ObjectiveTo identify the main chemical constituents of Anmeidan （AMD） and to explore the mechanism of AMD in regulating the extracellular signal-regulated kinase 1/2 （ERK1/2）/mitogen-activated protein kinase （MAPK）-interacting serine/threonine-protein kinase （MNK）/eukaryotic translation initiation factor 4E （eIF4E） signaling pathway to improve circadian rhythm disturbances in insomnia rats. MethodsThe main chemical constituents of AMD were identified using ultra-high-performance liquid chromatography-linear ion trap-electrostatic orbital trap mass spectrometry （UPLC-LTQ/Orbitrap/MS） in combination with reference standards. Sixty male Sprague-Dawley （SD） rats were randomly divided into control， model， melatonin， and AMD low-， medium-， and high-dose groups， with 10 rats in each group. Except for the control group， all rats were administered p-chlorophenylalanine via intraperitoneal injection to establish an insomnia model. The activity-rest rhythm of rats was assessed using the open field test and circadian rhythm test. Hematoxylin-eosin （HE） staining and Nissl staining were used to observe structural changes in hypothalamic neurons. Immunofluorescence， real-time quantitative polymerase chain reaction （Real-time PCR）， and Western blot analysis were employed to detect mRNA and protein expression levels of ERK1/2， MNK， and eIF4E in the hypothalamus. ResultsA total of 50 chemical components， including flavonoids， phenylpropanoids， triterpenoid saponins， alkaloids， and lignans， were identified in AMD. Compared with the control group， the model group exhibited significantly increased total distance traveled， average speed， central area residence time， and cumulative rearing time （P<0.01）， as well as prolonged cumulative activity time and total activity time in both light and dark phases （P<0.01）. Hypothalamic neurons in the model group were sparsely arranged， reduced in number， and exhibited nuclear disappearance or nucleolar rupture， with a significantly increased apoptosis index （P<0.01）. The cytoplasm appeared turbid， Nissl body staining was lighter， and the Nissl body apoptosis index was significantly increased （P<0.01）. The mRNA expression levels of ERK1/2， MNK， and eIF4E were significantly decreased （P<0.01）， along with a significant reduction in protein expression levels of ERK1/2， phosphorylated ERK1/2 （p-ERK1/2）， MNK， phosphorylated MNK （p-MNK）， eIF4E， and phosphorylated eIF4E （p-eIF4E） （P<0.01）. Compared with the model group， the total distance， average speed， central area residence time and body upright cumulative time of the AMD high-dose group were significantly reduced （P<0.01）. The total distance， average speed and body upright cumulative time of the AMD medium-dose group were significantly reduced （P<0.01）. The cumulative time of light activity and total time of activity in each dose group of AMD were significantly shortened （P<0.01）. The cumulative time of dark activity in the high-dose group of AMD was prolonged （P<0.01）. The neurons in the middle and high dose groups of AMD were closely arranged， the number of neurons increased， and the apoptosis index of hypothalamic cells decreased significantly （P<0.05， P<0.01）. The cytoplasm of the low， middle and high dose groups of AMD was clear， the color of Nissl body became darker， and the apoptosis index of Nissl body decreased significantly （P<0.01）. The expression of ERK1/2， MNK and eIF4E mRNA and protein in the hypothalamus of the middle and high dose groups of AMD increased significantly （P<0.05， P<0.01）. ConclusionAMD primarily contains 50 chemical constituents， including flavonoids， phenylpropanoids， and triterpenoid saponins. It exhibits a "synergistic enhancement" effect through multiple components and multiple pathways to improve insomnia. AMD ameliorates circadian rhythm disturbances in p-chlorophenylalanine-induced insomnia rats by upregulating ERK1/2/MNK/eIF4E signaling pathway-related proteins.

IKBKG is the essential modulator for nuclear factor-κB(NF-κB) signaling pathway, and mutations within this gene can lead to anhidrotic ectodermal dysplasia and immunodeficiency (EDA-ID). Here we report a male patient, who presented with mild frontal bossing, sparse hair, skin pigmentation, conical teeth, and recurrent infections involving bacteria, fungi, and viruses after one month of age, together with hypogammaglobulinemia. These symptoms were consistent with the phenotype of EDA-ID. Genetic analysis revealed a hemizygous mutation c.1249T > G (p.Cys417Gly) in exon 10 of the IKBKG gene in the patient. The mother of the patient was identified as heterozygous carriers of the same mutation. Immunological assessment revealed a significant reduction in memory B cells. The patient showed no skewed TCR diversity. PHA-induced T-cell proliferation was impaired. IFN-γ secretion by Th cells was significantly reduced after PHA stimulation. IκBα degradation rate retained the same in the patient. We summarized the clinical features of the patient and conducted immunological analysis, in order to increase pediatricians′awareness of this rare disease. For boys with early-onset recurrent infections together with ectodermal dysplasia, IKBKG mutation should be considered. In addition, genetic analysis is needed to exclude pseudogene interference and functional evaluations are required.

