PGG.SV: a whole-genome-sequencing-based structural variant resource and data analysis platform.

Structural variations (SVs) play important roles in human evolution and diseases, but there is a lack of data resources concerning representative samples, especially for East Asians. Taking advantage of both next-generation sequencing and third-generation sequencing data at the whole-genome level, we developed the database PGG.SV to provide a practical platform for both regionally and globally representative structural variants. In its current version, PGG.SV archives 584 277 SVs obtained from whole-genome sequencing data of 6048 samples, including 1030 long-read sequencing genomes representing 177 global populations. PGG.SV provides (i) high-quality SVs with fine-scale and precise genomic locations in both GRCh37 and GRCh38, covering underrepresented SVs in existing sequencing and microarray data; (ii) hierarchical estimation of SV prevalence in geographical populations; (iii) informative annotations of SV-related genes, potential functions and clinical effects; (iv) an analysis platform to facilitate SV-based case-control association studies and (v) various visualization tools for understanding the SV structures in the human genome. Taken together, PGG.SV provides a user-friendly online interface, easy-to-use analysis tools and a detailed presentation of results. PGG.SV is freely accessible via https://www.biosino.org/pggsv.

Mediation of metabolic syndrome in the association between long-term exposure to particulate matter and incident cardiovascular disease: Evidence from a population-based cohort in Chengdu.

BACKGROUND: Particulate matter (PM) exposure has been linked with cardiovascular disease (CVD) and metabolic syndrome (MetS), the latter characterized by concurrent multiple metabolic disorders. As a result, the mechanisms assumption from PM to CVD through MetS have emerged, thus requiring further epidemiological evidence. This cohort study aimed to assess whether MetS mediates the associations of PM with CVD risk. METHODS: This study included 14,195 participants from the Chengdu cohort of the China Multi-Ethnic Cohort (CMEC) study in 2018. The primary outcome of incident CVD diagnoses was identified using matched hospital records from the Health Information Center of Sichuan Province. Residence-specific levels of PM with aerodynamic diameters of ≤ 1 µm (PM1), ≤ 2.5 µm (PM2.5), and ≤ 10 µm (PM10) were estimated by spatiotemporal models. Causal mediation analyses were applied to evaluate the indirect effect of MetS. RESULTS: Increased exposure levels to PM were significantly associated with MetS and CVD. Mediation analyses indicated that the associations between PM exposure and CVD were mediated by MetS, with the proportion of multiple mediations being 19.3%, 12.1%, and 13.5% for PM1, PM2.5, and PM10, respectively. Further moderated mediation analyses suggested that male, overweight individuals, alcohol drinkers, and those suffering from indoor air pollution may experience more significant adverse effects from PM exposure on CVD via MetS than others. CONCLUSIONS: Our findings suggest that MetS partially mediates the association between long-term exposure to PM and CVD. These mediation effects appear to be amplified by demographic characteristics and unhealthy lifestyles.

Prognostic factor analysis in patients with early-stage nasopharyngeal carcinoma in the USA.

Aim: To evaluate independent risk factors specific for early-stage nasopharyngeal carcinoma (NPC). Methods: A total of 566 patients with early-stage NPC from 2004 to 2019 were identified using the Surveillance, Epidemiology and End Results database. Results: Older ages (70-79 and >80 years) were independent risk factors, with hazard ratios of 1.961 and 5.011, respectively. The hazard ratio for early-stage NPC in Asian and Pacific Islander residents (0.475) was lower than that for White residents. A tumor size <3 cm was a protective factor for overall and cancer-specific survival in the current study. Conclusion: In patients with early-stage NPC, age >70 years, race and tumor size were independent prognosticators for cancer-specific survival.

