ObjectiveTo observe the intervention effect of Fufang Kangjiaolv capsules on anxiety-like behaviors in the rat model of chronic restraint stress （CRS） and explore the mechanism underlying the anti-anxiety effect via the microbiota-gut-brain axis. MethodsRats were assigned into blank， model， positive drug （diazepam， 1 mg·kg^-1）， and low-， medium-， and high-dose （0.75， 1.5， 3 g·kg^-1， respectively） Fufang Kangjiaolv capsules groups. After 14 days of administration， the elevated plus maze test， open field test， light and dark box test， and marble burying test were performed. Hematoxylin-eosin staining was employed to observe the pathological changes in the hippocampus and colon of rats， and Nissl staining was conducted to observe the damage of hippocampal neurons. The gut microbiota was analyzed by 16S rRNA gene sequencing. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to determine the mRNA levels of zonula occludens-1 （ZO-1） and occludin in the colon of rats. The levels of tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and interleukin-1β （IL-1β） in the colon， serum， and hippocampus were determined by enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of ZO-1， occludin， nuclear factor-κB p65 （NF-κB p65） in the colon tissue and NF-κB p65 and brain-derived neurotrophic factor （BDNF） in the hippocampal tissue. ResultsCompared with the blank group， the model group showed reductions in the time and frequency ratio of rats entering the elevated plus maze， the time and frequency of rats entering the central area of the open field， the time of entering the open box， the times of passing through the light and dark box， and the number of unburied beads （P<0.05， P<0.01）. Compared with the model group， Fufang Kangjiaolv capsules ameliorated the anxiety of the model rats to varying degrees， and the high-dose group had the best effect， with increases in the proportions of time and frequency of rats entering the open arm in the elevated plus maze （P<0.05）， the number of rats entering the central area in the open field （P<0.05）， the time of entering the open box， the times of passing through the light and dark boxes， and the number of unburied beads （P<0.01）. Moreover， the high-dose group showed alleviated pathological damage of hippocampal neurons and colon. The results of 16S rRNA gene sequencing showed that the model group had increased relative abundance of Firmicutes， Deferribacterota， Romboutsia， and Phascolarctobacterium， while it had decreased relative abundance of Bavcteroidota and Lactobacillus. The drug administration groups showed increased relative abundance of Bavcteroidota， Bacteroides， norank f norank o Clostridia UCG-014， and Blautia and decreased relative abundance of Firmicutes and Deferribacterota. Compared with the blank group， the model group showed down-regulated protein and mRNA levels of ZO-1 and occludin in the colon （P<0.01）， elevated levels of TNF-α， IL-6， and IL-β in the colon， serum， and hippocampus （P<0.01）， up-regulated protein level of NF-κB p65 in the colon and hippocampus （P<0.01）， and down-regulated protein level of BDNF in the hippocampus （P<0.05）. Compared with the model group， high-dose Fufang Kangjiaolv capsules up-regulated the mRNA levels of ZO-1 and occludin in the colon （P<0.01）， lowered the levels of TNF-α， IL-6， and IL-β in the colon， serum， and hippocampus （P<0.01）， up-regulated the protein levels of ZO-1 （P<0.01） and occludin （P<0.05） in the colon， down-regulated the protein level of NF-κB p65 in the colon and hippocampus （P<0.05）， and up-regulated the protein level of BDNF in the hippocampus. ConclusionFufang Kangjiaolv capsules can reduce the anxiety-like behaviors in the rat model of CRS by regulating the gut microbiota disturbance， up-regulating the expression of tight junction proteins in the colon， repairing intestinal mucosal mechanical barrier， and down-regulating NF-κB/BDNF signaling pathway， thereby reducing peripheral and central inflammation. This study proves the hypothesis that Fufang Kangjiaolv capsules play an anti-anxiety role via the microbiota-gut-brain axis， providing a new idea for further research.