Objective To develop a group psychological training program to cultivate a sense of meaning in life for retired soldiers during the transitional period,and to explore the efficacy of the training.Methods A psychological training program for retired soldiers during the transitional period was developed based on Delphi method.A total of 64 retired soldiers who participated in a retirement training in transitional period were subjected through cluster sampling in a certain city in Shandong province and assigned into a training group(n=31)and a control group(n=33).The training group attended group psychological training based on the developed training program(twice a week,1.5～2 h each time,for 4 weeks),and the control group received routine adaptive training for retirement.Purpose in Life(PIL)Test,Chinese Perceived Stress Scale(CPSS)and Social Adaptability Scale(SAS)were used to survey the participants before(T1),and at the end of(T2)and 1 month after the training(T3).Results ① The results of 2 rounds of expert consultation showed that the coefficient of authority(Cr)was 0.89,and the Kendall's coefficient(ω)of the 2 rounds was 0.134 and 0.204,respectively(P＜0.001),with a coefficient of variation ranging from 0.05 to 0.14.The prepared group psychological training program consisted of 3 first-level indexes,6 second-level indexes,and 24 third-level indexes.② The results of the interaction between group and time after training showed that the training group had significantly higher total score and score of each dimension in sense of meaning in life,and increased total score of social adaptability,but lower total score of perceived stress when compared with the control group(all P＜0.001).③ The results of group main effect after training indicated that obviously higher total score and score of each dimension in sense of meaning in life(P＜0.01),elevated total score of social adaptability(P＜0.01),and reduced total score of perceived stress(P＜0.001)were observed in the training group than the control group.④ The results of the post-training time main effect revealed that the training group had notably higher total score and score of each dimension in sense of meaning in life(P＜0.05),increased total score of social adaptability(P＜0.001),and decreased total score of perceived stress(P＜0.001)when compared with the control group.Conclusion Our developed group psychological training can significantly improve the sense of meaning in life,reduce perceived stress and improve their social adaptation ability for retired soldiers during the transitional period.

Objective To examine the regulation of malignant progression of colorectal cancer by LINC00626 via the JAK1/STAT3/KHSRP signaling axis and its molecular mechanism.Methods 96 individuals diagnosed with colorectal cancer at our hospital during June 11,2021 and June 11,2023 were chosen as research subjects,and their cancerous tissue and nearby normal tissue were collected.Cultivate colorectal cancer cell lines(SW620,HCT116,HT29,DLD-1,LOVO,Caco-2)and normal colorectal cells(NCM460)in vitro,and detect the expression of LINC00626 and KHSRP in colorectal cancer tissue and cell lines using qRT-PCR.Screening out cell lines infected with lentivirus,SW620 and HCT116 cell lines were transfected with knockdown lentivirus and its control,while HT29 and DLD-1 cell lines were transfected with overexpressing lentivirus and its control,respectively.Select stable transfected cell lines for cell function experiments to detect proliferation,migration,and invasion abilities.Detection of the effect of LINC00626 on the growth and migration of colorectal cancer tumors in live mouse experiments.The expression level of KHSRP protein in stable labeled cells was determined using a western blot analysis.Rescue experimental research on the regulatory relationship between LINC00626 and KHSRP.Results qRT-PCR showed low expression of LINC00626 and high expression of KHSRP in colorectal cancer tissues and cell lines.Cell function experiments showed that compared with the sh-NC group,the sh-LINC00626 group promoted cell proliferation,migration,and invasion in SW620 and HCT116 cells,while the overexpression group showed the opposite.Cell rescue experiments showed that,LINC00626+KHSRP significantly reversed the promotion effects of knocking down LINC00626 on cell proliferation,migration,and invasion.In the nude mouse experiment,com-pared with the sh-NC group,the sh-LINC00626 group showed a significant increase in tumor volume and weight,cell proliferation rate,and the number of lung metastases from colorectal cancer in the nude mice;Overexpression results in the opposite.The signal pathway experiment revealed that relative to the sh-NC group,the expression levels of JAK1 and STAT3 mRNA in the sh-LINC00626 group were significantly increased,whereas the results in the overexpression group were the opposite.Conclusion LINC00626 suppression the malignant progression of colorec-tal cancer metastasis through the JAK1/STAT3/KHSRP signaling axis.

