Stimulating the expression of sphingosine kinase 1 (SphK1) is beneficial to reduce acrylamide-induced nerve cell damage.

Sphingosine kinase 1 (SphK1) is an important signaling molecule for cell proliferation and survival. However, the role of SphK1 in acrylamide (ACR)-induced nerve injury remains unclear. The purpose of this study was to investigate the role and potential mechanism of SphK1 in ACR-induced nerve injury. Liquid chromatography triple quadrupole tandem mass spectrometry (LC-MS/MS) and reverse transcription-quantitative PCR (RT-qPCR) were used to detect sphingosine 1-phosphate (S1P) content in serum and SphK1 content in whole blood from an occupational work group exposed to ACR compared to a non-exposed group. For in vitro experiments, SphK1 in human SH-SY5Y neuroblastoma cells was activated using SphK1-specific activator phorbol 12-myristate 13-acetate (PMA). Our research also utilized cell viability assays, flow cytometry, western blots, RT-qPCR and related protein detection to assess activity of the mitogen activated protein kinase (MAPK) signaling pathway. The results of the population study showed that the contents of SphK1 and S1P in the ACR-exposed occupational contact group were lower than in the non-exposed group. The results of in vitro experiments showed that expression of SphK1 decreased with the increase in ACR concentration. Activating SphK1 improved the survival rate of SH-SY5Y cells and decreased the apoptosis rate. Activating SphK1 in SH-SY5Y cells also regulated MAPK signaling, including enhancing the phosphorylation of extracellular signal-regulated protein kinases (ERK) and inhibiting the phosphorylation of c-Jun N-terminal kinase (JNK) and p38. These results suggest that activating SphK1 can protect against nerve cell damage caused by ACR.

A urinary metabolomic study from subjects after long-term occupational exposure to low concentration acrylamide using UPLC-QTOF/MS.

Because long-term occupational exposure to low concentrations of acrylamide (ACR) has the potential to cause neurological damage, it is important to identify biomarkers that can be used to evaluate this risk. In the present study, urine metabolomics of the ACR-exposed and non-exposed groups to identify potential metabolites was carried out using ultra high performance liquid chromatography coupled with quadrupole time of flight mass spectrometry. Serum biochemical indexes of the exposed and non-exposed groups were also determined. Principal component analysis showed a differential separation between exposed group and non-exposed group and a total of 7 metabolites were identified in positive and negative ionization modes; Area under curve of anthranilic acid, ß-guanidinopropionic acid and mesobilirubinogen were 0.980, 0.843 and 0.801 respectively and these metabolites showed high sensitivity and specificity. The 13 biochemical indexes were divided into three classes based on physiological functions. Only biomarkers of dysregulated liver function including alanine aminotransferase, aspartic transaminase, total bilirubin, direct bilirubin and triglyceride were significantly higher in the exposed group than in the non-exposed group. This study identifies important related metabolic changes in the bodies of workers after long-term occupational exposure to low concentration ACR and suggests new biomarkers of nervous system injury caused by ACR. The study also provides a sound basis for exploring the biochemical mechanisms and metabolic pathways of nervous system toxicity caused by ACR.

Three Chemosensory Proteins Involved in Chemoreception of Oedaleus asiaticus (Orthopera: Acridoidea).

Chemosensory proteins (CSPs) are thought to play roles in the insect olfactory system by binding and carrying hydrophobic odorants across the aqueous sensillar lymph. The band-winged grasshopper, Oedaleus asiaticus Bei-Bienko, is one of the most important grasshopper pests in northern China, but there is little information about its olfactory system. In order to investigate the olfactory functions of CSPs in this pest, three CSP genes (OasiCSP4, OasiCSP11 and OasiCSP12) were expressed in Escherichia coli, and the binding affinities of the three recombinant CSP proteins were measured for 16 volatiles from the host plant (Stipa krylovii), fecal material and body of live adult O. asiaticus using fluorescence competitive binding assays. To further verify their olfactory functions, RNA interference (RNAi) and electrophysiological recording were conducted. The three recombinant proteins displayed different degrees of binding to various volatiles in ligand-binding assays, with OasiCSP12 having higher binding affinities for more volatiles than OasiCSP4 and OasiCSP11. OasiCSP12 exhibited strong binding affinities (Ki < 20 µΜ) for five host plant volatiles and one volatile from the live body of adult O. asiaticus. The transcript levels of the three OasiCSP genes were significantly lower after silencing the individual genes by RNAi, which in turn reduced the EAG responses in adults of both sexes to most tested compounds. Our study indicates that these three OasiCSPs are involved in the detection of volatile semiochemicals, and may play important roles in finding host plants and in aggregation in O. asiaticus.

