AIM:To analyze differential proteins associated with the pathogenesis of dry eye(DE)using bioinformatics methods, in order to reveal their potential molecular mechanisms.METHODS: Articles published in PubMed and EMBASE databases from the inception of the database to August 31, 2023, that used proteomic methods to detect protein expression in clinical samples of dry eye were searched. Differential proteins were selected and further analyzed using the STRING database and Cytoscape software for hub gene screening and module analysis. Protein-protein interaction(PPI)analysis, gene ontology(GO)functional annotation, and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed.RESULTS: A total of 21 articles were included, identifying 74 differentially expressed proteins. The most frequently occurring differential proteins were calgranulin A(SA1008), lipocalin-1(LCN1), lysozyme C(LYZ), mammaglobin-B(SCGB2A1), proline-rich protein 4(PRR4), transferrin(TF), and calgranulinB(S100A9). The top 10 hub genes were serum albumin(ALB), tumor necrosis factor(TNF), interleukin 6(IL6), IL1B, IL8, matrix metalloproteinase 9(MMP9), alpha-1-antitrypsin(SERPINA1), IL10, complement component 3(C3), and lactotransferrin(LTF). Module analysis suggested MMP9 and PRR4 as seed genes. KEGG analysis showed that differential proteins were mainly enriched in the IL17 signaling pathway(61.9%).CONCLUSION: The results reveal potential molecular targets and pathways for DE and confirm the association between the pathogenesis of DE and inflammation. Further in-depth research is needed to confirm the significance of these biomarkers in clinical practice.

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

Objective To study on the traits,micro-traits and microscopic identification characteristics of common cyrtomium Chinese medicines,including Dryopteridis Crassirhizomatis Rhizoma,Osmundae Rhizoma which were recorded in 2020 edition of Chinese Pharmacopoeia and their three chaotic breeds(Woodwardia unigemmata(Makino)Nakai,Woodwardia japonica(L.F.)Smith and Matteuccia struthiopteris(L.)Todaro,providing a reference for the identification and reference of cyrtomium Chinese medicine.Methods Morphological and micro-character identification methods were used in the character identification,and the normal bright field and polarized dark field contrast observation methods were used in the microscopic identification of cross-sections,combined with extended depth of field imaging and large image stitching techniques,to obtain holographic color images and local feature high definition image.The Chiral labeling methods were used to identified the images.Results The image data of the characters,micro-characters and the normal light and polarized light holographic color image data of cross-section of common cyrtomium herbs and its chaotic varieties were obtained at the first time.The main identification feature retrieval tables and correlation tables of medicinal materials,traits,micro traits and microscopic characters were made.Conclusion Common cyrtomium herbs and its three chaotic varieties can be identified by using characters,micro-characters and microscopic identification comprehensively,especially the microscopic character under polarized light of the cross-section,which has obvious identification significance.

Objective To explore the predictive value of serum homo sapiens microRNA(hsa-miR)-30c-5p expression in patients with type 2 diabetes mellitus(T2DM)for microvascular complications.Methods A total of 205 T2DM patients admitted to Bazhong Central Hospital from May 2021 to September 2022 were selected as the diabetes group,and the diabetes group was further divided into diabetes with combined group(n=124)and non combined group(n=81)according to the microvascular complications of the patients.In addition,205 healthy people who underwent physical examination during the same period were selected as the control group.The expression of hsa-miR-30c-5p in serum was detected by reverse transcription-polymerase chain reaction(RT-PCR)and compared.The factors affecting microvascular complications were analyzed using multivariate logistic regression analysis,and receiver operating characteristic(ROC)curves were plotted to predict the value of serum hsa-miR-30c-5p expression in predicting microvascular complications in T2DM patients.Results The expression of serum hsa-miR-30c-5p in the combined group(0.58±0.06)and the non-combined group(0.72±0.08)were lower than that in the control group(0.89±0.21),and the differences were significant(t=16.038,7.079,all P=0.001).The combined group was lower than the non-combined group,and the difference was significant(t=14.289,P=0.001).The course of diabetes[(OR(95％CI):3.873(2.976～4.770)],uric acid[(OR(95％CI):2.125(1.211～3.040)]and glycosylated hemoglobin[(OR(95％CI):2.680(1.745～3.616)]were independent risk factors for microvascular complications in T2DM patients(all P＜0.05),while the time within the target range of glucose[(OR(95％CI):0.491(0.135～0.846)]and serum hsa-miR-30c-5p[(OR(95％CI):0.532(2.976～4.770)]were protective factors for microvascular complications in T2DM patients(all P＜0.05).The sensitivity,specificity and area under the curve(95％CI)of serum hsa-mir-30c-5p expression in predicting microvascular complications in T2DM patients were 81.45％,85.19％and 0.802(0.741～0.854),respectively.Conclusion The expression of serum hsa-miR-30c-5p in patients with T2DM is abnormally reduced,and serum hsa-miR-30c-5p is a protective factor for microvascular complications in patients with T2DM.It may have a certain predictive value for microvascular complications in patients with T2DM.

