Objective:To discuss the regulatory role of complement alternative pathway in mouse colorectal cancer(CRC)liver metastasis model based on bioinformatics methods,and to clarify its mechanism through experimental verification.Methods:Using"CRC liver metastasis"as the keyword,the GSE81558 dataset was retrieved from Gene Expression Omnibus(GEO)database,including normal colon tissue samples,CRC tissue samples and CRC liver metastasis tissue samples.Bioinformatics methods were used to analyze and screen differentially expressed genes(DEGs).Gene Ontology(GO)functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis were performed using R and Cytoscape software,and the results were visualized.Search Tool for the Retrieval of Interacting Genes/Proteins(STRING)database was used to evaluate protein-protein interactions(PPIs)of DEGs and construct PPI network.Twelve C57BL/6 mice were injected with SL4 tumor cells into spleen,and the liver tissues were collected at 0,7 and 14 d.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of complement pathway-related genes in liver metastatic foci.The CRC liver metastasis mouse model was used to verify the complement signaling pathway.The mice were divided into control group,factor B knockout group(FB-/-)and C4 factor knockout group(C4-/-),and there were 6 mice in each group.The liver weights of the mice were measured;HE staining was used to detect the percentage of metastatic area in liver tissue in control group and FB-/-group;immunohistochemistry was used to detect macrophage infiltration in liver tissue in control group and FB-/-group,and the percentage of macrophage infiltration was calculated.Results:The distances between normal colon tissue samples and CRC tissue samples,as well as between CRC tissue samples and CRC liver metastasis tissue samples were far,indicating significant differences between samples,allowing subsequent analysis of DEGs.A total of 1 908 DEGs were screened in the dataset comparing normal colon tissue samples and CRC tissue samples,including 771 up-regulated DEGs and 1 137 down-regulated DEGs.Twenty-three up-regulated DEGs and 100 down-regulated DEGs were identified in the dataset comparing CRC and CRC liver metastasis.The GO functional enrichment analysis results showed that compared with normal colon tissue samples,DEGs in CRC samples were mainly enriched in biological processes(BP)related to cell cycle and mitosis,including mitotic cell cycle process,cell division,response to hormone,mitotic nuclear division and response to lipid.Compared with CRC samples,the DEGs in CRC liver metastasis samples were mainly enriched in coagulation-related BP,including platelet degranulation,blood coagulation regulation,acute-phase response,hemostasis regulation and coagulation regulation.The KEGG pathway enrichment analysis results showed that compared with normal colon tissue samples,the DEGs in CRC tissue samples were mainly enriched in cell cycle and p53 signaling pathways.Compared with CRC tissue samples,the DEGs in CRC liver metastasis tissue samples were mainly enriched in complement,coagulation cascade and metabolism-related signaling pathways.The Hub genes identified in PPI network were related to blood proteins.The RT-qPCR results showed that compared with 0 d group,the mRNA expression level of complement related genes complement 1q(C1q)in liver metastatic foci tissue sampres in 7 d group was significantly decreased(P<0.05),the mRNA expression levels of complement 3(C3),complement 5(C5),FB,and factor D(FD)were significantly increased(P<0.05 or P<0.01),the mRNA expression levels of complement pathway-related genes C1q,complement 2(C2),C3,complement fragment 3a receptor(C3aR),C5,complement fragment 5a receptor(C5aR),decay-accelerating factor(DAF),FB and FD in liver metastatic foci tissue sampres in 14 d group were significantly increased(P<0.05 or P<0.01).Compared with control group,the liver weight of the mice in FB-/-group was significantly decreased(P<0.01),while there was no significant difference was observed in C4-/-group(P>0.05).The HE staining results showed that compared with control group,the liver metastatic foci in FB-/-mice were significantly decreased,and the percentage of metastatic area was decreased(P<0.01).The immunohistochemistry results showed that compared with control group,the macrophage infiltration in liver metastatic foci of the mice in FB-/-group was reduced,and the percentage of macrophage infiltration was decreased(P<0.01).Conclusion:Complement cascade is associated with CRC liver metastasis,and the alternative complement pathway regulates CRC liver metastasis,suggesting this pathway may serve as a potential therapeutic target for CRC liver metastasis.

Limb spasticity is a common complication after stroke,with clinical manifestations such as increased muscle tone,limb stiffness and pain,which reduces the quality of patients'daily life and increases the economic burden.Currently,scales,surface electromyography and biomechanical methods are mostly used to assess limb spasticity in clinical practice,but clinical scales rely on experience and are highly subjective;surface electromyography is prone to compensations and test errors;and biomechanical methods are more restrictive.Musculoskeletal ultrasound is not only capable of detecting muscle parameters and providing quantitative information such as muscle hardness and elasticity,but also has the unique advantages of high-resolution and real-time imag-ing,which has been increasingly used in clinical diagnosis and treatment.In this paper,we review the evidence related to musculoskeletal ultrasound and limb spasticity assessment,with the aim of exploring a simple and reasonable quantitative assessment method,which will provide some thoughts for the future clinical assessment of limb spasticity.

