Objective:To investigate the role of m.4435A>G and YARS2 c.572G>T(p.G191V)mutations in the development of essential hypertension.Methods:A hypertensive patient with m.4435A>G and YARS2 p.G191V mutations was identified from previously collected mitochondrial genome and exon sequencing data.Clinical data were collected,and a molecular genetic study was conducted in the proband and his family members.Peripheral venous blood was collected,and immortalized lymphocyte lines constructed.The mitochondrial transfer RNA(tRNA),mitochondrial protein,adenosine triphosphate(ATP),mitochondrial membrane potential(MMP),and reactive oxygen species(ROS)in the constructed lymphocyte cell lines were measured.Results:Mitochondrial genome sequencing showed that all maternal members carried a highly conserved m.4435A>G mutation.The m.4435A>G mutation might affect the secondary structure and folding free energy of mitochondrial tRNA and change its stability,which may influence the anticodon ring structure.Compared with the control group,the cell lines carrying m.4435A>G and YARS2 p.G191V mutations had decreased mitochondrial tRNA homeostasis,mitochondrial protein expression,ATP production and MMP levels,as well as increased ROS levels(all P<0.05).Conclusion:The YARS2 p.G191V mutation aggravates the changes in mitochondrial translation and mitochondrial function caused by m.4435A>G through affecting the steady-state level of mitochondrial tRNA and further leads to cell dysfunction,indicating that YARS2 p.G191V and m.4435A>G mutations have a synergistic effect in this family and jointly participate in the occurrence and development of essential hypertension.

Objective To investigate the association of neutrophil gelatinase-associated lipocalin(NGAL)and severity of coronary stenosis in patients with stable coronary artery disease(SCAD).Methods Totally 144 patients who underwent coronary angiography firstly due to suspected coronary artery disease were enrolled.The patients were divided into SCAD group(n=101)and non-CAD group(n=43),and the clinical data were compared.The SCAD group was fuether divided into three subgroups accoding to Gensini score:mild stenosis group(Gensini<16,n=32),moderate stenosis(16≤Gensini<30,n=33)and severe stenosis group(Gensini≥36,n=35).The clinical data of each group were compared.Odinal Logistic regression was used to analysis the influencing factors of severe stenosis.The receiver operating characteristic(ROC)curve was performed to evaluated the ability of NGAL in discriminating severe stenosis.Results Serum NGAL,high sensitive C-reactive protein(hs-CRP)level in SCAD group were higher than those in non-CAD group(P<0.01).Comparing with mild and moderate stenosis groups,NGAL and hs-CRP level were more higher in severe stenosis group(P<0.01).Multivariable Logistic regression analysis showed that NGAL and hs-CRP were independent risk factors for severe coronary stenosis(both P<0.05).Receiver operating characteristic(ROC)curve analysis showed that the areas under the curve of NGAL and hs-CRP were 0.771,0.702 respectively(both P<0.01).The difference between two AUCs was 0.069 which was insignificant(P>0.05).Conclusion Serum NGAL increased is closely associated with severe coronary stenosis in patients with SCAD,which has a certain ability of discriminating severity of coronary stenosis.

Objective To explore the function of short-chain dehydrogenases/reductases (SDRs) in safflower flavonoid, especially hydroxysafflower yellow A (HSYA) biosynthesis. Methods SDRs involved in HSYA biosynthesis pathway were screened based on safflower transcriptome database and metabolome database. The expression pattern was analyzed by qRT-PCR. The overexpression vector was constructed by seamless cloning technology, then genetically transformed to the Yunnan Weishan safflower strain by Agrobacterium gv3101. The transgenic T₂ generation plants were positively verified, and the gene expression of corolla SDRs was analyzed. The content of secondary metabolites was assayed by UPLC-Q-TOF/MS. Results Three SDRs genes named CtSDR1, CtSDR2 and CtSDR3 involved in HSYA biosynthesis pathway were screened. Their expression in safflower from high to low was corolla > leaf > stem > root. The expression level in corolla increased gradually with corolla development. qRT-PCR analysis of corolla with positive verification of genome insertion sequence showed that the transcription level of CtSDR3 in corolla of T₂ positive plants increased by 2~3 times compared with the blank control group, and the content of secondary metabolite HSYA increased by 7.1%~16.6% (P< 0.05). Conclusion CtSDR3 may be involved in the biosynthesis of flavonoids, especially HSYA, in safflower. It provides the support data for explaining the function of CtSDR3 in HSYA biosynthesis pathway.

