Phellopterin attenuates ovarian cancer proliferation and chemoresistance by inhibiting the PU.1/CLEC5A/PI3K-AKT feedback loop.

Ovarian cancer is known as the second leading cause of gynecologic cancer-associated deaths in women worldwide. Developing new and effective compounds to alleviate chemoresistance is an urgent priority in ovarian cancer. Here, we aimed to reveal the biological function and underlying mechanisms of phellopterin, a naturally sourced ingredient of Angelica dahurica, in ovarian cancer progression as well as evaluate the therapeutic potential of phellopterin in ovarian cancer patients. In this investigation, we found that phellopterin mitigated DNA replication and induced cell cycle arrest, apoptosis, and DNA damage, attenuating cell proliferation and chemoresistance of ovarian cancer. Interestingly, bioinformatics analyses of data from our RNA sequencing and The Cancer Genome Atlas ovarian cancer dataset suggested that phellopterin presented anti-cancer activities in ovarian cancer cells by modulating signals affecting ovarian cancer progression and identified phellopterin as a potential compound in improving ovarian cancer patients' prognosis. In addition, the C-Type Lectin Domain Containing 5A (CLEC5A) was demonstrated as a downstream effector of phellopterin and involved in a positive PU.1/CLEC5A/PI3K-AKT feedback loop. Interestingly, phellopterin might inactivate the positive feedback circuit to suppress ovarian cancer progression. Collectively, our investigation revealed that phellopterin mitigated ovarian cancer proliferation and chemoresistance through suppressing the PU.1/CLEC5A/PI3K-AKT feedback loop, and predicted phellopterin as a new and effective cytotoxic drug and CLEC5A as a potential target for the treatment of ovarian cancer.

Lipidomic and transcriptomic profiles of glycerophospholipid metabolism during Hemerocallis citrina Baroni flowering.

BACKGROUND: Hemerocallis citrina Baroni (daylily) is a horticultural ornamental plant and vegetable with various applications as a raw material in traditional Chinese medicine and as a flavouring agent. Daylily contains many functional substances and is rich in lecithin, which is mostly composed of glycerophospholipids. To study the comprehensive dynamic changes in glycerophospholipid during daylily flowering and the underlying signalling mechanisms, we performed comprehensive, time-resolved lipidomic and transcriptomic analyses of 'Datong Huanghua 6' daylily. RESULTS: Labelling with PKH67 fluorescent antibodies clearly and effectively helped visualise lipid changes in daylily, while relative conductivity and malonaldehyde content detection revealed that the early stages of flowering were controllable processes; however, differences became non-significant after 18 h, indicating cellular damage. In addition, phospholipase D (PLD) and lipoxygenase (LOX) activities increased throughout the flowering process, suggesting that lipid hydrolysis and oxidation had intensified. Lipidomics identified 558 lipids that changed during flowering, with the most different lipids found 12 h before and 12 h after flowering. Transcriptome analysis identified 13 key functional genes and enzymes in the glycerophospholipid metabolic pathway. The two-way orthogonal partial least squares analysis showed that diacylglycerol diphosphate phosphatase correlated strongly and positively with phosphatidic acid (PA)(22:0/18:2), PA(34:2), PA(34:4), and diacylglycerol(18:2/21:0) but negatively with phospholipase C. In addition, ethanolamine phosphotransferase gene and phospholipid-N-methyltransferase gene correlated positively with phosphatidylethanolamine (PE)(16:0/18:2), PE(16:0/18:3), PE(33:2), and lysophosphatidylcholine (16:0) but negatively with PE(34:1). CONCLUSIONS: Overall, this study elucidated changes in the glycerophospholipid metabolism pathway during the daylily flowering process, as well as characteristic genes, thus providing a basis for future studies of glycerophospholipids and signal transduction in daylilies.