Limb spasticity is a common complication after stroke,with clinical manifestations such as increased muscle tone,limb stiffness and pain,which reduces the quality of patients'daily life and increases the economic burden.Currently,scales,surface electromyography and biomechanical methods are mostly used to assess limb spasticity in clinical practice,but clinical scales rely on experience and are highly subjective;surface electromyography is prone to compensations and test errors;and biomechanical methods are more restrictive.Musculoskeletal ultrasound is not only capable of detecting muscle parameters and providing quantitative information such as muscle hardness and elasticity,but also has the unique advantages of high-resolution and real-time imag-ing,which has been increasingly used in clinical diagnosis and treatment.In this paper,we review the evidence related to musculoskeletal ultrasound and limb spasticity assessment,with the aim of exploring a simple and reasonable quantitative assessment method,which will provide some thoughts for the future clinical assessment of limb spasticity.

Chronic kidney disease(CKD),a major threat to global public health,exhibits disease progression closely linked to renal cellular senescence.This review outlines the key molecular mecha-nisms underpinning cellular senescence in CKD,including mitochondrial dysfunction,DNA damage response,senescence-associated secretory phenotype,and the central role of epigenetic regulation.Furthermore,the advances in traditional Chinese medicine(TCM)for mitigating renal senescence through multi-target strategies such as oxidative stress inhibition,anti-inflammatory modulation,and interventions in cell cycle arrest are summarized.There is evidence that active TCM compounds and formulas exert anti-senescence potential by modulating pathways including sirtuin 1/PTEN induced kinase 1 and Wnt/β-catenin signaling.However,clinical translation remains constrained by a poor knowledge of the mechanisms,challenges to dose standardization,and a lack of clinical validation.Future studies should integrate kidney-specific transgenic models with single-cell omics to resolve the cell hetero-geneity of senescence while developing novel delivery systems to enhance the targeting efficiency of TCM components so as to facilitate precision interventions in CKD.

Chronic kidney disease(CKD),a major threat to global public health,exhibits disease progression closely linked to renal cellular senescence.This review outlines the key molecular mecha-nisms underpinning cellular senescence in CKD,including mitochondrial dysfunction,DNA damage response,senescence-associated secretory phenotype,and the central role of epigenetic regulation.Furthermore,the advances in traditional Chinese medicine(TCM)for mitigating renal senescence through multi-target strategies such as oxidative stress inhibition,anti-inflammatory modulation,and interventions in cell cycle arrest are summarized.There is evidence that active TCM compounds and formulas exert anti-senescence potential by modulating pathways including sirtuin 1/PTEN induced kinase 1 and Wnt/β-catenin signaling.However,clinical translation remains constrained by a poor knowledge of the mechanisms,challenges to dose standardization,and a lack of clinical validation.Future studies should integrate kidney-specific transgenic models with single-cell omics to resolve the cell hetero-geneity of senescence while developing novel delivery systems to enhance the targeting efficiency of TCM components so as to facilitate precision interventions in CKD.

Objective To evaluate the mechanism of governor vessel electroacupuncture in rats with post-stroke limb spasm by observing the changes of glutathione antioxidant system-related factors.Methods A total of 60 SD rats were randomly divided into the normal group(n=12),sham operation group(n=12)and modeling group(n=36).The middle cerebral artery obstruction model was prepared by thread approach method in the modeling group,and 24 rats with successful modeling were randomly divided into the model group and the electroacupuncture group,with 12 rats in each group.At the 3rd day after modeling,the electroacupuncture group was treated with electroacupuncture at three acupoints of the governor vessel,namely,"Dazhui"(GV14),"Jizhong"(GV6)and"Houhui"(anteromedial of the transverse process of the sixth lumbar vertebra),for 30 min each time,once a day for 7 days.The neurological function of rats was assessed by Zea Longa neurological deficit score.The muscle tension of rats was detected by modified Ashworth dystonia rating and electrophysiological tracing method.The brain tissue water content was measured by the dry-wet weight method.The volume of cerebral infarction of rats was measured by the TTC staining method.The contents of glutathione(GSH),catalase(CAT),oxidized glutathione(GSSG),superoxide dismutase(SOD),and malondialdehyde(MDA)in the cortex of rats were detected by colorimetry.The protein and mRNA expressions of glutathione reductase(GR),glutamate cysteine ligase(GCL)C,GCLM,and glutathione peroxidase 4(GPX4)in the cortex of rats were measured by Western blotting and real-time PCR,respectively.Results Compared with rats in the normal and sham operation groups,the Zea Longa neurological deficit score,modified Ashworth dystonia rating,the volume of cerebral infarction,brain tissue water content,and GSSG and MDA contents in cortex were increased in the model group,the tension signal value and the proteins and mRNA expressions of GR,GCLC,GCLM,and GPX4 in cortex were decreased,and the contents of GSH,CAT,and SOD in cortex were decreased(P＜0.05).Compared with the model group,the Zea Longa neurological deficit score,modified Ashworth dystonia rating,the volume of cerebral infarction,brain tissue water content,and GSSG and MDA contents in cortex were decreased in the electroacupuncture group,the tension signal value and the proteins and mRNA expressions of GR,GCLC,GCLM,and GPX4 in cortex were increased,and the contents of GSH,CAT,and SOD in cortex were increased(P＜0.05).Conclusion Governor vessel electroacupuncture can improve the severity of post-stroke limb spasm in rats,and its mechanism may be related to the regulation of glutathione antioxidant system in cerebral cortex.

