Objective To investigate whether the pyroptosis-related Toll-like receptor 4(TLR4)signaling pathway influences the malignant transformation of actinic keratosis(AK)into cutaneous squamous cell carcinoma(SCC).Methods A total of 6 lesion tissue samples each from patients with AK and SCC,as well as 5 normal skin tissue samples from healthy subjects as controls,were collected from the First Affiliated Hospital of Kunming Medical University between August 2020 and August 2021.Quantitative PCR(qPCR)and Western blot analysis were performed to determine the mRNA and protein expression levels of TLR4 pathway-related factors,including TLR4,CPB1,NLRP3,IL-1β,and IL-18.TLR4/TUNEL double immunofluorescence staining was used to evaluate TLR4 expression and the level of cellular pyroptosis in tissue samples.In addition,Western blotting was performed to analyze the expression differences of pyroptosis-related core proteins(pro-caspase-1,cleaved caspase-1/p20,GSDMD,and cleaved N-terminal GSDMD)and TLR4 among the normal keratinocyte cell line HaCaT and SCC cell lines A431 and SCL-1.Results Quantitative PCR and Western blot results showed that the mRNA and protein expression levels of TLR4,CPB1,NLRP3,IL-1β,and IL-18 were significantly higher in SCC tissues than in AK and normal skin tissues(P<0.05).TLR4/TUNEL double immunofluorescence results revealed a progressive increasing trend in TLR4 expression and pyroptosis levels from normal skin to AK and further to SCC(P<0.05).Furthermore,in SCL-1 cells,the expression levels of pro-caspase-1,cleaved caspase-1/p20,cleaved N-terminal GSDMD,and TLR4 were significantly upregulated(P<0.05),whereas in A431 cells,only TLR4 expression was increased(P<0.05),and the levels of other pyroptosis-related proteins were downregulated compared to HaCaT cells(P<0.05).Conclusion The expression of the TLR4 signaling pathway gradually increased from AK to SCC,and its activation status varied among different cutaneous squamous cell carcinoma cell lines.

Objective To investigate the effect of Cyclocarya paliurus(Batal.)lljinskaja polysaccharides on insulin resistance in type 2 diabetic rats by regulating glucose transporter 4(GLUT4)translocation in islet and liver.Methods High-fat diet combined with low-dose streptozotocin(35 mg·kg-1)to induce type 2 diabetes model,all the rats were randomly divided into model control group,Cyclocarya paliurus polysaccharides groups(5,10 g·kg-1)and metformin group(0.25 g·kg-1),and treated for eight weeks(n=9 in each group).Fasting glucose and lipid were determined.Histopathology of rat islet and liver were observed by hematoxylin and eosin staining.Protein expressions of phosphorylated phosphoinositide-3-kinase(p-PI3K),phosphorylated serine-threonine kinase 1(p-Akt1),and GLUT4 in islet were measured by immunohistochemistry staining.GLUT4 translocation in the islet and liver was detected by immunofluorescence.Results Compared with the model control group,the Cyclocarya paliurus polysaccharides group and metformin group had declined fasting glucose levels and increased high-density lipoprotein(P＜0.05).The structure of the islets and liver was relatively complete.The content of p-PI3K,p-Akt1 and GLUT4 in the islet increased(P＜0.05).GLUT4 translocation in the liver and islet enhanced(P＜0.05).Conclusions Cyclocarya paliurus polysaccharides alleviate glucose and lipid metabolism disorders.The mechanism may lay in it activating protein expressions of p-PI3K,p-Akt1,and GLUT4 in islet cells.GLUT4 translocation to the islet and liver cell membrane are increased to regulate peripheral islet resistance.

