ObjectiveTo explore the effects of astragalin （AST） on autophagy and apoptosis of astrocytes in the L_4-5 dorsal horn of the spinal cord in mice with inflammatory pain induced by complete Freund's adjuvant （CFA）. MethodsTwenty-four male C57BL/6 mice， aged six months， were randomly assigned to four groups： control group， saline group， CFA model group， and CFA+AST group， six mice in each group. The inflammatory pain model was established by injection of 10 µL CFA into the right lateral malleolus fossa. The saline group were injected with an equal amount of normal saline at the same site. The inflammatory pain mice in CFA+AST group were further treated with AST （60 mg/kg） intraperitoneally once a day for 21 consecutive days. Multiplex immunofluorescence staining was used to detect the coexpression of autophagy-related factors including ATG 12 and Beclin-1， apoptosis-related factors including Cleaved-Caspase3 and Caspase9， and the astrocyte marker such as GFAP in the L_4-5 spinal dorsal horn of the mice in each group. Western blot was used to examine the protein expression levels of autophagy-related proteins（ATG12， Beclin-1） and apoptosis-related proteins（Caspase 3， Caspase 9） in the L_4-5 spinal dorsal horn of mice. ResultsImmunofluorescent staining showed that in the L_4-5 dorsal horn of the spinal cord， the fluorescence intensity of ATG12 （P<0.000 1） and Beclin-1 （P<0.000 1） was significantly increased， while that of Cleaved-Caspase 3 （P<0.001） and Caspase 9 （P<0.000 1） was decreased in the CFA+AST group when compared to the CFA model group. Furthermore， AST could inhibit the activation of astrocytes. Western blot further confirmed that AST significantly upregulated the expression of ATG12 （P<0.000 1） and Beclin-1 （P<0.000 1） in the L_4-5 spinal cord of CFA mice， and downregulated the expression of Caspase 3 （P<0.01） and Caspase 9 （P<0.001）. ConclusionsAST promotes autophagy of astrocytes and inhibits their apoptosis in the L_4-5 spinal dorsal horn of CFA mice.

OBJECTIVE: The investigation of the correlation between ecological factors and the genetic characteristics or metabolites of plants offers valuable insights into the regional causes of genetic and metabolic diversity. Here, Gastrodia elata, a medicinal plant, is employed as a model to explore the environmental factors that influence its genetic characteristics and metabolic accumulations. METHODS: A total of 23 G. elata populations from six cultispecies and 11 cultivated regions were selected based on the predictions of the global geographic information system. The genetic characteristics of these populations were evaluated using highly polymorphic simple sequence repeat markers. Additionally, the metabolic accumulations and antioxidant capacity of mature tubers were measured employing colorimetry and high performance liquid chromatography (HPLC). Ecological data of each region were obtained from the WorldClim-global climate database and harmonized world soil database. To assess the influence of ecological factors on the genetic characteristics and metabolic profiles of G. elata, Pearson's correlation analysis was conducted. RESULTS: Genetic variation among G. elata populations exceeded that within populations. Genetic diverisity, distance and structure manifested regional and species-specific patterns. Metabolic profiling and antioxidant capacity exhibited regional variations. Notably, the Lueyang region demonstrated that a content range of total polysaccharide, total protein, and phenolic glycosides was 9.34%-189.67% higher than the average. Similarly, in the Hubei region, total phenolic content, p-hydroxybenzyl alcohol content, and antioxidant indicators were observed to be higher than the average levels, by 106.57%, 136.47% and 12.50%-91.14%, respectively. Furthermore, ecological factors had a significant comprehensive impact on G. elata genetic characteristics (r > 0.256 and P < 0.05). Multivariate metabolite accumulations in G. elata were influenced by dominant ecological factors. Temperature notably impacted the accumulation of total protein (|r| > 0.528 and P < 0.05). Moisture, encompassing precipitation and soil content, significantly affected the production of phenolic glycosides (|r| > 0.503 and P < 0.05). CONCLUSION The genetic characteristics of G. elata manifested regional and species-specific patterns, with the metabolic accumulations and antioxidant capacity of mature tubers exhibited regional variations. Specifically, multivariate ecological factors comprehensively influenced genetic characteristics. Temperature and moisture played pivotal roles in regulating the accumulations of proteins and phenolic glycosides, respectively. These findings underscore the significant impact of ecological factors on the shaping of G. elata, highlighting their crucial role in enhancing the quality of Chinese medicinal materials.