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Objective: To investigate the status of medication adherence and various types of risky behaviors of schizophrenia patients in a certain area of western China and to explore accordingly the correlation between the two. Methods: A total of 292 667 patients with schizophrenia were enrolled in a follow-up survey between 2006 and 2018. In addition, based on the outcome-wide analysis strategy, a multivariate Cox proportional risk regression model was used to estimate and compare the impact of medication adherence on different types of risky behaviors in schizophrenia patients. Results: In this 13-year prospective cohort, 65 175 patients (31.4%) showed good medication adherence, while 142 394 patients (68.6%) showed poor medication adherence. The incidence rates of various risky behaviors during the follow-up period were as follows, minor nuisances, 12.25%, violation of the Law of the People's Republic of China on Penalties for Administration of Public Security (APS law), 3.82%, violation of criminal law, 0.94%, suicide completed, 0.28%, self-harm, 1.42%, and attempted suicide, 0.82%. Schizophrenia patients who had poor medication adherence had higher risks of committing violence against others and self-inflicted injury compared to patients with good medication adherence did, with the associated effects being minor nuisances (hazard ratio [HR]=1.31, 95% confidence interval [CI]: 1.27-1.35), violation of APS law (HR=1.47, 95% CI: 1.38-1.56), violation of criminal law (HR=1.17, 95% CI: 1.05-1.31), and self-harm (HR=1.43, 95% CI: 1.32-1.56), respectively, while the risk of suicide completed is lower in schizophrenia patients with poor medication adherence than that in patients with good medication adherence (HR=0.56, 95% CI: 0.47-0.66). There was no statistically significant association between attempted suicide and medication adherence. Conclusion: There are variations in the direction and strength of the association between medication adherence and different types of risky behaviors and further research is needed to elucidate the mechanisms of the association.

MVDINET: A Novel Multi-Level Enzyme Function Predictor With Multi-View Deep Interactive Learning.

As a class of extremely significant of biocatalysts, enzymes play an important role in the process of biological reproduction and metabolism. Therefore, the prediction of enzyme function is of great significance in biomedicine fields. Recently, computational methods for predicting enzyme function have been proposed, and they effectively reduce the cost of enzyme function prediction. However, there are still deficiencies for effectively mining the discriminant information for enzyme function recognition in existing methods. In this study, we present MVDINET, a novel method for multi-level enzyme function prediction. First, the initial multi-view feature data is extracted by the enzyme sequence. Then, the above initial views are fed into various deep specific network modules to learn the depth-specificity information. Further, a deep view interaction network is designed to extract the interaction information. Finally, the specificity information and interaction information are fed into a multi-view adaptively weighted classification. We compressively evaluate MVDINET on benchmark datasets and demonstrate that MVDINET is superior to existing methods.

Transcriptional-profile changes in the medial geniculate body after noise-induced tinnitus.

Tinnitus is a disturbing condition defined as the occurrence of acoustic hallucinations with no actual sound. Although the mechanisms underlying tinnitus have been explored extensively, the pathophysiology of the disease is not completely understood. Moreover, genes and potential treatment targets related to auditory hallucinations remain unknown. In this study, we examined transcriptional-profile changes in the medial geniculate body after noise-induced tinnitus in rats by performing RNA sequencing and validated differentially expressed genes via quantitative polymerase chain reaction analysis. The rat model of tinnitus was established by analyzing startle behavior based on gap-pre-pulse inhibition of acoustic startles. We identified 87 differently expressed genes, of which 40 were upregulated and 47 were downregulated. Pathway-enrichment analysis revealed that the differentially enriched genes in the tinnitus group were associated with pathway terms, such as coronavirus disease COVID-19, neuroactive ligand-receptor interaction. Protein-protein-interaction networks were established, and two hub genes (Rpl7a and AC136661.1) were identified among the selected genes. Further studies focusing on targeting and modulating these genes are required for developing potential treatments for noise-induced tinnitus in patients.

Associations of Long-Term Exposure to Fine Particulate Constituents With Cardiovascular Diseases and Underlying Metabolic Mediations: A Prospective Population-Based Cohort in Southwest China.

