Various functions of detoxification enzymes against insecticides in Nilaparvata lugens selected by toxicity assays and RNAi methods.

The brown planthopper (BPH), Nilaparvata lugens is a devastating agricultural pest of rice, and they have developed resistance to many pesticides. In this study, we assessed the response of BPH nymphs to nitenpyram, imidacloprid, and etofenprox using contact and dietary bioassays, and investigated the underlying functional diversities of BPH glutathione-S-transferase (GST), carboxylesterase (CarE) and cytochrome P450 monooxygenase (P450) against these insecticides. Both contact and ingestion toxicity of nitenpyram to BPH were significantly higher than either imidacloprid or etofenprox. Under the LC50 concentration of each insecticide, they triggered a distinct response for GST, CarE, and P450 activities, and each insecticide induced at least one detoxification enzyme activity. These insecticides almost inhibited the expression of all tested GST, CarE, and P450 genes in contact bioassays but induced the transcriptional levels of these genes in dietary bioassays. Silencing of NlGSTD2 expression had the greatest effect on BPH sensitivity to nitenpyram in contact test and imidacloprid in dietary test. The sensitivities of BPH to insecticide increased the most in the contact test was etofenprox after silencing of NlCE, while the dietary test was nitenpyram. Knockdown of NlCYP408A1 resulted in BPH sensitivities to insecticide increasing the most in the contact test was nitenpyram, while the dietary test was imidacloprid. Taken together, these findings reveal that NlGSTD2, NlCE, and NlCYP408A1 play an indispensable role in the detoxification of the contact and ingestion toxicities of different types of insecticides to BPH, which is of great significance for the development of new strategies for the sucking pest control.

Effect of green flickering light on myopia development and expression of M1 muscarinic acetylcholine receptor in guinea pigs.

AIM: To investigate the effects of green flickering light on refractive development and expression of muscarinic acetylcholine receptor (mAChR) M1 in the eyes of guinea pigs. METHODS: Thirty guinea pigs (15-20 days old) were randomly divided into three groups (n=10/group). Animals in group I were raised in a completely closed carton with green flickering light illumination. Those in group II were kept in the open top closed carton under normal natural light. Guinea pigs were raised in a sight-widen cage under normal natural light in group III. The refractive status and axial length were measured before and after 8 weeks' illumination. Moreover, total RNA extracted from retinal, choroidal, and scleral tissues were determined by real-time reverse transcription polymerase chain reaction (RT-PCR). The expressions of the receptor M1 were also explored in the retina, choroid, and sclera using immunohistochemistry. RESULTS: There was a remarkable reduction in refractive error and increase in axial length after 8-weeks' green flickering light stimulation (P<0.001). The expression of M1 receptor mRNA in sclera and retina in myopia group were remarkably lower than that in group II and III (P<0.01). Significant reduced expression of M1 receptor stimulated by green flickering light in retina and sclera tissues were also observed (P<0.05). However, there was no M1 receptor expression in choroid in 3 groups. CONCLUSION: Myopia can be induced by 8 weeks' green flickering light exposure in the animal model. M1 receptor may be involved causally or protectively in myopia development.

Genetically confirmed familial hypercholesterolemia in outpatients with hypercholesterolemia.

BACKGROUND: Familial hypercholesterolemia (FH) is an autosomal dominant disorder of lipoprotein metabolism which can lead to premature coronary heart disease (pCHD). There are about 3.8 million potential FH patients in China, whereas the clinical and genetic data of FH are limited. METHODS: Dutch Lipid Clinic Network (DLCN) criteria was used to diagnose FH in outpatients with hypercholesterolemia. Resequencing chip analysis combined with Sanger sequencing validation were used to identify mutations in the definite FH patients according to DLCN criteria. In silico analysis was conducted in mutations with previously unknown pathogenicity. Then, the novel mutant receptors were transfected into human embryo kidney 293 (HEK-293) cells. The binding and the internalization activities of the mutant receptors were analyzed by flow cytometry. RESULTS: The prevalence of definite FH in outpatients with hypercholesterolemia in this study is 3.2%. Using genetic testing, one homozygous FH (HoFH), one heterozygous FH (HeFH) and three compound heterozygous FH patients were confirmed. Eight mutations in low-density lipoprotein receptor (LDLR) gene were identified, in which c.357delG was a novel mutation and co-segregated with the FH phenotype. Bioinformatic analysis confirmed that c.357delG was a pathogenic mutation. Furthermore, when compared with the wild-type LDLRs by flow cytometry analysis, the binding and internalization activities of c.357delG mutant LDLRs were reduced by 35% and 49%, respectively. CONCLUSIONS: This study identified eight LDLR gene mutations in five patients with definite FH, in which c.357delG is a novel pathogenic mutation. These findings increase our understanding of the genetic spectrum of FH in China.