Objective To explore the effect of circadian rhythm genes on flavonoids biosynthesis in safflower and its molecular mechanism. Methods Based on the transcriptome and metabolomic database of safflower corolla, we screened the circadian rhythm genes that correlate with biosynthesis of flavonoids in safflower. qPCR was used to quantify the expressions of circadian rhythm genes in different flowering stages at different time points in a single day. LC-MS was performed to determine the accumulation of flavonoids. The correlation between them was analyzed as well. Yeast Two-Hybrid experiment was used to verify the interactive proteins of these genes. Results Seven circadian rhythm genes PRR1, PRR2, ELF3, FT, PHYB, GI and ZTL were obtained. PRR1 gene was positively correlated with flavonoids accumulation (r≥0.7). The full length of PRR1 is 3 201 bp, encoding 421 amino acids, which is highly homologous with rice OsPRR73 gene and named as CtPRR1 (GenBank accession number: MW492035). CtPRR1 was mainly expressed in flowers, and the expression level increased in the daytime and declined in the evening gradually. Correspondingly, the content of flavonoids showed an opposite variation. Both of them displayed a circadian rhythm with a negative correlation (r≥−0.7). In addition, 2 heat shock proteins along with 3 AP2 transcription factors interacting with CtPRR1 protein were obtained via Yeast Two-Hybrid experiment. Conclusion CtPRR1 negatively regulated the safflower flavonoids accumulation in a circadian rhythm way, which may be affected by these interacting proteins.

Objective A state quo survey of laboratory animal resource in Guangdong province is performed to provide reference data for government management decision-making and market assessment of laboratory animals. Methods We used questionnaires focusing on the laboratory animal facilities with authorization by Guangdong Province Government, which mainly included the production and use of laboratory animals,the qualification of employees and the facilities space. Results The total production and use of laboratory animals(except for the eggs)had been increasing in the last four years. 1.57 million laboratory animals were produced and 0.754 million laboratory animals were used in 2016. There were 2352 employees,roughly the same as in 2015. The facilities space for breeding was 121008 m2,and for animal experiment was 73470 m2,which were rising in the past three years. Conclusions In order to reinforce the industry development of laboratory animals in Guangdong province,some suggestions were given in our study,such as facilitating the application of superiority resource including non-human primate and aquatic laboratory animals,supporting the standardization production of several scarce mice and rats, improving relevant employees' overall level and constructing laboratory animal facilities sharing platform.

Objective: To investigate the association between etheno-DNA adduct and the promoter of DNA methylation levels of cyclin dependent kinase inhibitor 2A (P16), Ras association domain family 1 (RASSF1A) and O-6-methylguanine-DNA methyltransferase (MGMT) in workers with occupational exposure to diesel engine exhaust (DEE). Methods: We recruited 124 diesel engine testing workers as DEE exposure group and 112 water pump operator in the same area as control group in Henan province in 2012 using cluster sampling. The demographic data were obtained by questionnaire survey; urine after work and venous blood samples were collected from each subject. The urinary etheno-DNA adducts were detected using UPLC-MS/MS, including 1,N6-etheno-2'-deoxyadenosine (εdA) and 3,N4-etheno-2'-deoxycytidine(εdC). The DNA methylation levels of P16, RASSF1A, and MGMT were evaluated using bisulfite-pyrosequencing assay. The percentage of methylation was expressed as the 5-methylcytosine (5mC) over the sum of cytosines (%5mC). Spearman correlation and multiple linear regression were applied to analyze the association between etheno-DNA adducts and DNA methylation of P16, RASSF1A, and MGMT. Results: The median (P₂₅-P₇₅) of urinary εdA level was 230.00 (98.04-470.91) pmol/g creatinine in DEE exposure group, and 102.10 (49.95-194.48) creatinine in control group. The level of εdA was higher in DEE exposure group than control group (P<0.001). DNA methylation levels of P16, RASSF1A and MGMT were 2.04±0.41, 2.19 (1.94-2.51), 2.22 (1.94-2.46)%5mC in exposure group, and 2.19±0.40, 2.41 (2.11-2.67), 2.44 (2.15-2.91)%5mC in control group. DNA methylation levels were lower in exposure group (P values were 0.005, 0.002 and 0.001, respectively). Spearman correlation analysis showed that DNA methylation levels of P16, RASSF1A, and MGMT were negative associated with urinary εdA level (r values were -0.155, -0.137, and -0.198, respectively, P<0.05). No significant correlation was observed between the εdC level and any measured DNA methylation levels (P>0.05) . Multiple linear regression confirmed the negative correlation between εdA and DNA methylation levels of P16, RASSF1A, and MGMT in non-smoking group (β (95%CI) was -0.068 (-0.132--0.003), -0.082 (-0.159--0.004) and -0.048 (-0.090--0.007), P values were 0.039, 0.039 and 0.024, respectively). Moreover, εdC was negative associated with DNA methylation level of MGMT in non-smoking group (β (95%CI) was -0.094 (-0.179--0.008), P=0.032). Conclusion DEE exposure could induce the increased of εdA and decreased of DNA methylation levels of P16, RASSF1A and MGMT.

