Objective : To explore the mechanism of isorhamnetin (Iso) in the treatment of oral squamous cell carcinoma (OSCC) using network pharmacology and molecular docking methods and to verify it in vitro. Methods : The key targets were obtained by constructing the PPI protein interaction network based on the common intersection targets of Iso-OSCC. At the same time, gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the related signaling pathways of the intersection targets. Iso and core targets were also analyzed through molecular docking and visualization. Colony formation assay and Transwell assay were used to identify the effect of Iso on the proliferation and invasion of Cal-27 cells. Western blot was used to analyze the regulatory effects of different concentrations of Iso on estrogen receptor-1 (ESR1), phosphoinositide-3-kinase regulatory subunit-1 (PIK3R1), Src tyrosine kinase (SRC), and phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling pathway proteins. Results: A total of 269 potential intersection targets of Iso-regulated OSCC were obtained. According to the degree obtained by topological analysis, PIK3R1, AKT1, SRC, ESR1, and other core targets were screened out. KEGG analysis showed that 165 signaling pathways were enriched in the intersection targets of Iso-OSCC, among which the PI3K/AKT signaling pathway played an important role in the treatment of OSCC with Iso. Molecular docking results showed that the absolute value of binding energy between target proteins PIK3R1, AKT1, SRC, ESR1, and Iso was high. After Cal-27 cells were treated with Iso, the number of cell colony formations, the number of transmembrane cells, and the expression of PIK3R1, ESR1, SRC, p-PI3K, and p-AKT were negatively correlated with the increase in Iso concentration (P < 0.05). Conclusion Iso can inhibit PI3K/AKT signal transduction and influence the expression of PIK3R1, AKT1, SRC, and ESR1 proteins, thereby inhibiting the occurrence and development of OSCC.

Objective: To understand the home reading-writing levels among children and adolescents in Shanghai after school, and to explore its association with screening myopia, so as to provide a scientific basis for the prevention and control of myopia. Methods: From April to December 2024, 641 primary and middle school students were recruited from 2 urban schools and 1 rural school in Shanghai to participate in the survey. An illuminance meter was used to measure the illuminance of home reading-writing activities after school. Screening myopia was determined through visual acuity examination and refractive detection under non ciliary muscle paralysis conditions among children and adolescents. A binary Logistic regression model was used to analyze the association between home reading-writing illuminance and screening myopia. Results: The detection rate of screening myopia among children and adolescents in Shanghai was 59.9%. The median home reading-writing illuminance after school was 340.9(112.2, 753.5) lx, and 45.4% was found of less than 300 lx. The family illuminance in the primary school stage [432.0 (136.9, 837.0) lx] was higher than that in the junior high school stage [113.1(53.7, 375.1) lx], and main urban area group [503.9 (212.6, 969.5) lx] was higher than that in the rural group [141.6 (53.7, 416.9) lx], the differences were statistically significant (Z=-7.56, -9.95,both P<0.05). The results of Logistic regression analysis showed that compared with the family illuminance of 150-500 lx, children and adolescents with family illuminance<150 and >500 lx had increased risks of screening myopia detection[OR(95%CI)=1.56(1.01-2.42), 1.74(1.15-2.62),both P<0.05]. Conclusions The home reading-writing illuminance after school is suboptimal. Both excessively low and high home reading-writing illuminance levels are associated with screen-detected myopia. It is necessary for children and adolescents to improve lighting conditions during evening reading-writing activities, and strengthen health education according to different regions and school stages.