Objective To establish a method of LC-MS/MS for determining cordycepin(Cor)and 3′-deoxyinosine(3′-Deo)concentration in rat plasma,and to study their pharmacokinetics in rats.Methods Protein was precipitated with methanol using 2-chloadenosine(2-Chl)as an internal standard.The chromatography was performed on Kinetex C18(3 mm×100 mm,2.6 μm,Phenomenex,USA)with gradient elution in aqueous(5 mmol·L-1 ammonium acetate)-methanol solution as mobile phase.ESI ion source was used for mass spectrometry,and positive ion multiple reaction monitoring(MRM)was used for scanning detection.The pharmacokinetics of Cor and 3′-Deo after oral administration of Cor(10 mg·kg-1)were studied in rats.Results Cor at 0.5-100 ng·mL-1 and 3′-Deo at 1-200 ng·mL-1 had good linearity,and the lower limits of quantification were 0.5 and 1 ng·mL-1,respectively.After oral administration of Cor in rats,the plasma concentration of Cor was low,which was mainly converted into the metabolite 3′-Deo.The Cmax of Cor and 3′-Deo were(5.4±3.4)and(142.0±50.0)ng·mL-1,and AUC0-360min min were(658.4±459.3)and(18 034.9±4 981.1)ng·min·mL-1,respectively.Conclusion The method is simple,sensi-tive,and accurate,which is suitable for determining Cor and 3′-Deo concentration in plasma and the pharmacokinetic study.

Objective: To understand the current situation and influencing factors of sexual assault among middle school students, and to provide a basis for promoting healthy psychosocial environment for children and adolescents. Methods: A questionnaire survey was conducted among 1 471 middle school students in one city of Sichuan Province from April to May 2020 using multi stage stratified cluster sampling. Results: The proportion of middle school students who have been raped, indecent assault and sexual assault was 1.6%, 3.7% and 9.4%, respectively. About 23.8% of them had suffered two or more types of sexual assault; 75.1 % to 82.1% of sexual assault occurred between the ages of 9 and 15. Between 36.3% and 62.3% of sexual assault subjects were acquaintances of the same generation. Pornography exposure was the same risk factor for the three types of sexual assault (rape: OR= 9.93, 95%CI =3.09-27.57; sexual obscenity: OR=7.83, 95%CI =3.95-15.53; sexual harassment: OR=5.22, 95%CI= 3.52- 7.73, P <0.01). Low gender identity was the same risk factor for both suffering sexual obscenity and sexual harassment (sexual obscenity: OR=2.37, 95%CI =1.31-4.29; sexual harassment: OR=1.73, 95%CI=1.16-2.58, P <0.01). The long term absence of mothers was a risk factor for suffering sexual rape among middle school students ( OR=3.10, 95%CI =1.31-7.30), as well as father s sex education was a risk factor for suffering sexual obscenity among middle school students ( OR=2.52, 95%CI = 1.26 - 5.03 )( P <0.01). Conclusion Pornography exposure is the same risk factor for all types of sexual assault among middle school students. Low gender identity is the same risk factor for indecent assault and sexual harassment. Sexual education capabilities of families and schools should be improved, and attention should be paid to the gender identity and sexual mental health of middle school students.

Abstract: Objective To investigate the prevalence and influencing factors of depressive symptoms among primary and middle school students in Xihu District, Hangzhou City, so as to provide insights into depression control among primary and middle school students. Methods: Students were sampled from two primary schools, two junior high schools, two ordinary high schools and one vocational high school in Xihu District using a stratified cluster random sampling method from September to November, 2021. Participants' demographics, diet, exercise, sleep and campus bullying were collected using questionnaire surveys. The depressive symptoms were evaluated using the Chinese version of the Center for Epidemiologic Studies-Depression Scale (CES-D), and factors affecting the depressive symptoms were identified among primary and middle school students using a multivariable logistic regression model. Results: A total of 1 518 students were investigated, including 518 primary school students (34.12%), 363 junior high school students (23.91%), 463 ordinary high school students (30.50%) and 174 vocational high school students (11.46%). Of all participants, there were 805 males (53.03%). The prevalence of depressive symptoms was 38.87% among the participants, and was 36.10%, 42.42%, 42.33% and 30.46% among primary school, junior high school, ordinary high school and vocational high school students, respectively. The multivariable logistic regression analysis showed that gender (OR=1.601, 95%CI: 1.289-1.989), type of school (junior high school, OR=1.299, 95%CI: 0.974-1.734; ordinary high school, OR=1.351, 95%CI: 1.025-1.780; vocational high school, OR=0.862, 95%CI: 0.585-1.272), frequency of fresh vegetable intake (less than once daily, OR=0.454, 95%CI: 0.304-0.679; once and more daily, OR=0.359, 95%CI: 0.235-0.548), duration spent on the internet (0.5 to 1 h daily, OR=1.447, 95%CI: 1.044-2.006; 1 h and longer daily, OR=1.456, 95%CI: 1.072-1.978), sufficient sleep (OR=0.525, 95%CI: 0.371-0.744), suffering from campus bullying (OR=1.992, 95%CI: 1.491-2.661) were associated with the development of depressive symptoms. Conclusions The prevalence of depressive symptoms is high among primary and middle school students in Xihu District. Gender, type of school, dietary behaviors, sleep duration and campus bullying are factors affecting the development of depressive symptoms.

