The HPLC-MS/MS method was developed to profile the dynamics of abscisic acid (ABA) and ABA-glucose ester (ABA-GE) after cloning glycosyltransferase enzyme family gene AtUGT71C5 into Arabidopsis thaliana. By constructing over-expression lines (OE) and down-expression lines (DN), we acquired mutant strains to analyze the function of AtUGT71C5. The multiple-reaction monitoring (MRM) was used for quantitative determination in negative mode. The transition was m/z 263.1-153.0 for ABA ([M-H]t), m/z 425.1-263.0 for ABA-GE ([M-H]t), and m/z 321.0-152.0 for chloramphe-nicol. The linear range was 0.8684-217.1 ng/mL for ABA and 0.3920-196.0 ng/mL for ABA-GE. The accuracy was 88.0-109.0% for ABA and 86.6-113.0% for ABA-GE; the inter-day and intra-day precisions were less than 5.4%for ABA and 8.9%for ABA-GE, respectively. This method is simple and sensitive enough for determination of ABA and ABA-GE in A. thaliana leaves. All the evidence confirmed the speculation that AtUGT71C5 can mediate abscisic acid homeostasis.

OBJECTIVEStatins are still underused for the prevention of cardiovascular disease (CVD) in China. Hence, we conducted a systemic review on the pharmacology, clinical efficacy, and adverse events of atorvastatin, as well as on patient adherence.

DATA SOURCESWe conducted a systemic search in PubMed with the following keywords: "atorvastatin" (Supplementary concept) or "atorvastatin" (All field) and ("China" [AD] or "China" [all field] or "Chinese" [All field]).

STUDY SELECTIONClinical or basic research articles on atorvastatin were included.

RESULTSAtorvastatin is a reversible and competitive inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A reductase, decreasing the de novo cholesterol synthesis. The pharmacokinetics of atorvastatin among Chinese is similar to those in Caucasians, and several gene polymorphisms have proved to be associated with the metabolism of atorvastatin in the Chinese population. Several international multiple-center randomized control trials have demonstrated the benefit of atorvastatin for primary and secondary prevention of CVD. None of them, however, included the Chinese, and current evidence in the population is still inadequate, due to the small sample size, low study quality, short study duration, and the use of surrogate endpoints instead of clinical endpoints. The overall incidence of adverse events observed with atorvastatin did not increase in the 10-80 mg dose range, and was similar to that observed with placebo and in patients treated with other statins, which makes atorvastatin well-tolerated in the Chinese population. Moreover, high patient adherence was observed in clinical studies.

CONCLUSIONSBased on the current available evidence, there is no significant difference between Chinese and non-Chinese population in term of pharmacology and clinical efficacy/safety. High-quality evidence is still needed to support the use of atorvastatin in high-risk Chinese population.

Atorvastatin Calcium ; Cardiovascular Diseases ; drug therapy ; China ; Heptanoic Acids ; therapeutic use ; Humans ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; therapeutic use ; Pyrroles ; therapeutic use

OBJECTIVETo assess the association between single nucleotide polymorphisms (SNPs) of melanocortin-1 receptor gene (MC1R) and freckles in Chinese Han population from Chengdu.

METHODSTwenty randomly selected samples were used to select SNPs of the MC1R gene through DNA sequencing. Pyrosequencing in combination with DNA pooling technique was used to assess allelic frequencies of the selected SNPs in 111 individuals with freckles and 124 normal controls. Representative SNPs were selected based on their functional implications and minimum allele frequency (MAF> 0.05). Genotype of the SNPs were determined with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or pyrosequencing.

RESULTSBased on results of DNA sequencing and pyrosequencing, 4 SNPs (rs2228479, rs885479, rs33932559 and rs2228478) were selected to determine the genotype for each sample. Comparison of genotypic and allelic frequencies of the 4 SNPs with χ (2) test has found no significant difference between the two groups (P> 0.05). For rs33932559, the frequencies of T allele were respectively 90.09% and 91.94% for individuals with freckles and normal controls. For rs2228479 and rs2228478, the frequencies of G and A allele were both about 77%. For rs885479, the frequency of T allele was about 60%. None of the above 3 SNPs showed a significant difference between the two groups in terms of allelic or genotypic frequencies.

CONCLUSIONNo association between the selected SNPs of MC1R gene has been found with development of freckles for the selected Chinese Han population from Chengdu.

Adult ; Alleles ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; China ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Melanosis ; genetics ; Middle Aged ; Polymorphism, Single Nucleotide ; Receptor, Melanocortin, Type 1 ; genetics ; Young Adult

OBJECTIVETo study the changes of the contralateral testicular histology and germ cell apoptosis after unilateral testicular torsion (UTT) and to determine whether the contralateral testis is injured or not.

METHODSSixty SD male rats were divided into control group (12 rats) and experimental group(48 rats). The former underwent sham operation of the left testis under general anaesthesia. The latter underwent left testis torsion(720 degrees) for 6 h, and then 4 of them were sacrificed and the other 44 were subdivided into the torsed testis untwisted group (22 rats) and the torsed testis removal group (22 rats), 7-8 rats were sacrificed and both testes (twisted and untwisted) were removed 1 day, 1 week and 4 weeks after surgery. All testes underwent histological and germ cell apoptosis examination.