Objective To investigate the role of JAK1/STAT3/KHSRP axis in mediating the regulatory effect of LINC00626 on progression of esophagogastric junction adenocarcinoma.Methods We collected surgical tumor and adjacent tissue specimens from 64 patients with esophagogastric junction adenocarcinoma and examined the expression levels of LINC00626 and KHSRP.qRT-PCR was used to detect the expressions of LINC00626 and KHSRP in 6 esophageal adenocarcinoma cell lines(OE-19,TE-7,Bic-1,Flo-1,SK-GT-4,and BE-3)and a normal esophageal epithelial cell line(HET-1A).OE-19 and TE-7 cell lines with stable LINC00626 knockdown and FLO-1 and SK-GT-4 cells stably overexpressing LINC00626 were constructed by lentiviral transfection,and the changes in proliferation,migration and invasion of the cells were evaluated using Cell Counting Kit-8(CCK-8)assay and Transwell migration/invasion assay.The expressions of KHSRP and JAK/STAT pathway proteins in the transfected cells were detected with Western blotting.The effects of LINC006266 knockdown and overexpression on subcutaneous tumor formation and lung metastasis of OE-19 and FLO-1 cell xenografts were tested in nude mice.Results The expression levels of LINC00626 and KHSRP were significantly increased in esophagogastric junction adenocarcinoma tissues and in esophageal adenocarcinoma cells.LINC00626 knockdown obviously inhibited the proliferation,migration and invasion of esophageal adenocarcinoma cells in vitro and decreased their tumor formation and lung metastasis abilities in nude mice,while overexpression of LINC00626 produced the opposite effects.In esophageal adenocarcinoma cells,LINC0626 knockdown significantly decreased and LINC00626 overexpression strongly enhanced the phosphorylation of JAK1 and STAT3.Conclusion High LINC00626 expression promotes esophageal-gastric junction adenocarcinoma metastasis by activating the JAK1/STAT3/KHSRP signal axis.

Objective To explore the mechanism underlying the protective effect of Lonicerae japonicae flos(LJF)extract against doxorubicin(DOX)-induced liver injury in mice.Methods Network pharmacology methods were used to obtain the intersection genes between LJF targets and disease targets,based on which the protein-protein interaction(PPI)network was constructed using STRING database for screening the core targets using Cytoscape software.DAVID database was used for bioinformatics analysis,and the core components and core targets were verified using molecular docking study.In a mouse model of DOX-induced liver injury,the effect of LJF extract on liver pathologies,serum levels of ALT and AST,and hepatic expressions of HYP,ROS,TNF-α,IL-6,COL-IV and P53 proteins were evaluated using HE and Masson staining,ELISA,and Western blotting.Results We identified 12 core targets from 43 intersection genes involving cancer pathway,IL-17 signaling pathway,and TNF signaling pathways.Molecular docking study suggested that 10 core components of LJF could bind to different core targets.The mice with DOX-induced liver injury showed elevated serum AST and ALT levels with obvious liver injury and fibrosis,increased ROS content,and enhanced expressions of TNF-α,IL-6,HYP,COL-IV and P53 proteins in the liver tissue.All these changes in the mouse models were significantly alleviated by treatment with LJF extract,suggesting obviously lowered levels of oxidative stress,inflammation and fibrosis in the liver tissues.Conclusion LJF extract is capable of alleviating DOX-induced liver injury in mice by downregulating Trp53,TNF and IL-6 to reduce liver oxidative stress,inflammation and fibrosis.

Objective To investigate the role of JAK1/STAT3/KHSRP axis in mediating the regulatory effect of LINC00626 on progression of esophagogastric junction adenocarcinoma.Methods We collected surgical tumor and adjacent tissue specimens from 64 patients with esophagogastric junction adenocarcinoma and examined the expression levels of LINC00626 and KHSRP.qRT-PCR was used to detect the expressions of LINC00626 and KHSRP in 6 esophageal adenocarcinoma cell lines(OE-19,TE-7,Bic-1,Flo-1,SK-GT-4,and BE-3)and a normal esophageal epithelial cell line(HET-1A).OE-19 and TE-7 cell lines with stable LINC00626 knockdown and FLO-1 and SK-GT-4 cells stably overexpressing LINC00626 were constructed by lentiviral transfection,and the changes in proliferation,migration and invasion of the cells were evaluated using Cell Counting Kit-8(CCK-8)assay and Transwell migration/invasion assay.The expressions of KHSRP and JAK/STAT pathway proteins in the transfected cells were detected with Western blotting.The effects of LINC006266 knockdown and overexpression on subcutaneous tumor formation and lung metastasis of OE-19 and FLO-1 cell xenografts were tested in nude mice.Results The expression levels of LINC00626 and KHSRP were significantly increased in esophagogastric junction adenocarcinoma tissues and in esophageal adenocarcinoma cells.LINC00626 knockdown obviously inhibited the proliferation,migration and invasion of esophageal adenocarcinoma cells in vitro and decreased their tumor formation and lung metastasis abilities in nude mice,while overexpression of LINC00626 produced the opposite effects.In esophageal adenocarcinoma cells,LINC0626 knockdown significantly decreased and LINC00626 overexpression strongly enhanced the phosphorylation of JAK1 and STAT3.Conclusion High LINC00626 expression promotes esophageal-gastric junction adenocarcinoma metastasis by activating the JAK1/STAT3/KHSRP signal axis.