BACKGROUND: The health effects of particulate matter with an aerodynamic diameter ≤2.5 µm (PM2.5) might differ depending on compositional variations. Little is known about the joint effect of PM2.5 constituents on metabolic syndrome and cardiovascular disease (CVD). This study aims to evaluate the combined associations of PM2.5 components with CVD, identify the most detrimental constituent, and further quantify the mediation effect of metabolic syndrome. METHODS AND RESULTS: A total of 14 427 adults were included in a cohort study in Sichuan, China, and were followed to obtain the diagnosis of CVD until 2021. Metabolic syndrome was defined by the simultaneous occurrence of multiple metabolic disorders measured at baseline. The concentrations of PM2.5 chemical constituents within a 1-km2 grid were derived based on satellite- and ground-based detection methods. Cox proportional hazard models showed that black carbon, organic matter (OM), nitrate, ammonium, chloride, and sulfate were positively associated with CVD risks, with hazard ratios (HRs) ranging from 1.24 to 2.11 (all P<0.05). Quantile g-computation showed positive associations with 4 types of CVD risks (HRs ranging from 1.48 to 2.25, all P<0.05). OM and chloride had maximum weights for CVD risks. Causal mediation analysis showed that the positive association of OM with total CVD was mediated by metabolic syndrome, with a mediation proportion of 1.3% (all P<0.05). CONCLUSIONS: Long-term exposure to PM2.5 chemical constituents is positively associated with CVD risks. OM and chloride appear to play the most responsible role in the positive associations between PM2.5 and CVD. OM is probably associated with CVD through metabolic-related pathways.

Does awareness of diabetic status increase risk of depressive or anxious symptoms? Findings from the China Multi-Ethnic cohort (CMEC) study.

INTRODUCTION: People with diabetes mellitus (DM) have increased risk of depressive symptoms (DS) or anxious symptoms (AS). This study explores whether awareness of DM will contribute to prevalence of DS or AS. METHODS: The baseline data including 81,717 adults from Southwest China was analyzed. DS and AS were assessed using PHQ-2 and GAD-2. Exposures were defined as 1) having self-reported physician diagnosis of diabetes (self-reported DM), 2) no prior diagnosis of diabetes but meeting diagnostic criteria (newly diagnosed DM), 3) having self-reported physician diagnosis or meeting criteria of non-diabetic diseases (non-diabetic patients), 4) healthy participants. Generalized linear mixed models were used to assess impact of presence and awareness of DM on DS or AS, adjusting for regional and individual related factors. RESULTS: The prevalence of DS in self-reported DM, newly diagnosed DM, non-diabetic patient and healthy participants was 7.08 %, 4.30 %, 5.37 % and 3.17 %. The prevalence of AS was 7.80 %, 5.77 %, 6.37 % and 3.91 %. After adjusting for related factors, compared with healthy participants, self-reported DM and non-diabetic patients were associated with DS [AORDS, self-reported = 1.443(1.218,1.710), AORDS, nondiabetic patients = 1.265(1.143,1.400)], while the association between newly diagnosed DM and DS was not statistically significant. The associations between self-reported DM, newly diagnosed DM, non-diabetic patients and AS were all statistically significant. LIMITATIONS: DS and AS were assessed through self-report and may suffer recall or information bias. CONCLUSIONS: The association between awareness of diabetes and DS/AS suggests to pay attention to distinguish between self-reported and newly diagnosed DM and screening for DS and AS in diabetic population.

Study on tinnitus-related electroencephalogram microstates in patients with vestibular schwannomas.

Tinnitus is closely associated with cognition functioning. In order to clarify the central reorganization of tinnitus in patients with vestibular schwannoma (VS), this study explored the aberrant dynamics of electroencephalogram (EEG) microstates and their correlations with tinnitus features in VS patients. Clinical and EEG data were collected from 98 VS patients, including 76 with tinnitus and 22 without tinnitus. Microstates were clustered into four categories. Our EEG microstate analysis revealed that VS patients with tinnitus exhibited an increased frequency of microstate C compared to those without tinnitus. Furthermore, correlation analysis demonstrated that the Tinnitus Handicap Inventory (THI) score was negatively associated with the duration of microstate A and positively associated with the frequency of microstate C. These findings suggest that the time series and syntax characteristics of EEG microstates differ significantly between VS patients with and without tinnitus, potentially reflecting abnormal allocation of neural resources and transition of functional brain activity. Our results provide a foundation for developing diverse treatments for tinnitus in VS patients.