AIM:To comprehend the mechanism by which ginsenoside Rc protects against cerebral ischemia-reperfusion injury(CIRI),with a particular emphasis on pyroptosis.METHODS:The C57BL/6 mice were randomly di-vided into 6 groups:sham group,middle cerebral artery occlusion/reperfusion(MCAO/R)group,ginsenoside Rc+MCAO/R group(Rc group),AMP-activated protein kinase(AMPK)agonist acadesine/5-aminoimidazole-4-carboxamide-1-β-D-ribofuranoside(AICAR)+MCAO/R group(agonist group),and ginsenoside Rc+MCAO/R+AMPK inhibitor Compound C group(Rc+inhibitor group).The mice in agonist group were given 500 mg/kg of AICAR intraperitoneally,while those in Rc+inhibitor group were given 20 mg/kg of Compound C intraperitoneally.Ginsenoside Rc was gavaged into the mice in Rc and Rc+inhibitor groups at a dose of 40 mg/kg once per day for 7 d after modeling,while the mice in sham and MCAO/R groups got the same volume of purified water.With the use of the Zea-Longa score,we determined which mice had neu-rological abnormalities.TTC staining was employed for assessing the cerebral infarct amount in mice,and the dry wet weight technique was utilized for determining the degree of cerebral edema.Moreover,HE staining was used to observe pathological alterations in the brain,and Western blot and RT-qPCR were applied for detecting the expression of AMPK,nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3),caspase-1 and gasdermin-D(GSDMD)in the brains of mice.Lastly,ELISA was employed for measuring the levels of inflammatory factors,interleukin-1β(IL-1β)and IL-18.RESULTS:Brain edema,infarct volume and neurological impairments were all diminished in agonist and Rc groups.Additionally,they demonstrated neuronal damage inhibition.The ratio of p-AMPK/AMPK and AMPK mRNA ex-pression in mouse brain tissues was elevated in both Rc and agonist groups.They showed decreased mRNA and protein levels of NLRP3,caspase-1 and GSDMD,as well as the levels of IL-1β and IL-18.Compared with Rc group,there were remarkable decreases in p-AMPK/AMPK ratio and AMPK mRNA expression in the brain tissue of mice in Rc+inhibitor group(P<0.05).The mRNA and protein levels of NLRP3,caspase-1 and GSDMD,and the IL-1β and IL-18 expression levels were significantly increased(P<0.05).Moreover,the neurological deficiency scores and infarct volume were in-creased,and the degrees of cerebral edema and neuronal pathological damage were enhanced(P<0.05).CONCLU-SION:Ginsenoside Rc may inhibit NLRP3/caspase-1/GSDMD-mediated pyroptosis by activating AMPK pathway,thereby reducing CIRI in mice.