AIM: To observe related influence of eye-use behaviors on the onset and progression of myopia among primary school student in Minhang district, Shanghai under the background of “double reduction”, and evaluate the impact of eye-use habit and behavior on the myopia development in children and adolescents.METHODS:A stratified cluster sampling method was adopted to select study subjects for longitudinal observation. Random sampling was conducted among grade 1-4 students from three primary schools in Minhang District, Shanghai from September to December 2021 after the implementation of the “double reduction” policy, with a 2-year observation period. Annual vision screenings were performed to obtain uncorrected visual acuity, non-cycloplegic refraction data, and spherical equivalent(SE). Biometric measurements were taken to acquire axial length(AL)and corneal curvature(CR). Questionnaires were administered to investigate the eye-use behaviors of participants. Taking myopia ≤-0.50 D as the diagnostic criterion, myopia incidence were calculated. SE and AL changes were used to evaluate myopia progression. Logistic regression analysis was employed to identify risk factors for myopia, and analyze the relationship between eye-use behaviors and myopia development or progression.RESULTS:A total of 2 986 children complete a follow-up for 2 a. The baseline prevalence of poor vision was 32.74%, increasing to 38.92% and 43.73% at 1 and 2 a follow-up, respectively, demonstrating a yearly upward trend. Taking uncorrected visual acuity combined with non-cycloplegic refraction as clinical screening criteria for myopia, the incidence was 24.72%, 27.60%, and 29.64% at baseline, 1, and 2 a follow-ups, respectively. These showed no statistical significant difference compared to the diagnostic standard of cycloplegic refraction SE(23.98%, 27.03%, and 29.17%; all P>0.05). Logistic regression analysis found that daily outdoor and sleep duration were protective factors against myopia(OR=0.54, 0.84, all P<0.05), while age, parental myopia, duration of offline education, online education, electronic device usage, and poor visual posture were identified as risk factors(OR=2.72, 1.54, 1.72, 1.12, 1.47, 1.26, all P<0.001). Further analysis of myopia progression in baseline myopic children over 2 a revealed that age, parental myopia, baseline SE, duration of offline/online education, electronic device usage, and poor visual posture may contribute to rapid myopia progression.CONCLUSION:Under the background of “double reduction”, the eye-use behavior among primary school students in Minhang District, Shanghai has an important impact on the occurrence and development of myopia. In order to effectively prevent and control myopia, it is necessary to further strengthen the education and guidance of eye-use behavior.

AIM: To compare the effectiveness of foldable capsular body(FCB)with traditional scleral buckling(SB)in the treatment of experimental retinal detachment animal models.METHODS: After successfully establishing rhegmatogenous retinal detachment(RRD)animal models, 24 New Zealand white rabbits were randomly divided into three groups(RRD models group, SB group, and FCB group), with 8 rabbits in each group. The FCB and SB groups underwent SB and FCB surgeries for the RRD animal models, while the RRD models group only consists of RRD models without any surgical intervention during the follow-up period. The follow-up duration was 3 mo. Wide-field neonatal fundus imaging system and ophthalmic B-ultrasound were used to assess the fundus conditions before and after surgery. The Icare® TONOVET Plus tonometer was utilized to evaluate intraocular pressure changes before and after surgery. The Eaton and Draize scoring systems were selected to monitor postoperative inflammatory reactions.RESULTS: The retinal reattachment rates in the FCB and SB groups were 87.5% and 75.0%, respectively, with no statistically significant difference between the groups(P>0.05). The intraocular pressure in both the FCB and SB groups increased postoperatively compared to preoperative levels(P<0.01), and there were no significant differences in intraocular pressure at any time points during the follow-up period between the groups(P>0.05). The intraocular pressure in the RRD models group remained at a low level throughout the follow-up period. The average surgical time for the FCB group was 16.87±2.29 min, which was shorter than 46.25±4.74 min in the SB group(t=-15.166, P<0.001). According to the Eaton and Draize scoring systems, the FCB group had lower grades of conjunctival hyperemia and edema in the early postoperative period compared to the SB group, indicating milder inflammatory reactions(P<0.05).CONCLUSION: Both FCB and SB are effective in treating experimental RRD. Compared to SB, FCB is simpler to operate, and also has a shorter surgical time and milder postoperative inflammatory reactions.

Objective:To investigate the relationship between CBCT imaging changes of condyle and clinical features,and related risk factors in patients with temporomandibular disorders(TMD).Methods:453 patients with TMD were enrolled and underwent CBCT scan for bilateral temporomandibular joints(TMJ),3D reconstruction of the TMJs was analyzed.Logistic regression analysis was performed to investigate the relationship between condylar bone changes and TMD clinical features.Results:Patients＜18 years old were more likely to have condylar bone changes than the adults.The symptoms of pain and restricted mouth opening were more likely to be detected in the condylar bone change group(n=133)than in the normal condylar bone group(n=320).The incidence of brux-ism in the normal condylar bone group was higher than that in the condylar bone change group.Univariate logistic regression analysis showed that only bruxism(OR=0.550),pain(OR=1.844)and mouth restriction(OR=2.024)were included in the regression equa-tion.Multivariate logistic regression analysis showed that,due to the protective effect of bruxism,the OR value of pain decreased from 1.844 to 1.791,and the OR value of mouth restriction decreased from 2.024 to 1.847.Conclusion:The condylar bone change in TMD patients more likely occur in puberty or patients with pain and restricted mouth opening.Bruxism may be a protective factor in the occurrence of condylar bone changes in TMD patients.