Transcriptional Profile Changes after Noise-Induced Tinnitus in Rats.

Tinnitus is an unpleasant symptom characterized by detective hearing without the actual sound input. Despite numerous studies elucidating a variety of pathomechanisms inducing tinnitus, the pathophysiology of tinnitus is not fully understood. The genes that are closely associated with this subtype of the auditory hallucination that could be utilized as potential treatment targets are still unknown. In this study, we explored the transcriptional profile changes of the auditory cortex after noise-induced tinnitus in rats using high throughput sequencing and verification of the detected genes using quantitative PCR (qPCR). Tinnitus models were established by analyzing startle behaviors through gap pre-pulse inhibition (PPI) of the acoustic startle. Two hundred and fifty-nine differential genes were identified, of which 162 genes were up-regulated and 97 genes were down-regulated. Analysis of the pathway enrichment indicated that the tinnitus group exhibited increased gene expression related to neurodegenerative disorders such as Huntington's disease and Amyotrophic lateral sclerosis. Based on the identified genes, networks of protein-protein interaction were established and five hub genes were identified through degree rank, including Fos, Nr4a1, Nr4a3, Egr2, and Egr3. Therein, the Fos gene ranked first with the highest degree after noise exposure, and may be a potential target for the modulation of noise-induced tinnitus.

The Comorbidity of Depression and Anxiety Symptoms in Tinnitus Sufferers: A Network Analysis.

OBJECTIVE: Sufferers of tinnitus, especially of the prolonged type, frequently suffer from comorbid depression and anxiety. From the perspective of the network model, this comorbidity is thought to be an interacting system of these two symptoms. In our study, we conducted a network analysis of depression and anxiety comorbidity in tinnitus sufferers, aiming to identify the central and bridge symptoms and make informed suggestions for clinical interventions and psychotherapy. METHOD: A total of 566 tinnitus sufferers were enrolled in our study. The Patient Health Questionnaire-9 (PHQ-9) and the Generalized Anxiety Disorder 7-Item Questionnaire (GAD-7) were selected to evaluate depression and anxiety symptoms, respectively, followed by network analysis to construct the interacting networks. RESULTS: The findings identified six edges of strongest regularized partial correlations in this network. Of these, three were depression symptoms and three were anxiety symptoms. The anxiety symptoms "Unable to control worry" and "Relaxation difficulty" and the depression symptom "Feeling depressed or hopeless" had the highest expected influence centrality. The analysis results also revealed three bridge symptoms: "Afraid something awful might happen", "Feeling of worthlessness", and "Trouble concentrating". As for "Suicidal ideation", the direct relations between this symptom and "Afraid something awful might happen" and "Feeling depressed or hopeless" were the strongest. CONCLUSIONS: The central and bridge symptoms of the interacting network of depression and anxiety symptoms in tinnitus sufferers can be considered a significant transdiagnostic intervention target for the management of this comorbidity. In particular, clinical prevention and psychotherapy should be implemented, targeting the symptoms that have the strongest associations with suicidal ideation.

The mediation role of blood lipids on the path from air pollution exposure to MAFLD: A longitudinal cohort study.