OBJECTIVES: This study aimed to explore the therapeutic effect and mechanism of ginsenoside Rb3 on experimental periodontitis and bone resorption in rats. METHODS: Male SD rats were randomly divided into a control group, a ligation group, an Rb3 group, and a doxycycline (Dox) group for in vivo experiments. A periodontitis model was established by ligating the maxillary second molar, and samples were collected after 3 weeks of drug treatment. Micro-CT assessment of alveolar bone resorption was performed, and hematoxylin-eosin (HE) staining was used to observe pathological changes in periodontal and visceral tissues. Tartrate resistant acid phosphatase (TRAP) staining was applied to detect the formation of osteoclasts in periodontal tissues, and enzyme-linked immunosorbent assay (ELISA) was adopted to detect the serum levels of interleukin (IL)-6, IL-8, immunoglobulin (Ig)M, and IgG. Quantitative polymerase chain reaction (qPCR) was employed to detect the expression of factors related to gingival inflammation and osteoclast formation. Immunofluorescence staining was used to detect phospho-extracellular signal-regulated kinase (p-ERK) expression. In vitro experiments were conducted by pretreating RAW264.7 cells with drugs and adding lipopolysaccharides (LPS) stimulation from Porphyromonas gingivalis (P. gingivalis). IL-1β and IL-6 mRNA expression was detected by qPCR, and Western blot was used to detect the effect of Rb3 on the mitogen-activated protein kinases (MAPKs) signaling pathway. RESULTS: Compared with the control group, the ligation group showed significant periodontitis and bone resorption. Compared with the ligation group, the Rb3 group showed a decrease in alveolar bone resorption and osteoclast formation; p-ERK/ERK ratio, IL-1β, IL-6, and nuclear factor of activated T cells (NFATc1) mRNA levels and downstream gene expression in periodontal tissues; serum IL-6, IL-8, IgG, and IgM levels. Rb3 reduced IL-8 and IL-1β mRNA expression levels and p-ERK/ERK and p-p38 MAPK/p38 MAPK ratios in RAW264.7 cells induced by P. gingivalis LPS stimulation. CONCLUSIONS Rb3 inhibits inflammation and bone resorption in experimental periodontitis in rats. Compared with Dox, Rb3 has better effects in inhibiting pro-inflammatory factors and osteoclast gene expression and may exert anti-inflammatory effects by activating the MAPK signaling pathway.
Animals ; Ginsenosides/therapeutic use* ; Rats, Sprague-Dawley ; Male ; Periodontitis/pathology* ; Rats ; Osteoclasts/drug effects* ; Interleukin-1beta/metabolism* ; Interleukin-6/blood* ; Mice ; Alveolar Bone Loss ; Interleukin-8/blood* ; Immunoglobulin G/blood* ; RAW 264.7 Cells ; Transcription Factors

Objective:To analyze the effect of long noncoding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) on the radiosensitivity of cervical cancer cells and explore its regulatory role in the miR-455-3p/ nuclear factor-κB (NF-κB) signaling pathway.Methods:The expression levels of lncRNA SNHG16 and miR-455-3p in human normal cervical epithelial cells H8, human cervical cancer cells SiHa, and radioresistant cervical cancer cells SiHa-R were detected by real-time reverse transcription polymerase chain reaction. SiHa-R cells were transfected separately, and then given a single dose of 4 Gy X-ray irradiation and continued to be cultured for subsequent experiments. The cells in each group were named siRNA-NC, siRNA-SNHG16 (interfering lncRNA SNHG16), NC mimic, miR-455-3p mimic (overexpressing miR-455-3p), siRNA-SNHG16+inhibitor NC, and siRNA-SNHG16+miR-455-3p inhibitor groups, respectively. The survival fraction of SiHa-R cells was detected by clone formation assay. The apoptosis rate of SiHa-R cells was analyzed by flow cytometry. The expression levels of apoptotic proteins [cysteine-containing aspartate-specific protease (Caspase)-3, Caspase-9, Bax] and NF-κB signaling pathway related proteins [NF-κB p65, phosphorylated (p)-NF-κB p65, p-IκB (inhibitory protein of NF-κB)] were measured by Western blot. The targeting relationship between lncRNA SNHG16 and miR-455-3p was determined by dual luciferase reporter gene assay. Comparison among different groups was conducted by one-way ANOVA, and paired comparison was carried out by LSD t-test. Comparison between two groups was performed by t-test. Results:Compared with H8 cells, the expression levels of lncRNA SNHG16 were increased in SiHa and SiHa-R cells, and SiHa-R cells had a higher level than SiHa cells. The expression levels of miR-455-3p were decreased in SiHa and SiHa-R cells, and SiHa-R cells had a lower level than SiHa cells (all P<0.001). Compared with the siRNA-NC group, the survival fraction of SiHa-R cells in the siRNA-SNHG16 group was decreased, the radiosensitization ratio (SER) was 1.571 (>1), the apoptosis rate and levels of Caspase-3, Caspase-9, and Bax proteins were increased, while the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). lncRNA SNHG16 could target miR-455-3p. Compared with the NC mimic group, miR-455-3p level in the miR-455-3p mimic group was increased, cell survival fraction was decreased, the SER was 1.826 (>1), the apoptosis rate and the levels of Caspase-3, Caspase-9, Bax proteins were increased, and the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). Inhibition of miR-455-3p expression could weaken the effect of interfering with lncRNA SNHG16 expression on SiHa-R cell apoptosis, radiotherapy sensitivity, and NF-κB signaling pathway (all P<0.001). Conclusions:Interference with lncRNA SNHG16 expression could induce the apoptosis of cervical cancer cells and enhance their radiation sensitivity by regulating the miR-455-3p/NF-κB signaling pathway.