Objective To explore the mechanism underlying the protective effect of Lonicerae japonicae flos(LJF)extract against doxorubicin(DOX)-induced liver injury in mice.Methods Network pharmacology methods were used to obtain the intersection genes between LJF targets and disease targets,based on which the protein-protein interaction(PPI)network was constructed using STRING database for screening the core targets using Cytoscape software.DAVID database was used for bioinformatics analysis,and the core components and core targets were verified using molecular docking study.In a mouse model of DOX-induced liver injury,the effect of LJF extract on liver pathologies,serum levels of ALT and AST,and hepatic expressions of HYP,ROS,TNF-α,IL-6,COL-IV and P53 proteins were evaluated using HE and Masson staining,ELISA,and Western blotting.Results We identified 12 core targets from 43 intersection genes involving cancer pathway,IL-17 signaling pathway,and TNF signaling pathways.Molecular docking study suggested that 10 core components of LJF could bind to different core targets.The mice with DOX-induced liver injury showed elevated serum AST and ALT levels with obvious liver injury and fibrosis,increased ROS content,and enhanced expressions of TNF-α,IL-6,HYP,COL-IV and P53 proteins in the liver tissue.All these changes in the mouse models were significantly alleviated by treatment with LJF extract,suggesting obviously lowered levels of oxidative stress,inflammation and fibrosis in the liver tissues.Conclusion LJF extract is capable of alleviating DOX-induced liver injury in mice by downregulating Trp53,TNF and IL-6 to reduce liver oxidative stress,inflammation and fibrosis.

Objective:To construct the training program system for hospice care volunteers and provide reference for the training of hospice care volunteers in China.Methods:The training program system for hospice care volunteers was initially determined by using the method of literature analysis and investigation, and 16 experts were consulted by two rounds of letters using the method of expert inquiry from May to July 2022, and finally the training program system was established.Results:The effective recovery rate of the two rounds of expert consultation questionnaire was 100%, the expert authority coefficient was 0.88, and the Kendall coordination coefficient was 0.141, 0.131 (both P<0.05). The final training program system for hospice care volunteers contained 7 first-class indicators including training objectives, training objects, training contents, training methods, training hours, training resources and training evaluation, 27 second-class indicators and 92 third-class indicators. Conclusions:The training program system for hospice care volunteers constructed in this study has high reliability and scientificity, and has a good guiding role and reference value for the training of hospice care volunteers.

Objective:To investigate the clinical and genetic characteristics, pathogenesis and treatment strategy of congenital nephrogenic diabetes insipidus(CNDI)combined with hyperuricemia.Methods:The clinical manifestations and laboratory data of an infant patient diagnosed as CNDI with hyperuricemia and his family members were collected and retrospectively analyzed. Whole exome sequencing(WES)was applied to detect the proband′s genome variation of each exon and suspected variants of AVPR2 and ABCG2 were verified by PCR-Sanger sequencing of members from his pedigree. Furthermore, we retrospectively collected the serum uric acid levels of patients(≤14-year-old) with CNDI in the First Affiliated Hospital of Zhengzhou University from January 2015 to January 2022.Results:The proband was clinically diagnosed with CNDI and the rest of the family members had no symptoms of polydipsia or polyuria. In addition to the proband, his father was also suffered from hyperuricemia. WES showed that the proband carried a hemizygous AVPR2 gene variation(p.S331R)and a heterozygous ABCG2 gene variation(p.N308K). The former was X-linked recessive inheritance from his mother, and the latter was autosomal dominant inheritance from the father. Fraction excretion of uric acid(FEUA)of the proband and his father with hyperuricemia were 3.1% and 2.7%, respectively. Twelve children(≤14-year-old)were diagnosed with CNDI from the respective study. Among all the cases, 11 patients were male and 1 was female, ranging from 3-month to 14-year-old. Five patients were accompanied with hyperuricemia.Conclusion:Children with CNDI may be complicated with hyperuricemia, and the regimen of hydrochlorothiazide combined with benzbromarone is effective. The pathogenicity of the AVPR2 gene variation(p.S331R)and ABCG2 gene variation(p.N308K)in this pedigree needs to be further studied.