Second-hand smoke prevalence in 252 regions of South Korea in three exposure locations.

PURPOSE: The association between regional contextual factors and second-hand smoke (SHS) prevalence is important, but is unclear. This study investigated the changes of SHS prevalence from 2011 to 2013 and explored the correlations of SHS prevalence and regional contextual factors by exposure location. METHODS: The data were obtained from the 2011 and 2013 Korean Community Health Survey and the Development of Health Indicators for Community Health Ranking report. A t-test was used to examine and compare the SHS prevalence in 2011 and 2013 by exposure location (home, workplace, and public places). A correlation analysis and linear regression were used to investigate the impacts of the regional variables on SHS prevalence by location. RESULTS: The prevalence of SHS in all three locations had a decreased trend overall, but remained high in public locales. There were clear differences in the prevalence of SHS and its change by region between 2011 and 2013. The SHS prevalence in the workplace and public places had increased in the high social and economic characteristic regions, compared to the other regions in 2 years. The SHS had an increased trend in regions featuring a high level of socioeconomic development. It was observed that regional factors affecting SHS prevalence differed in the three locations. CONCLUSION: The differences and changes of regional SHS prevalence by location were influenced by specific social contextual factors of the particular region. Local government initiatives regarding special SHS protective measures or tailored regulations, according to specific regional status and location, are recommended, with attention to high socioeconomic regions in particular.

Acute myopathy following intra-muscular injection of compound betamethasone: A case report.

RATIONALE: We report a case of acute steroid myopathy in a patient with eczema receiving one dose of intra-muscular injection of Compound betamethasone. PATIENT CONCERNS: Acute steroid myopathy (ASM) is usually caused by exogenous corticosteroids, and typically, occurs with therapy using intravenous corticosteroids at high doses. DIAGNOSES: The patient was considered as a diagnosis of acute steroid myopathy. INTERVENTIONS: The patient was treated with non-steroid anti-inflammatory drug and other symptomatic therapy. OUTCOMES: ASM was gradually improved after 2 weeks symptomatic treatment and completely recovered after one-month treatment. LESSONS: The diagnosis of steroid myopathy is a clinical diagnosis based on characteristic symptoms. Higher dose of steroids, especially fluorinated steroids, for longer periods of time increases the risk of steroid-induced myopathy.

The use of targeted exome sequencing in genetic diagnosis of young patients with severe hypercholesterolemia.

Familial hypercholesterolemia (FH) is an autosomal dominant disorder. Although genetic testing is an important tool for detecting FH-causing mutations in patients, diagnostic methods for young patients with severe hypercholesterolemia are understudied. This study compares the target exome sequencing (TES) technique with the DNA resequencing array technique on young patients with severe hypercholesterolemia. A total of 20 unrelated patients (mean age 14.8 years) with total cholesterol > 10 mmol/L were included. 12 patient samples were processed by DNA resequencing array, 14 patient samples were processed by TES, and 6 patient samples were processed by both methods. Functional characterization of novel mutations was performed by flow cytometry. The mutation detection rate (MDR) of DNA resequencing array was 75%, while the MDR of TES was 100%. A total of 27 different mutations in the LDLR were identified, including 3 novel mutations and 8 mutations with previously unknown pathogenicity. Functional characterization of c.673delA, c.1363delC, p.Leu575Phe and p.Leu582Phe variants found that all of them are pathogenic. Additionally, 7 patients were diagnosed with Heterozygous FH (HeFH) in which lipid levels were significantly higher than common HeFH patients. This data indicates that TES is a very efficient tool for genetic diagnosis in young patients with severe hypercholesterolemia.