Background and purpose:Polo-like kinase 4(PLK4)is a cell cycle regulatory protein,which is closely related to the occurrence and development of a variety of malignant tumors.The aim of this study was to investigate the role of PLK4 in the proliferation,invasion and apoptosis of oral squamous cell carcinoma(OSCC).Methods:Gene Expression Profiling Interactive Analysis 2(GEPIA2)and the University of Alabama at Birmingham Cancer Data Analysis Portal(UALCAN)were used to analyze the expression of PLK4 in head and neck squamous cell carcinoma and surrounding normal tissues.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot were employed to evaluate the mRNA and protein expression of PLK4 in OSCC cells.We further analyzed PLK4,phosphoinositide 3-kinase(PI3K),phosphorylated phosphoinositide 3-kinase(p-PI3K),protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),survivin,and cyclin D1 protein expression.The effects of PLK4 on cell proliferation,apoptosis and invasion were analyzed by cell counting kit-8(CCK-8)assay,flow cytometry and transwell assay.In addition,12 4-week-old SPF BALB/c female nude mice were divided into sh-NC group(n=6)and sh-PLK4 group(n=6)by random number table method.The sh-NC/sh-PLK4 cells were injected into the right anterior armpit of nude mice for subcutaneous tumor formation.The body weight,tumor volume and tumor weight of the two groups of nude mice were observed,and the stripped tumor tissues were analyzed by H-E staining.The animal experiments were approved by the Animal Experiment Center of Shandong Second Medical University(No:2024SDL840).Results:The results of GEPIA2 and UALCAN online databases showed that PLK4 was highly expressed in OSCC compared with normal tissues.In addition,PLK4 was highly expressed in OSCC cell lines(HN6,Cal-27,SCC-4,SCC-9,SCC-25)compared with oral mucosal epithelial cells(P＜0.05).Western blot results showed that the expression levels of PI3K/AKT signaling pathway proteins p-PI3K,p-AKT,cyclin D1 and survivin were decreased after PLK4 knockdown and increased after PLK4 overexpression in OSCC cells(P＜0.05).The cell proliferation activity and the number of transmembrane cells were positively correlated with the decrease and increase of PLK4 expression(P＜0.05),while the cell apoptotic rate was negatively correlated(P＜0.05),indicating that cell proliferation and apoptosis were both affected.In addition,compared with the sh-NC group,the tumor volume and weight of the nude mice in the sh-PLK4 group were decreased(P＜0.05),while there was no significant difference in the body weight of the nude mice between the two groups(P＞0.05).Moreover,the nuclear atypia of tumor tissues in sh-PLK4 group was lower than that in sh-NC group(P＜0.05).Conclusion:PLK4 can regulate the invasion,proliferation and apoptosis of OSCC cells,potentially through the activation of PI3K/AKT signaling pathway.