Objective To study the effects of positive urine glucose on maternal pregnancy.Methods A total of 1 338 pregnant women were tested urine glucose.On the basis of their urine-glucose level,two groups were devided:experimental,68 cases of posi-tive urine-glucose gravidas,control group,199 cases of negative urine-glucose gravidas.Analyze on the outcome of pregnancy of the two groups.Results Compared between positive group and control group,the incidence of died(abnormal)fetus,gestational hyper-tension,abnormal amniotic fluid,fetal distress,and fetal macrosomia were not statistically different(P >0.05),but the incidence of premature rupture of membranes was statistically different (P <0.05 ).Conclusion Positive glucose urine of gravidas might in-crease the risk of premature rupture of membranes,positive urine glucose detected during pregnancy should be highly valued.

Background Measurement of corneal thickness is important for the early diagnosis and treatment of some eye disorders,including corneal diseases and refractive errors.However,the corneal parameters from schoolage population are rarely reported.Objective The aim of this survey was to characterize the central corneal thickness (CCT),minimum corneal thickness (MCT) and paracentral corneal thickness in healthy Chinese schoolage population.Methods A cross-sectional study was designed in this study.Children aged 7 to 15 years with the diopter of-3.00 D to +3.00 D were recruited from two primary schools in Baoshan district in Shanghai based on random cluster sampling under the approval of Shanghai First People's Hospital and informed consent of child custodian.Routine examinations were firstly performed to determine the healthy participants.CCT (within 2 mm range away the corneal vertex),MCT and paracentral corneal thicknesses (2 to 5 mm zone away the cornea vertex in superior,inferior,nasal and temporal quadrants) were then measured by RTVue Fourier optical coherence tomography (OCT) for the comparison between both eyes and different gender.The subjects were grouped into the 7-9,10-12 and 13-15 years groups,and the correlations between age and CCT,MCT and paracental corneal thicknesses were analyzed.The coordinate position of the thinnest cornea was determined.Results A total of 147 children were enrolled in the study.The mean CCT value of the right eyes was (537.77±29.33) μm,and that of the left eyes was (539.22±29.16) μm,showing a significant difference between them (t =-3.21,P =0.00).The paracentral corneal thicknesses of the right and left eyes were (565.52±30.11) μm and (568.42±31.07) pm in the superior quadrant,and those in the temporal quadrant were (549.01 ±30.46) μm and (547.24±30.23) μm,with significant differences between them (t =-2.47,P =0.01 ; t =2.12,P =0.04).No significant difference was found in the CCT,MCT,paracentral corneal thicknesses from various quadrants (all at P＞0.05).In addition,no considerably correlation was seen between age and CCT,MCT and paracentral corneal thickness (all at P＞0.05).The thinnest cornea area was located in the inferotemporal region in 40.82％ right eyes and 57.82％ left eyes.The distance of thinnest cornea area away corneal vertex was (0.62±0.33)mm in the right eyes and (0.91±0.63)mm in the left eyes,with a significant difference between them (t =-5.17,P =0.00).Conclusions The central,superior and temporal corneal thicknesses are significantly different between the right and the left eyes among healthy Chinese school-age children,but corneal thickness change is not associated with age or gender.The thinnest corneal area does not locate at the vertex.

Objective To explore the status and risk factors of the dyslipidemia among health examination population of Lanzhou for providing the intervention measures .Methods According to the stratified cluster random sampling method ,4 505 health exami-nation individuals were recruited for the study from 5 hospitals in the Lanzhou region through questionnaire ,biochemical analysis . Results Prevalence of dyslipidemia of the population was 45 .79% ,high TG was the main type .The level of serum TC ,TG ,HDL-C and LDL-C were (5 .27 ± 1 .08) ,(1 .74 ± 1 .38) ,(1 .41 ± 0 .43) and (2 .83 ± 0 .82)mmol/L .The prevalence was 53 .49% in male , and 34 .93% in female .The prevalence was higher among the group of 35- <45 years old for male and 55- <65 years old for fe-male .The level of HDL-C was low among young people .There was aggregation of risk factors among the participants with dyslipi-demia .Non-conditional logistic regression analysis showed that the risk factors were age (OR=1 .701) ,overweight (OR=5 .560) , abdominal obesity(OR=2 .398) ,smoking(OR=0 .545) ,intake of greasy diet(OR=5 .313) ,sleep quality(OR=2 .005) and diastolic blood pressure(OR=3 .061) .Conclusion Lipid disorders becomes a serious problem in the health examination population ,measures such as rational diet ,weight control ,sleep improvement ,pressure control and quiting smoking must be taken .

10.3969/j.issn.2095-4344.2013.25.005