mice kidney tissue by RT-PCR technique, cloned into plasmid pGEM-T easy, then subcloned into plasmid pcDNA3.1+. After identification of DNA sequence, the recombinant plasmid pmACE2 was transfected into Cos7 cells with lipofectin reagent. The transient expression of ACE2 molecule was detected by SDS-PAGE. Sequence analysis was conducted with CLUSTALX program. Tissue distribution of ACE2 in mice was detected by RT-PCR.

RESULTSA fragment about 2.6 kb was amplified and the recombinant plasmid pmACE2 was confirmed by two-enzyme digesting and DNA sequencing. The cloned DNA sequence was consistent with that previously reported, except for 3 variations: A701G, T1102C and T1330C. SDS-PAGE proved that expression of a soluble, truncated products form of ACE2 was a glycoprotein of approximately 80 kD in Cos7 cells. The predicted mice ACE2 sequence contained an N-terminal signal sequence (amino acid residues 1-18), a single HHEMGHIQ zinc-binding domain (amino acid residues 373-380) and C-terminal membrane anchor (amino acid residues 738-765). Mice ACE2 showed 84 % identity with that of human, and 90 % identity with that of rat. Expression of ACE2 was the greatest in lungs, hearts and kidneys, and moderate levels were also detected in testes and livers.

CONCLUSIONMice ACE2 gene has been cloned and successfully expressed in vitro. The tissue-specific expression of ACE2 in different species is not identical.

Amino Acid Sequence ; Animals ; Base Sequence ; Carboxypeptidases ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Gene Expression ; Kidney ; metabolism ; Lung ; metabolism ; Male ; Mice ; Molecular Sequence Data ; Myocardium ; metabolism ; Peptidyl-Dipeptidase A ; Sequence Analysis ; Tissue Distribution

BACKGROUNDMutations in the cardiac sodium channel gene (SCN5A) may lead to a broad spectrum of familial arrhythmias, including long QT syndrome (LQTS), idiopathic ventricular fibrillation (IVF), and isolated cardiac conduction diseases. Recent studies have shown that polymorphisms in the SCN5A gene also play an important role in the manifestation of disorders involving cardiac excitability. In this study, we investigated the polymorphisms of the SCN5A gene in Han Chinese and its relation to Brugada syndrome (BS).

METHODSGenomic DNA was isolated from 120 unrelated healthy volunteers and 48 unrelated Brugada syndrome patients by means of standard procedures. All exons including the putative splicing sites of the SCN5A gene were amplified by PCR and sequenced directly or after subcloning using an ABI Prism 377 DNA sequencer.

RESULTSA total of 5 single nucleotide polymorphisms (SNPs) were identified in the Han Chinese population, including 3 novel ones: G87A(A29A), 4245 + 82A > G, and G6174A. The allele frequencies of each SNP in the Han Chinese population were as follows: G87A (A29A) 27.5%, A1673G (H558R) 10.4%, 4245 + 82A > G 32.8%, C5457T (D1819D) 41.3%, and G6174A 44.9%. S1102Y and 10 other SNPs identified in other ethnic populations were not detected in this study. There was no significant difference in the allele frequency of A1673G (H558R) between different ethnic populations (all P > 0.5). On the other hand, the allele frequency of C5457T (D1819D) among Han Chinese was similar to its frequency among Japanese (P > 0.5), but higher than that among Americans (P < 0.005). The allele G1673 (R558) was over-represented in BS patients compared to controls (P < 0.005), but there was no significant difference in genotype frequencies at this locus. There were also no differences in either the allele or genotype frequencies of the 4 other identified SNPs when comparing BS patients with healthy controls.

CONCLUSIONSThe distribution of SCN5A SNPs may vary between different ethnicities. The polymorphism of A1673G might be associated with BS and may contribute to a susceptibility to BS in Han Chinese.

Case-Control Studies ; China ; ethnology ; Gene Frequency ; Humans ; NAV1.5 Voltage-Gated Sodium Channel ; Polymorphism, Single Nucleotide ; Sodium Channels ; genetics ; Syndrome ; Ventricular Fibrillation ; genetics

OBJECTIVETo investigate whether puerarin can augment endothelial progenitor cells (EPCs) numbers, promote EPC proliferation, migration and adhesion.

METHODTotal mononuclear cells (MNCs) were isolated from peripheral blood by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin-coated culture dishes. After 7 days culture, attached cells were stimulated with puerarin (to make a series of final concentrations: 0. 1, 0.5, 1, 3 mmol x L(-1)) or vehicle control for the respective time points (6, 12, 24, 48 h). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation, migration and in vitro vasculogenesis activity were assayed with MT assay, modified Boyden chamber assay and in vitro vasculogenesis kit, respectively. EPCs adhesion assay was performed by replating those on fibronectin-coated dishes, then adherent cells were counted.

RESULTIncubation of isolated human MNCs with puerarin dose increased the number of EPCs, maximum at 3 mmol x L(-1), 24 hours (approximately 1-fold increase, P < 0.01). In addition, puerarin also promoted EPC proliferative, migratory, adhesive and in vitro vasculogenesis capacity.

CONCLUSIONPuerarin can augment the number of EPCs with enhanced functional activity.

Cell Adhesion ; drug effects ; Cell Division ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Humans ; Isoflavones ; isolation & purification ; pharmacology ; Neovascularization, Physiologic ; drug effects ; Plants, Medicinal ; chemistry ; Pueraria ; chemistry ; Stem Cells ; cytology ; drug effects ; Time Factors ; Veins ; cytology

OBJECTIVETo investigate the application of the Chinese Urological Association (CUA) Guidelines on Prostatitis and its effects on the clinical practice patterns of diagnosing and treating chronic pelvic pain syndrome (CPPS) among Chinese urologists and andrologists.

METHODSWe conducted a questionnaire investigation on the application of the CUA Guidelines on Prostatitis among the urologists and andrologists of 173 hospitals in 21 cities of China, and performed statistical analyses on all the eligible questionnaires collected.

RESULTSOf the 1 056 questionnaires distributed, 851 (80.6%) were eligible, of which 71.6% were from the urologists or andrologists in grade 3 hospitals, 80.7% of them with senior or intermediate professional titles and 97.5% had studied the CUA Guidelines. Most of the subjects agreed that Type III prostatitis is a clinical syndrome, whose diagnosis should exclude other conditions with similar symptoms, and whose treatment should aim at relieving pain, alleviating urination symptoms and improving the quality of life. Those who had and those who had not studied the CUA Guidelines differed in their viewpoints on CPPS as illustrated in the book. In clinical practice, the most common treatment options for CPPS were psychological therapy (80.7%), medication (80.4%) and life style adjustment (79.6%), and the most frequently used drugs were phytotherapy (80.0%), alpha-blockers (68.9%) and antibiotics (61.0%).

CONCLUSIONCUA Guidelines on Prostatitis has gained a nationwide application and promoted the standardization of the management of CPPS in China.

Humans ; Male ; Pelvic Pain ; diagnosis ; therapy ; Physicians ; Practice Guidelines as Topic ; Prostatitis ; diagnosis ; therapy ; Surveys and Questionnaires