Background and purpose:Polo-like kinase 4(PLK4)is a cell cycle regulatory protein,which is closely related to the occurrence and development of a variety of malignant tumors.The aim of this study was to investigate the role of PLK4 in the proliferation,invasion and apoptosis of oral squamous cell carcinoma(OSCC).Methods:Gene Expression Profiling Interactive Analysis 2(GEPIA2)and the University of Alabama at Birmingham Cancer Data Analysis Portal(UALCAN)were used to analyze the expression of PLK4 in head and neck squamous cell carcinoma and surrounding normal tissues.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot were employed to evaluate the mRNA and protein expression of PLK4 in OSCC cells.We further analyzed PLK4,phosphoinositide 3-kinase(PI3K),phosphorylated phosphoinositide 3-kinase(p-PI3K),protein kinase B(AKT),phosphorylated protein kinase B(p-AKT),survivin,and cyclin D1 protein expression.The effects of PLK4 on cell proliferation,apoptosis and invasion were analyzed by cell counting kit-8(CCK-8)assay,flow cytometry and transwell assay.In addition,12 4-week-old SPF BALB/c female nude mice were divided into sh-NC group(n=6)and sh-PLK4 group(n=6)by random number table method.The sh-NC/sh-PLK4 cells were injected into the right anterior armpit of nude mice for subcutaneous tumor formation.The body weight,tumor volume and tumor weight of the two groups of nude mice were observed,and the stripped tumor tissues were analyzed by H-E staining.The animal experiments were approved by the Animal Experiment Center of Shandong Second Medical University(No:2024SDL840).Results:The results of GEPIA2 and UALCAN online databases showed that PLK4 was highly expressed in OSCC compared with normal tissues.In addition,PLK4 was highly expressed in OSCC cell lines(HN6,Cal-27,SCC-4,SCC-9,SCC-25)compared with oral mucosal epithelial cells(P＜0.05).Western blot results showed that the expression levels of PI3K/AKT signaling pathway proteins p-PI3K,p-AKT,cyclin D1 and survivin were decreased after PLK4 knockdown and increased after PLK4 overexpression in OSCC cells(P＜0.05).The cell proliferation activity and the number of transmembrane cells were positively correlated with the decrease and increase of PLK4 expression(P＜0.05),while the cell apoptotic rate was negatively correlated(P＜0.05),indicating that cell proliferation and apoptosis were both affected.In addition,compared with the sh-NC group,the tumor volume and weight of the nude mice in the sh-PLK4 group were decreased(P＜0.05),while there was no significant difference in the body weight of the nude mice between the two groups(P＞0.05).Moreover,the nuclear atypia of tumor tissues in sh-PLK4 group was lower than that in sh-NC group(P＜0.05).Conclusion:PLK4 can regulate the invasion,proliferation and apoptosis of OSCC cells,potentially through the activation of PI3K/AKT signaling pathway.

Objective:To analyze the effect of long noncoding RNA (lncRNA) small nucleolar RNA host gene 16 (SNHG16) on the radiosensitivity of cervical cancer cells and explore its regulatory role in the miR-455-3p/ nuclear factor-κB (NF-κB) signaling pathway.Methods:The expression levels of lncRNA SNHG16 and miR-455-3p in human normal cervical epithelial cells H8, human cervical cancer cells SiHa, and radioresistant cervical cancer cells SiHa-R were detected by real-time reverse transcription polymerase chain reaction. SiHa-R cells were transfected separately, and then given a single dose of 4 Gy X-ray irradiation and continued to be cultured for subsequent experiments. The cells in each group were named siRNA-NC, siRNA-SNHG16 (interfering lncRNA SNHG16), NC mimic, miR-455-3p mimic (overexpressing miR-455-3p), siRNA-SNHG16+inhibitor NC, and siRNA-SNHG16+miR-455-3p inhibitor groups, respectively. The survival fraction of SiHa-R cells was detected by clone formation assay. The apoptosis rate of SiHa-R cells was analyzed by flow cytometry. The expression levels of apoptotic proteins [cysteine-containing aspartate-specific protease (Caspase)-3, Caspase-9, Bax] and