Sympathetic cues via the adrenergic signaling critically regulate bone homeostasis and contribute to neurostress-induced bone loss, but the mechanisms and therapeutics remain incompletely elucidated. Here, we reveal an osteoclastogenesis-centered functionally important osteopenic pathogenesis under sympatho-adrenergic activation with characterized microRNA response and efficient therapeutics. We discovered that osteoclastic miR-21 was tightly regulated by sympatho-adrenergic cues downstream the β2-adrenergic receptor (β₂AR) signaling, critically modulated osteoclastogenesis in vivo by inhibiting programmed cell death 4 (Pdcd4), and mediated detrimental effects of both isoproterenol (ISO) and chronic variable stress (CVS) on bone. Intriguingly, without affecting osteoblastic bone formation, bone protection against ISO and CVS was sufficiently achieved by a (D-Asp₈)-lipid nanoparticle-mediated targeted inhibition of osteoclastic miR-21 or by clinically relevant drugs to suppress osteoclastogenesis. Collectively, these results unravel a previously underdetermined molecular and functional paradigm that osteoclastogenesis crucially contributes to sympatho-adrenergic regulation of bone and establish multiple targeted therapeutic strategies to counteract osteopenias under stresses.
Adrenergic Agents/pharmacology* ; Apoptosis Regulatory Proteins/pharmacology* ; Bone Diseases, Metabolic/metabolism* ; Humans ; Liposomes ; MicroRNAs/genetics* ; Nanoparticles ; Osteoclasts ; Osteogenesis/physiology* ; RNA-Binding Proteins/pharmacology*

Per-and polyfluoroalkylated substances (PFASs) are a group of persistent organic pollutants that are widespread in the environment and organisms. Given their unique hydrophobicity, oil-repellence, and chemical stability, PFASs are widely used in various industrial and commercial products. PFASs can accumulate and be biomagnified through the food chain, and its toxic effects have posed a certain threat to human health. The response of the immune system to PFASs exposure is one of the most sensitive human health effects, and has attracted remarkable attention from related scientists and organizations. We summarized international and domestic epidemiological studies on the associations between exposure to PFASs and immune system, including immunosuppression and immunoenhancement. We also reviewed experimental evidence of PFASs on immune system from perspectives of immune organs, immune cells, and cytokines. Furthermore, the possible mechanisms of peroxisome proliferator-activated receptor-α (PPAR-α)-dependent, nuclear factor-κB (NF-κB)-activated, and mitochondrial apoptosis pathways were summarized. While the relationships between PFASs and immune-related diseases in human are not yet conclusive, accumulative epidemiological studies provide evidence for associations between PFASs and reduced immune response to vaccination in children. In addition, previous studies mainly focus on the immunotoxicity of traditional PFASs, and our understanding of the molecular mechanism of the effects of PFASs on immune system is still in its infancy. Therefore, it is necessary to further explore the immunotoxicity of new PFASs and associated mechanism.

