OBJECTIVE To investigate the regulatory effects of tofacitinib combined with methotrexate （MTX） on gut microbiota and the clinical efficacy of this regimen in patients with rheumatoid arthritis （RA）. METHODS A retrospective analysis was conducted on the clinical data of 182 patients with RA admitted to Xingtai People’s Hospital from January 2022 to June 2025. The patients were divided into a control group （ n ＝88， treated with MTX monotherapy） and an observation group （ n ＝94， treated with tofacitinib combined w ith MTX） based on their treatment regimen. Gut microbiota abundance， inflammatory and immunological indicators [C-reactive protein （CRP）， erythrocyte sedimentation rate （ESR）， rheumatoid factor （RF）， and anti-cyclic citrullinated peptide antibody （anti-CCP） ] ， clinical efficacy indicators [American College of Rheumatology 20% response rate （ACR20）， Disease Activity Score in 28 Joints （DAS28）， and Health Assessment Questionnaire （HAQ） score ] ， and adverse reactions during treatment were compared between the two groups before and after 12 weeks of treatment. RESULTS After treatment， the abundance of Lactobacillus and Bifidobacterium were significantly increased in both groups compared with before treatment， whereas the abundances of Enterococcus and Enterobacter ， as well as the levels of CRP， ESR， RF， anti-CCP， DAS28 score， and HAQ score， were significantly decreased （ P ＜0.05）. The degree of improvement in the observation group was significantly greater than that in the control group （ P ＜0.05）. The ACR20 response rate in the observation group was significantly higher than that in the control group （81.91% vs. 56.82%， P ＜0.05）. There was no statistically significant difference in the incidence of adverse reactions between the two groups （ P ＞0.05）， and the main adverse reactions were gastrointestinal reactions and abnormal liver function. CONCLUSIONS Tofacitinib combined with MTX can effectively improve gut microbiota balance in patients with RA by increasing the abundance of probiotics and reducing the abundance of opportunistic pathogenic bacteria， thereby improving immune and inflammatory status. In addition， this combination regimen can enhance clinical efficacy， reduce disease activity， and improve functional status， with a favorable safety profile.

Objective To explore the level of energy balance related protein antibody,adropin,and its correlation with collateral circulation status in elderly patients with AMI.Methods A total of 193 elderly AMI patients admitted to our department from February 2022 to February 2024 were enrolled,and based on their collateral circulation status,they were divided into a good circulation group(121 cases)and a poor circulation group(72 cases).The level of adropin was determined.Multivariate logistic regression analysis was employed to determine the influencing factors of poor collateral circulation in elderly AMI patients.Results The poor circulation group had significantly larger proportion of hypertension,higher fasting blood glucose(FBG),and larger red blood cell distribution width(RDW),but lower adropin level and mean platelet volume(MPV)when com-pared with the good circulation group(P＜0.05,P＜0.01).Multivariate logistic regression analy-sis showed that adropin,FBG,RDW,MPV,and hypertension were all influencing factors for poor collateral circulation in elderly AMI patients(P＜0.05,P＜0.01).The AUC value of adropin,FBG,RDW,MPV,and hypertension was 0.810,0.762,0.761,0.715 and 0.563,respectively in pre-dicting poor collateral circulation.Among them,adropin level had the highest predictive value(P＜0.01).Conclusion The decrease in adropin level in elderly AMI patients is closely associated with poor collateral circulation,and it is a predictive factor for collateral circulation.

