This study aims to reveal the effect and mechanism of Gentianella turkestanorum total extract(GTI) in ameliorating non-alcoholic steatohepatitis(NASH). UPLC-Q-TOF-MS was employed to identify the chemical components in GTI. SwissTarget-Prediction, GeneCards, OMIM, and TTD were utilized to screen the targets of GTI components and NASH. The common targets shared by GTI components and NASH were filtered through the STRING database and Cytoscape 3.9.0 to identify core targets, followed by GO and KEGG enrichment analysis. AutoDock was used for molecular docking of key components with core targets. A mouse model of NASH was established with a methionine-choline-deficient high-fat diet. A 4-week drug intervention was conducted, during which mouse weight was monitored, and the liver-to-brain ratio was measured at the end. Hematoxylin-eosin staining, Sirius red staining, and oil red O staining were employed to observe the pathological changes in the liver tissue. The levels of various biomarkers, including aspartate aminotransferase(AST), alanine aminotransferase(ALT), hydroxyproline(HYP), total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione(GSH), in the serum and liver tissue were determined. RT-qPCR was conducted to measure the mRNA levels of interleukin 1β(IL-1β), interleukin 6(IL-6), tumor necrosis factor α(TNF-α), collagen type I α1 chain(COL1A1), and α-smooth muscle actin(α-SMA). Western blotting was conducted to determine the protein levels of IL-1β, IL-6, TNF-α, and potential drug targets identified through network pharmacology. UPLC-Q-TOF/MS identified 581 chemical components of GTI, and 534 targets of GTI and 1 157 targets of NASH were screened out. The topological analysis of the common targets shared by GTI and NASH identified core targets such as IL-1β, IL-6, protein kinase B(AKT), TNF, and peroxisome proliferator activated receptor gamma(PPARG). GO and KEGG analyses indicated that the ameliorating effect of GTI on NASH was related to inflammatory responses and the phosphoinositide 3-kinase(PI3K)/AKT pathway. The staining results demonstrated that GTI ameliorated hepatocyte vacuolation, swelling, ballooning, and lipid accumulation in NASH mice. Compared with the model group, high doses of GTI reduced the AST, ALT, HYP, TC, and TG levels(P<0.01) while increasing the HDL-C, SOD, and GSH levels(P<0.01). RT-qPCR results showed that GTI down-regulated the mRNA levels of IL-1β, IL-6, TNF-α, COL1A1, and α-SMA(P<0.01). Western blot results indicated that GTI down-regulated the protein levels of IL-1β, IL-6, TNF-α, phosphorylated PI3K(p-PI3K), phosphorylated AKT(p-AKT), phosphorylated inhibitor of nuclear factor kappa B alpha(p-IκBα), and nuclear factor kappa B(NF-κB)(P<0.01). In summary, GTI ameliorates inflammation, dyslipidemia, and oxidative stress associated with NASH by regulating the PI3K/AKT/NF-κB signaling pathway.
Animals ; Non-alcoholic Fatty Liver Disease/genetics* ; Mice ; Network Pharmacology ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Chromatography, High Pressure Liquid ; Liver/metabolism* ; Mice, Inbred C57BL ; Humans ; Mass Spectrometry ; Tumor Necrosis Factor-alpha/metabolism* ; Disease Models, Animal ; Molecular Docking Simulation