ObjectiveTo investigate the potential value of thyroid hormones in cerebrospinal fluid in predicting autism-like behaviors induced by hypothyroidism in pregnant rats. MethodsTwelve pregnant Wistar rats were randomly divided into a control group and a hypothyroidism group （hypothyroidism model group）. Offspring from both groups had serum and cerebrospinal fluid levels of thyroid-stimulating hormone （TSH）， total triiodothyronine （TT3）， total thyroxine （TT4）， free triiodothyronine （FT3）， and free thyroxine （FT4） levels in serum and cerebrospinal fluid. Ultrasonic vocalization tests were conducted on postnatal day 2 （P2）， day 7 （P7）， and day 14 （P14）， while behavioral tests using the three-box social interaction test were performed on day 21（P21）. ResultsCompared with the control group， free T4 （FT4） levels in the cerebrospinal fluid of the hypothyroidism group were significantly reduced during the developmental period （P0-P21； P2： P<0.05； P7： P<0.05； P14： P<0.01； P21： P<0.01）， with no statistical difference between the two groups only at P0 （P>0.05）. In the ultrasonic vocalization （USV） tests， the number and duration of USVs in offsprings from the hypothyroidism group were significantly reduced compared with those of the control group on P2， P7 and P14： for USV counts （P2： P<0.05； P7： P<0.001； P14： P<0.01）； for USV duration （P2： P<0.05； P7： P<0.001； P14： P<0.001）. In the three-box social tests， offsprings of the hypothyroidism group showed significantly reduced sniffing time with unfamiliar rats at P21 compared to the control group （all P<0.001）. The FT4 levels in cerebrospinal fluid had a significantly positive correlation with USV counts （P7： r=0.883， P<0.05； P14： r=0.902， P<0.05） and sniffing time with unfamiliar rats （r=0.814， P<0.01）. ConclusionMeasuring free T4 in cerebrospinal fluid can predict autism-like behaviors in offsprings of rats induced by hypothyroidism during pregnancy.