Objective:To investigate whether long non-coding RNA(lncRNA) AW112010 can improve insulin resistance in aging adipocytes through the miR-204/POU2F2 signaling pathway.Methods:In vivo experiment: C57BL/6 mice were divided into young control group(4 months old) and aging model group(18 months old) based on body weight. The expression levels of AW112010, miR-204-5p, POU2F2, aging related indicators(p16, p21), and insulin signaling pathway genes [insulin receptor(INSR), insulin receptor substrate 1(IRS1), phosphatidylinositol kinase(PI3K), protein kinase B(AKT)] in epididymal adipose tissue were detected using real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting. In vitro experiment: Using adriamycin(ADR) to induce 3T3-L1 aging adipocyte model, β-gal staining was used to observe cellular senescence, and miR-204 inhibitor and miR-204 mimic small interfering RNA were successfully constructed and transfected into 3T3-L1 adipocytes. Results:RT-qPCR and Western blot results showed that compared with the young group, the expression of AW112010 in the adipose tissue of aging mice was increased, while the expression of miR-204-5p was decreased. The expressions of POU2F2, p16, and p21 in the adipose tissue of aging mice were increased, while the expressions of INSR, IRS1, PI3K, GLUT4 mRNA and protein were decreased. The β-gal stainging results showed that the number of 3T3-L1 senescent adipocytes induced by ADR was significantly increased, and the expression levels of AW112010, POU2F2, p16, and p21 in ADR-induced senescent adipocytes were increased compared with the control group, while the expression levels of miR-204-5p, INSR, IRS1, PI3K, GLUT4 were decreased, and remaining glucose in the culture medium was increased. Compared with control, overexpression of miR-204 resulted in decreased expressions of aging indicators p16, p21, and target gene POU2F2 while the expressions of INSR and GLUT4 were increased.Conclusion:Upregulation of lncRNA AW112010 in adipocytes of aging mice may induce insulin resistance by targeting miR-204-5p/POU2F2/IRS1.

ObjectiveBy starting with the combination of Os Draconis， Bupleuri Radix， and Ostreae Concha， the role of mineral medicine Os Draconis in the combination of the Bupleuri Radix-containing tri-herbal medicines was preliminarily explored from the perspective of supramolecular system formation. Method① The appearance and Tyndall phenomenon of single decoction of Os Draconis， Bupleuri Radix， and Ostreae Concha， as well as co-decoction of Bupleuri Radix-Os Draconis， Bupleuri Radix-Os Draconis-Ostreae Concha， and Bupleuri Radix-Ostreae Concha were observed， and the average particle size， dispersion coefficient， and Zeta potential of suspension particles in each decoction were determined. The micromorphology of supramolecular structures was observed by scanning electron microscope （SEM）. ② The pH of different compatibility systems， liquid viscosity coefficient， liquid surface tension， freeze-dried powder yield rate， and other physical properties were determined， and the interaction of different compatibility systems was detected by infrared absorption spectroscopy （FTIR） and UV-visible spectrophotometry （UV-Vis）. ③ The composition and content difference of different compatible systems were determined by high-performance liquid chromatography （HPLC） and ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry （UPLC-Q-TOF-MS）. ResultCompared with the single decoction， the co-decoction had more obvious turbidity and Tyndall phenomenon. The particles in the co-decoction suspension were smaller and more evenly distributed， and the Zeta potential was reduced， indicating a more stable system. Under SEM， Bupleuri Radix was irregularly lamellar， and Bupleuri Radix-Os Draconis and Bupleuri Radix-Os Draconis-Ostreae Concha were mainly spherical nanoparticles. Bupleuri Radix-Ostreae Concha was irregularly lamellar， with a small number of spherical nanoparticles. The pH of the single decoction of Bupleuri Radix and co-decoction increased， and the viscosity coefficient increased. The liquid surface tension decreased. The freeze-dried powder yield rate of the Bupleuri Radix-Os Draconis co-decoction was the highest， followed by Bupleuri Radix-Ostreae Concha decoction and Bupleuri Radix-Os Draconis-Ostreae Concha decoction， and the yield rate of Bupleuri Radix single decoction was the lowest. The main change of FTIR was the stretching vibration of -OH， and the co-decoction moved to the low-frequency direction obviously. UV-Vis showed that the maximum absorption occurred at 295.8 nm for all groups， and the absorption intensity was different （Bupleuri Radix-Os Draconis>Bupleuri Radix-Os Draconis-Ostreae Concha>Bupleuri Radix-Ostreae Concha>Bupleuri Radix）. The components of Bupleuri Radix were used as the indexes， and the content of methanol extract determined by HPLC was higher than that of water extract， and the components of Bupleuri Radix single decoction were mainly saikosaponin a （SSa） and saikosaponin c （SSc）， which were slightly higher after co-decoction compatibility. UPLC-Q-TOF-MS could identify 37 compounds in both single decoction and co-decoction. ConclusionThe combination of Bupleuri Radix， Os Draconis， and Ostreae Concha can form a smaller， more uniform， and stable nano-sized supramolecular system， which is conducive to the dissolution of the main components of Bupleuri Radix， and the Os Draconis contributes the most in this process.