BACKGROUND & AIMS: Recent cross-sectional studies found that exposure to ambient air pollution (AP) was associated with an increased risk of metabolic dysfunction-associated fatty liver disease (MAFLD). The alternation of blood lipids may explain the association, but epidemiological evidence is lacking. We aimed to examine whether and to what extent the association between long-term exposure to AP and incident MAFLD is mediated by blood lipids and dyslipidemia in a prospective cohort. METHODS: We included 6350 participants from the China Multi-Ethnic Cohort (CMEC, baseline 2018-2019, follow-up 2020-2021). Three-year average (2016-2018) of AP (PM1, PM2.5, PM10, NO2), blood lipids (TC, LDL-C, HDL-C, TG with their combinations) and incident MAFLD for each individual were assessed chronologically. Linear and logistic regression was used to assess the associations among AP, blood lipids, and MAFLD, and the potential mediation effects of blood lipids were evaluated using causal mediation analysis. RESULTS: A total of 744 participants were newly diagnosed with MAFLD at follow-up. The odds ratios of MAFLD associated with a 10 µm increase in PM1, PM2.5, and NO2 were 1.35 (95 % CI: 1.14, 1.58), 1.34 (1.10, 1.65) and 1.28 (1.14, 1.44), respectively. Blood lipids are important mediators between AP and incident MAFLD. LDL-C (Proportion Mediated: 6.9 %), non-HDL (13.4 %), HDL-C (20.7 %), LDL/HDL (30.1 %), and dyslipidemia (6.5 %) significantly mediated the association between PM2.5 and MAFLD. For PM1, the indirect effects were similar to those for PM2.5, with a larger value for the direct effect, and the mediation proportion by blood lipids was less for NO2. CONCLUSION: Blood lipids are important mediators between AP and MAFLD, and can explain 5 %-30 % of the association between AP and incident MAFLD, particularly cholesterol-related variables, indicating that AP could lead to MAFLD through the alternation of blood lipids. These findings provided mechanical evidence of AP leading to MAFLD in epidemiological studies.

Association of long-term exposure to ambient PM2.5 and its constituents with gut microbiota: Evidence from a China cohort.

Accumulating animal experiments and epidemiological studies have found that exposure to fine particulate matter (PM2.5) is associated with altered gut microbiota (GM). However, it is unclear what kind of role the PM2.5 constituents play in the PM2.5-GM association. Therefore, this study aimed to investigate the association of long-term exposure to PM2.5 and its constituents (PMcons) with GM. This study included 1583 participants from a cohort in Southwest China. Satellite remote sensing and chemical transport modelling were used to determine the yearly average concentrations of PMcons. GM data were derived from 16 s sequencing based on stool samples. Generalized propensity score weighting regression and Bayesian Kernel Machine Regression (BKMR) were used to estimate the individual and joint association of exposure to PMcons with the Shannon index. The weighted correlation analysis was used to estimate the association of PMcons with the composition of GM. The result showed that an interquartile range increase of 3-year average black carbon (BC), ammonium, nitrate, organic matter (OM), sulfate, and soil particles (SOIL) were negatively associated with Shannon index with mean difference (95 % confidence interval) being -0.144 (-0.208, -0.080), -0.141 (-0.205, -0.078), -0.126 (-0.184, -0.068), -0.117 (-0.172, -0.062), -0.153 (-0.221, -0.085), and - 0.153 (-0.222, -0.085). BKMR indicated joint exposure to PMcons was associated with decreased Shannon index, and BC had the largest posterior inclusion probability (0.578). Weighted correlation analyses indicated PMcons were associated with decreased Bacteroidetes (r = -0.204, P < 0.001 for PM2.5) and increased Proteobacteria (r = 0.273, P < 0.001 for PM2.5). These results revealed that long-term exposure to PMcons was associated with GM. BC was the most important constituent in the association, indicating that the source of BC should be controlled to mitigate the negative effects of PM2.5 on GM.

Extended high-frequency audiometric analyses of tinnitus patients exhibiting normal hearing on conventional pure-tone audiometry.