Epigenetic modification-mediated inhibition of SOSTDC1 expression promotes malignant biological behaviors in cervical cancer cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：探究含硬化蛋白域蛋白1（SOSTDC1）对宫颈癌细胞恶性生物学行为的调控及其分子机制。方法：收集2020年8月至2022年5月间在福建省肿瘤医院活检或手术切除的53例宫颈癌组织和相应的癌旁组织标本，免疫组化法检测SOSTDC1蛋白在宫颈癌组织及相应癌旁组织中的表达，qPCR法检测正常宫颈细胞、宫颈癌细胞中SOSTDC1 mRNA表达；将SOSTDC1过表达慢病毒（OE-sostdc1）和对照空病毒（NC）感染宫颈癌细胞SiHa及CaSki，将其分为SiHa-OE-sostdc1、SiHa-NC、CaSki-OE-sostdc1、CaSki-NC组，采用WST-1法、细胞集落形成实验、Transwell实验和WB法检测转染各组SiHa及CaSki细胞的增殖、集落形成、迁移和侵袭能力和BMP、Wnt/β-catenin信号途径相关蛋白及上皮-间充质转化（EMT）相关蛋白的表达。用DNA甲基化酶抑制剂5-氮杂2'-脱氧胞苷（5'-Aza-CdR）处理宫颈癌细胞后采用qPCR和WB法检测SOSTDC1 mRNA及蛋白的表达变化，用甲基化特异性PCR（MSP）检测5例配对宫颈癌组织与癌旁组织中SOSTDC1基因启动子区甲基化水平，同时qPCR检测其SOSTDC1 mRNA水平。结果：与癌旁组织比较，SOSTDC1蛋白在宫颈癌组织中呈低表达（P<0.01），且与淋巴结转移与FIGO分期有关联（均P<0.05）；与正常宫颈HUCEC细胞比较，SOSTDC1 mRNA在宫颈癌C33A、HeLa、SiHa、CaSki细胞中均呈低表达（均P<0.01）。过表达SOSTDC1显著抑制SiHa及CaSki细胞的增殖、迁移和侵袭能力（均P<0.05）。WB法结果检测显示，过表达SOSTDC1显著抑制SiHa及CaSki细胞中磷酸化Smad、Dvl2/3、β-catenin、VIM、N-cadherin、Snail蛋白的表达（均P<0.05），5'-Aza-CdR处理后的SiHa及CaSki细胞中SOSTDC1 mRNA和蛋白水平均显著增加（均P<0.05），MSP检测结果显示，相较于癌旁组织，宫颈癌组织中SOSTDC1基因启动子区呈高度甲基化，且SOSTDC1 mRNA水平降低（P<0.01）。结论：SOSTDC1在宫颈癌组织中呈低表达且与肿瘤的恶性进展关联，其表达下调与其基因启动子区高度甲基化有关，过表达SOSTDC1可能通过阻断BMP及Wnt/β-catenin信号通路从而抑制SiHa、CaSki细胞的增殖、侵袭和迁移能力。

Prevalence of multiple sclerosis in Guangzhou, China: A population-based case-finding prospective study.

BACKGROUND: Multiple sclerosis (MS) is rare in China, and the prevalence previously reported may be biased. Currently, few studies that have investigated the prevalence of MS in China based on the latest diagnostic criteria. METHODS: Through a population-based survey from August 8, 2021 to December 31, 2021, we calculated the prevalence of multiple sclerosis in 18,676,605 residents of Guangzhou, China. MS patients were identified through the health insurance system of the Guangzhou Health Insurance Bureau, and we surveyed 17 large tertiary hospitals using a case-finding approach. All MS patients were diagnosed according to the 2017 McDonald criteria. RESULTS: A total of 143 patients in the resident population of Guangzhou were diagnosed with MS, with a crude prevalence of 0.77 per 100,000 (95% confidence interval (CI): 0.65-0.90), and the prevalence was higher in in females (1.14/100,000) than in males (0.44/100,000). The age-adjusted prevalence was 0.92 per 100,000 (95% CI: 0.77-1.10). The prevalence peaked at the age of 25-29 years (2.86/100,000) for both males and females (1.44/100,000 and 4.42/100,000, respectively). CONCLUSIONS: This is the first study to report the prevalence of MS in Guangzhou, China, according to the criteria. Our study shows that the prevalence of MS in Guangzhou is lower than that in other cities in China.

ReLU-FCM trained by quasi-oppositional bare bone imperialist competition algorithm for predicting employment rate.

Fuzzy cognitive maps (FCMs) are a powerful tool for system modeling, which can be used for static and dynamic analysis. However, traditional FCMs are usually learned by gradient-based methods, and the adopted sigmoid nonlinear activation function frequently causes gradient saturation. These two shortcomings set a limit on the modeling accuracy. To overcome those problems, we propose in this paper a new FCM with two improvements. First, the rectified linear unit (ReLu) activation function is adopted to replace the sigmoid function. Second, a newly proposed quasi-oppositional bare bone imperialist competition algorithm (QBBICA) is used to learn the FCM. The improved FCM is used to predict the employment rate of graduates from Liren College, Yanshan University. Experimental results show that the improved FCM is effective in employment rate prediction.