The effect of aspirin eugenol ester(AEE)on the metabolism of rats was investigated to provide theoretical references for the clinical rational use of the drug.Firstly,the appropriate con-centration of AEE suspension was prepared.Wistar rats were randomly divided into three groups:the normal group,the AEE low-dose group(18 mg/kg),and the AEE high-dose group(72 mg/kg).The rats in the dosing group were dosed once daily,and the Wistar rats in the normal group were dosed once daily with an equal volume of 0.5％sodium carboxymethylcellulose solution.The feces and urine were collected after 7 days of continuous gavage,and the feces and urine were ana-lyzed by ultra-performance liquid chromatography-quadrupole time-of-flight tandem mass spec-trometry(UPLC-QTOF-MS/MS)for non-targeted metabolomics and Metabo Analyst 5.0 was used for metabolic pathway enrichment.The results showed that the dose of AEE selected in this experiment was not toxic to the growth of rats.The results of the metabolomics study found that 10 and 8 differential metabolites were identified in rat feces and urine,respectively,involving meta-bolic pathways such as phenylalanine,tyrosine and tryptophan biosynthesis,phenylalanine metabo-lism,steroid hormone biosynthesis,biosynthesis of unsaturated fatty acids,aminosugar and nucleo-tide sugar metabolism,fatty acid biosynthesis,and β-alanine metabolism.AEE had no significant effect on the body weight of rats(P＞0.05),but AEE could affect the metabolism of rat organ-ism.Fecal metabolites were mainly involved in metabolic pathways including unsaturated fatty acid biosynthesis,tyrosine metabolism,fatty acid biosynthesis,and steroid hormone biosynthesis;urina-ry metabolites were mainly involved in metabolic pathways including purine metabolism,fatty acid biosynthesis,arginine,and proline metabolism.Therefore,the metabolic effects of AEE on rats are mainly closely related to the regulation of lipid metabolism,amino acid metabolism,and energy metabolism.The results of this experiment can provide some references for the efficacy and clinical application of AEE in animals.

Objective:To explore any effects of combining virtual reality training with aromatherapy in caring for Alzheimer′s disease patients in a nursing home.Methods:Fifty nursing home residents with Alzheimer′s were divided at random into an observation group and a control group, each of 25. Both groups received routine rehabilitation, while the observation group was additionally given 45 minutes of virtual reality training combined with aromatherapy, 3 times a week for 6 months. Both groups′ cognition was then evaluated using the MMSE and an Alzheimer′s cognition assessment scale (ADAS-cog). Psycho-behavioral symptoms were quantified using the Alzheimer′s disease pathological behavior scale (BEHAVE-AD). Motor functioning was quantified using the timed up and go test (TUGT), the 30-second sit-to-stand test (30sCST), the 30-second arm curl test (30sACT) and the sit-and-reach test (CSRT). Ability in the activities of daily living (ADL) and life quality were quantified using the activity of daily living scale and of the quality of life scale for Alzheimer′s disease (QOL-AD) before and after the intervention.Results:After the intervention the average MMSE, ADAS-cog, BEHAVE-AD and ADL scores of both groups had improved significantly, with the average improvement in the observation group significantly greater than that in the control group. The TUGT, 30sCST, 30sACT and CSRT results of both groups were also significantly better, with those of the observation group again significantly superior, on average, to the control group′s results. The average QOL-AD score in the observation was significantly improved after the intervention, and was then significantly better than the control group′s average.Conclusions:Virtual reality training combined with aromatherapy can significantly improve the cognition, psycho-behavioral symptoms, activity in daily living, motor functioning and life quality of Alzheimer′s patients in a nursing home. It is worthy of promotion and application in nursing homes.

Objective:To observe any effect of virtual reality (VR) training on the cognitive functioning and functional fitness of nursing home residents with subjective cognitive decline (SCD).Methods:Fifty-six of such residents were randomly divided into an observation group and a control group, each of 28. Both groups received health education and routine care, but the observation group was additionally provided with 45 minutes of VR training three times a week for 6 months. The training included Baduanjin, magic, flying bird, supermarket shopping, gravity ball and gym episodes. Both groups′ cognition was evaluated using the subjective cognitive decline questionnaire (SCD-Q), the Montreal cognitive assessment (MoCA), the Rivermead Behavioural Memory Test (second edition) (RBMT-Ⅱ), a digit symbol substitution test (DSST), an animal fluency test (AFT) and trail-making test A-B (TMT A-B). Functional fitness was quantified using the 8-foot up-and-go test (8UGT), a 30-second arm curl test (30sACT), a 30-second chair stand test (30sCST), a back scratching test (BST), the sit-and-reach test (CSRT) and a 2-minute step test (2MST) before and after the 6-month intervention.Results:After the intervention, the average SCD-Q, MoCA, RBMT-Ⅱ, DSST, TMT-A, and TMT-B scores of the observation group were significantly better than before the intervention, and significantly better than the control group′s averages. And except for the back scratching their functional results were also significantly better, on average, than those of the control group.Conclusions:VR training can effectively improve the cognition and functional fitness of nur-sing home residents with SCD. Such training is worthy of promotion and wider application in nursing homes.