BACKGROUND: While hearing loss is the greatest risk factor associated with developing tinnitus, some tinnitus patients exhibit no hearing loss on conventional pure-tone audiometry (PTA). OBJECTIVES: This study was developed to assess whether tinnitus patients with normal hearing as measured via conventional PTA would exhibit differences from normal controls upon extended high-frequency (EHF) audiometric evaluation. METHODS: In total, 102 tinnitus patients were separated into unilateral and bilateral tinnitus groups. Age- and sex-matched controls without tinnitus were enrolled. RESULTS: No significant differences were observed when comparing EHF audiometry results in the 9-14 kHz range between controls and tinnitus patients, with only left-sided tinnitus ears exhibiting higher hearing thresholds than contralateral ears at 9, 10, and 14 kHz. Relative to normal controls, the hearing thresholds in the 2-8 kHz range for tinnitus ears were significantly increased. CONCLUSIONS AND SIGNIFICANCE: Relative to controls, tinnitus patients with normal hearing did not exhibit any significant hearing loss in the EHF range. Unexpectedly, tinnitus patients with normal hearing exhibited significant hidden hearing loss in the conventional frequency range rather than in the EHF range. For patients with normal hearing, it appears to be unnecessary to conduct EHF examinations to detect hearing loss in the EHF range.

Association between acute kidney injury and long-term mortality in patients with aneurysmal subarachnoid hemorrhage: A retrospective study.

Background: Though acute kidney injury (AKI) in the context of aneurysmal subarachnoid hemorrhage (aSAH) worsens short-term outcomes, its impact on long-term survival is unknown. Aim: We aimed to evaluate the association between long-term mortality and AKI during hospitalization for aSAH. Methods: This was a retrospective study of patients who survived >12 months after aSAH. All patients were evaluated at West China Hospital, Sichuan University, between December 2013 and June 2019. The minimum follow-up time was over 1 year. the maximum follow-up time was about 7.3 years. AKI was defined by the KDIGO (The Kidney Disease Improving Global Outcomes) guidelines, which stratifies patients into three stages of severity. The primary outcome was long-term mortality, which was analyzed with Kaplan-Meier curves and Cox proportional hazards models. Results: During this study period, 238 (9.2%) patients had AKI among 2,592 patients with aSAH. We confirmed that AKI during care for aSAH significantly increased long-term mortality (median 4.3 years of follow-up) and that risk increased with the severity of the kidney failure, with an adjusted hazard ratio (HR) of 2.08 (95% CI 1.49-2.89) for stage 1 AKI, 2.15 (95% CI 1.05-4.43) for stage 2 AKI, and 2.66 (95% CI 1.08-6.53) for stage 3 AKI compared with patients without AKI. Among patients with an AKI episode, those with renal recovery still had increased long-term mortality (HR 1.96; 95% CI 1.40-2.74) compared with patients without AKI but had better long-term outcomes than those without renal recovery (HR 0.51, 95% CI 0.27-0.97). Conclusions: Among 12-month survivors of aSAH, AKI during their initial hospitalization for aSAH was associated with increased long-term mortality, even for patients who had normal renal function at the time of hospital discharge. Longer, multidisciplinary post-discharge follow-up may be warranted for these patients.

Pan-cancer analysis identifies NT5E as a novel prognostic biomarker on cancer-associated fibroblasts associated with unique tumor microenvironment.