Circular RNA CDK6 suppresses cervical cancer proliferation and metastasis by sponging miR-449a.

Cervical cancer is a malignant tumor that severely threatens female health. Recently, more and more studies indicated that circRNA could function as a tumor activator or suppressor in cervical cell development. Therefore, we aimed to study the effect of circRNA CDK6 (circCDK6) on the development and biological behavior of cervical cancer. We used quantitative real-time PCR (qRT-PCR) to examine the circCDK6 expression level in cervical cancer cell lines. RNA-fluorescence in situ hybridization confirmed the location of circCDK6 and miR-449a in HeLa and CaSki cells, respectively. Then, the biological function of silencing circCDK6 in cellular proliferation, metastasis, and Epithelial-Mesenchymal Transition (EMT)-related process was determined. We also performed RNA Binding Protein Immunoprecipitation (RIP) and Dual-luciferase reporter assay to determine the relationship between the circCDK6 and miR-449a. Finally, the results showed that circCDK6 level remarkably increased in several cervical cancer cells, especially in Hela and CaSki cells. The miR-449a was further confirmed to be a potential target of circCDK6, and its expression increased by silencing circCDK6. The circCDK6 participated in tumorigenesis and cancer progression and might serve as a tumor suppressive factor in cervical cell progression via Epithelial-MesenchymalTransition (EMT) process by regulating miR-449a.

[Screening and confirmation of pesticide residues in substitutional tea such as orange peel, lotus leaf, pueraria lobata and Pangdahai].

OBJECTIVE: The screening method was established for pesticide residues in substitutional teas such as orange peel, lotus leaf, pueraria lobata and Pangdahai using data base. The method for analyzing of 26 pesticide residues was confirmed in four substitutional teas with gas chromatography-tandem mass spectrometry(GC-MS/MS) and dispersive solid-phase extraction(d-SPE) for sample preparation. METHODS: The 26 types of pesticides were selected as target compounds, including screened out, commonly used in substitutional tea planting and forbidden in agriculture planting. The samples extracted with acetonitrile, and purified by adsorbent(e. g PSA, GCB, C_(18)), the purification solutions were separated on DB-5 MS UI column((15 m+PUU+15 m)×0. 25 mm×0. 25 µm) with programmed temperature and determined by GC-MS/MS in multi-reaction monitoring(MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity(r≥0. 995)in concentration range(2-100 µg/L) for 26 kinds of pesticides. The limits of quantification(LOQ) were 3-40 µg/kg. The average recoveries range were 71. 9%-114. 4% in three add levels of 10, 50, 200 µg/kg, and relative standard deviations of 1. 0%-16. 0%(n=6). In the four matrixes, the pesticides were detected concentrating on orange peel, and there were kinds of residues. CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of pesticide screening and simultaneous analysis of multiple pesticide residues in orange peel, lotus leaf, pueraria lobata and Pangdahai.

Parental attitudes and willingness to donate children's biospecimens for congenital heart disease research: a cross-sectional study in Shanghai, China.

OBJECTIVES: To assess attitudes and willingness of parents of children with congenital heart disease (CHD) regarding donating biospecimens for future CHD research, and to identify factors associated with biospecimen donation. DESIGN: Face-to-face cross-sectional survey data were analysed using logistic regression. SETTING: Cardiothoracic Surgery Inpatient Department, Shanghai Children's Medical Centre. PARTICIPANTS: Parents of children attending the cardiothoracic surgery inpatient department at Shanghai Children's Medical Center, 1 March-31 December 2016. PRIMARY AND SECONDARY OUTCOME MEASURES: Willingness and motivation regarding donating children's biospecimens, and ethical and legal considerations concerning children's future willingness to donate. RESULTS: Of 550 parents, 508 completed the questionnaire (response rate=92.4%). Overall, 69.1% (n=351) were willing to donate their children's biospecimens for medical research. Multivariate analysis indicated higher education level (college/graduate degree: OR 2.435, 95% CI 1.221 to 4.857, p=0.012; high school: OR 1.827, 95% CI 1.190 to 2.804, p=0.006) and children's hospitalisation history (OR 1.581; 95% CI 1.069 to 2.338, p=0.022) were positively associated with willingness to donate. The most common motivation for donation was potential benefit to other children with CHD (81.2%, n=285). The main barriers to donation were physical discomfort to their children (52.3%, n=54) and concerns about personal privacy (47.1%, n=48). Most parents (86.0%, n=302) wanted to be informed of research results using their children's donated biospecimens, and 34.8% (n=177) believed that children aged 10-18 years had the right to consent independently to research participation. CONCLUSIONS: Nearly 70% of the parents in this study were willing to donate their children's biospecimens for future CHD research. Parents' education level and children's hospitalisation history influenced willingness to donate. Most parents wanted to receive the research results related to their children's biospecimens.

