ObjectiveTo investigate the effect of icariin （ICA） on steroid-induced ferroptosis in bone microvascular endothelial cells （BMECs）. MethodsRat BMECs were selected and treated with 500 mg·L^-1 hydrocortisone for 1.5 h to establish a ferroptosis model of BMECs. The experimental cells were divided into a blank group， hormone group （500 mg·L^-1 hydrocortisone）， ICA group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA）， and ferroptosis agonist group （500 mg·L^-1 hydrocortisone + 34 mg·L^-1 ICA + 2.7 mg·L^-1 erastin）. Cell viability was detected by CCK-8. The levels of ferrous ion， glutathione （GSH）， malondialdehyde （MDA）， superoxide dismutase （SOD）， and reactive oxygen species （ROS） were detected by related kit species. The ferroptosis-related proteins， such as glutathione peroxidase 4（GPX4）， ferritin light chain （FTL）， and transferrin receptor protein1 （sTfR） were detected by Western blot， as well as autophagy-related proteins including microtubule-associated protein 1 light chain 3B （LC3B）， Beclin1， B-cell lymphoma-2 （Bcl-2）， and Caspase-3. Results500 mg·L^-1 hydrocortisone intervention for 1.5 h could effectively induce ferroptosis in BMECs， and ferroptosis levels could reach a peak as the intervention continued. In terms of cellular antioxidant capacity， compared with those in the blank group， the cell vitality， GSH in the hormone group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions were significantly increased （P<0.01）. Compared with those in the hormone group， the cell viability， GSH were significantly increased， and the levels of ROS， SOD， MDA， and ferrous ions were decreased in the ICA group （P<0.01）. Compared with those in the ICA group， the cell vitality， GSH in the ferroptosis agonist group decreased significantly， and the levels of ROS， SOD， MDA， and ferrous ions increased significantly （P<0.01）. In terms of the relationship between ferroptosis and autophagy， compared with the blank group， the hormone group had significantly increased expression levels of LC3B， sTfR， Beclin1， and FTL and significantly decreased expression levels of GPX4 （P<0.01）. Compared with the hormone group， The ICA group had significantly decreased expression levels of LC3B， sTfR， and FTL and significantly increased expression levels of Beclin 1 and GPX4 （P<0.01）. Compared with those in the ICA group， the expression levels of LC3B， sTfR， and FTL increased in the rapamycin group， and those of Beclin 1 and GPX4 decreased （P<0.01）. In terms of cell ferroptosis and apoptosis，compared with the blank group， the hormone group had significantly increased expression levels of FTL， sTfR and Caspase-3 and significantly decreased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with the hormone group， the ICA group had significantly decreased expression levels of FTL， sTfR and Caspase-3 and significantly increased expression levels of GPX4， and Bcl-2 （P<0.01）. Compared with those in the ICA group， the expression levels of FTL， sTfR and Caspase-3 in the ferroptosis agonist group were increased， and the expression levels of GPX4， and Bcl-2 were decreased （P<0.01）. In terms of cell function，compared with that in the blank group， the ability of cell migration and tube formation was significantly decreased in the hormone group （P<0.01）. Compared with that in the hormone group， the cell migration and tube formation ability in the ICA group were significantly increased （P<0.01）. ConclusionFerroptosis is involved in steroid-induced damage in BMECs. ICA can inhibit steroid-induced ferroptosis in BMECs， and the mechanism may be associated with the inhibition of ferroptosis by regulating autophagy.