Objective:To discuss the effect of microRNA(miR)-30c-5p on the proliferation,migration,and invasion of the human prostate cancer cells(LNCap),and to clarify its possible mechanism.Methods:The LNCap cells were divided into LNCap group(without plasmid transfection),miR-30c-5p mimic group(transfected with miR-30c-5p mimic),mimic NC group(transfected with miR-30c-5p mimic NC),sh-DNA damage inducible transcript 4(DDIT4)group(transfected with sh-DDIT4),sh-NC group(transfected with sh-DDIT4 NC),miR-30c-5p mimic+pc-DNA3.1-NC group(co-transfected with miR-30c-5p mimic and pc-DNA3.1 empty vector),and miR-30c-5p mimic+pc-DNA3.1-DDIT4 group(co-transfected with miR-30c-5p mimic and pc-DNA3.1-DDIT4 over-expression plasmid).The RWPE-1 cells were cultured normally.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-30c-5p and DDIT4 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of DDIT4 protein in the cells in various groups;CCK-8 method was used to detect the proliferation rates of the LNCap cells in various groups;Transwell assay was used to detect the numbers of the invasion LNCap cells in various groups;Scratch assay was used to detect the scratch healing rates of LNCap cells in various groups;dual-luciferase reporter assay was used to detect the targeting relationship between miR-30c-5p and DDIT4.In the in vivo tumor formation experiment,18 male BALB/c nude mice were divided randomly into blank group,agomiR-NC group(transfected with agomiR-30c-5p NC),and agomiR-30c-5p group(transfected with agomiR-30c-5p);there were six mice in each group.The mice in agomiR-NC group and agomiR-30c-5p group were subcutaneously injected with LNCap cells,while the mice in blank group were given an equal volume of physiological saline.The volumes of tumor of the mice in various groups were detected.HE staining was used to observe the morphology of prostate cancer tissue the mice of in various groups;RT-qPCR method and immunofluorescence staining were used to detect the expression levels of miR-30c-5p and DDIT4 mRNA and the fluorescence intensities of DDIT4 protein in prostate cancer tissue of the mice in various groups.Results:The In vitro prostate cancer cell experiment results showed that compared with RWPE-1 cells,the expression level of miR-30c-5p in the prostate cancer LNCap cells was decreased(P<0.01),and the expression levels of DDIT4 mRNA and protein were increased(P<0.05 or P<0.01).After 48 of transfection,compared with LNCap group and mimic NC group,the expression level of miR-30c-5p in the LNCap cells in miR-30c-5p mimic group was increased(P<0.01).Compared with LNCap group and sh-NC group,the expression level of DDIT4 mRNA in the LNCap cells in sh-DDIT4 group was decreased(P<0.01).Compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of miR-30c-5p in The LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 mRNA in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 protein in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.05).The CCK-8 method results showed that compared with LNCap group and mimic NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the proliferation rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The Transwell assay results showed that compared with LNCap group and mimic NC group,the number of the invasion LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the number of invasion LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the number of the invasion LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The scratch assay results showed that compared with LNCap group and mimic NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the scratch healing rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The dual-luciferase reporter assay results showed that compared with the LNCap cells co-transfected with WT-DDIT4 and mimic NC,the luciferase activity of the LNCap cells co-transfected with WT-DDIT4 and miR-30c-5p mimic was decreased(P<0.01).The in vivo nude mouse tumor formation experiment results showed that on the 3 rd,6 th,9 th,12 th,and 15th days after cell injection,compared with blank group and agomiR-NC group,the tumor volumes of the nude mice in agomiR-30c-5p group were decreased(P<0.05).The HE staining results showed that in prostate cancer tissue of the mice in blank group and agomiR-NC group,the cell nuclei were enlarged,and nucleoli were prominent and deformed.In the mice in agomiR-30c-5p group,some regions of prostate cancer tissues results showed neatly arranged cells with normally shaped nuclei.The RT-qPCR and immunofluorescence staining showed that compared with agomiR-NC group,the expression level of miR-30c-5p in prostate cancer tissue of the mice in agomiR-30c-5p group was increased(P<0.01).Compared with blank group and agomiR-NC group,the expression level of DDIT4 mRNA in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).DDIT4 protein was mainly expressed in the cytoplasm.Compared with blank group and agomiR-NC group,the fluorescence intensity of DDIT4 protein in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).Conclusion:The expression level of miR-30c-5p in the prostate cancer LNCap cells is decreased,and it inhibits the proliferation,migration,and invasion of the prostate cancer cells by targeting downregulation of DDIT4,thereby participating in the occurrence and development of prostate cancer.

Objective:To obtain the normal reference range of hemodynamic ultrasound parameters after pediatric liver transplantation through big data query and statistical analysis, and compare their changes with age.Methods:The clinical liver transplantation ultrasound imaging database software V1.0 independently developed by Tianjin First Central Hospital was used to query the ultrasound hemodynamic parameters of 0-14 years old pediatric patients after parental liver transplantation from December 2012 to December 2022, including portal vein diameter (PVD) on the 1st day, 7th day, 1st month, 6th month, 1st year, 5th year, and 10th year after surgery. The changes in ultrasound blood flow parameters such as portal vein velocity (PVV), hepatic artery peak velocity (S), hepatic artery end diastolic velocity (D), and left hepatic vein velocity (LHVV) with postoperative time in different age groups were analyzed, the differences between age groups were compared, and statistical analysis was perform to obtain the 95% reference value range for each parameter.Results:A total of 731 pediatric patients aged 0-14 years who underwent parental liver transplantation were included in this study, with a total of 5 283 monitoring results. The ultrasound hemodynamic parameters PVV, hepatic artery S, D, and LHVV were highest at 7th day after surgery, and gradually showed a sustained and slightly decreasing trend with the prolongation of postoperative time. At the same time, there was no statistically significant difference in PVV, hepatic artery S, D, and LHVV among different age groups (all P>0.05). Conclusions:This study obtains the trend of ultrasound hemodynamic parameters after pediatric liver transplantation with prolonged follow-up time, and compared them among different age groups.After pediatric liver transplantation, there is no significant change in liver hemodynamic parameters with increasing age. Obtaining reference value ranges for various parameters in different age groups is of great clinical significance for early detection and diagnosis of postoperative vascular complications. Pediatric liver transplantation is more complex than adult liver transplantation, and in clinical monitoring, more attention should be paid to the dynamic changes of transplanted liver blood flow, combining with individual patient status, to provide imaging support for clinical diagnosis.