[Determination of pesticide residues in substitutional tea by dispersive solid-phase extraction and gas chromatography-triple quadrupole mass spectrometry].

OBJECTIVE: The method was established for determination 14 kinds of psticide residues in substitutional tea using gas chromatography-triple quadrupole mass spectrometry( GC-MS/MS) for instrument method and dispersive solid-phase extraction( dSPE) for sample preparation. METHODS: The samples extracted with acetonitrile, and purified by N-propyl ethlene diamine( PSA), the extracts were separated on DB-5MS column( 30 m × 0. 25 mm, 0. 25 µm) with programmed temperature and determined by GCMS/MS in multi-reaction monitoring( MRM) model. The external standard method was applied to quantify the pesticides. RESULTS: The method showed a good linearity( r ≥0. 999) in certain ranges for 14 kinds of pesticides. The limits of quantification( LOQ)were 4-40 µg/kg. The average recoveries range were 65. 6%-116. 6% in three add levels of 20, 40 and 200 µg/kg, and relative standard deviations( RSD S) of 1. 5%-15. 7%( n = 6). CONCLUSION: This method is simple, fast, sensitive, selective and can satisfy the request of simultaneous analysis of multiple pesticide residues in substitutional tea.

Transcriptome profiling of cancer and normal tissues from cervical squamous cancer patients by deep sequencing.

Cervical cancer is the fourth leading cause of cancer mortality in women worldwide. High­risk human papillomavirus infection is a major cause of cervical cancer. A previous study revealed the role of different oncogenes and tumor suppressors in cervical cancer initiation and progression. However, the complicated genetic network regulating cervical cancer remains largely unknown. The present study reported transcriptome sequencing analysis of three cervical squamous cell cancer tissues and paired normal cervical tissues. Transcriptomic analysis revealed that 2,519 genes were differently expressed between cervical cancer tissues and their corresponding normal tissues. Among these, 236 differentially expressed genes (DEGs) were statistically significant, including many DEGs that were novel in cervical cancer, including gastrulation brain homeobox 2,5­hydroxytryptamine receptor 1D and endothelin 3. These 236 significant DEGs were highly enriched in 28 functional gene ontology categories. The Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis suggested involvement of these DEGs in multiple pathways. The present study provides a transcriptome landscape of cervical cancer in Chinese patients and an improved understanding of the genetic regulatory network in cervical cancer tumorigenesis.

Association of coronary artery calcium with bone mineral density in postmenopausal women.

OBJECTIVES: Atherosclerosis and osteoporosis (OP) are common diseases in elderly individuals and may share common pathogenetic mechanisms. The aim of this study was to investigate the association between bone mineral density (BMD) and coronary artery calcium (CAC) in postmenopausal women. METHODS: In this cross-sectional study, 186 postmenopausal women 50-80 years of age were included. BMD of the spine and femoral neck was measured by dual-energy X-ray absorptiometry. The coronary artery calcium score (CACS) was measured by multidetector computed tomography. RESULTS: The study included postmenopausal women aged 65.6±7.3 years, 109 of whom (58.6%) showed CAC. Thirty-three (17.7%) of the patients were found to have OP in the lumbar spine and 83 (44.6%) had osteopenia, whereas in the femoral neck, 26 patients (14.0%) had OP and 87 patients (46.8%) had osteopenia. The mean CACS was 57.6±108.3 in normal status, 89.7±143.5 in OP, and 156.4±256.9 in osteopenia at the spine (P<0.05). The mean CACS was 43.2±89.9 in normal status, 126.9±180.3 in OP, and 198.2±301.2 in osteopenia at the femoral neck (P<0.05). Multivariable logistic regression analysis showed that BMD was an independent marker for an increased risk of developing CAC in postmenopausal women. The multiple regression model showed that T-scores were the independent predictors of CACS. CONCLUSION: BMD identified on images from dual-energy X-ray absorptiometry were strongly related to multidetector computed tomography measures of CAC. This low-cost, minimal radiation technique used widely for OP screening is a promising marker of generalized coronary atherosclerosis.