Objective:To investigate the neuroprotective effect of vanillin on experimental autoimmune encephalomyelitis(EAE)rats by regulating C-X-C motif chemokine ligand 12(CXCL12)/chemokine(C-X-C motif)receptor(CXCR4)signaling path-way.Methods:A total of 50 rats were injected with 400 μl guinea pig spinal cord and water in oil mixture of complete Freund's adju-vant to establish EAE rat model,and were divided into model group,low-dose vanillin group(50 mg/kg),medium-dose vanillin group(100 mg/kg),high-dose vanillin group(200 mg/kg)and positive drug group(5 mg/kg prednisone acetate),another 10 rats were only injected with the mixture of the same amount of normal saline and complete Freud adjuvant as control group,since the first day of mod-eling,all rats were given corresponding drugs by gavage for 16 consecutive days,and after modeling,rats in each group were scored for neurological function every day;HE and LFB staining were used to observe the pathology and demyelination of spinal cord in rats;levels of serum TNF-α,IL-1β and IL-6 were detected by ELISA;immunohistochemistry was used to detect expressions of CD68 and Iba-1 in spinal cord of rats;Western blot was used to detect expressions of CXCL12 and CXCR4 proteins in spinal cord of rats.Results:Compared with control group,rats in model group showed obvious infiltration of inflammatory cells,a large number of inflammatory cells gathered around the small blood vessels,and the structure of myelin sheath in spinal cord of rats was abnormal,accompanied by a large number of myelin sheath loss,the neurological function score,inflammatory infiltration score and demyelination score of spinal cord,serum TNF-α,IL-1β and IL-6 levels,spinal cord CD68 and Iba-1 proteins,CXCL12 and CXCR4 proteins expressions in-creased obviously(P<0.05);compared with model group,inflammatory cell infiltration and myelin sheath loss of rats in low,medium and high doses groups and positive drug group were obviously alleviated,neurological function score,inflammatory infiltration score and demyelination score of spinal cord,serum TNF-α,IL-1β and IL-6 levels,and expressions of CD68 and Iba-1 proteins in spinal cord decreased obviously(P<0.05),while expressions of CXCL12 and CXCR4 proteins in spinal cord further increased(P<0.05).Conclusion:Vanillin inhibits inflammatory response and alleviates nerve injury in EAE rats,and its mechanism may be related to acti-vation of CXCL12/CXCR4 signaling pathway.

Objective:To investigate the neuroprotective effect of vanillin on experimental autoimmune encephalomyelitis(EAE)rats by regulating C-X-C motif chemokine ligand 12(CXCL12)/chemokine(C-X-C motif)receptor(CXCR4)signaling path-way.Methods:A total of 50 rats were injected with 400 μl guinea pig spinal cord and water in oil mixture of complete Freund's adju-vant to establish EAE rat model,and were divided into model group,low-dose vanillin group(50 mg/kg),medium-dose vanillin group(100 mg/kg),high-dose vanillin group(200 mg/kg)and positive drug group(5 mg/kg prednisone acetate),another 10 rats were only injected with the mixture of the same amount of normal saline and complete Freud adjuvant as control group,since the first day of mod-eling,all rats were given corresponding drugs by gavage for 16 consecutive days,and after modeling,rats in each group were scored for neurological function every day;HE and LFB staining were used to observe the pathology and demyelination of spinal cord in rats;levels of serum TNF-α,IL-1β and IL-6 were detected by ELISA;immunohistochemistry was used to detect expressions of CD68 and Iba-1 in spinal cord of rats;Western blot was used to detect expressions of CXCL12 and CXCR4 proteins in spinal cord of rats.Results:Compared with control group,rats in model group showed obvious infiltration of inflammatory cells,a large number of inflammatory cells gathered around the small blood vessels,and the structure of myelin sheath in spinal cord of rats was abnormal,accompanied by a large number of myelin sheath loss,the neurological function score,inflammatory infiltration score and demyelination score of spinal cord,serum TNF-α,IL-1β and IL-6 levels,spinal cord CD68 and Iba-1 proteins,CXCL12 and CXCR4 proteins expressions in-creased obviously(P<0.05);compared with model group,inflammatory cell infiltration and myelin sheath loss of rats in low,medium and high doses groups and positive drug group were obviously alleviated,neurological function score,inflammatory infiltration score and demyelination score of spinal cord,serum TNF-α,IL-1β and IL-6 levels,and expressions of CD68 and Iba-1 proteins in spinal cord decreased obviously(P<0.05),while expressions of CXCL12 and CXCR4 proteins in spinal cord further increased(P<0.05).Conclusion:Vanillin inhibits inflammatory response and alleviates nerve injury in EAE rats,and its mechanism may be related to acti-vation of CXCL12/CXCR4 signaling pathway.