In recent decades,the advent of coronaviruses has engendered multiple large-scale infectious outbreaks,leading to profound human casualties and posing a significant menace to global public health.At the crux of the coronavirus lifecycle,the assembly and release phases emerge as pivotal processes,necessitating meticulous investigation into the underlying mechanisms.This paper systematically expounds upon the intricate assembly and packaging mechanisms intrinsic to coronaviruses,scrutinizes the diverse release pathways employed by these viruses,and delves into the discourse surrounding targeted antiviral drugs.The comprehensive insights encapsulated in this exposition serve as valuable reference points for formulating effective strategies in the prevention and treatment of coronavirus infections.

Objective:To investigate the risk factors of intraspinal cement leakage in the treatment of osteoporotic vertebral compression fracture by percutaneous vertebroplasty (PVP).Methods:From January 2018 to December 2021, 156 patients with OVCF who received surgical treatment in Beijing Tongzhou Integrated Traditional Chinese and Western Medicine Hospital were enrolled in this retrospective case-control study. The postoperative CT imaging results were analyzed, and the patients were divided into two groups according to the occurrence of intraspinal cement leakage: leakage group ( n=28) and non-leakage group ( n=128). Measurement data was expressed as mean±standard deviation ( ± s), and t-test was used for comparison of visual analogue score (VAS) between groups; the count data was expressed as the number of cases and percentage (%); univariate and multivariate Logistic regression analysis were used to evaluate the risk factors of postoperative intraspinal cement leakage. Results:All the patients were treated by PVP successfully, without obvious adverse reactions and serious complications occurred during and after the operation. Univariate Logistic regression analysis showed that higher bone mineral density ( P=0.005), OVCF combined with posterior vertebral wall injury ( P<0.001) and higher bone cement dosage ( P=0.013) were the risk factors leading to intraspinal cement leakage. Multivariate Logistic regression analysis showed that higher bone mineral density ( P=0.009, 95% CI: 0.152-0.762, OR=0.340), OVCF combined with posterior vertebral wall injury ( P=0.001, 95% CI: 2.134-15.780, OR=5.803), and higher bone cement dosage ( P=0.005, 95% CI: 1.175-2.505, OR=1.715) were the independent risk factors of intraspinal cement leakage. Conclusion:Intraspinal cement leakage was common complication after PVP. Higher bone mineral density, OVCF combined with posterior vertebral wall injury, and higher bone cement dosage were the independent risk factors affecting intraspinal cement leakage.

Objective To investigate the correlation between abdominal obesity and autoimmune thyroid disease in the view point of helper T cells and cytokines.Methods Clinical and laboratory data were collected from 108 pa-tients with Hashimoto's thyroiditis(HT)plus abdominal obesity and 122 patients of Hashimoto's thyroiditis without abdominal obesity who visited the People's Hospital of Xinjiang Uygur Autonomous Region and also from the control population.Abdominal circumference was measured,and patients in the HT patients were grouped according to whether they were abdominally obese or not.The thyroglobulin antibody(TgAb)and thyroid peroxidase antibody(TPOAb)were detected,and the ratio of helper T cells and related cytokines were detected by flow cytometry and enzyme-linked immunosorbent assay.Results The abdominal circumference of the TgAb-positive group was higher than that of the TgAb-negative group(P＜0.05).Correlation analysis suggested that abdominal circumference was significantly and positively correlated with TgAb and IL-4 but negatively correlated with Th1.After correcting for gender and age,and abdominal obesity and IL-4 were risk factors for TgAb antibody positivity OR=3.080(95％CI:1.022-9.284)and OR=1.296(95％CI:1.022-9.284),both with P＜0.05.Conclusions Abdominal obesity may be an influential factor in TgAb antibody positivity,with larger abdominal circumference having higher TgAb antibody titers,lower Th1 levels,and higher IL-4 levels.Visceral adiposity may exacerbate autoimmune dam-age of thyroid tissue by disruption of helper T cell pathway.