Intranasal immunization with a peptide conjugated to Salmonella flagellin induces both systemic and mucosal peptide-specific antibody responses in mice.

In this study, the mucosal adjuvant activity of Salmonella flagellin as a carrier in a conjugate of EXP153-rFliC was investigated. EXP153-rFliC was made by conjugation of a synthetic B-cell epitope peptide derived from Plasmodium falciparum exported protein-1(EXP153) to recombinant phase 1 flagellin of Salmonella enterica serovar Typhimurium expressed in Escherichia coli (rFliC), and used to immunize BALB/c mice via intranasal instillation. It was found that robust EXP153-specific serum IgG antibodies were induced without additional adjuvant. EXP153-specific sIgA antibodies were also induced, these being detected in bronchoalveolar, nasal, vaginal and intestinal washes. These observations demonstrate that Salmonella flagellin as a carrier is an effective mucosal adjuvant in that its conjugated peptide induces antibody responses.

Ameliorative effects of Qingfei Tongluo formula on experimental mycoplasmal pneumonia in mice.

Mycoplasma pneumoniae pneumonia (MPP) is a common disease in children. Qingfei Tongluo formula (QTF) has been used for the treatment of MPP clinically, but the chemical constituents and mechanism involved remain unclear. This study aimed to analyze the main chemical constituents and to explore the possible mechanism of action associated with QTF treatment of MPP. Liquid chromatography-mass spectrometry was employed to identify the compounds contained in the QTF extract. A BALB/c mouse model of MP infection was established. After treatment with QTF (0.85 and 1.70 g/kg) for 3 days, hematoxylin and eosin staining was performed in lung tissues for histological examination. Inflammatory cytokines were detected by ELISA. Western blot analysis was used for detecting phosphorylated proteins involved in MAPK and nuclear factor (NF)-κB signaling pathways. In the mouse model, a large amount of pulmonary interstitial infiltration of lymphocytes and plasmacytes were seen as well as bronchus and vasodilation congestion. Following QTF treatment, inflammation was alleviated significantly compared with the model group. Inflammatory cytokines [interleukin (IL)-6, transforming growth factor-ß1, IL-8, IL-1ß and tumor necrosis factor-α] in bronchoalveolar lavage fluid were decreased dramatically. In addition, we found that QTF inhibited activation of phosphorylation of JNK, ERK and NF-κB. In conclusion, QTF alleviates MPP inflammation possibly via inhibitory activation of MAPK/NF-κB pathways, which can act as a new agent for MPP treatment.

Preliminary findings on the reliability and validity of the Cantonese Birmingham Cognitive Screen in patients with acute ischemic stroke.

BACKGROUND: There are no currently effective cognitive assessment tools for patients who have suffered stroke in the People's Republic of China. The Birmingham Cognitive Screen (BCoS) has been shown to be a promising tool for revealing patients' poststroke cognitive deficits in specific domains, which facilitates more individually designed rehabilitation in the long run. Hence we examined the reliability and validity of a Cantonese version BCoS in patients with acute ischemic stroke, in Guangzhou. METHOD: A total of 98 patients with acute ischemic stroke were assessed with the Cantonese version of the BCoS, and an additional 133 healthy individuals were recruited as controls. Apart from the BCoS, the patients also completed a number of external cognitive tests, including the Montreal Cognitive Assessment Test (MoCA), Mini Mental State Examination (MMSE), Albert's cancellation test, the Rey-Osterrieth Complex Figure Test, and six gesture matching tasks. Cutoff scores for failing each subtest, ie, deficits, were computed based on the performance of the controls. The validity and reliability of the Cantonese BCoS were examined, as well as interrater and test-retest reliability. We also compared the proportions of cases being classified as deficits in controlled attention, memory, character writing, and praxis, between patients with and without spoken language impairment. RESULTS: Analyses showed high test-retest reliability and agreement across independent raters on the qualitative aspects of measurement. Significant correlations were observed between the subtests of the Cantonese BCoS and the other external cognitive tests, providing evidence for convergent validity of the Cantonese BCoS. The screen was also able to generate measures of cognitive functions that were relatively uncontaminated by the presence of aphasia. CONCLUSION: This study suggests good reliability and validity of the Cantonese version of the BCoS. The Cantonese BCoS is a very promising tool for the detection of cognitive problems in Cantonese speakers.