RESULTSThere were significant histological changes in the contralateral testis, and the germ cell apoptosis was increased greatly in the contralateral testis.

CONCLUSIONSUTT can cause contralateral testicular injury, whose mechanism may be related to reperfusion, and torsed testis removal can prevent or reduce damage to the contralateral testis.

Animals ; Apoptosis ; Germ Cells ; pathology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; pathology ; Spermatic Cord Torsion ; pathology ; Testis ; pathology

This study was aimed to investigate the effects of sodium valproate (VPA) on the proliferation and regulation of histone acetylation of multiple myeloma cell line U266. U266 cells were treated with VPA. Cell proliferation was determined by CCK-8 assay, and cell cycle were analyzed by flow cytometry (FCM). The expression level of HDAC1 mRNA was detected by RT-PCR, and the protein levels of HDAC1 and histone H3, H4 acetylation was detected by Western blot. The results showed that the VPA inhibited the proliferation of U266 cells in concentration-and time-dependent manners.After exposure to different concentrations of VPA for 48 hours, the proportion of G(0)/G(1) cells increased, while the proportion of S phase cells decreased. The cell cycle was arrested obviously in G(0)/G(1) phase (p < 0.05). The expression of HDAC1 mRNA was inhibited, and the protein level of HDAC1 was down-regulated, while the histone H3/H4 acetylation was up-regulated in U266 cells. It is concluded that the VPA can inhibit cell proliferation of U266 and induce G(0)/G(1) phase arrest. The increase of histone H3/H4 acetylation resulting from inhibiting HDAC1 by VPA might be considered as a possible mechanism.
Acetylation ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Histone Deacetylase 1 ; metabolism ; Histone Deacetylase Inhibitors ; pharmacology ; Histones ; metabolism ; Humans ; Multiple Myeloma ; metabolism ; Valproic Acid ; pharmacology

BACKGROUNDTrichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dermatophytosis caused by T. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin ®, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection.

METHODSThe reconstructed human epidermis (RHE) was infected by inoculating low-dose (400 conidia) and high-dose (4000 conidia) T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy (SEM) observation.

RESULTSThe histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of T. rubrum infection in an intuitionistic way.

CONCLUSIONSWe established the T. rubrum infection model on RHE in vitro successfully. It is a promising model for further investigation of the mechanisms involved in T. rubrum infection.

Animals ; Disease Models, Animal ; Epidermis ; microbiology ; Humans ; Keratinocytes ; cytology ; Tissue Culture Techniques ; Trichophyton ; pathogenicity

OBJECTIVETo analyze the cardiac manifestations of patients with primary and secondary Sjogren syndrome.

METHODSClinical data (clinical manifestations, serologic measurements, echocardiogram) of 396 patients with Sjogren syndrome who admitted to our hospital from 2004--2007 were retrospectively analyzed. Patients with congenital, rheumatic and coronary heart diseases, hypertension and diabetes (n = 221) and patients with incomplete clinic data (n = 51) were excluded.

RESULTSA total of 124 cases were included in this analysis (mean age 47.4 years old; 5 males; average disease duration 85.5 months). Cardiac involvement in Sjogren syndrome is usually asymptomatic. Pericardial effusion (PE) were evidenced in 20.2%, left ventricular diastolic dysfunction (LVDD) in 13.7%, pulmonary artery hypertension (PAH) in 12.9%, left atrium enlargement/in 7.3%, mitral insufficiency in 4.8%, aortic dilation in 5.6%, tricuspid insufficiency in 3.2%, left ventricular enlargement in 2.4% and left ventricular systolic dysfunction in 0.8% patients by echocardiography examinations. Patients with PE had significantly lower CH50, C3, C4 levels and significantly higher C reactive protein level (CRP) and SSA positive rate than patients without PE (all P < 0.05). The serum level of CRP was significantly associated with PE (OR 0.976, 95% CI 0.956 - 0.997, P < 0.05). Age is positively correlated to LVDD (OR 0.884, 95% CI 0.811 - 0.964, P < 0.005). The gammaglobulin level is significantly higher in the PAH group than that in the non-PAH group (P < 0.05).

CONCLUSIONSCardiac involvement is not rare in patients with Sjogren syndrome. PE, LVDD and PAH are usual cardiac manifestations in these patients. The serum level of CRP is positively related to PE in these patients with Sjogren syndrome.

Adult ; Echocardiography, Doppler, Color ; Female ; Humans ; Hypertension, Pulmonary ; diagnostic imaging ; etiology ; Male ; Middle Aged ; Pericardial Effusion ; diagnostic imaging ; etiology ; Retrospective Studies ; Sjogren's Syndrome ; complications ; diagnostic imaging ; Ventricular Dysfunction, Left ; diagnostic imaging ; etiology

BACKGROUNDPrevious studies reported interleukin-27 (IL-27), interferon-γ (IFN-γ), or adenosine deaminase (ADA) alone plays a helpful role in diagnosing tuberculous pleural effusion (TPE). The present study aims at comparing the diagnostic accuracy of pleural IL-27, IFN-γ, and ADA, and investigate the diagnostic accuracy of the combination of IL-27, IFN-γ, or/and ADA for differentiating TPE from pleural effusions with the other etiologies.