Objective To observe the efficacy of different dosage of midazolam oral solution in re-lieving anxiety in children undergoing laparoscopic high ligation of the hernia sac with oblique inguinal her-nia.Methods A total of 120 children,93 males and 27 females,aged 2-6 years,78-120 cm in height and 11-25 kg in weight,ASA physical statusⅠ orⅡ,were selected to perform laparoscopic high ligation of inguinal oblique hernia sac under general anesthesia.According to random number table method,the chil-dren were divided into three groups:the oral midazolam 0.25 mg/kg group(group M1),0.5 mg/kg group(group M2),and 0.75 mg/kg group(group M3)30 minutes before anesthesia,40 children in each group.Modified Yale preoperative anxiety scale-short form(mYPAS-SF)was recorded at premedication,parental separation,and immediate induction of anesthesia.Induction compliance checklist(ICC)score,recovery time,extubation time,PACU residence time,pediatric anesthesia emergence delirium scale(PAED)and the modified face,legs,activity,cry and consolability scale(FLACC)30 minutes after operation were also recorded.Results Compared with before taking medication,mYPAS-SF scores in groups M2 and M3 at parental separation and immediate induction of anesthesia were significantly decreased(P＜0.05).Com-pared with group M1,mYPAS-SF scores at parental separation and immediate induction of anesthesia and ICC scores at immediate induction of anesthesia were significantly lower in groups M2 and M3(P＜0.05),the recovery time,extubation time and PACU resident time in groups M2 and M3 were significantly pro-longed,PAED score was decreased significantly within 30 minutes after operation(P＜0.05).Compared with group M2,the awakening time and extubation time in group M3 were significantly prolonged.(P＜0.05).Conclusion Oral midazolam 0.5 mg/kg or 0.75 mg/kg 30 minutes before anesthesia can effectively alleviate the preoperative anxiety of children,improve the degree of cooperation in anesthesia in-duction,and reduce the occurrence of postoperative agitation,the recovery time and extubation time pro-longed in children with oral midazolam 0.75 mg/kg.Therefore,an oral solution of midazolam 0.5 mg/kg was a more appropriate dose for preoperative antianxiety regimen in children.

The purpose of this study was to enrich the genomic information and provide a basis for further development and utilization of Polygonatum kingianum. The fresh rhizome of P. kingianum was used as the experimental material, and the full-length transcriptome was sequenced by PacBio Sequel platform. The final measured polymerase reads were 1 120 485, with a total of 77.73 GB of data. In NR database, 41 864 homologous sequence alignments were aligned to 5 species; 40 506 were annotated in the KOG database and classified into 26 categories based on their functionality; 69 060 GO annotated 32 functional groups divided into 3 categories: cellular components, molecular functions, and biological processes; 45 779 annotations were added to 145 metabolic pathways in the KEGG database. Among them, there are 127 identified transcripts related to polysaccharide synthesis in the glycolysis/gluconeogenesis metabolic pathway, 144 in the starch and sucrose metabolic pathway, 85 in the amino acid sugar and nucleotide sugar metabolic pathway, and 69 in the fructose and mannose metabolic pathway. In addition, 1 781 transcription factors were detected, distributed in 37 transcription factor families; 180 293 SSR loci were detected, among which single base repeats accounted for the most, accounting for 30.48% of all base repeats, and at least five base repeats, accounting for only 0.14% of single base repeats. The results of this study provide important data supporting for enriching the genomic information of P. kingianum and exploring its effective biosynthetic pathway.

Objective Little is known about the association between whole-blood nicotinamide adenine dinucleotide(NAD+)levels and nabothian cysts.This study aimed to assess the association between NAD+levels and nabothian cysts in healthy Chinese women. Methods Multivariate logistic regression analysis was performed to analyze the association between NAD+levels and nabothian cysts. Results The mean age was 43.0±11.5 years,and the mean level of NAD+was 31.3±5.3 μmol/L.Nabothian cysts occurred in 184(27.7%)participants,with single and multiple cysts in 100(15.0%)and 84(12.6%)participants,respectively.The total nabothian cyst prevalence gradually decreased from 37.4%to 21.6%from Q1 to Q4 of NAD+and the prevalence of single and multiple nabothian cysts also decreased across the NAD+quartiles.As compared with the highest NAD+quartile(≥34.4 μmol/L),the adjusted odds ratios with 95%confidence interval of the NAD+Q1 was 1.89(1.14-3.14)for total nabothian cysts.The risk of total and single nabothian cysts linearly decreased with increasing NAD+levels,while the risk of multiple nabothian cysts decreased more rapidly at NAD+levels of 28.0 to 35.0 μmol/L. Conclusion:Low NAD+levels were associated with an increased risk of total and multiple nabothian cysts.