Comparative Analysis of Transcriptome Responses to Cold Stress in Galeruca daurica (Coleoptera: Chrysomelidae).

Galeruca daurica (Joannis) has become a new insect pest in the Inner Mongolia grasslands since 2009, and its larvae and eggs have strong cold tolerance. To get a deeper insight into its molecular mechanisms of cold stress responses, we performed de novo transcriptome assembly for G. daurica by RNA-Seq and compared the transcriptomes of its larvae exposed to five different temperature treatments (-10, -5, 0, 5, and 25°C for 1 h and then recovered at 25°C for 1 h), respectively. Compared with the control (25°C), the numbers of differentially expressed genes (DEGs) decreased from 1,821 to 882, with the temperature declining from 5 to -10°C. Moreover, we obtained 323 coregulated DEGs under different low temperatures. Under four low temperatures (-10, -5, 0, and 5°C), a large number of genes were commonly upregulated during recovery from cold stresses, including those related to cuticle protein, followed by cytochrome P450, clock protein, fatty acid synthase, and fatty acyl-CoA reductase; meanwhile, lots of genes encoding cuticle protein, RNA replication protein, RNA-directed DNA polymerase, and glucose dehydrogenase were commonly downregulated. Our findings provide important clues for further investigations of key genes and molecular mechanisms involved in the adaptation of G. daurica to harsh environments.

The Complete Mitochondrial Genome of the Plant Bug Lygus pratensis Linnaeus (Hemiptera: Miridae).

Lygus pratensis is a phytophagous pest responsible for yield losses in Bt alfalfa and other economic crops in Northwestern China. To better characterize Miridae at the genomic level, the complete mitochondrial (mt) genome of L. pratensis was sequenced and analyzed in this study. The mt genome was amplified via the polymerase chain reaction to generate overlapping fragments. These fragments were then sequenced, spliced, and analyzed to include the examination of nucleotide composition, codon usage, compositional biases, protein-coding genes (PCGs), and RNA secondary structures. Phylogenetic relationships between L. pratensis and other species in different Heteroptera families were also examined. The mt genome was found to be a typical circular genome with a length of 16,591 bp and a total AT content of 75.1%, encoded for 13 PCGs, 22 transfer RNAs (tRNAs), 2 ribosomal RNAs (lrRNA and srRNA), and a noncoding control region. The nucleotide composition of the entire mt genome was heavily biased toward A and T. All of the tRNAs were predicted to have classic clover leaf structures, but three of the tRNAs (tRNAAsn, tRNAHis, tRNAHis) were missing the TΨC loop. The control region (2,017 bp), which was found to be located between 12S and tRNAIle, contained three tandem repeat elements. Phylogenetic analyses showed that L. pratensis is closely related to the other three examined Lygus bugs, and that it is a sister group to Apolygus and Adelphocoris. This study confirms the usability of the mt genome in phylogenesis studies pertaining to the Lygus genus, within Miridae.

Increased Chondrocyte Apoptosis in Kashin-Beck Disease and Rats Induced by T-2 Toxin and Selenium Deficiency.