Salmonella flagellin is a potent carrier-adjuvant for peptide conjugate to induce peptide-specific antibody response in mice.

As an agonist to innate immune system, Salmonella flagellin has been proven to be a potent adjuvant either admixed or genetically fused with antigens and applied to a variety of vaccines against infectious diseases. However, relatively little is known about its carrier-adjuvant effect for conjugate vaccine. Conjugation is an effective approach often used to make haptens such as some peptides and polysaccharides immunogenic and in some cases used to make poor immunogens more immunogenic. In the current study, Salmonella flagellin was tested for its carrier-adjuvant effect in a peptide conjugation. The recombinant Salmonella flagellin (rFliC) purified from Escherichia coli was firstly modified by maleimide groups, then coupled with a synthetic peptide (EXP153:CDNNLVSGP) that is a B-cell epitope derived from Plasmodium falciparum exported protein-1 to generate the conjugate of EXP153-rFliC. Bioactivity assay showed that both chemical modification and conjugation did not apparently impair the TLR5-ligand activity of rFliC. EXP153-rFliC was used to immunize BALB/c mice via subcutaneous route, and the sera obtained from immunized mice were examined by ELISA and IFA. While no detectable antibody responses were induced by the peptide admixed with rFliC, the robust peptide-specific antibody responses were observed in mice immunized with the peptide conjugated to rFliC in the absence of any additional adjuvant. The immune sera induced by the conjugate recognized the native protein of malaria parasite. The data obtained from this study demonstrate the carrier-adjuvant activity of Salmonella flagellin in peptide conjugate immunization and indicate its promising application for conjugate vaccine research and development.

Intensive Atorvastatin Therapy Attenuates the Inflammatory Responses in Monocytes of Patients with Unstable Angina Undergoing Percutaneous Coronary Intervention via Peroxisome Proliferator-Activated Receptor γ Activation.

Periprocedural myocardial injury is a prognostically important complication of percutaneous coronary intervention (PCI). However, it still remains unclear whether and how intensive atorvastatin therapy attenuates the unfavorable inflammatory responses of monocytes associated with PCI. The aim of the study was to investigate the impact of intensive atorvastatin therapy on inflammatory responses of monocytes in Chinese patients with unstable angina who received PCI in order to explore the potential anti-inflammatory mechanism. Ninety-six patients with unstable angina were randomly assigned to atorvastatin 80 mg (intensive) or atorvastatin 20 mg (conventional) treatment at a 1:1 ratio. Creatine kinase MB (CK-MB), cTnI, hs-CRP, and IL-6 were assessed, and circulating CD14(+) monocytes were simultaneously obtained using CD14 MicroBeads 2 h before and 24 h after PCI. Plasma levels of CK-MB, cTnI, hs-CRP, and IL-6 were higher in the conventional dose group versus those in the intensive dose group following PCI. Furthermore, intensive atorvastatin treatment markedly reduced the expressions and responses of Toll-like receptor 2 (TLR2), TLR4, and CCR2 of CD14(+) monocytes versus the conventional dose group and significantly increased the activated peroxisome-proliferator-activated receptor (PPAR) γ in the CD14(+) monocytes post-PCI. Notably, the changes in responses of TLR2, TLR4, and CCR2 of CD14(+) monocytes between the two groups were all reversed by PPARγ antagonist and augmented by PPARγ agonist. In conclusion, a single high (80 mg) loading dose of atorvastatin reduced the inflammatory response in Chinese patients with unstable angina following PCI. The anti-inflammatory role of intensive atorvastatin was possibly due to attenuation of inflammatory response in monocytes via PPARγ activation.

[Potential role of CRELD1 gene in the pathogenesis of atrioventricular septal defect].