Isocitrate dehydrogenase 1 (IDH1) R132H is the most common mutated gene in grade II-III gliomas and oligodendrogliomas. Instead of activating telomerase (a reverse transcriptase which using RNA as a template to extend telomere length), the majority of IDH1^R132H mutant glioma maintain telomere length through an alternative mechanism that relies on homologous recombination (HR), which is known as alterative lengthening of telomere (ALT).The phenotype of ALT mechanism include: ALT associated promyelocytic leukemia protein (PML) bodies (APBs); extrachromosomal telomeric DNA repeats such as C- and T-loops; telomeric sister chromatid exchange (T-SCE), etc. The mechanism of ALT activation is not fully understood. Recent studies have shown that mutation IDH1 contributes to ALT phenotype in glioma cells in at least three key ways. Firstly, the IDH1^R132H mutation mediates RAP1 down-regulation leading to telomere dysfunction, thus ensuring persistent endogenous telomeric DNA damage, which is important for ALT activation. Spontaneous DNA damage at telomeres may provide a substrate for mutation break-induced replication (BIR)‑mediated ALT telomere lengthening, and it has been demonstrated that RAP1 inhibits telomeric repeat-containing RNA, transcribed from telomeric DNA repeat sequences (TERRA) transcription to down-regulate ALT telomere DNA replication stress and telomeric DNA damage, thereby inhibiting ALT telomere synthesis. Similarly, in ALT cells, knockdown of telomere-specific RNaseH1 nuclease triggers TERRA accumulation, which leads to increased replication pressure. Overexpression of RNaseH1, on the other hand, attenuates the recombination capacity of ALT telomeres, leading to telomere depletion, suggesting that RAP1 can regulate the level of replication pressure and thus ALT activity by controlling TERRA expression. Secondly, the IDH1^R132H also alters the preference of the telomere damage repair pathway by down-regulating XRCC1, which inhibits the alternative non-homologous end joining (A-NHEJ) pathway at telomeres and alters cellular preference for the HR pathway to promote ALT. Finally, the IDH1^R132H has a decreased affinity for isocitric acid and NADP+ and an increased affinity for α ketoglutarate (α‑KG) and NADPH, so that the mutant IDH1^R132H catalyzes the hydrogenation of α‑KG to produce 2-hydroxyglutarate (2-HG)in a NADPH-dependent manner. Because 2-HG is structurally similar to α‑KG, which maintains the trimethylation level of H3k9me3 by competitively inhibiting the activity of the α‑KG-dependent histone demethylase KDM4B, and recruits heterochromatin protein HP1α to heterochromatinize telomeres, and promote ALT phenotypes in cooperation with the inactivating of ATRX. In addition, it has been shown that APBs contain telomeric chromatin, which is essentially heterochromatin, and HP1α is directly involved in the formation of APBs. Based on these studies, this article reviews the mechanism of IDH1^R132H mediated telomere dysfunction and the preference of DNA repair pathway at telomeres in cooperate with ATRX loss to promote ALT, which may provide references for clinical targeted therapy of IDH1^R132H mutant glioma.

ObjectiveCoronavirus is a class of long-standing pathogens, which are enveloped single-stranded positive-sense RNA viruses. The genome all encodes 4 structural proteins: spike protein (S), nucleocapsid protein (N), membrane protein (M), and envelope protein (E). The nucleocapsid protein (NP) serves as a key structural component of coronaviruses, playing a vital function in the viral life cycle. NP acts as an RNA-binding protein, with a critical role in identifying specific sequences within the viral genome RNA, facilitating the formation of ribonucleoprotein (RNP) complexes with viral RNA to stabilize the viral genome and contribute to viral particles assembly. The NP consists of two primary structural domains, the N-terminal domain (NTD) and the C-terminal domain (CTD). The NTD is primarily responsible for RNA binding, whereas the CTD is involved in polymerization. The N protein demonstrated to trigger the host immune response and to modulate the cell cycle of infected cells by interacting with host proteins. The NP, one of the most abundant protein in coronaviruses, is essential in understanding the pathogenic mechanism of coronaviruses through its interaction with host factors, which response for determining the virus pathogenicity. HCoV-229E is a widely distributed coronavirus that typically causes mild upper respiratory tract diseases, accounting for a significant portion of common cold cases. However, its pathogenicity is notably lower compared to other coronaviruses like MERS-CoV, SARS-CoV, and SARS-CoV-2. The exact molecular mechanism behind remains unexplained, and how HCoV-229E N protein influences virus replication, host antiviral immunity, and pathogenesis need to be further explored. MethodsProximity labeling-mass spectrometry technique and bioinformatics analysis were used to screen for potential host factors interacting with the NP of human coronavirus 229E (HCoV-229E). In this study, a recombinant adenovirus Ad-V5-NP^HCoV-229E-TurboID was constructed to express the fusion protein of HCoV-229E NP and biotin ligase (TurboID). A549 cells were infected with the Ad-V5-NP^HCoV-229E-TurboID. After 30 min biotin treatment, NP interacting proteins were labeled with biotin by biotin ligase, and subsequently isolated with streptavidin cross-linked magnetic beads. The potential interacting proteins were identified using label-free proteomic mass spectrometry and further validated through immunoprecipitation and immunofluorescence assays. ResultsWe identified a total of 584 potential interacting proteins. Gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis highlighted the enrichment of glycogen synthase kinase (GSK)3A and GSK3B in the glycolysis/gluconeogenesis pathway, indicating HCoV-229E NP connection to diabetes through aberrant activity. Moreover, SARS-CoV-2 infection can exacerbate hyperglycemia and metabolic dysregulation in diabetic individuals by activating the ACE2 receptor. Moreover, SARS-CoV-2 was observed to cause potentially harm to pancreatic β‑cells and leading to insulin deficiency, which not only worsens the condition of diabetic patients but also raises the possibility of new-onset diabetes in non-diabetic individuals. We demonstrated that GSK3A and GSK3B interacted with NP of HCoV-229E, suggesting that the NP may engage in various coronavirus pathogenic processes by interacting with GSK3. ConclusionThese findings suggest that proximity labeling-mass spectrometry technique is a valuable tool for identifying virus-host interaction factors, and lay the foundation for future investigations into the mechanisms underlying coronavirus replication, proliferation, and pathogenesis.