Background: Ecto-5'-nucleotidase (NT5E) encodes the cluster of differentiation 73 (CD73), whose overexpression contributes to the formation of immunosuppressive tumor microenvironment and is related to exacerbated prognosis, increased risk of metastasis and resistance to immunotherapy of various tumors. However, the prognostic significance of NT5E in pan-cancer is obscure so far. Methods: We explored the expression level of NT5E in cancers and adjacent tissues and revealed the relationship between the NT5E expression level and clinical outcomes in pan-cancer by utilizing the UCSC Xena database. Then, correlation analyses were performed to evaluate the relationship between NT5E expression and immune infiltration level via EPIC, MCP-counter and CIBERSORT methods, and the enrichment analysis were employed to identify NT5E-interacting molecules and functional pathways. Furthermore, we conducted single-cell analysis to explore the potential role of NT5E on single-cell level based on the CancerSEA database. Meanwhile, gene set enrichment analysis (GSEA) in single-cell level was also conducted in TISCH database and single-cell signature explorer was utilized to evaluate the epithelial-mesenchymal transition (EMT) level in each cell type. Results: The expression level of NT5E was aberrant in almost all cancer types, and was correlated with worse prognosis in several cancers. Notably, NT5E overexpression was related to worse overall survival (OS) in pancreatic adenocarcinoma (PAAD), head and neck squamous cell carcinoma (HNSC), mesothelioma (MESO), stomach adenocarcinoma (STAD), uveal melanoma (UVM) and cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) (p < 0.01). NT5E-related immune microenvironment analysis revealed that NT5E is associated positively with the degree of infiltration of cancer-associated fibroblasts (CAFs) and endothelial cells in most cancers. Enrichment analysis of cellular component (CC) demonstrated the critical part of NT5E played in cell-substrate junction, cell-substrate adherens junction, focal adhesion and external side of plasma membrane. Finally, single-cell analysis of NT5E illuminated that EMT function of CAFs was elevated in basal cell carcinoma (BCC), skin cutaneous melanoma (SKCM), HNSC and PAAD. Conclusion: NT5E could serve as a potential prognostic biomarker for cancers. The potential mechanism may be related to the upregulated EMT function of CAFs, which provides novel inspiration for immunotherapy by targeting CAFs with high NT5E expression.

Appropriate Reference Genes for RT-qPCR Normalization in Various Organs of Anemone flaccida Fr. Schmidt at Different Growing Stages.

Anemone flaccida Fr. Schmidt is a traditional medicinal herb in southwestern China and has multiple pharmacological effects on bruise injuries and rheumatoid arthritis (RA). A new drug with a good curative effect on RA has recently been developed from the extract of A. flaccida rhizomes, of which the main medicinal ingredients are triterpenoid saponins. Due to excessive exploitation, the wild population has been scarce and endangered in a few of its natural habitats and research on the cultivation of the plant commenced. Studies on the gene expressions related to the biosynthesis of triterpenoid saponins are not only helpful for understanding the effects of environmental factors on the medicinal ingredient accumulations but also necessary for monitoring the herb quality of the cultivated plants. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) as a sensitive and powerful technique has been widely used to detect gene expression across tissues in plants at different stages; however, its accuracy and reliability depend largely on the reference gene selection. In this study, the expressions of 10 candidate reference genes were evaluated in various organs of the wild and cultivated plants at different stages, using the algorithms of geNorm, NormFinder and BestKeeper, respectively. The purpose of this study was to identify the suitable reference genes for RT-qPCR detection in A. flaccida. The results showed that two reference genes were sufficient for RT-qPCR data normalization in A. flaccida. PUBQ and ETIF1a can be used as suitable reference genes in most organs at various stages because of their expression stabilitywhereas the PUBQ and EF1Α genes were desirable in the rhizomes of the plant at the vegetative stage.

The effects of perceived discrimination and city identity on the social adaptation of migrant children in public and private schools.

This study investigated the effect of migrant children's perceived discrimination on their social adaptation and the moderating effect of their city identity. In addition, differences in the relationships among the aforementioned variables were compared across two school types in terms of public schools and privately operated migrant schools. A cluster sampling method was used to survey 410 junior high school students in China using the Social Adaptation Assessment Questionnaire, the Perceived Discrimination Questionnaire for Individuals, and the City Identity Questionnaire as measures. Results showed that perceived discrimination significantly and negatively affected the social adaptation of migrant children. The relationship between perceived discrimination and social adaptation was moderated by the city identity of migrant children in private schools but not by those in public schools. Under the condition of high or medium city identity, discrimination negatively predicts social adaptation; under the condition of low city identity, however, discrimination does not predict social adaptation. In conclusion, perceived discrimination is an important factor regarding the social adaptation of migrant children, and their relationships vary by different types of city identity and schools.