Objective To screen and verify the genes that play key role in the occurrence and development of esophageal cancer by u-sing bioinformatics and real-time fluorescence quantitative PCR(qRT-PCR)methods to find new markers for diagnosis of esophageal cancer(ESCA).Methods Using the TCGA database and Wayne plot analysis,the cross genes between the differentially expressed genes of ESCA and the genes which have the most significant impacts on disease-free survival(DFS)rate in esophageal cancer patients were preliminarily identified.Following conducting protein-protein interaction(PPI)network analysis on the overlapping genes,GO and KEGG functional analysis was performed to screen the potential key genes as the diagnostic markers of esophageal cancer.qRT-PCR was used to quantitatively analyze the expression of mRNA of the key gene in peripheral blood.Statistical analysis was con-ducted based on the clinico-pathological characteristics of the patients to determine its potential value as a new diagnostic marker for e-sophageal cancer.Results After overlapping of differentially expressed genes of ESCA and disease-free survival genes in the TCGA database,39 upregulated genes and 20 downregulated genes were found to be differentially expressed,all of which affected disease-free survival rate.After conducting PPI network analysis,15 upregulated genes with core interactions were identified,and the downregulat-ed genes did not form any interaction network.Further enrichment analysis of these 15 core interacting genes through GO and KEGG,revealed that fibronectin 1(FN1)may be a potential biomarker for ESCA diagnosis.The qRT-PCR results showed that compared with the healthy control group,the mRNA expression level of FN1 in the peripheral blood of esophageal cancer patients was significantly ele-vated.After analyzing the clinical characteristics of patients,it was found that the patients with poor differentiation and high clinico-pathological staging had significantly increased peripheral blood FN1 mRNA levels.The model with FN1 mRNA expression levels can distinguish esophageal cancer patients from healthy individuals.Conclusion FN1 mRNA may be a potential non-invasive diagnostic biomarker for esophageal cancer.

Objective To investigate the epidemiological characteristics of Mycoplasma pneumoniae(MP)infection in children in Nanjing Integrated Traditional Chinese and Western Medicine Hospital before and after the COVID-19 outbreak,and provide an experi-mental basis for the prevention and treatment of MP infection.Methods The clinical data of 17 976 children visited Nanjing Integrated Traditional Chinese and Western Medicine Hospital from January 2019 to November 2023 due to respiratory tract infections were retro-spectively analyzed.The levels of serum specific MP-IgM in the children were detected by the direct luminescence immunoassay,and the detection rates of MP-IgM in different genders,seasons,and ages before and after the COVID-19 epidemic were analyzed using the chi square test to explore the epidemiological characteristics of MP infection.Results The total detection rate of serum MP-IgM in 17 976 children with respiratory tract infections was 28.45%(5 114/17 976).Among them,the total detection rate of serum MP-IgM in female children(31.69%,2 672/8 432)was significantly higher than that in male children(25.59%,2 442/9 544,χ2=81.89,P＜0.001).The detection rate of serum MP-IgM was highest in 2019(34.35%,1 415/4 119),followed by in 2023(30.11%,2 409/8 001)and in 2020-2022(22.03%,1 290/5 856),with statistically significant difference(χ2=19.95,P＜0.001).The detection rate of serum MP-IgM was highest in autumn(33.16%,1 683/5 075),followed by in summer(28.61%,1 053/3 681),winter(27.65%,1 826/6 604),and spring(21.10%,552/2 616),with statistically significant difference(χ2=126.90,P＜0.001).Among different age groups,the detection rate of serum MP-IgM was highest in the age group of 7-9 years old(35.83%,1 190/3 321),fol-lowed by 4-6 years old(28.06%,1 882/6 707),1-3 years old(26.55%,1 493/5 623),10-18 years old(23.64%,486/2 056),and＜1 year old(23.42%,63/269),with statistically significant difference(χ2=126.11,P＜0.001).Conclusion MP is a common pathogen of respiratory tract infections in children,especially in the age range of 7-9 years old,with female children having a higher in-cidence than male children,and the peak incidence in autumn.The effective prevention and control measures during the COVID-19 ep-idemic have reduced the detection rate of MP-IgM,which may provide certain experimental basis for the control and prevention of MP infection transmission and other respiratory diseases.