Affecting Factors of Secondhand Smoke Exposure in Korea: Focused on Different Exposure Locations.

Exposure to secondhand smoke (SHS) not only can cause serious illness, but is also an economic and social burden. Contextual and individual factors of non-smoker exposure to SHS depend on location. However, studies focusing on this subject are lacking. In this study, we described and compared the factors related to SHS exposure according to location in Korea. Regarding individual factors related to SHS exposure, a common individual variable model and location-specific variable model was used to evaluate SHS exposure at home/work/public locations based on sex. In common individual variables, such as age, and smoking status showed different relationships with SHS exposure in different locations. Among home-related variables, housing type and family with a single father and unmarried children showed the strongest positive relationships with SHS exposure in both males and females. In the workplace, service and sales workers, blue-collar workers, and manual laborers showed the strongest positive association with SHS exposure in males and females. For multilevel analysis in public places, only SHS exposure in females was positively related with cancer screening rate. Exposure to SHS in public places showed a positive relationship with drinking rate and single-parent family in males and females. The problem of SHS embodies social policies and interactions between individuals and social contextual factors. Policy makers should consider the contextual factors of specific locations and regional and individual context, along with differences between males and females, to develop effective strategies for reducing SHS exposure.

Characterization of the unique Chinese W483X mutation in the low-density lipoprotein-receptor gene in young patients with homozygous familial hypercholesterolemia.

BACKGROUND: Recent guidelines suggest that more attention should be focused on children with homozygous familial hypercholesterolemia (HoFH). China may have 3.8 million potential FH patients, but there are limited data focused on HoFH children. OBJECTIVE: We systematically analyzed the characteristic phenotype and the relationship between the genotype and the phenotype in HoFH children with the unique Chinese W483X mutation in the low-density lipoprotein (LDL)-receptor gene. METHODS: A systematic retrospective analysis of the lipid and cardiovascular characteristics of HoFH patients in the atherosclerosis clinic of Beijing Anzhen Hospital was performed. The W483X mutation was confirmed using DNA sequencing of the patients and their parents. RESULTS: Two HoFH and 9 compound heterozygous patients (mean age = 14.7 years) with 2 novel mutations, Q254X and c.1363delC, were found. In total, 81.8% of the patients were from southern China. All the patients had xanthoma, and the average TC and LDL-C levels were 16.8 and 14.4 mmol/L, respectively. Echocardiography showed that 63.6% of the patients had aortic calcification, and 54.5% had mild regurgitation of the aortic valve. The coronary flow velocity reserve had a mean value of 2.12, and the cIMT was 0.17 cm. The follow-up period was between 3 months and 8 years. Although all the patients began the lipid-lowering treatment, 2 patients died because of severe cardiovascular disease. The LDL-C levels of 6 patients were slightly decreased by approximately 21% and remained far from the target values, and the other 3 patients' LDL-C levels increased by 13%. CONCLUSIONS: The results suggest that younger HoFH patients with W483X mutations had a severe phenotype and should receive more aggressive treatment.

The distribution and characteristics of LDL receptor mutations in China: A systematic review.

Familial hypercholesterolemia (FH) is a common and serious dominant genetic disease, and its main pathogenic gene is the low-density lipoprotein receptor (LDLR) gene. This study aimed to perform a systematic review of LDLR mutations in China. Using PubMed, Embase, Wanfang (Chinese), the Chinese National Knowledge Infrastructure (Chinese), and the Chinese Biological and Medical database (Chinese), public data were limited to December 2014. The Medical Subject Headings terms and the following key words were used: "familial hypercholesterolemia", "Chinese", "China", "Hong Kong", and "Taiwan". A total of 74 studies including 295 probands with 131 LDLR mutations were identified. Most of the mutations were located in exon 4 of LDLR and approximately 60% of the mutations were missense mutations. Thirty new mutations that were not recorded in the LDLR databases were found. In silico analysis revealed that most of the mutations were pathogenic. The primary LDLR mutations were C308Y, H562Y, and A606T, and all of the mutations had functional significance. Prevalence data suggest that there are nearly 3.8 million FH patients in China, although reported numbers are much smaller, suggesting that FH is widely misunderstood. This systematic review provides information that is specific to China for inclusion in the international FH database.