Pancreatic cancer is one of the most lethal malignant tumors. While the existing surgical resection and chemotherapy have failed to improve the prognosis of patients well, it is urgent to find safer and more effective therapies to meet the clinical therapeutic needs. Antibody drug conjugate (ADC) is a class of targeted antitumor agents that combines monoclonal antibodies with small molecule cytotoxic drugs via chemical linkers. ADC provided wider therapeutic windows and enhanced pharmacokinetic/pharmacodynamic properties which depended on their characteristics of high selectivity, high efficacy and low toxicity. In recent years, the successful application of ADC in the treatment of multiple tumors has aroused a research upsurge in ADC for pancreatic cancer. This review summarizes the structure and mechanism of ADC and the current status of ADC for pancreatic cancer in order to provide some references for the future development of ADC for pancreatic cancer.

Objective:To deeply analyze the effect path of shared decision making(SDM)level on the quality of family doctor contracted service for multimorbidity among the elderly population,and to provide reference for promoting the health management of chronic diseases,improving the patient's medical experience and the quality of contracted service.Methods:A multi-stage random sampling method was used to conduct a questionnaire survey on 599 elderly patients with multimorbidity and their corresponding 63 contracted doctors in Wuhan and Guiyang.The hierarchical regression model was used to analyze the effect of SDM level on the quality of contracted services.Results:The score of SDM in elderly patients with multimorbidity in the central and western regions of China was 32.07±6.46,and the score of family doctor contracted service quality was 95.20±8.73,among which the scores of economy and vertical continuity were lower.The results of hierarchical regression model analysis showed that the level of SDM had a positive impact on the quality of contracted services(β=0.369,P＜0.05),and family doctors had a positive moderating effect on the degree of trust in contracted patients(β=0.548,P＜0.05).Conclusion:The level of SDM and the quality of contract service for elderly patients with multimorbidity in the central and western regions of China are at a medium level and still need to be improved.Shared decision-making significantly improves the quality of contracted service.The degree of trust in contracted patients strengthens the positive effect of SDM on service quality.In the future,we should strengthen the SDM,standardize the referral service of patients with chronic diseases,strengthen the communication and trust between doctors and patients,and effectively improve the quality of family doctor contracted service.

End-stage patients experience unbearable pain because of refractory symptoms.Palliative sedation is a form of palliative care which relieves patients' agony by lowering their consciousness.Standard palliative sedation can help patients die with dignity.It is distinct from euthanasia and does not alter the survival of patients.Sufficient palliative care is the premise of palliative sedation.Repeated and detailed clinical evaluation,as well as multidisciplinary involvement,is necessary for the standardized implementation of palliative sedation.Here,we proposed the standard process and specifications of palliative sedation in Peking Union Medical College Hospital.Furthermore,we reported a case of palliative sedation for an advanced cancer patient with refractory delirium and living pain to demonstrate its application in clinical practice.
Humans ; Anesthesia ; Pain ; Hospitals ; Palliative Care ; Universities