OBJECTIVE: To investigate chondrocyte apoptosis and the expression of biochemical markers associated with apoptosis in Kashin-Beck disease (KBD) and in an established T-2 toxin- and selenium (Se) deficiency-induced rat model. METHODS: Cartilages were collected from the hand phalanges of five patients with KBD and five healthy children. Sprague-Dawley rats were administered a selenium-deficient diet for 4 weeks prior to T-2 toxin exposure. The apoptotic chondrocytes were observed by terminal deoxynucleotidyl transferase dUTP nick end labeling staining. Caspase-3, p53, Bcl-2, and Bax proteins in the cartilages were visualized by immunohistochemistry, their protein levels were determined by Western blotting, and mRNA levels were determined by real-time reverse transcription polymerase chain reaction. RESULTS: Increased chondrocyte apoptosis was observed in the cartilages of children with KBD. Increased apoptotic and caspase-3-stained cells were observed in the cartilages of rats fed with normal and Se-deficient diets plus T-2 toxin exposure compared to those in rats fed with normal and Se-deficient diets. Caspase-3, p53, and Bax proteins and mRNA levels were higher, whereas Bcl-2 levels were lower in rats fed with normal or Se-deficiency diets supplemented with T-2 toxin than the corresponding levels in rats fed with normal diet. CONCLUSION: T-2 toxin under a selenium-deficient nutritional status induces chondrocyte death, which emphasizes the role of chondrocyte apoptosis in cartilage damage and progression of KBD.

Dibromoacetic Acid Induces Thymocyte Apoptosis by Blocking Cell Cycle Progression, Increasing Intracellular Calcium, and the Fas/FasL Pathway in Vitro.

Dibromoacetic acid (DBAA), a haloacetic acid found in drinking water as a disinfection by-product, can cause many adverse effects, including immunotoxicity. In a previous study, we confirmed that DBAA can induce obvious immunotoxicity in mice but that the underlying mechanisms are not clearly understood. In our current study, we confirmed that DBAA induced cytotoxicity and apoptosis in thymocytes isolated from mice by a range of DBAA concentrations (0, 5, 10, 20, or 40 µM). The data showed that DBAA exposure led to a significant decrease in proliferative responses to T-cell mitogens and obvious inhibition in the production of cytokines interleukin-2 and interleukin-4. We found obvious morphological changes of apoptosis in thymocytes and observed the percentage of apoptotic thymocytes to increase significantly as the DBAA concentration increased. Further investigation showed that DBAA can cause G0/G1 arrest in cell cycle analysis, increase intracellular calcium ([Ca(2+)]i) levels, increase the expression of Fas/FasL proteins, and decrease the expression of Bcl-2 protein. It is concluded that in vitro exposure to DBAA can lead to marked cytotoxicity and apoptosis among thymocytes, and the mechanism involved is strongly related to blocking cell cycle progression, increasing intracellular calcium, and increasing Fas/FasL expressions.

Supercooling capacity and cold hardiness of band-winged grasshopper eggs (Orthoptera: Acrididae).

The band-winged grasshopper, Oedaleus asiaticus Bei-Bienko, is one of the most dominant and economically important grasshopper species in the steppe grasslands and farming-pastoral ecotone in northern China. It is a univoltine species and overwinters as eggs in soil. The cold hardiness of its eggs was examined in the laboratory. Water content in soil significantly affected the supercooling points (SCPs), water content and fat content of prediapause eggs. With the increase of water content in soil, the SCP, and water content of prediapause eggs rose whereas the fat content declined. There was a significant relationship between the SCP and water content or fat content of prediapause eggs. The SCPs of prediapause and diapause eggs varied from -7.6 to -28.4°C and the SCPs of eggs 30 d after oviposition could be divided into two groups. The means of high SCP group (-11.0 to -11.9°C) were much higher than those of low SCP group (-21.8 to -21.9°C), and the majority belonged to the latter (90.48-93.33%). The SCPs of prediapause eggs and early-diapause eggs 30 d after oviposition were significantly higher than those of deep-diapause eggs 60 d after oviposition. The survival rates of diapause eggs were significantly different among different temperature treatments. The survival rate was higher than 88% at greater than -20°C and declined significantly to 57% at -25°C, and suddenly dropped to zero at -30°C. The lower lethal temperature (Ltemp50) for 12 h exposure was -25.3°C and the lower lethal time (Ltime50) at -20°C was 32.8 d. As the mean SCPs of diapause eggs were similar to their Ltemp50, the SCP of eggs can be considered as a good indicator of cold hardiness for O. asiaticus and that this grasshopper is a freeze-intolerant insect.

[Function disorder assessment on patients with mild psychiatric impairment due to road traffic accidents].

OBJECTIVE: To explore the methods to assess the function disorder of patients with mild psychiatric impairment due to road traffic accidents. METHODS: In contrast to that of the patient with physical impairment, the function disorder of the patient with mild psychiatric impairment was assessed by the standard assessment instruments, such as the Barthel index of ADL (BI), Functional Activities Questionnaire (FAQ), Social Disability Screening Schedule (SDSS), Activity of Daily Living Scale (ADL), Physical Self-maintenance Scale (PSMS), and Instrumental Activities of Daily Living Scale (IADL). RESULTS: Except BI, the each total score of FAQ, SDSS, ADL, PSMS, and IADL correlated well with the rank of psychiatric impairment. The difference of each total score of rating scales among different physical impairment rank was not statistically significant. At the impairment rank of level 10, the difference of each total score of FAQ and ADL was not significant between the two groups; each total score of BI, SDSS, and IADL in psychiatric impairment was higher than that of physical impairment, while the total score of PSMS in psychiatric impairment was lower than that of physical impairment. At the level 9, except PSMS, the each total score of BI, FAQ, SDSS, IADL and ADL in psychiatric impairment was higher than that of physical impairment. At the level 8, except BI and PSMS, the each total score of FAQ, SDSS, IADL and ADL in psychiatric impairment was higher than that of physical impairment. CONCLUSION: The different criteria for assessment of mild psychiatric and physical impairment may exist, and appropriate indexes for assessment between physical and psychiatric impairment need to be further elucidated.

[Posture evoked response detecting in normal lower limb muscle and its influencing factors].

OBJECTIVE: To analyze the latency of posture evoked response of normal lower limb muscle in different stimulations and explore its influencing factors. METHODS: The normal lower limb was induced to produce postural evoked response by the dynamic posturography through two kinds of perturbations, the supporting surface rotation stimulation (Toes-up and Toes-down) and the horizontal perturbation stimulation (Forward and Backward). The latencies of tibialis anterior muscle and gastrocnemius muscle were recorded by surface electromyography acquisition system. The differences of the left and right limb, gender and height on the latency of postural evoked response were analyzed. RESULTS: (1) Under the Toes-up and Backward perturbation, the latency of tibialis anterior muscle was longer than gastrocnemius muscle; under the Toes-down and Forward perturbation, the latency of gastrocnemius muscle was longer than tibialis anterior muscle. (2) The latencies of left limb and right limb had no significant difference. (3) The latency in male was longer than that in female. (4) The latency gradually increased with the increase of height. CONCLUSION: In the postural evoked response, different perturbations, gender and height have significant impacts on the latency of posture evoked response of lower limb muscle. However, the effect of height and gender should be not considered referring to the same individual.

[Characteristics of computed dynamic posturography of patients with peripheral vertigo].

OBJECTIVE: To quantify the posture control ability of patients with peripheral vertigo by computed dynamic posturography (CDP). METHODS: Ninety-one subjects diagnosed with peripheral vertigo by caloric test were divided into unilateral vestibular dysfunction group (unilateral group) and bilateral vestibular disorder dysfunction group (bilateral group). CDP tests including sensory organization test (SOT) and motor control test (MCT) were performed on all subjects. The CDP results were compared with the normal data by t-test. RESULTS: In unilateral group, there was no statistically significant difference in SOT compared with normal data (P > 0.05). In bilateral group, the composite equilibrium score was lower than the normal data, especially during the moving of the platform and the changes of visual stimulation (P < 0.05). In MCT test, the latencies of both groups showed no statistically significant difference compared with normal data (P > 0.05). CONCLUSION: During the non-acute period of peripheral vertigo, patients could maintain static equilibrium. Compared with normal people, dynamic equilibrium function is normal in patients with unilateral vestibular dysfunction, but declined in patients with bilateral vestibular dysfunction.

Stature estimation based on the length of tibia and fibula measured by digital X-ray in Chinese Han teenagers.

OBJECTIVE: To derive regression formulae for stature estimation using forensic radiography from the tibia and fibula of Chinese Han teenagers in Sichuan Province. METHODS: To construct equations, measurements were conducted on the training sample (412 adults, 201 males and 211 females). The whole length of the fibula and four measurements of tibia were determined using CR radiography, rectified through theoretical magnification. The regression formulae were relatively constructed to the real stature measured in an erect position. Through using the testing sample (40 adults) for the regression formulae, the reliability of the regression formulae was assessed. RESULTS: The range of correlation coefficients of four measurements for tibia was 0.880-0.895 in the sex-unknown group, 0.869-0.893 in the male, and 0.845-0.855 in the female. The five measurements were found to be better correlated with stature in the male than in the female. CONCLUSION: The digital X-ray of the tibia and fibula for stature estimation is proved to be effective in forensic individual identification; therefore, these equations can be of great assistance to the stature estimation of the contemporary Chinese Han teenagers.

[Forensic medical identification of 355 cases with hearing impairment].

OBJECTIVE: To analyze the difference of subjective hearing threshold and objective hearing threshold, and to discuss the importance of standard for hearing evaluation in forensic medicine. METHODS: Three hundred and fifty-five cases (387 ears) of forensic medical identification with hearing impairment were retrospectively analyzed including the items entrusted and hearing test results. All cases were collected from 2004 to 2012 in the forensic science center. RESULTS: In the 387 ears, 218 ears (56.3%) were evaluated the degree of disability and 106 ears (27.4%) were identified the degree of damage. In the disability degree evaluation, the subjective hearing threshold and the objective hearing threshold were significant different in 120 ears (55.0%), while in damage degree evaluation, the subjective hearing threshold and the objective hearing threshold were significant different in 69 ears (65.1%). CONCLUSION: Because of camouflaging or exaggerating the hearing impairment by the wounded, the subjective hearing threshold can't accurately assess the existence and the degree of hearing impairment. In the forensic identification, auditory brainstem response, 40 Hz auditory event related potential and auditory steady-state response should be combined in the application to evaluate the hearing impairment for the wounded in order to ensure the reliability of the evaluation of hearing impairment.

[Comparisons among three frequency-specific auditory evoked potentials in normal hearing adults].

OBJECTIVE: To provide supports for the application of auditory evoked potential (AEP) in the evaluation of behavioral threshold, by studying the difference and relevance between the pure tone audiometry (PTA) and three frequency-specific auditory evoked potentials, including 40 Hz auditory event related potentials (40 Hz AERP), tone burst auditory brainstem response (Tb-ABR) and auditory steady-state response (ASSR). METHODS: Three frequency-specific AEP and PTA thresholds were recorded at speech frequency (0.5, 1, 2, 4 kHz) from thirty-four adults with normal hearing (68 ears). Then, the relationship between the AEP thresholds and PTA thresholds were analyzed respectively. RESULTS: There were good correlations between three frequency-specific AEP thresholds and PTA thresholds at speech frequency. However, the difference of thresholds between each frequency-specific AEP and PTA was not same. The difference of thresholds were the smallest and the relevance were the best between 40 Hz AERP and PTA at 0.5 kHz, and between ASSR and PTA at 2, 4 kHz. At 1 kHz, there were not statistical difference between ASSR, 40 Hz AERP and PTA, while the relevance of 40 Hz AERP was better than ASSR. CONCLUSION: Different methods should be chosen to assess the objective behavioral thresholds at different frequency.

An Osteoporosis Susceptibility Allele at 11p15 Regulates SOX6 Expression by Modulating TCF4 Chromatin Binding.

Osteoporosis is an age-related complex disease clinically diagnosed with bone mineral density (BMD). Although several genomewide association studies (GWASs) have discovered multiple noncoding genetic variants at 11p15 influencing osteoporosis risk, the functional mechanisms of these variants remain unknown. Through integrating bioinformatics and functional experiments, a potential functional single-nucleotide polymorphism (SNP; rs1440702) located in an enhancer element was identified and the A allele of rs1440702 acted as an allelic specificities enhancer to increase its distal target gene SOX6 (~600 Kb upstream) expression, which plays a key role in bone formation. We also validated this long-range regulation via conducting chromosome conformation capture (3C) assay. Furthermore, we demonstrated that SNP rs1440702 with a risk allele (rs1440702-A) could increase the activity of the enhancer element by altering the binding affinity of the transcription factor TCF4, resulting in the upregulation expression of SOX6 gene. Collectively, our integrated analyses revealed how the noncoding genetic variants (rs1440702) affect osteoporosis predisposition via long-range gene regulatory mechanisms and identified its target gene SOX6 for downstream biomarker and drug development. © 2022 American Society for Bone and Mineral Research (ASBMR).

Efforts to eliminate schistosomiasis in Hubei province, China: 2005-2018.

BACKGROUND: The Hubei province is one of the most schistosomiasis-epidemic-prone provinces in China. A series of strategies were adopted by the government to curb the rebound schistosomiasis endemic status that has prevailed since the early 2000s. This study aimed to elucidate the trends of schistosomiasis transmission and to appraise the effectiveness of the integrated control strategy in lake and marshland areas. METHODS: Surveillance data of schistosomiasis in the Hubei province between 2005 and 2018 were analyzed, including conventional health control measures, integrated strategies, and measures that focused on the infection source. According to the local annual plan for schistosomiasis control in endemic counties, previous measures were human and snail control and surveillance. Residents aged 6-65 years were screened by an immunological detection method called indirect hemagglutination assay (IHA) after the transmission season each year. All residents who tested positive were then asked to provide a fecal sample for examination by the miracidium hatching technique (MHT) to detect the presence of schistosomes. Moreover, systematic snail surveys were conducted as a part of the combined environmental sampling method. The latter included integrated strategies and measures that focused on the infection source. Bovine stool samples were also collected and concurrently assessed using the MHT by the agriculture department, river-hardening slope protection was constructed by the water conservancy department, and forestation promotion was conducted by the forest department. The effectiveness of the integrated control strategy was assessed using two indicators of resident and livestock infection rates and three indicators of snail epidemics across all endemic areas. RESULTS: From 2005 to 2018, a total of 28. 46 million and 2. 05 million residents were assessed by immunological (IHA) and etiological (MHT) detection techniques, respectively. Snail surveys and molluscicide application were performed in 2. 26 hectares and 0. 37 hectares, respectively. Moreover, 2. 60 million bovines were assessed by etiological detection techniques (MHT). The river-hardening slope protection project was implemented in 503 places, and 46 thousand hectares in endemic areas underwent environmental modification. Forestation was implemented at an area of 0. 15 million hectares. Between 2005 and 2018, the epidemic indicators, including resident and livestock infection rates and the infested areas and infection rate of snails, all presented downward trends. The resident infection rate decreased from 3. 78% in 2005 to 0% in 2016, which persisted through 2018. The livestock infection rate decreased from 5. 63% in 2005 to 0% in 2013, which also persisted through 2018. From 2005 to 2018, the snail-inhabited area was slightly reduced, but the area of infected snails decreased to 0 in 2012; this persisted through 2018. All counties met the goal for schistosomiasis infection control, transmission control, and disruption of schistosomiasis activity in 2008, 2013, and 2018 separately. That means the goal has been achieved in each stage. CONCLUSIONS: The decline of the schistosomiasis epidemic rate demonstrates that the Chinese government was successful in meeting its public health goal in Hubei province. In the next decade, precision interventions must be implemented in endemic counties with a relatively low epidemic status to achieve the goals of the Outline of the Healthy China 2030 Plan. A similar strategy can be applied in other countries to eliminate schistosomiasis globally.

[Identifying malingering symptoms of balance disorders by computerized dynamic posturography].

OBJECTIVE: To develop an objective criteria with computerized dynamic posturography (CDP) for identification of non-organic or malingering symptoms of balance disorders. METHODS: One hundred and four normal subjects were instructed to feign balance disturbance after naturally performing on CDP. Paired t-test was used to determine if there was significant difference of CDP equilibrium scores between normal and malingering performance. The CDP records including raw curve of 50 subjects were critically evaluated by previously published seven criteria in indicating malingering performance. If the sensitivity of a criterion was greater than 90%, it would be accepted as our laboratory's identifying criterion. The CDP records of remaining 54 subjects were used to test the sensitivity and specificity of the accepted criteria. RESULTS: There were statistically significant differences of CDP equilibrium scores between normal and malingering performance (P < 0.01). The mean equilibrium scores of malingering performance were lower than that of normal performance. The sensitivity and specificity of our criteria for identifying malingerers were 92.5% and 98.2%, respectively. CONCLUSION: It is necessary to identify malingering performance before analyzing the CDP equilibrium scores. Our identifying criteria of malingerers seems to be quite useful and accurate.

[Application of slow vertex response in auditory threshold prediction for subjects with hearing loss].

OBJECTIVE: To study the value of slow vertex response (SVR) in the evaluation of hearing loss by comparing the hearing thresholds acquired with SVR and pure tone audiometry (PTA). METHODS: Twenty-five subjects (40 ears) with sensorineural hearing loss were tested by PTA and SVR. According to the thresholds of PTA, these ears were subdivided into mild, moderate and severe hearing loss groups, and rank sum test was performed on the thresholds of SVR and PTA for all the hearing loss groups. Then, the correlation between PTA thresholds and SVR thresholds was analyzed and the mathematical models were established for predicting behavioral thresholds by the thresholds of SVR. RESULTS: At four test frequencies (0.5, 1, 2 and 4kHz), the thresholds of SVR had high correlations with thresholds of PTA. Four liner regression equations were established, and the correlation coefficient(r) were 0.971, 0.976, 0.957 and 0.928, respectively (P < 0.05). Back substitution test showed that the liner regression equations would be an easy method for estimating the behavior thresholds. CONCLUSION: The behavioral threshold can be well judged and evaluated by the liner regression equations established with SVR thresholds.

RBP4 Is Associated With Insulin Resistance in Hyperuricemia-Induced Rats and Patients With Hyperuricemia.

Objective: Hyperuricemia (HUA) is strongly associated with abnormal glucose metabolism and insulin resistance (IR). However, the precise molecular mechanism of HUA-induced IR is still unclear. Retinol binding protein 4 (RBP4) has been shown to induce IR in type 2 diabetes mellitus. This study was designed to clarify the relationship between RBP4 and HUA-induced IR and its potential mechanisms. Methods: Patients with HUA were collected to detect the levels of plasma RBP4 and clinical biochemical indicators. Rats were fed with 10% high yeast and oteracil potassium (300 mg/kg) via intraperitoneal injection once daily for eight weeks, and gavage with adenine (100 mg/kg) once daily from the fifth week to induce the HUA model. Glucose consumption testing was performed to determine the capacity of glucose intake and consumption in 3T3-L1 adipocytes. Real-time polymerase chain reaction (RT-PCR) and western blot were used to detect the mRNA and protein level of RBP4 and insulin receptor substrate-phosphatidylinositol 3-kinase-active protein kinase (IRS/PI3K/Akt) signaling pathway-related proteins. Results: The levels of plasma RBP4 in both HUA patients and HUA rat models were significantly higher than that in the control groups. The level of plasma RBP4 was positively correlated with plasma uric acid, creatinine, fasting insulin, IR index, total cholesterol and triglyceride levels in patients with HUA. In HUA rats, the level of plasma RBP4 was positively correlated with plasma uric acid, IR index, and triglycerides. HUA rats also exhibited IR. After inhibition of RBP4 expression, the phosphorylation levels of the IRS/PI3K/Akt signaling pathway were increased, and IR was significantly improved. Conclusion: HUA induced IR both in vitro and in vivo. RBP4 may be involved in HUA-induced IR by inhibiting IRS/PI3K/Akt phosphorylation. Our findings may provide a new insight for the treatment of IR caused by HUA.

An Intronic Risk SNP rs12454712 for Central Obesity Acts As an Allele-Specific Enhancer To Regulate BCL2 Expression.

Genome-wide association studies (GWAS) have reproducibly associated the single nucleotide polymorphism (SNP) rs12454712 with waist-to-hip ratio adjusted for BMI (WHRadjBMI), but the functional role underlying this intronic variant is unknown. Integrative genomic and epigenomic analyses supported rs12454712 as a functional independent variant. We further demonstrated that rs12454712 acted as an allele-specific enhancer regulating expression of its located gene BCL2 by using dual-luciferase reporter assays and clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Specifically, the rs12454712-C allele can bind transcription factor ZNF329, which efficiently elevates the enhancer activity and increases BCL2 expression. Knocking down Bcl2 in 3T3-L1 cells led to the downregulation of adipogenic differentiation marker genes and increased cell apoptosis. A significant negative correlation between BCL2 expression in subcutaneous adipose tissues and obesity was observed. Our findings illustrate the molecular mechanisms behind the intronic SNP rs12454712 for central obesity, which would be a potential and promising target for developing appropriate therapies.