Identification of the gene defect responsible for severe hypercholesterolaemia using whole-exome sequencing.

Familial hypercholesterolaemia (FH) is a serious genetic metabolic disease. We identified a specific family in which the proband had typical homozygous phenotype of FH, but couldn't detect any mutations in usual pathogenic genes using traditional sequencing. This study is the first attempt to use whole exome sequencing (WES) to identify the pathogenic genes in Chinese FH. The routine examinations were performed on all parentage members, and WES on 5 members. We used bioinformatics methods to splice and filter out the pathogenic gene. Finally, Sanger sequencing and cDNA sequencing were used to verify the candidate genes. Half of parentage members had got hypercholesterolaemia. WES identified LDLR IVS8[-10] as a candidate mutation from 222,267 variations. The Sanger sequencing showed proband had a homozygous mutation inherited from his parents, and this loci were cosegregated with FH phenotype. The cDNA sequencing revealed that this mutations caused abnormal shearing. This mutation was first identified in Chinese patients, and this homozygous mutation is a new genetic type of FH. This is the first time that WES was used in Chinese FH patients. We detected a novel genetic type of LDLR homozygous mutation. WES is powerful tools to identify specific FH families with potentially pathogenic gene mutations.

Risk Factors of Cardiovascular Disease and Their Related Socio-Economical, Environmental and Health Behavioral Factors: Focused on Low-Middle Income Countries- A Narrative Review Article.

BACKGROUND: In order to decrease the burden of cardiovascular disease (CVD), social determinants for CVD risk factors have been extensively studied in developed countries. However, few studies about them have been performed in low-middle-income countries. This study describes factors related to CVD risk factors in low-middle-income countries at a national level. METHODS: Data were assembled from international databases for 47 low-middle-income countries and were collected from various sources including WHO, World Bank, and previous studies. Coefficient estimates between male and female CVD risk factor prevalence and each independent variable were calculated via linear regression. RESULTS: Statistically significant inverse associations were observed between adult literacy rate and systolic blood pressure, blood glucose. Pump price for gasoline was negatively associated with blood glucose also. Associations for female unemployment, adult literacy rate, paved roads and urban population, alcohol and western diet were positively associated with CVD risk factors. Unemployment, urban population and alcohol were positively associated with CVD risk factors in males. CONCLUSION: The effectiveness of intervention program for the prevention of cardiovascular disease in populations in developing countries should be explored, and more attention should be given to women.

Perception of Lay People Regarding Determinants of Health and Factors Affecting It: An Aggregated Analysis from 29 Countries.

BACKGROUND: This study aimed to evaluate the perception of lay people regarding determinants of health at global level and factors affecting it. METHODS: Data was collected from International Social Survey Program (ISSP) and World Bank website. Multilevel regression analysis was done and lay people's perception regarding health behavior, environment, poverty and genes as health determinants was assessed. Various socio demographic factors were used as independent variables. RESULTS: The highest percentage of people agreed environment as determinant of health. An inverse relationship was observed between GNI quartiles and an individual's agreement with poverty, health behavior, and environment as health determinant. There was a significant negative association of females with health damaging behavior (P<0.05) and positive association with environment and genes (P<0.05) as health determinants. Elderly people agreed with poverty as determinant of health (P<0.05). GNI was negatively related to environment (P<0.05) and poverty (P<0.05) as health determinant. CONCLUSION: The common public is now becoming aware of a broadened concept of health and people belonging to different backgrounds have different perceptions regarding determinants of health. Our results show that highest percentage of people agreed with environment as determinant of health, which is consistent with scientific view of increased burden of disease, caused by environmental factors. Thus, tailored health programs and policies that address an individual's specific problems are likely to induce a change in behavior and attitude, hence decreasing the disease burden.