Objective:To investigate effect of andrographolide(AG)on neuroinflammation in young epileptic rats by regulating cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA)/cAMP response element binding protein(CREB)signaling pathway.Methods:SPF grade young SD rats were randomly divided into Control group,Model group,low-dose AG group(AG-L,125 mg/kg),high-dose AG group(AG-H,250 mg/kg)and high-dose AG+PKA inhibitor H-89 group(AG-H+H-89,250 mg/kg AG+2 mg/kg H-89),with 12 rats in each group.Epilepsy model of young rats was established by intraperitoneal injection of kainic acid(KA).Morris water maze test was used to detect learning and memory functions of rats.ELISA was used to detect levels of TNF-α,IL-10,malondialdehyde(MDA),superoxide dismutase(SOD)and cAMP in hippocampus of rats in each group.Histomorphology of hippo-campus was detected by Nissl staining.TUNEL test was usd to determine apoptosis rate of neurons in rat hippocampus.PKA and CREB mRNA expressions in hippocampus of rats in each group were detected by RT-qPCR.Western blot was used to detect protein expressions of PKA,Bax,Caspase-3,CREB,p-CREB and brain derived neurotrophic factor(BDNF)in hippocampus of rats in each group.Results:Compared with control group,escape latency,TNF-α and MDA levels in hippocampus,apoptosis rate of nerve cells,Bax and Caspase-3 protein expressions in Model group were obviously increased(P<0.05),target quadrant residence time,SOD,IL-10,cAMP levels,PKA,CREB mRNA and protein expressions,BDNF protein expression in hippocampus were decreased obviously(P<0.05),hippocampal tissue showed pathological damage and a large number of Nissl bodies were lost.Compared with Model group,cor-responding indexes of rats in AG-H group were contrary to the above(P<0.05),loss of Nissl corpuscles was reduced.H-89 alleviated improvement of AG on neuroinflammation in young epileptic rats.Conclusion:AG may reduce neuroinflammation in young epileptic rats by activating cAMP/PKA/CREB signaling pathway.

Objective To systematically evaluate the risk factors for persistent cough after lung resection, providing a theoretical basis for preventing persistent postoperative cough. Methods The Cochrane Library, Web of Science, EMbase, PubMed, Chinese Biomedical Literature Database, Wanfang, CNKI, and VIP databases were searched for studies related to risk factors for persistent cough after lung resection. The search period was from database inception to March 30, 2023. Two researchers independently screened the literature, extracted data, and performed quality assessment. RevMan 5.3 software was used for meta-analysis. Results A total of 17 articles with 3 698 patients were included. Meta-analysis results showed that females [OR=3.10, 95%CI (1.99, 4.81), P<0.001], age [OR=1.72, 95%CI (1.33, 2.21), P<0.001], right-sided lung surgery [OR=2.36, 95%CI (1.80, 3.10), P<0.001], lobectomy [OR=3.40, 95%CI (2.47, 4.68), P<0.001], upper lobectomy [OR=8.19, 95%CI (3.87, 17.36), P<0.001], lymph node dissection [OR=3.59, 95%CI (2.72, 4.72), P<0.001], bronchial stump closure method [OR=5.19, 95%CI (1.79, 16.07), P=0.002], and postoperative gastric acid reflux [OR=6.24, 95%CI (3.27, 11.91), P<0.001] were risk factors for persistent cough after lung resection, while smoking history was a protective factor against postoperative cough [OR=0.59, 95%CI (0.45, 0.77), P<0.001]. In addition, the quality of life score of patients with postoperative cough decreased compared with that before surgery [MD=1.50, 95%CI (0.14, 2.86), P=0.03]. Conclusion Current evidence suggests that females, age, right-sided lung surgery, lobectomy, upper lobectomy, lymph node dissection, bronchial stump closure method (stapler closure), and postoperative gastric acid reflux are independent risk factors for persistent postoperative cough in lung resection patients, while smoking history may be a protective factor against postoperative cough. This provides evidence-based information for clinical medical staff on how to prevent and reduce persistent postoperative cough in patients and improve their quality of life in the future.