NF-κB signaling pathway related proteins [NF-κB p65, phosphorylated (p)-NF-κB p65, p-IκB (inhibitory protein of NF-κB)] were measured by Western blot. The targeting relationship between lncRNA SNHG16 and miR-455-3p was determined by dual luciferase reporter gene assay. Comparison among different groups was conducted by one-way ANOVA, and paired comparison was carried out by LSD t-test. Comparison between two groups was performed by t-test. Results:Compared with H8 cells, the expression levels of lncRNA SNHG16 were increased in SiHa and SiHa-R cells, and SiHa-R cells had a higher level than SiHa cells. The expression levels of miR-455-3p were decreased in SiHa and SiHa-R cells, and SiHa-R cells had a lower level than SiHa cells (all P<0.001). Compared with the siRNA-NC group, the survival fraction of SiHa-R cells in the siRNA-SNHG16 group was decreased, the radiosensitization ratio (SER) was 1.571 (>1), the apoptosis rate and levels of Caspase-3, Caspase-9, and Bax proteins were increased, while the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). lncRNA SNHG16 could target miR-455-3p. Compared with the NC mimic group, miR-455-3p level in the miR-455-3p mimic group was increased, cell survival fraction was decreased, the SER was 1.826 (>1), the apoptosis rate and the levels of Caspase-3, Caspase-9, Bax proteins were increased, and the levels of p-NF-κB p65 and p-IκB proteins were decreased (all P<0.001). Inhibition of miR-455-3p expression could weaken the effect of interfering with lncRNA SNHG16 expression on SiHa-R cell apoptosis, radiotherapy sensitivity, and NF-κB signaling pathway (all P<0.001). Conclusions:Interference with lncRNA SNHG16 expression could induce the apoptosis of cervical cancer cells and enhance their radiation sensitivity by regulating the miR-455-3p/NF-κB signaling pathway.

Objective: The study aims to analyze the prevalence and associated factors of myopia among 4 to 9 grade students in Guangdong Province in 2022, so as to provide a scientific basis for targeted intervention measures for myopia in children and adolescents. Methods: From September to October 2022, stratified cluster random sampling was used to select 29 095 of 4 to 9 grade students from Guangzhou, Jiangmen, and Meizhou in Guangdong Province for myopia screening and questionnaire surveys. The Chisquare test was applied to compare the differences between groups, and multivariable Logistic stepwise regression analysis was used to analyze factors associated with myopia. Results: The myopia detection rate of 4 to 9 grade students was 61.7%, with detection rates of 51.5% for 4 to 6 grade primary school students and 71.95% for 7 to 9 grade junior high school students. Multivariable Logistic regression analysis showed that higher myopia rates were detected among girls (OR=1.39, 95%CI=1.30-1.49), students with one (OR=1.82, 95%CI=1.69-1.96) or both parents having myopia (OR=2.86, 95%CI=2.56-3.18), and indoor sedentary time >6 h(OR=1.28, 95%CI=1.17-1.39) in the 4 to 6 grade. Lower myopia rates were detected in the county (OR=0.92, 95%CI=0.86-0.99) and outdoors at recess activities (OR=0.88, 95%CI=0.81-0.95). Meanwhile, higher myopia rates were detected among girls (OR=1.84, 95%CI=1.69-1.99), students with one (OR=1.87, 95%CI=1.71-2.04) or both parents having myopia (OR=3.03, 95%CI=2.63-3.50), and indoor sedentary time >6 h/d (OR=1.11, 95%CI=1.01-1.23) in the 7 to 9 grade. Lower myopia rates were detected in the county (OR=0.74, 95%CI=0.68-0.80), outdoors at recess activities (OR=0.83, 95%CI=0.76-0.91), and outdoor activity time ≥2 h/d(OR=0.87, 95%CI=0.80-0.95)(P<0.05). Conclusions The detection rate of myopia among 4 to 9 grade students in Guangdong Province is relatively high. Place of recess activities, daily outdoor activity and indoor sedentary duration are associated with myopia. Therefore, targeted intervention measures should be adopted, such as appropriately increasing outdoor activity to reduce the occurrence of myopia among primary and middle school students.