Use of targeted exome sequencing in genetic diagnosis of Chinese familial hypercholesterolemia.

Familial hypercholesterolemia is an autosomal dominant inherited disease characterized by elevated plasma low-density lipoprotein cholesterol (LDL-C). It is mainly caused by mutations of the low-density lipoprotein receptor (LDLR) gene. Currently, the methods of whole genome sequencing or whole exome sequencing for screening mutations in familial hypercholesterolemia are not applicable in China due to high cost. We performed targeted exome sequencing of 167 genes implicated in the homozygous phenotype of a proband pedigree to identify candidate mutations, validated them in the family of the proband, studied the functions of the mutant protein, and followed up serum lipid levels after treatment. We discovered that exon 9 c.1268 T>C and exon 8 c.1129 T>G compound heterozygous mutations in the LDLR gene in the proband derived from the mother and father, respectively, in which the mutation of c.1129 T>G has not been reported previously. The mutant LDL-R protein had 57% and 52% binding and internalization functions, respectively, compared with that of the wild type. After 6 months of therapy, the LDL-C level of the proband decreased by more than 50% and the LDL-C of the other family members with heterozygous mutation also reduced to normal. Targeted exome sequencing is an effective method for screening mutation genes in familial hypercholesterolemia. The exon 8 and 9 mutations of the LDLR gene were pedigree mutations. The functions of the mutant LDL-R protein were decreased significantly compared with that of the wild type. Simvastatin plus ezetimibe was proven safe and effective in this preschool-age child.

[Research progression of LDLR mutations in Chinese Familial hypercholesterolemia].

Familial hypercholesterolemia (FH), one monogenic autosomal dominant disease, mainly results from genetic defects in the low-density lipoprotein receptor (LDLR) gene, which leads to the reduction or absence of cell surface LDLR, disorder of cholesterol metabolism, and cholesterol deposition in different tissues and organs. FH is a common metabolic disease clinically characterized by the presence of xanthomas and premature coronary heart disease. To date, about 1 741 variants have been identified in gene LDLR, among which 108 variants were identified in Chinese FH patients. To better understand the features of LDLR gene mutations and help to FH diagnosis and therapy, this review provides a comprehensive overview of LDLR gene mutations in Chinese FH patients.

Transient versus prolonged hyperlocomotion following lateral fluid percussion injury in mongolian gerbils.

Posttraumatic hyperactivity is a neurobehavioral symptom commonly seen in patients after traumatic brain injury (TBI). No useful animal model has yet been established for evaluation of this important symptom. We induced either mild (MILD, 0.7-0.9 atm) or moderate (MOD, 1.3-1.6 atm) lateral fluid percussion injury (LFPI) in Mongolian gerbils. Open-field and T-maze tests were used during a 7-day period after the trauma. All animals were perfusion fixed for histopathological examinations. Transient locomotor hyperactivity was found with a peak at 6 hr after injury in the MILD animals, whereas MOD animals showed prolonged and severe hyperlocomotion throughout the 7-day posttrauma period (P < 0.0001). Interestingly, the temporal profile of the posttraumatic hyperactivity was similar to that of the working memory deficit in both injury groups. Histological examination revealed significant neural tissue damages, including cortical necrosis, white matter rarefaction, and neuronal loss in the hippocampus in the ipsilateral hemisphere of the MOD animals, vs. only negligible changes in the MILD animals. Correlation analysis revealed that the volume of white matter lesions was significantly correlated with both posttraumatic hyperactivity (r = 0.591, P < 0.01) and working memory deficit (r = -0.859, P < 0.0001). Taken together, our findings confirm the successful reproduction of posttraumatic hyperactivity following experimental TBI. The posttraumatic hyperlocomotion probably shared pathomechanisms common to those of cognitive dysfunction caused by LFPI, supporting the speculation from previous studies that some neurobehavioral abnormities intimately correlate with TBI-induced cognitive dysfunction. Histopathologically, significant involvement of white matter damage in the posttraumatic functional deficits was indicated.