BACKGROUND:Numerous scholars have previously researched certain greater tuberosity fractures and the procedures used to treat them.Few researchers,however,have studied the comminuted split fracture of the greater tuberosity of the humerus(Liu-Gang type IV)with rotator cuff tear in great detail. OBJECTIVE:To compare the clinical therapeutic effect of open repair position modified calcaneal plate combined with suture anchors and proximal humeral internal locking system(PHILOS)plate in the treatment of comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV). METHODS:Case data of 30 patients with comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV)from May 2012 to May 2022 were retrospectively analyzed.They were divided into the modified calcaneal plate combined with suture anchor group(group A)and the PHILOS with#2 Johnson group(group B),with 15 cases in each group.Intraoperative blood loss,surgical time,and incision length of all patients were recorded.Pain visual analog scale score,Constant-Murley score,as well as shoulder joint abduction,forward flexion,external rotation,and dorsal expansion activities during the last follow-up(>1 year)were evaluated. RESULTS AND CONCLUSION:(1)The surgical incision length and operation time were shorter,and blood loss was less in group A than those in group B(P<0.05).(2)No significant difference in visual analog scale score and Constant-Murley score was detected between the two groups(P>0.05).(3)During the last follow-up,forward flexion in group A was better than that in group B(P<0.05).No significant difference in abduction,external rotation,and dorsal expansion was determined between group A and group B(P>0.05).(4)In terms of complications,there was 1 case of shoulder joint pain and discomfort in group A(7%),2 cases of subacromial impingement syndrome,2 cases of upward movement of nodules,and 2 cases of shoulder joint pain(40%)in group B.There were significant differences in complication rates between the two groups(P=0.031).(5)In summary,the modified calcaneal plate combined with suture anchors in the treatment of comminuted fracture of split-type greater tuberosity of humerus combined with rotator cuff tears(Liu-Gang type IV)could better restore the forward flexion function of the shoulder joint and has a small incision,less blood loss,shorter operation time and fewer complications.

Objective Genotypes(G)1,3,and 5 of the Japanese encephalitis virus(JEV)have been isolated in China,but the dominant genotype circulating in Chinese coastal areas remains unknown.We searched for G5 JEV-infected cases and attempted to elucidate which JEV genotype was most closely related to human Japanese encephalitis(JE)in the coastal provinces of China. Methods In this study,we collected serum specimens from patients with JE in three coastal provinces of China(Guangdong,Zhejiang,and Shandong)from 2018 to 2020 and conducted JEV cross-neutralization tests against G1,G3,and G5. Results Acute serum specimens from clinically reported JE cases were obtained for laboratory confirmation from hospitals in Shandong(92 patients),Zhejiang(192 patients),and Guangdong(77 patients),China,from 2018 to 2020.Seventy of the 361 serum specimens were laboratory-confirmed to be infected with JEV.Two cases were confirmed to be infected with G1 JEV,32 with G3 JEV,and two with G5 JEV. Conclusion G3 was the primary infection genotype among JE cases with a definite infection genotype,and the infection caused by G5 JEV was confirmed serologically in China.

Objective This study aims to explore the complex relationship between social engagement and depressive symptoms among older adults in China,focusing particularly on the moderating role of marital status. Methods This study used data from the latest Chinese Longitudinal Healthy Longevity Survey(CLHLS).The analysis used the latent class analysis to delineate personality clusters and hierarchical linear regression,supplemented by the PROCESS macro,to investigate the effects of social engagement and marital status on depressive symptoms. Results The analysis encompassed 7,789 respondents(mean age:82.53[s=11.20]years),with 54%female.The personality analysis categorized participants into four clusters,with the majority(77.60%)classified as Confident Idealists,who exhibited the lowest levels of depressive symptoms.Hierarchical linear regression analysis yielded several significant findings:Higher levels of social engagement were significantly associated with fewer depressive symptoms(t=-7.932,P<0.001,B=-0.463).Marital status was a significant factor;married individuals reported fewer depressive symptoms compared to their unmarried counterparts(t=-6.368,P<0.001,B=-0.750).There was a significant moderating effect of marital status on the relationship between social engagement and depressive symptoms(t=-2.092,P=0.037,B=-0.217). Conclusion This study demonstrates that,among Chinese older adults,both social engagement and marital status significantly influence depressive symptoms.Higher social engagement,particularly in other activities like doing household chores,gardening,reading newspapers or books,and playing cards or Mahjong,is associated with fewer depressive symptoms,especially among married individuals.