Background Previous studies have confirmed that nicotine exposure is an independent risk factor for miscarriage, but it is not clear whether nicotine causes unexplained recurrent spontaneous abortion (URSA) through oxidative stress. Objective To explore potential mediating effect of oxidative stress on the relationship between nicotine exposure and URSA. Methods Using a 1∶1 matched case-control study, 88 patients with URSA visiting Beijing Obstetrics and Gynecology Hospital affiliated to Capital Medical University from April to October in 2018 were selected as the case group, and 88 pregnant women without adverse pregnancy outcomes and seeking induced abortion in the outpatient clinic of the same hospital were selected as the control group. The levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 8-iso-prostaglandin F2α (8-iso-PGF2α) in urine were determined by enzyme-linked immunosorbent assay, and the level of urinary nicotine was determined by gas chromatography-mass spectrometry. Conditional logistic regression was used to analyze the associations of nicotine, 8-OHdG, and 8-iso-PGF2α with the risk of URSA. Multiple linear regression was used to analyze the association of nicotine with 8-OHdG and 8-iso-PGF2α. The potential mediating effect of oxidative stress on URSA after nicotine exposure was explored by dichotomous mediating model. Results The median concentrations (creatinine corrected) of nicotine, 8-OHdG, and 8-iso-PGF2α in urine of the case group were 7.78, 4.84, and 44.10 μg·g⁻¹, respectively, while those of the control group were 6.48, 3.34, and 29.39 μg·g⁻¹, respectively. The concentrations of nicotine, 8-OHdG, and 8-iso-PGF2α in urine of the case group were all higher than those of the control group (P < 0.05). The results of conditional logistic regression model showed that after adjusting selected confounding factors, compared with the Q₁ groups of nicotine and 8-iso-PGF2α, the OR (95%CI) values of URSA in the Q₄ groups were 4.20 (1.33-13.29) and 6.25 (1.66-23.59), respectively. Compared with the Q₁ group of 8-OHdG, the OR (95%CI) values of URSA in the Q₁, Q₂, and Q₃ groups were 5.47 (1.43-20.93), 4.24 (1.28-14.07), and 6.36 (1.82-22.28), respectively. The results of multiple linear regression showed that after adjusting confounding factors, there was a positive correlation between urinary nicotine and 8-OHdG in both the case group and the control group, and the b (95%CI) values were 0.76 (0.67-0.86) and 0.81 (0.67-0.95) respectively; there was a positive correlation between urinary nicotine and 8-iso-PGF2α in both the case group and the control group, and the b (95%CI) values were 0.65 (0.55-0.75) and 0.76 (0.64-0.87), respectively. The results of dichotomous mediating analysis showed that the mediating effect of 8-iso-PGF2α and its 95%CI on the relationship between nicotine exposure and URSA was 1.518 (0.749-2.311). Conclusion Internal nicotine exposure is a risk factor for URSA and is positively correlated with oxidative stress, and it may lead to URSA through lipid peroxidation damage.

Objective:The study is to examine association of sleep duration and cognitive impairment in the older adults aged 65 years and older in China.Methods:We analyzed data from 2017-2018 wave of Chinese Longitudinal Healthy Longevity Survey (CLHLS). A total of 14 966 participants were included in the analysis. Data with respect to socioeconomic status, community involvement, behavior pattern, diet, life style, family structure, disease condition, mental health and cognitive function were collected. Cognitive function was measured with Mini-mental State Examination (MMSE). We conducted generalized linear mixed models to examine associations of sleep duration with cognitive impairment, and subgroup analyses of sex and age were conducted.Results:Among 14 966 participants, the percentage of participants aged 65 to 79 years, 80 to 89 years, 90 to 99 years and 100 years and older was 5 148 (4.40%), 3 777 (25.24%), 3 322 (22.20%) and 2 719 (18.16%), respectively. A total of 2 704 participants reported sleep duration of 5 h and less, and 3 883 reported 9 h and more, accounting for 18.94% and 27.19%, respectively. In total, 3 748 were defined with cognitive impairment, accounting for 25.04%. The results of generalized linear mixed models showed that both short (≤5 h) and long (≥ 9 h) sleep duration were associated with cognitive impairment compared with sleep duration of 7 h, with OR(95% CI) of 1.35(1.09-1.68) and 1.70(1.39-2.07), respectively. The association of sleep duration with cognitive impairment was more obvious in males and individuals aged 65 to 79 years old. Conclusion:Short or long sleep duration was responsible for increased risk of cognitive impairment in older Chinese.