Objective:To explore the clinical manifestations and genetic etiology of a Chinese pedigree affected with Familial focal epilepsy with variable foci (FFEVF).Methods:A FFEVF pedigree presented at the Department of Medical Genetics of Linyi Maternal and Child Health Care Hospital on March 14, 2023 was selected as the study subject. The proband was subjected to whole exome sequencing (WES). Candidate variants were verified by Sanger sequencing of the proband and other affected members and bioinformatic analysis. This study was approved by Medical Ethics Committee of the Linyi Maternal and Child Health Care Hospital (Ethics No. QTL-YXLL-2023048).Results:WES revealed that the proband has harbored a heterozygous c. 1642C>T (p.Arg548Cys) missense variant in exon 15 of the NPRL3 gene. Sanger sequencing confirmed that the variant was inherited from the proband′s father, and multiple members of the pedigree had also harbored the same variant. Based on guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as variant of unknown significance (PM2_supporting + PP3). Conclusion:The clinical phenotype of FFEVF patients caused by variants of NPRL3 gene is extensive, and the patients may present with neurological abnormality of autism spectrum disorder in addition to seizures.

Objective:To investigate the protective effects of an anti-inflammatory mixture on acute lung injury (ALI) induced by sepsis in rats, as well as its possible mechanisms.Methods:A total of 40 Sprague-Dawley (SD) rats were randomly divided into the sham group, septic ALI model group (model group), 3-methyladenine (3-MA) control group, and anti-inflammatory mixture pretreatment group, with 10 rats in each group. Cecal ligation and perforation (CLP) was performed to reproduce a septic ALI model. The rats in the sham group only underwent opening and closing the abdomen without perforation and ligation. Both groups were given saline gavage and intraperitoneal injection for 3 consecutive days before surgery. The 3-MA control group was given intraperitoneal injection of saline and autophagy inhibitor 3-MA 15 mg/kg for 3 consecutive days before modeling. The anti-inflammatory mixture pretreatment group was given 8.8 mL/kg of anti-inflammatory mixture by gavage [the composition of anti-inflammatory mixture: rhubarb 15 g (after the next), coptis chinensis 15 g, baical skullcap root 12 g, magnoliae cortex 12 g, dahurian patrinia herb 30 g] and saline intraperitoneal injection for 3 consecutive days before modeling. The rats in each group were anesthetized 24 hours after surgery and died due to abdominal aortic blood collection. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of serum inflammatory cytokines interleukins (IL-1β and IL-6). Lung tissue was taken and then the bronchoalveolar lavage fluid (BALF) was collected, and the levels of IL-1β and IL-6 were detected by ELISA. Lung wet/dry weight (W/D) ratio was measured. After hematoxylin-eosin (HE) staining, the histopathological changes of the lungs were observed under light microscopy. Western blotting was used to detect the expression of autophagy markers microtubule-associated protein 1 light chain 3-Ⅱ/Ⅰ (LC3-Ⅱ/Ⅰ) and Beclin-1 protein in lung tissue. Autophagosomes in lung tissue were observed with transmission electron microscopy.Results:Compared with the sham group, the rats in the model group exhibited severe destruction of lung tissue structure, with significant infiltration of inflammatory cells, the lung W/D ratio and the levels of IL-1β and IL-6 in serum and BALF were significantly increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein were down-regulated, the autophagosomes were more. The rats in the 3-MA control group exhibited more severe lung tissue injury as compared with the model group, the lung W/D ratio and the levels of inflammatory cytokines in serum and BALF were further increased, the expressions of LC3-Ⅱ/Ⅰ and Beclin-1 protein still showed a decrease tendency as compared with the sham group, and the autophagosomes were less than that in the model group. Compared with the model group, the anti-inflammatory mixture pretreatment group showed milder lung tissue injury with a minimal amount of inflammatory cell infiltration, the lung W/D ratio was significantly reduced (7.07±1.02 vs. 11.33±1.85, P < 0.05), the levels of IL-1β and IL-6 in both serum and BALF were significantly decreased [IL-1β (ng/L): 26.04±3.86 vs. 40.83±5.46 in serum, 17.75±2.02 vs. 26.86±4.32 in BALF; IL-6 (ng/L): 91.28±10.15 vs. 129.44±13.05 in serum, 76.06±7.51 vs. 120.91±7.47 in BALF, all P < 0.05], and the ratio of LC3-Ⅱ/Ⅰ and Beclin-1 protein expression were significantly increased [LC3-Ⅱ/Ⅰ ratio: 1.23±0.02 vs. 0.60±0.02, Beclin-1 protein (Beclin-1/GAPDH): 2.37±0.33 vs. 0.62±0.05, both P < 0.05]. Furthermore, an increase in the number of autophagosomes was observed. Conclusion:The anti-inflammatory mixture improves lung injury in rats with sepsis induced by CLP and reduce inflammation levels, potentially through upregulation of Beclin-1-mediated autophagy.