OBJECTIVE: To screen potential mutation of the CRELD1 gene in congenital atrioventricular septal defect (AVSD) and explore its functional implications. METHODS: Fragments encompassing the 11 coding exons of CRELD1 gene, including at least 50 bp of flanking intronic regions, were amplified with PCR and subjected to DNA sequencing. Results of sequencing were compared with predicted sequence from the GenBank database. Eukaryotic expression vector pcDNA3.1CRELD1 containing the mutational sequence was constructed. Western blotting and real-time fluorescent quantitative reverse transcription polymerase chain reaction (FQ RT-PCR) was applied to examine the expression of CRELD1, Tenascin C and Aggrecan. RESULTS: C857G was identified in a girl with an isolated partial AVSD. The mutation has resulted in a substitution of Alanine for Proline at amino acid 286 in the first cbEGF domain. Western blotting and FQ RT-PCR confirmed that the P286R missense mutation has been a gain-of-function mutation. Compared with the unloaded control, the Aggrecan mRNA expression was downregulated for both wild-type and mutant type samples (t=140.27 vs. 26.36, P < 0.01). The downregulation was more significant in mutant type (t=25.69, P=0.002). There was no significant difference of the Tenascin C expression between wild-type and the unload control (t=1.167, P> 0.05), whilst the Tenascin C expression was up-regulated in mutant type (t=6.66, P=0.022). CONCLUSION: Mutation of the CRELD1 gene may increase the risk for AVSD rather than being directly causative. The P286R mutation of CRELD1 can downregulate the expression of Aggrecan and upregulates the expression of Tenascin C protein, both of which are crucial to extracellular matrix in the formation of the atrioventricular septum. The P286R mutation of CRELD1 may be correlated to the occurrence of AVSD.

Novel risk loci for rheumatoid arthritis in Han Chinese and congruence with risk variants in Europeans.

OBJECTIVE: To investigate differences in genetic risk factors for rheumatoid arthritis (RA) in Han Chinese as compared with Europeans. METHODS: A genome-wide association study was conducted in China with 952 patients and 943 controls, and 32 variants were followed up in 2,132 patients and 2,553 controls. A transpopulation meta-analysis with results from a large European RA study was also performed to compare the genetic architecture across the 2 ethnic remote populations. RESULTS: Three non-major histocompatibility complex (non-MHC) loci were identified at the genome-wide significance level, the effect sizes of which were larger in anti-citrullinated protein antibody (ACPA)-positive patients than in ACPA-negative patients. These included 2 novel variants, rs12617656, located in an intron of DPP4 (odds ratio [OR] 1.56, P = 1.6 × 10(-21) ), and rs12379034, located in the coding region of CDK5RAP2 (OR 1.49, P = 1.1 × 10(-16) ), as well as a variant at the known CCR6 locus, rs1854853 (OR 0.71, P = 6.5 × 10(-15) ). The analysis of ACPA-positive patients versus ACPA-negative patients revealed that rs12617656 at the DPP4 locus showed a strong interaction effect with ACPAs (P = 5.3 × 10(-18) ), and such an interaction was also observed for rs7748270 at the MHC locus (P = 5.9 × 10(-8) ). The transpopulation meta-analysis showed genome-wide overlap and enrichment in association signals across the 2 populations, as confirmed by prediction analysis. CONCLUSION: This study has expanded the list of alleles that confer risk of RA, provided new insight into the pathogenesis of RA, and added empirical evidence to the emerging polygenic nature of complex trait variation driven by common genetic variants.

Influence of genes and the environment in familial congenital heart defects.

The present study aimed to investigate genetic and environmental factors involved in the pathogenesis of congenital heart disease (CHD). A total of 61 familial pedigrees with CHD were analyzed, and 134 patients out of 761 family members had a diagnosis of CHD confirmed. The present study revealed that the prevalence of CHD in first­degree relatives (55/249, 22.0%) was significantly higher than that in second­degree relatives (18/526, 3.4%). Additionally, the recurrence rate of CHD in families in which the patient's mother (12/61) or sister (15/61) had CHD were significantly higher than in cases with the father (6/61) or brother (4/61) having CHD. The subtypes of CHD with increased risk of recurrence were ventricle septal defect (VSD) and atrial septal defect (ASD), followed by patent ductus arteriosus and tetralogy of fallot (TOF). In the 21 sets of twins among the 61 familial pedigrees analyzed, the concordance of both twins affected by CHD in identical and dizygotic twins was 94.4% (17/18) and 33.3% (1/3), respectively. Identical subtypes of CHD were identified in 10 out of 21 sets of twins. Of note, the following pattern was identified in three sets of the twins: One twin had TOF, while the other one had VSD. A risk factor survey revealed that threatened abortion in early pregnancy was associated with familial CHD. In conclusion, genetic factors may have important roles in the development of CHD, and TOF and VSD may have similar molecular mechanisms. Threatened abortion in early pregnancy is a novel environmental factor that may be specific in Chinese females with CHD.