Objective:To compare the effects of standard cognitive behavioral therapy for insomnia (CBT-I) and enhanced cognitive behavioral therapy for insomnia(CBT-I Plus) in patients with chronic insomnia disorder comorbid anxiety or depressive symptoms.Methods:This prospective study included 148 patients with chronic insomnia disorder and anxiety/depression symptoms who were treated at the Sleep Disorder clinic of Shanghai Mental Health Center between July 2020 and August 2023. Participants (56 males, 92 females; aged 18-65 years, mean age 35.08±10.30 years) were randomly assigned in a 1∶2 ratio to the CBT-I group ( n=54) or CBT-I Plus group ( n=94). The CBT-I Plus group received additional treatments targeting anxiety and depressive symptoms. Treatment lasted 8 weeks, with assessment conducted at baseline, weeks 2, 4, and 8. Depression severity was measured using the 17-item Hamilton Depression Rating Scale (HAMD 17), anxiety severity with the Hamilton Anxiety Scale (HAMA), and sleep quality with the Pittsburgh Sleep Quality Index (PSQI). Paired sample t-tests were used to evaluate within-group changes, repeated-measures ANOVA compared treatment effects between groups, and ANCOVA was employed to adjust for confounding variables. Results:Significant reductions in PSQI, HAMD 17, and HAMA scores were observed in both groups after treatment: CBT-I group: PSQI ((14.15±2.54) vs. (7.50±3.35), t=13.25), HAMD 17 ((14.70±4.09) vs. (7.40±4.61), t=9.33), and HAMA ((14.94±4.11) vs. (5.56±3.67), t=12.38) (all P<0.001).CBT-I Plus group: PSQI ((14.87±3.01) vs. (7.19±3.86), t=18.75), HAMD 17 ((16.84±3.91) vs. (6.84±4.79), t=17.42), and HAMA ((15.57±3.93) vs. (6.10±4.57), t=18.39) (all P<0.001). After adjusting for HAMD 17 scores and medication use, no statistically significant between-group differences were observed in changes in PSQI, HAMD 17, and HAMA scores ( P>0.05). A significant time-by-group interaction was found for the PSQI daytime dysfunction subscale ( F=4.87, P<0.01). Conclusion:Both CBT-I and CBT-I Plus improve sleep and emotional symptoms in patients with chronic insomnia disorder and comorbid anxiety/depression symptoms. However, CBT-I Plus has no significant advantages over standard CBT-I. Further studies are needed to refine the timing and content of interventions.