Objective:To investigate the eye movement characteristics of non-emotional pictures and emotion-al pictures in college students with different self-rating levels of depression.Methods:The Baker Depression Ques-tionnaire(BDI-2)was used to select 20 college students in high score group(BDI-2 score≥ 18 points)and 20 col-lege students in low score group(BDI-2 score ≤2 points).In experiment 1 the different finding tasks were used to investigate the difference of eye movement features between the two groups when viewing non-emotional pic-tures.In experiment 2 the expression recognition tasks were used to investigate the difference of the eye movement features between the two groups when viewing emotional pictures.Results:The results of experiment 1 showed that when completing the judgment task,the fixation times of each interest area of non-emotional images was smaller in the high score group than in the low score group(P＜0.01).The results of experiment 2 showed that the reaction time was longer to sad faces than to happy faces in the high score group(P＜0.05),and the correct rate to sad faces was higher in the high score group than in the low score group(P＜0.05).For the eye interest area,the first fixation arrival time was earlier in the high score group than in the low score group(P＜0.001),and the first fixa-tion time and total fixation time were shorter in the high score group than in the low score group(Ps＜0.01).Con-clusion:College students with high self-rating level of depression show a decrease of interest when viewing non-e-motional pictures,and show an advantage in processing negative emotional information and avoiding eye gaze when viewing emotional pictures.

Purpose To investigate the clinical pathologic features of five SMARCA4-deficient non-small lung cancers(SMARCA4-dNSCLCs).Methods Five cases of SMARCA4-dNSCLC was underwent by HE,immunohistochemical staining,and molecular detection,analyzed their clinicopathological char-acteristics and reviewed relevant literatures.Results All 5 ca-ses were male,and mean age was 66 years.Five patients had a history of smoking,three patients were treated with cough and blood in sputum as the first symptom,one was treated with a history of pulmonary tuberculosis combined with limb mobility disorder,and one was diagnosed with pulmonary nodules by physical examination.Under microscopic observation,tumor cells were poorly differentiated,with solid nest sheet distribu-tion,some with glandular structure,tumor cells had abundant e-osinophilic or transparent cytoplasm,vacuolar nuclear chroma-tin,nucleoli was visible,and nuclear mitosis was common.In-flammatory cell infiltration and sheet of necrosis were seen in the stroma.Immunohistochemical staining showed 5/5 diffuse ex-pression of CK(AE1/AE3)and CK7,5/5 loss expression of BRG1,1/5 diffuse expression of p40 and CK5/6,and Ki67 proliferating index ranged from 20％to 90％.FISH tests showed that 4/4 SMARCA4 genes missed.Five patients were followed up for 2-15 months,3 patients died and 2 patients survived.Conclusions SMARCA4-dNSCLC can have extensive morphologi-cal features,high degree of malignancy,and complicated treat-ment.BRG1 deficiency is helpful for diagnosis.Deepening the understanding of SMARCA4-dNSCLC can help the clinical cor-rect choice of treatment strategies and accurately evaluate patient prognosis.