Objective To investigate the prevalence of allergic and infectious diseases in children， and to assess the influence of indoor and outdoor environmental factors on these two common diseases in children. Methods A questionnaire was used to investigate the prevalence of allergic and infectious diseases in 140 children of 7 years old in Laizhou Bay， Shandong Province. Logistic regression was used to analyze the associations between indoor and outdoor environmental factors and children’s allergic and infectious diseases， respectively. Results The prevalence of previous eczema and other allergic diseases for the past year in children was 37.9% and 15.0%， respectively， and the prevalence of infectious diseases for the past year was 35.7%. As for allergic diseases， eye irritation due to outdoor air （ OR =2.977； 95% CI ： 1.407‒6.296） and nose irritation due to outdoor air （ OR =5.147； 95% CI ： 1.272‒20.827） were the risk factors for previous eczema in children. Indoor musty taste increased the risks of urticaria （ OR =4.306； 95% CI ： 1.062‒17.454） and previous eczema （ OR =3.853； 95% CI ： 1.080‒13.743）. The use of cockroach killers indoors increased the risk of rhinitis （ OR =6.102； 95% CI ：1.297‒28.697）. As for infectious diseases， having outdoor pollution sources increased the risk of gastrointestinal infection （ OR =4.937； 95% CI ： 1.050‒23.216）， and the use of mosquito coils and clothing mothproofing agents increased the risks of respiratory （ OR =6.333； 95% CI ： 1.397‒28.714） and gastrointestinal infections （ OR =3.218； 95% CI ： 1.074‒9.644）， respectively. However， we did not find associations between indoor passive smoking and allergic or infectious diseases. Conclusion Except outdoor passive smoking， all the other indoor and outdoor environmental factors increase the risks of children’s allergies and infectious diseases.

Objective To observe humoral and cellular immune responses induced in mice by a TSOL18 recombinant Mycobacterium smegmatis (MS) vaccine of Taenia solium.Methods Sixty specific pathogen-free (SPF) Kunming mice (18-22 g,half male and half female) were divided into 3 groups by random number table method,20 mice in each group.One group was administered with recombinant MS-TSOL18 vaccine,one group was administered with MS as control,and one group was administered phosphate buffer saline (PBS) as control.Kunming mice were vaccinated once every two weeks for 2 times through intragastric administration.At 0,2,4,6,and 8 weeks after immunization,blood sample was collected and serum was separated.The levels of IgG and IgG2a in serum were detected using enzyme linked immunosorbent assay (ELISA).The spleen was separated to prepare spleen suspension.The proliferation of spleen lymphocyte with the stimulation of a specific antigen was detected by CCK-8.The levels of interleukin (IL)-2 and IL-4 in spleen cell culture supernatant with the stimulation of a specific antigen were detected by ELISA.Results Compared with MS control group (IgG:0.160 ± 0.019,0.187 ± 0.038,0.193 ± 0.050,0.170 ± 0.005;IgG2a:0.213 ± 0.010,0.198 ± 0.012) and PBS control group (IgG:0.159 ± 0.015,0.184 ± 0.029,0.191 ± 0.025,0.165 ± 0.018;IgG2a:0.198 ± 0.032,0.178 ± 0.025),the levels of specific IgG (0.310 ± 0.034,0.391 ± 0.029,0.443 ± 0.030,0.373 ± 0.021) in recombinant MS-TSOL18 vaccine group increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),the levels of specific IgG2a (0.446 ± 0.056,0.339 ± 0.026) in recombinant MS-TSOL18 vaccine group increased at 6 and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of spleen lymphocyte proliferation increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th week.After antigen stimulation,compared with MS and PBS control groups,the levels of IL-2 and IL-4 in spleen lymphocyte culture supernatant increased at 2,4,6,and 8 weeks after immunization (P ＜ 0.05),and reached the highest level by the 6th and 4th weeks,respectively.Conclusion The recombinant MS-TSOL18 vaccine of Taenia solium might induce mice to produce humoral and cellular immune responses.

Objectives To explore the relationships between the current physical exercise behaviors and psychological factors of middle school students,so as to provide scientific evidence for improving health education and carrying out targeted health intervention measures.Methods The Multi-stage stratified sampling and cluster sampling method was adopted to choose 3600 students from 12 junior and senior middle schools in Hangzhou,Wuhan and Xi'an.The date was collected using the adolescent physical exercise behavior-psychological measurement scale(Chinese version).The structural equation model was built to explore the relationships between psychological factors(including the change of strategy,decision balance positive effect,decision balance negative effect and self-efficiency)and physical exercise.Results The change of strategy has biggest effect on physical exercise behaviors,with the standardized direct effect value of 0.344.The standardized direct effect value of decision balance negative effect,decision balance positive effect and self-efficiency were 0.071,0.093 and 0.169 respectively.The negative correlation among the four dimensions was significant statistically.Conclusion Psychological factors have direct effects on the change of the behaviors,and the change of the strategy has the largest impact on physical exercise behaviors of middle school students.The four psychological factors are closely correlated with one another,which is important in formulating middle school sports programs to promote the physical quality of middle school students.