This study aims to explore the medication rules and mechanisms of treating chronic renal failure(CRF) by Jinling medical school based on data mining, network pharmacology, and experimental validation systematically and deeply. Firstly, the study selected the papers published by the inherited clinicians in Jinling medical school in Chinese journals using the subject headings named "traditional Chinese medicine(TCM) + chronic renal failure", "TCM + chronic renal inefficiency", or "TCM + consumptive disease" in China National Knowledge Infrastructure, Wanfang, and VIP Chinese Science and Technology Periodical Database and screened TCM formulas for treating CRF according to inclusion and exclusion criteria. The study analyzed the frequency of use of single TCM and the four properties, five tastes, channel tropism, and efficacy of TCM used with high frequency and performed association rule and clustering analysis, respectively. As a result, a total of 215 TCM formulas and 235 different single TCM were screened, respectively. The TCM used with high frequency included Astragali Radix, Rhei Radix et Rhizoma, Salviae Miltiorrhizae Radix et Rhizoma, Poria, and Atractylodis Macrocephalae Rhizoma(top 5). The single TCM characterized by "cold properties, sweet flavor, and restoring spleen channel" and the TCM with the efficacy of tonifying deficiency had the highest frequency of use, respectively. Then, the TCM with the rules of "blood-activating and stasis-removing" and "diuretic and dampness-penetrating" appeared. In addition, the core combination of TCM [(Hexin Formula, HXF)] included "Astragali Radix, Rhei Radix et Rhizoma, Poria, Salviae Miltiorrhizae Radix, and Angelicae Sinensis Radix". The network pharmacology analysis showed that HXF had 91 active compounds and 250 corresponding protein targets including prostaglandin-endoperoxide synthase 2(PTGS2), PTGS1, sodium voltage-gated channel alpha subunit 5(SCN5A), cholinergic receptor muscarinic 1(CHRM1), and heat shock protein 90 alpha family class A member 1(HSP90AA1)(top 5). Gene Ontology(GO) function analysis revealed that the core targets of HXF predominantly affected biological processes, cellular components, and molecular functions such as positive regulation of transcription by ribonucleic acid polymerase Ⅱ and DNA template transcription, formation of cytosol, nucleus, and plasma membrane, and identical protein binding and enzyme binding. Kyoto Encyclopedia of Genes and Genomes(KEGG) analysis revealed that CRF-related genes were involved in a variety of signaling pathways and cellular metabolic pathways, primarily involving "phosphatidylinositol 3-kinase(PI3K)-protein kinase B(Akt) pathway" and "advanced glycation end products-receptor for advanced glycation end products". Molecular docking results showed that the active components in HXF such as isomucronulatol 7-O-glucoside, betulinic acid, sitosterol, and przewaquinone B might be crucial in the treatment of CRF. Finally, a modified rat model with renal failure induced by adenine was used, and the in vivo experimental confirmation was performed based on the above-mentioned predictions. The results verify that HXF can regulate mitochondrial autophagy in the kidneys and the PI3K-Akt-mammalian target of rapamycin(mTOR) signaling pathway activation at upstream, so as to alleviate renal tubulointerstitial fibrosis and then delay the progression of CRF.
Data Mining ; Drugs, Chinese Herbal/chemistry* ; Network Pharmacology ; Humans ; Kidney Failure, Chronic/metabolism* ; Medicine, Chinese Traditional ; China

OBJECTIVES: To investigate the expression of apolipoprotein C1 (APOC1) in papillary thyroid carcinoma (PTC) and its effects on proliferation and apoptosis of PTC cells. METHODS: The expression level of APOC1 in PTC and its impact on prognosis were analyzed using GEPIA 2 and Kaplan-Meier databases. Immunohistochemistry (IHC) and Western blotting were used to detect the expression of APOC1 in PTC and adjacent tissues and in 3 PTC cell lines and normal thyroid Nthyori 3-1 cells. In TPC-1 and BCPAP cells, the effect of Lipofectamine 2000-mediated transfection with APOC1 siRNA or an APOC1-overexpressing plasmid on cell growth and colony formation ability were examined by observing the growth curves and using colony-forming assay. The changes in cell cycle and apoptosis of the transfected cells were analyzed with flow cytometry. RT-qPCR and Western blotting were used to detect the changes in expressions of P21, P27, CDK4, cyclin D1, Bcl-2, Bax, caspase-3 and caspase-9 and the key proteins in the JAK2/STAT3 signaling pathway. RESULTS: APOC1 expression was significantly higher in PTC tissues and the 3 PTC cell lines than in the adjacent tissues and Nthyori 3-1 cells, respectively. In TPC-1 and BCPAP cells, APOC1 knockdown obviously reduced cell proliferative activity, increased the percentage of G0/G1 phase cells, lowered the percentages of S and G2 phase cells, promoted cell apoptosis, and downregulated mRNA and protein expression levels of CDK4, cyclin D1 and Bcl-2 and the protein levels of p-JAK2 and p-STAT3. APOC1 overexpression in the cells produced the opposite effects on cell proliferation, apoptosis, cell cycle and the mRNA and protein expressions. The application of AG490, a JAK2 inhibitor, strongly attenuated APOC1 overexpression-induced activation of the JAK2/STAT3 signaling pathway in BCPAP cells. CONCLUSIONS APOC1 overexpression promotes proliferation and inhibits apoptosis of PTC cells possibly by activating the JAK2/STAT3 signaling pathway and accelerating cell cycle progression.