Objective:To explore the value of ultra-short echo time magnetization transfer (UTE-MT) techniques for quantitatively dynamic monitoring of anterior patellar tendon (patellar tendon, quadriceps tendon) changes in amateur marathon runners before and after competition.Methods:Between October 2020 and January 2021, 23 amateur marathoners in Zhuhai, aged 28-50 (40±6) years, were prospectively recruited. Three-dimensional UTE-MT and dual-echo UTE-T 2* sequence scans of bilateral knee joints were performed before, 48 hours and 4 weeks after the marathon running, respectively. Another 5 non-running volunteers were recruited for verification of sequence stability. UTE-magnetization transfer ratio (MTR) and UTE-T 2* value of the patellar tendon, quadriceps tendon, and 3 tendon-bone insertion points (patellar tendon-tibial insertion point, patellar tendon-patellar insertion point, and quadriceps tendon-patellar insertion point) were measured independently on sagittal images of the knee joint by 2 radiologists. The stability of the 2 serial measurements and consistency tests between the 2 radiologists were assessed with a two-way mixed intraclass correlation coefficient (ICC). Repeated-measures analysis of variance was used to compare the differences in UTE-MTR and UTE-T 2* values of the prepatellar tendon before and after the marathon running. Results:Both UTE-MT and dual-echo UTE-T 2* sequence measurements had good stability, with ICC values of 0.98 and 0.92, respectively. Measurements of UTE-MTR and UTE-T 2* value of the patellar tendon, quadriceps tendon, and the 3 tendon-bone insertion points by the 2 radiologists were in good agreement (ICC>0.80). Forty-eight hours after the marathon running, the UTE-MTR of the patellar tendon, quadriceps tendon, and the 3 tendon-bone insertion points decreased, and UTE-MTR of the patellar tendon continued to decrease 4 weeks after the race, while UTE-MTR of other regions increased. Only the difference in UTE-MTR for the patellar tendon was statistically significant ( F=7.46, P=0.001) among pre-marathon (0.34±0.04), 48 h after the race (0.32±0.04), and 4 weeks after the race (0.31±0.04). UTE-T 2* value was mildly elevated in all regions at 48 h after the marathon running, but the differences among the three points were not statistically significant ( P>0.05). Conclusion:The UTE-MT has better reproducibility and inter-rater reliability. The UTE-MT can be used to monitor the dynamic changes of the prepatellar tendon before and after marathon exercise, where the UTE-MTR of the patellar tendon consistently decreases after marathon exercise.

Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on postoperative patient-controlled intravenous analgesia in pediatric patients undergoing lower extremity orthopedic surgery.Methods:Sixty-eight pediatric patients of both sexes, aged 3-15 yr, of American Society of Anesthesiologists physical status Ⅰor Ⅱ, undergoing elective lower extremity orthopedic surgery under general anesthesia, were divided into 2 groups ( n=34 each) by the random number table method: TEAS group (group T) and control group (group C). In group T, the bilateral Hegu and Neiguan acupoints were stimulated starting from 10 min before induction of anesthesia until the end of procedure, with the frequency of disperse-dense wave of 2/10 Hz, and the current intensity was gradually adjusted to the maximum intensity (10-15 mA) that children could tolerate. In group C, the electrodes were applied to the same acupoints, but electrical stimulation was not applied. The severity of pain was assessed by the Faces Pain Scale-Revised scale immediately after returning to the ward and at 2, 24 and 48 h after operation. The emergence agitation was evaluated using the Pediatric Anesthesia Emergence Delirium scale. The intraoperative consumption of propofol and remifentanil and time to extubation after stopping administration were recorded. The time to first pressing of patient-controlled analgesia (PCA), effective pressing times of PCA on 1st and 2nd days after surgery and postoperative adverse reactions such as postoperative nausea and vomiting, pruritus, drowsiness, and respiratory depression were recorded. Results:Compared with group C, the Faces Pain Scale-Revised scale scores were significantly decreased immediately after returning to the ward and at 2, 24 and 48 h after operation, the incidence of emergence agitation and intraoperative consumption of remifentanil were decreased, the time to extubation was shortened, the time to first pressing of PCA was prolonged, and the effective pressing times of PCA on 1st and 2nd days after surgery were decreased ( P<0.05). There was no significant difference in the intraoperative consumption of propofol and incidence of postoperative adverse reactions between the two groups ( P>0.05). Conclusions:TEAS can effectively enhance the effect of postoperative patient-controlled intravenous analgesia in pediatric patients undergoing lower extremity orthopedic surgery.