METHODSThe concentrations of IL-27, IFN-γ and ADA were simultaneously determined in pleural fluids and sera from 40 patients with TPE; 26 with malignant pleural effusion, seven with infectious pleural effusion, and eight with transudative pleural effusion by enzyme linked immunosorbent assay and colorimetric method. The corresponding biochemical indexs were also simultaneously determined.

RESULTSThe concentrations of pleural IL-27 and IFN-γ in the tuberculous group were significantly higher than those in the malignant, infectious, and transudative groups. The concentrations of ADA in TPE were significantly higher than those in MPE or transudative effusions, while much lower than those in infectious effusions. Among these three biomarkers, IL-27 was the most effective for TPE diagnosis, with the cut off value of 900.8 ng/L. IL-27 had a high sensitivity of 95% and specificity of 97.6% for differential diagnosis of TPE from non-TPEs. Combinations of IL-27, IFN-γ and ADA measurements further increased the sensitivity or specificity up to 100%.

CONCLUSIONSCompared to non-TPEs, IL-27, IFN-γ and ADA all simultaneously increased in TPE; and among these three rapid detection methods, IL-27 appeared to be the best for distinguishing tuberculous from non-TPEs, especially from MPE. Combinations of the three markers (IL-27, IFN-γ and ADA) yielded the highest sensitivity and specificity. These findings suggest that the applications of a new biomarker, IL-27, alone or with IFN-γ and ADA, may contribute to more efficient diagnosis strategies in the management of tuberculous pleurisy.

Adenosine Deaminase ; blood ; metabolism ; Aged ; Female ; Humans ; Interferon-gamma ; blood ; metabolism ; Interleukin-27 ; blood ; metabolism ; Male ; Middle Aged ; Pleural Effusion ; blood ; metabolism ; Tuberculosis, Pleural ; blood ; diagnosis ; metabolism

OBJECTIVETo study the antitumour effects of sodium valproate (VPA) on the proliferation, differentiation and cell cycle of Molt-4 cell and to investigate its demethylation mechanisms.

METHODSAfter Molt-4 cells trated with VPA at different concentrations, cell viability and growth curve were assessed by MTT assay. Cell cycle changes were analyzed by flow cytometry. The expression level of p15, DNA methyltransferase 1 (DNMT-1), DNMT3A and 3B mRNA were detected by RT-PCR and the methylation level was detected by hn-MSPCR.

RESULTSVPA significantly inhibited the proliferation of Molt-4 cells. After 48 h culture with 5.0 mmol/L VPA, the percentages of Molt-4 cells in G(0)/G(1) phase was (66.87 ± 3.31)% and in S phase was (8.47 ± 2.56)%, while in control group, the cells in G(0)/G(1) phase increased and in S phase decreased significantly. The p15 gene in Molt-4 cells failed to express due to its hypermethylation. The expression level of p15 gene mRNA increased significantly after exposure to VPA for 48 h. As compared with control group, the expression of DNMT-1 was down-regulated in a dose-dependent manner. The expression level of DNMT3B decreased at 10.0 mmol/L concentration.

CONCLUSIONVPA has a demethylation effect on p15 INK4B gene by inhibiting the DNMT-1 and DNMT3B gene activities to recover p15 gene activity, which arrests Molt-4 cell in G(0)/G(1) phase.

Cell Cycle ; drug effects ; Cell Line, Tumor ; DNA Methylation ; drug effects ; RNA, Messenger ; genetics ; Valproic Acid ; pharmacology

OBJECTIVETo study the relation of the detection rates of the novel influenza virus A/H1N1 RNA in clinically confirmed patients in the 2009 pandemic with the age distribution of the patients and the disease course.

METHODSA total of 151 clinical patients with H1N1 infection were enrolled in this study, from whom 833 dynamic throat swab samples were obtained for detecting the H1N1 RNA using real-time PCR. A statistical analysis of the age distribution was performed among the patients with different disease courses. Chi-square for trend test was used to study the correlation between the detection rates of H1N1 RNA and the time of disease onset.

RESULTSThe majority of patients were young with their ages ranging from 10 to 20 years (57.26%) and 20 to 30 years (22.18%). Chi-square for trend test revealed that the positivity rates of the throat swabs in the patients decreased with the prolongation of the disease course (chi(2)=9.784, P=0.002).

CONCLUSIONMost of the H1N1 patients are young within the age range of 10-30 years, and the longest disease course can exceed 10 days. The positivity rates of throat swabs from the H1N1 patients decreases with the prolongation of the disease course.

Adolescent ; Adult ; Age Factors ; Child ; China ; epidemiology ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; Influenza, Human ; epidemiology ; virology ; Male ; Pharynx ; virology ; RNA, Viral ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult