Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective To compare the application effects of plankton multiplex polymerase chain reac-tion-capillary electrophoresis(PCR-CE),SYBR Green Ⅰ real-time quantitative PCR(qPCR)and microwave digestion-vacuum filtration-automated scanning electron microscopy(MD-VF-Auto SEM)in the diagnosis of drowning.Methods Lung,liver and kidney tissues from 212 drowned corpses and 30 non-drowned corpses were examined respectively by the three drowning-related plankton testing methods,and the detection rates of plankton in each tissue by three methods were compared.Results In drowned corpses,the total detection rates of PCR-CE,qPCR,and MD-VF-Auto SEM were 93.9%,96.2%,and 95.3%,respectively,with no statistically significant difference(P>0.05).The detection rate of lung tissue by MD-VF-Auto SEM(100%)was higher than those of PCR-CE and qPCR(P<0.05),and there was no significant difference in the detection rates of the three methods in liver or kidney tissues(P>0.05).In non-drowning corpses,a small number of diatoms(less than 10 cells/10 g)were detected by MD-VF-Auto SEM method,only in liver and kidney tissues,while the other two methods yielded negative results for all tissues.Conclusion All three methods have good efficacy in the examination of drowned corpses.The MD-VF-Auto SEM method directly observes diatom morpho-logical characteristics through scanning electron microscopy,and the qualitative and quantitative analy-ses are intuitive and accurate.It has great advantages in the examination of difficult degradation samples.The PCR-CE method and qPCR method have a low sample demand(0.5 g),are easy to operate and have short detection time(4-7 h).They are easy to be applied in the grassroots depart-ments and are suitable for the rapid determination of drowned corpses in routin cases.The combina-tion of the two DNA methods with the MD-VF-Auto SEM method can increase the detection rate of plankton,ensuring the reliability of examination results.This combined use is of significant importance in the application of drowning diagnosis.

Aim To explore the therapeutic effect and mechanism of Qingjie Huagong decoction in modulating PI3K/AKT/NF-κB signaling pathway in inflammatory response of acute pancreatitis(AP)mice.Methods Twenty-four mice were randomly divided into Blank group,Model group,Ustekin group,and Qingjie Hua-gong decoction group,with six mice in each group.The AP model was prepared by using rain frogin.Serum α-AMS,PNLP,IL-1β,IL-6,IL-8,IL-18,and TNF-α lev-els were detected by ELISA;the pancreatic pathology was detected by HE staining;the expressions of PI3K,AKT,and NF-κB-related proteins and mRNAs were de-tected by immunohistochemistry,Western blot,and RT-qPCR.Results Compared with the blank group,the model group showed obvious pathological damage to the pancreas,with significantly higher serum α-AMS,PN-LP,IL-1β,IL-6,IL-8,IL-18,and TNF-α levels(P＜0.01),and significantly higher levels of PI3K,AKT,and NF-κB-related proteins and mRNA expression(P＜0.01).Compared with the model group,both the Qingjie Huagong decoction group and the ustekin group improved the histopathological changes in the pancreas of AP mice,decreased the serum α-AMS,PNLP,IL-1β,IL-6,IL-8,IL-18,and TNF-α levels,and down-reg-ulated the expression levels of pancreatic PI3K,AKT,NF-κB-related proteins and mRNA(P＜0.05 or P＜0.01).Conclusion Qingjie Huagong decoction may inhibit the inflammatory response and protect pancreat-ic tissues by regulating the expression of PI3K/AKT/NF-κB signaling pathway.

OBJECTIVE: To investigate the association between long-term exposure to ambient fine particulate matter (PM _2.5) and its constituents and the risk of incident type 2 diabetes mellitus (T2DM), and to examine the modification roles of overweight status. METHODS: This prospective study included 27,507 adults living in rural China. The annual mean residential exposure to PM _2.5 and its constituents was estimated using a satellite-based statistical model. Cox models were used to estimate the risk of T2DM associated with PM _2.5 and its constituents. Stratified analysis quantified the role of overweight status in the association between PM _2.5 constituents and T2DM. RESULTS: Over a median follow-up of 9.4 years, 3,001 new T2DM cases were identified. The hazard ratio ( HR) for a 10 μg/m ³ increase in ambient PM _2.5 was 1.30 (95% confidence interval [ CI]: 1.17, 1.45). Among the constituents, the strongest association was observed with black carbon. Being overweight significantly modified the association between certain constituents and the risk of T2DM. Participants who were overweight and exposed to the highest quartile of PM _2.5 constituents had the highest risk of T2DM ( HR: 2.46, 95% CI: 2.04, 2.97). CONCLUSIONS Our findings indicate that PM _2.5 was associated with an increased risk of T2DM, with black carbon potentially being the primary contributor. Being overweight appeared to enhance the association between PM _2.5 and T2DM. This suggests that controlling both PM _2.5 exposure and overweight status may reduce the burden of T2DM.
Humans ; Diabetes Mellitus, Type 2/chemically induced* ; China/epidemiology* ; Particulate Matter/analysis* ; Overweight/epidemiology* ; Female ; Male ; Middle Aged ; Rural Population ; Air Pollutants/analysis* ; Adult ; Prospective Studies ; Environmental Exposure/adverse effects* ; Aged ; Risk Factors

Aim To explore the therapeutic effect and mechanism of Qingjie Huagong decoction in modulating PI3K/AKT/NF-κB signaling pathway in inflammatory response of acute pancreatitis(AP)mice.Methods Twenty-four mice were randomly divided into Blank group,Model group,Ustekin group,and Qingjie Hua-gong decoction group,with six mice in each group.The AP model was prepared by using rain frogin.Serum α-AMS,PNLP,IL-1β,IL-6,IL-8,IL-18,and TNF-α lev-els were detected by ELISA;the pancreatic pathology was detected by HE staining;the expressions of PI3K,AKT,and NF-κB-related proteins and mRNAs were de-tected by immunohistochemistry,Western blot,and RT-qPCR.Results Compared with the blank group,the model group showed obvious pathological damage to the pancreas,with significantly higher serum α-AMS,PN-LP,IL-1β,IL-6,IL-8,IL-18,and TNF-α levels(P＜0.01),and significantly higher levels of PI3K,AKT,and NF-κB-related proteins and mRNA expression(P＜0.01).Compared with the model group,both the Qingjie Huagong decoction group and the ustekin group improved the histopathological changes in the pancreas of AP mice,decreased the serum α-AMS,PNLP,IL-1β,IL-6,IL-8,IL-18,and TNF-α levels,and down-reg-ulated the expression levels of pancreatic PI3K,AKT,NF-κB-related proteins and mRNA(P＜0.05 or P＜0.01).Conclusion Qingjie Huagong decoction may inhibit the inflammatory response and protect pancreat-ic tissues by regulating the expression of PI3K/AKT/NF-κB signaling pathway.

Objective:To screen novel diagnostic marker or therapeutic target for multiple myeloma(MM).Methods:Sel1L,SPAG4,KCNN3 and PARM1 were identified by bioinformatics method based on GEO database as high expression genes in MM.Their RNA and protein expression levels in bone marrow mononuclear cells from myeloma cell lines U266,NCI-H929,MM.1s,RPMI8226 and leukemia cell line THP1,as well as 31 MM patients were evaluated by RT-PCR and Western blot,respectively.Meanwhile,5 samples of bone marrow from healthy donors for allogeneic hematopoietic stem cell transplantation were employed as controls.Results:Compared with leukemia cell line THP1,the expression levels of KCNN3,PARM1 and Sel1L mRNA were significantly increased in myeloma cell lines U266,NCI-H929 and MM.1s,while PARM1 was further increased in myeloma cell lines 8226.Western blot showed that the 4 genes were all expressed in the 4 myeloma cell lines.Compared with healthy controls,the expression levels of Sel1L,SPAG4,KCNN3 and PARM1 mRNA were significantly higher in MM patients(all P＜0.05).Western blot showed that the 4 genes were all expressed in MM patients,and the protein expression level of Sel1L and KCNN3 were significantly different compared with healthy donors(all P＜0.01).Conclusion:Sel1L,SPAG4,KCNN3 and PARM1 may be potential diagnostic markers and therapeutic targets for MM.