Humans ; Apoptosis ; Cell Proliferation ; STAT3 Transcription Factor/metabolism* ; Signal Transduction ; Janus Kinase 2/metabolism* ; Thyroid Neoplasms/pathology* ; Thyroid Cancer, Papillary ; Cell Line, Tumor ; Carcinoma, Papillary

Microalgae possess high photosynthetic efficiency, robust adaptability, and substantial biomass, serving as excellent biological resources for large-scale cultivation. Malic enzyme (ME), a ubiquitous metabolic enzyme in living organisms, catalyzes the decarboxylation of malate to produce pyruvate, CO₂, and NAD(P)H, playing a role in multiple metabolic pathways including energy metabolism, photosynthesis, respiration, and biosynthesis. In this study, we identified the Scenedesmus quadricauda malic enzyme gene (SqME) and its biological functions, aiming to provide excellent target genes for the genetic improvement of higher plants. Based on the RNA-seq data from S. quadricauda under the biofilm cultivation mode with high CO₂ and light energy transfer efficiency and small water use, a highly expressed gene (SqME) functionally annotated as ME was cloned. The physicochemical properties of the SqME-encoded protein were systematically analyzed by bioinformatics tools. The subcellular localization of SqME was determined via transient transformation in Nicotiana benthamiana leaves. The biological functions of SqME were identified via genetic transformation in Nicotiana tabacum, and the potential of SqME in the genetic improvement of higher plants was evaluated. The ORF of SqME was 1 770 bp, encoding 590 amino acid residues, and the encoded protein was located in chloroplasts. SqME was a NADP-ME, with the typical structural characteristics of ME. The ME activity in the transgenic N. tabacum plant was 1.8 folds of that in the wild-type control. Heterologous expression of SqME increased the content of chlorophyll a, chlorophyll b, and total chlorophyll by 20.9%, 26.9%, and 25.2%, respectively, compared with the control. The transgenic tobacco leaves showed an increase of 54.0% in the fluorescence parameter NPQ and a decrease of 30.1% in Fo compared with the control. Moreover, the biomass, total lipids, and soluble sugars in the transgenic tobacco leaves enhanced by 20.5%, 25.7%, and 9.5%, respectively. On the contrary, the starch and protein content in the transgenic tobacco leaves decreased by 22.4% and 12.2%, respectively. Collectively, the SqME-encoded protein exhibited a strong enzymatic activity. Heterologous expressing of SqME could significantly enhance photosynthetic protection, photosynthesis, and biomass accumulation in the host. Additionally, SqME can facilitate carbon metabolism remodeling in the host, driving more carbon flux towards lipid synthesis. Therefore, SqME can be applied in the genetic improvement of higher plants for enhancing photosynthetic carbon fixation and lipid accumulation. These findings provide scientific references for mining of functional genes from S. quadricauda and application of these genes in the genetic engineering of higher plants.
Nicotiana/genetics* ; Photosynthesis/physiology* ; Malate Dehydrogenase/biosynthesis* ; Plant Leaves/genetics* ; Scenedesmus/enzymology* ; Carbon Cycle/genetics* ; Lipid Metabolism/genetics* ; Plants, Genetically Modified/metabolism*