Objective:To evaluate the role of neuronal nitric oxide synthase (nNOS)-nitric oxide synthase 1 adaptor protein (NOS1AP) coupling in remifentanil-induced hyperalgesia in rats.Methods:Forty clean-grade healthy adult male Sprague-Dawley rats, weighing 240-260 g, aged 2-3 months, were divided into 4 groups ( n=10 each) using a random number table method: control group (group C), remifentanil group (group R), nNOS-NOS1AP inhibitor ZLc002 group (group C+ Z) and remifentanil + ZLc002 group (group R+ Z). Normal saline was intravenously infused at a rate of 0.1 ml·kg -1·min -1 for 60 min in C group. Remifentanil was intravenously infused at a rate of 1.0 μg·kg -1·min -1 for 60 min in R group. ZLc002 10 mg/kg was intraperitoneally injected for 3 consecutive days, and then normal saline 0.1 ml·kg -1·min -1 and remifentanil 1.0 μg·kg -1·min -1 were intravenously infused for 60 min in C+ Z group and R+ Z group. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured at 24 h before intravenous infusion and 6, 24 and 48 h after intravenous infusion (T 0-3). All the rats were sacrificed after the last measurement of pain thresholds, and the L 4-6 segments of the spinal cord were removed for determination of the expression of nNOS, NOS1AP and Dexamethasone-induced Ras-related protein 1 (Dexras1) protein and mRNA using the real-time polymerase chain reaction. Nitrosylated proteins were extracted by biotin conversion for determination of the expression of nNOS, NOS1AP and total and nitrosylated Dexras1 (by Western blot) and co-expression of nNOS-NOS1AP (by co-immunoprecipitation). The content of NO in the spinal cord was measured. Results:Compared with group C, the MWT was significantly decreased, and the TWL was shortened at T 1-3, the expression of nNOS and NOS1AP protein and mRNA was up-regulated, the co-expression of nNOS-NOS1AP and NO production were increased, and the expression of nitrosylated Dexras1 was up-regulated in group R ( P<0.05), and no significant change was found in each aforementioned parameter in group C+ Z ( P>0.05). Compared with group R, the MWT was significantly increased, and the TWL was prolonged at T 1-3, the co-expression of nNOS-NOS1AP and NO production were decreased, the expression of nitrosylated Dexras1 was down-regulated ( P<0.05), and no significant change was found in the expression of nNOS and NOS1AP protein and mRNA in group R+ Z ( P>0.05). There were no significant differences in total Dexras1 protein and mRNA expression among the four groups ( P>0.05). Conclusions:The mechanism by which remifentanil induces hyperalgesia may be related to up-regulating the expression of nNOS and NOS1AP in the spinal cord, promoting interaction between nNOS and NOS1AP and mediating NO generation and Dexras1 nitrosylation modification in rats.

Background Individuals may experience significant alterations in sleep hygiene during the major public health emergencies,consequently impacting their sleep quality.Objective To explore the relationship between sleep quality and sleep hygiene among adult residents during the major public health emergencies,so as to provide references for improving the sleep quality of residents during such a period.Methods A sample of 1 364 adult residents were enrolled as the research subjects from February 20 to 29,2020.All participants were asked to complete self-administered questionnaire to obtain basic-demographic information and sleep hygiene.Pittsburgh Sleep Quality Index(PSQI)was applied to assess sleep quality.Residents were classified into poor sleepers with PSQI score≥8 and good sleepers defined as PSQI score<8.Binary Logistic regression analysis was conducted to identify factors associated with sleep quality.Radar chart was used to visualize and compare the sleep hygiene between poor sleepers and good sleepers.Results According to PSQI score,891(65.32%)residents were good sleepers,while 473(34.68%)residents were poor sleepers.Comparison revealed that age(χ2=3.887),past medical history(χ2=27.938),awareness rate of importance of sleeping before major public health emergencies(χ2=4.337),impact of sleep quality on quality of life during the major public health emergencies(χ2=178.138),frequency of staying up late during the major public health emergencies(χ2=139.390),compensatory sleep behaviors(χ2=39.257),impact of sleep problems on daytime functioning(χ2=285.879),change of bedtime(χ2=63.031),sleep latency(χ2=168.672),wake-up time(χ2=59.221),changes in sleep duration(χ2=172.332),time spent in the bedroom(χ2=23.071),and sum of money spent on improving sleep environment(χ2=58.584)yielded statistical difference between poor sleepers and good sleepers(P<0.05 or 0.01).Logistic regression analysis denoted that past medical history(OR=1.680,95%CI:1.185～2.382),negative impact of sleep quality on quality life(OR=4.181,95%CI:2.722～6.422),staying up late 3 to 4 times per week(OR=3.145,95%CI:1.497～6.605),staying up late almost every day(OR=4.271,95%CI:1.970～9.260),negative impact of sleep problems on daytime functioning(OR=7.169,95%CI:5.188～9.907),prolonged sleep latency(OR=2.836,95%CI:2.019～3.982)and shortened sleep duration(OR=3.518,95%CI:2.144～5.772)were risk factors of poor sleep quality.The sum of money spent on improving sleep environment following the major public health emergencies≤500 RMB(OR=0.334,95%CI:0.134～0.830)was related to the incidence rate of poor sleep quality.Radar chart showed that poor sleepers were characterized by extravagant concerns,excessive cleanliness and poor sleep hygiene practices during the major public health emergencies,and poor sleepers were more likely to stay up late due to stress and emotional issues.Conclusion Some residents are facing poor sleep quality during the major public health emergencies,and poor sleep hygiene practice also contributes to poor sleep quality.

Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Objective To explore the influencing factors of type 2 diabetes mellitus(T2DM)complicated with metabolic syndrome(MS)and investigate the correlation between the level of 25(OH)D3 and T2DM combined with MS.Methods 592 in patients with T2DM admitted to the Department of Endocrinology of Shenzhen Second People's Hospital were selected from January to December 2020.They were divided into simple T2DM group(T2DM,n=194)and T2DM combined with MS group(MS,n=398).Results Compared with the T2DM group,the MS group had lower 25(OH)D3 levels[26.30(20.50,32.39)vs 24.20(20.16,29.47),P<0.05].Spearman correlation analysis showed that 25(OH)D3 was negatively correlated with MS,FPG,TG,LDL-C,and 24 hUAlb(P<0.05 or P<0.01).Logistic regression analysis showed that 25(OH)D3 was a protective factor for T2DM combined with MS.Conclusions Patients with T2DM and MS have lower 25(OH)D3 levels.25(OH)D3 is a protective factor for T2DM combined with MS.

Hearing loss is one of the common radiotherapy-induced complications of head and neck tumors, including nasopharyngeal carcinoma. These side reactions can be classified into acute or delayed types, which affect all structures of the auditory organs, resulting in conductive, sensorineural or mixed hearing loss. Up to 40% of patients develop acute middle ear side effects during radical radiotherapy, while approximately 1/3 develop late sensorineural hearing loss. The total radiation dose and tumor site appear to be the most important factors associated with the risk of hearing loss. The mechanisms of conductive and sensorineural hearing impairment are different. New radiotherapy techniques (three-dimensional conformal radiotherapy, intensity modulated radiotherapy, proton therapy) enable better dose distribution, lower dose to non-target organs, and gradually increase the feasibility of protecting normal tissues. The present article illustrates recent progress in radiotherapy-induced hearing loss, specially focusing on the occurrence, the mechanisms and related factors of ear toxicity, detection and diagnosis, and treatment.

Objective:Based on the specific cleavage and non-specific "trans-cleavage" activities of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein(CRISPR/Cas13), we established a visually amplification-free rapid detection technique of 2019-nCoV nucleic acid. This technique is easily processed with a low detection limit and good specificity.Methods:According to the 2019-nCoV gene sequence, specific CRISPR RNAs were screened and designed by bioinformatics analysis, and then synthesized as universal signal-strained RNA transcription targets in vitro to establish and optimize the reaction system. Moreover, the 2019-nCoV pseudoviral nucleic acid was used as a standard substance to evaluate the detection limit. A total of 65 positive samples were collected from various 2019-nCoV variants, while 48 negative samples included other clinically common respiratory pathogens, such as influenza A virus, influenza B virus, human parainfluenza virus, Klebsiella pneumonia, etc. All samples were tested by quantitative PCR (qPCR), digital PCR, and the method established in this study. The sensitivity and specificity of the newly established method were analyzed and evaluated. Results:With the newly established technique, the detection time for 2019-nCoV nucleic acid could be minimized to 6 minutes. In addition, the detection limit was 14 copies/μl when assisted by the displaying instrument, whereas it increased to 28 copies/μl with the naked eye. This technique had a sensitivity and specificity of 98.5% (66/67) and 100% (46/46) respectively, showing no statistically significant difference compared to the gold standard qPCR( P=1). Conclusions:This study has successfully established a CRISPR/Cas13a-based visually rapid detection technique for 2019-nCoV nucleic acid. This technique offers the advantages of a simple process, convenient operation, low environmental operating requirements, a detection limit close to qPCR, and a strong potential for on-site testing applications.