Objective:To investigate the levels of diphtheria-specific binding antibodies and neutralizing antibodies in mice immunized with pneumococcal polysaccharide conjugate vaccine using diphtheria toxoid as a carrier.Methods:NIH mice were immunized with one batch of diphtheria, tetanus and acellular pertussis combined vaccine, absorbed (DTaP-1) or three different batches of 13-valent pneumococcal polysaccharide conjugate vaccine (PCV13, containing diphtheria toxoid vector) at three dilutions (5-, 10- and 20-fold dilution). Serum samples were collected to test for diphtheria-specific antibody titers and diphtheria potencies of the vaccines. Another three batches of DTaP vaccine and three batches of tetanus toxoid, reduced diphtheria toxoid and acellular pertussis combined vaccine (Tdap) were used to immunize NIH mice. Serum samples were collected and the diphtheria potencies were detected. Statistical analysis was performed using one-way analysis of variance.Results:At the 5-fold and 10-fold dilutions, the titers of diphtheria-specific antibodies induced by three batches of PCV13 vaccine were all lower than those by DTaP-1 vaccine (all P<0.001), while there was no statistically significant difference at the 20-fold dilution ( P>0.05). The diphtheria potencies of the DTaP-1 vaccine and the three batches of PCV13 vaccine were 100.5, 76.2, 64.5, and 62.0 IU/ml, respectively. The diphtheria potencies of another three batches of DTaP vaccine were 82.5, 83.6, and 79.9 IU/ml, respectively, and those of three batches of Tdap vaccine were 10.3, 12.2, and 12.9 IU/ml, respectively. There was no statistically significant difference in diphtheria potency between DTaP vaccine and PCV13 vaccine( P>0.05), while there was a statistically significant difference between Tdap vaccine and the PCV13 vaccine ( P<0.001). Conclusions:The pneumococcal polysaccharide conjugate vaccine with diphtheria toxoid has good diphtheria immunogenicity and can induce the production of higher levels of diphtheria-specific binding antibodies and protective neutralizing antibodies in vivo. Pneumococcal capsular polysaccharide exerts an immune enhancement effect on diphtheria toxoid. The relevant results provide valuable guidance for determining carrier protein dosage in bacterial polysaccharide conjugate vaccines, planning vaccine co-administration, and selecting the dosage of diphtheria toxoid antigen in the research and development of combined vaccines.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Aim To explore the effects of D-limonene on the steatosis of primary mouse hepatocytes and its potential mechanism of action.Methods Oleic acid-induced steatosis in primary mouse hepatocytes was used as a model to observe the effects of D-limonene on cell viability,cellular lipid content,and intracellular expression of proteins such as AMP-activated protein kinase(AMPK),acetyl-coenzyme A carboxylase 1(ACC1),and carnitine palmitoyl transferase 1A(CPT1A).Results It was found that a low dose of D-limonene could effectively enhance the viability of primary mouse hepatocytes.When oleic acid at a con-centration of 300 μmol·L-1 successfully induced steatosis in primary mouse hepatocytes,D-limonene re-duced the lipid content of the cells,and D-limonene up-regulated the cellular AMPK expression level,down-regulated the cellular ACC1 and fatty acid synthetase(FAS)expression levels,which in turn promoted the overexpression of CPT1A.Conclusions D-limonene has the effect of reducing lipid deposition in primary mouse hepatocytes,and the mechanisms may be related to the activation of AMPK,the inhibitions of ACC1 and FAS,and the up-regulation of CPT1A protein expres-sion level.

OBJECTIVES: To investigate the relationship between the polygenic risks for various psychiatric disorders and clinical and neuropsychological characteristics in children with attention-deficit/hyperactivity disorder (ADHD). METHODS: Using a cross-sectional design, 285 children with ADHD and 107 healthy controls were assessed using the Child Behavior Checklist, the Behavior Rating Inventory of Executive Function for parents, the Wechsler Intelligence Scale for Children, Fourth Edition, and the Cambridge Neuropsychological Test Automated Battery. Blood samples were collected for genetic data. Polygenic risk scores (PRSs) for various psychiatric disorders were calculated using the PRSice-2 software. RESULTS: Compared with the healthy controls, the children with ADHD displayed significantly higher PRSs for ADHD, major depressive disorder, anxiety disorder, and obsessive-compulsive disorder (P<0.05). In terms of daily-life executive function, ADHD-related PRS was significantly correlated with the working memory factor; panic disorder-related PRS was significantly correlated with the initiation factor; bipolar disorder-related PRS was significantly correlated with the shift factor; schizophrenia-related PRS was significantly correlated with the inhibition, emotional control, initiation, working memory, planning, organization, and monitoring factors (P<0.05). The PRS related to anxiety disorders was negatively correlated with total IQ and processing speed index (P<0.05). The PRS related to obsessive-compulsive disorder was negatively correlated with the processing speed index and positively correlated with the stop-signal reaction time index of the stop-signal task (P<0.05). CONCLUSIONS PRSs for various psychiatric disorders are closely correlated with the behavioral and cognitive characteristics in children with ADHD, which provides more insights into the heterogeneity of ADHD.
Humans ; Attention Deficit Disorder with Hyperactivity/genetics* ; Child ; Male ; Female ; Cross-Sectional Studies ; Neuropsychological Tests ; Multifactorial Inheritance ; Adolescent ; Mental Disorders/etiology* ; Executive Function ; Genetic Risk Score