Objective The objective of this study was to establish a mouse model of allergic rhinoconjunctivitis and investigate the role of the NLRP3 signaling pathway in allergic rhinoconjunctivitis.Methods Thirty-three female C57 mice(SPF)were randomLy divided into 3 groups:the control group,the experimental group,and the NLRP3-/-group.On days 0,4,7,14,and 21,the experimental group and NLRP3-/-group received a 0.2 mL intraperitoneal injection of medicine containing OVA(100 μg)and adjuvant Al(OH)3(4 mg),respectively.After an interval of 3 days,each eye and nose were dosed with 10 μL of 5%OVA for five consecutive days a week to induce allergic symptoms.During sensitization and excitation stages,the control group was replaced with an equiva-lent amount of PBS.Ocular and nasal symptoms were observed and scored.The levels of OVA-specific IgE,IL-4,IL-17,and IL-18 in serum were measured using ELISA,while changes in palpebral conjunctiva and nasal mucosa were assessed by hematoxylin-eosin staining.The expression of NLRP3 mRNA in conjunctival tissue and nasal mucosa was determined using real-time PCR analysis.Statistical analysis was performed using SPSS17.0 software with P<0.05 considered as statistically significant difference.Results The experimental group and NLRP3-/-group exhibited induced nasal and ocular allergic symptoms.In the experimental group,the duration of nasal allergy symptoms was(10.500±1.080)days,while the duration of eye allergy symptoms was(20.300±2.058)days.In the NLRP3-/-group,the duration of nasal allergy symptoms was(13.400±1.955)days,and for eye allergy symp-toms it was(20.900±2.132)days.The duration of nasal allergies in the NLRP3-/-group significantly exceeded that in the experimental group(P<0.05),whereas there were no significant differences observed in eye allergy durations between these two groups(P>0.05).Levels of OVA-specific IgE,IL-4,and IL-17 were significantly higher in both the experimental and NLRP3-/-groups compared to those in the control group(P<0.05).Additionally,serum IL-18 content increased significantly in the experimental group when compared with both control and NLRP3-/-groups(P<0.05).Conjunctival tissue lesions as well as nasal mucosa damage were evident in both experimental and NLRP3-/-groups.mRNA expression levels of NLRP3 within conjunctival tissue and nasal mucosa from the experimental group showed a significant increase when compared to those from both control and NLRP3-/-groups(P<0.05).Conclusion Allergic rhinoconjunctivitis pathogenesis is influenced by various factors;however,the involvement of NLPR3 signaling pathway promotes its development.

A Clinical pharmacist was fully involved in the anticoagulation drug treatment management process of a 104-year-old patient with a large area of cerebral infarction combined with chronic kidney disease stage Ⅴ and secondary deep vein thrombosis.After the patient was diagnosed with deep vein thrombosis,the clinical pharmacist comprehensively analyzed the patient's super-advanced age,history of atrial fibrillation,large area of cerebral infarction,extremely poor kidney function,deep vein thrombosis,and high bleeding risk indicated by the HAS-BLED score.They worked with the clinical doctor to develop an individualized anticoagulation treatment strategy for the patient.At the beginning of the treatment,warfarin was given to the patient at a daily dose of 1.25 mg,and the patient's coagulation indicators and kidney function were dynamically rechecked.The patient's blood creatinine level did not show significant changes throughout the anticoagulation treatment process.On the 8th day of medication,the patient's INR was 2.47,and the clinical pharmacist suggested adjusting the Warfarin to an alternate-day dose of 1.25 mg and 0.625 mg.Subsequently,the patient's INR was 2.41,and the condition improved,leading to discharge.Throughout the anticoagulation drug management process,the clinical pharmacist participated in the clinical decision-making for anticoagulant drug selection,provided professional medication guidance,and pharmacological monitoring to ensure the safe clinical use of drugs for special populations.

We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.
Female ; Humans ; Uterine Cervical Neoplasms/drug therapy* ; Prospective Studies ; Quality of Life ; Neoplasm Staging ; Chemoradiotherapy ; Chemotherapy, Adjuvant/adverse effects* ; Adjuvants, Immunologic ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use* ; Retrospective Studies

Renal interstitial fibrosis (RIF) is the crucial pathway in chronic kidney disease (CKD) leading to the end-stage renal failure. However, the underlying mechanism of Shen Qi Wan (SQW) on RIF is not fully understood. In the current study, we investigated the role of Aquaporin 1 (AQP1) in SQW on tubular epithelial-to-mesenchymal transition (EMT). A RIF mouse model induced by adenine and a TGF-β1-stimulated HK-2 cell model were etablished to explore the involvement of AQP 1 in the protective effect of SQW on EMT in vitro and in vivo. Subsequently, the molecular mechanism of SQW on EMT was explored in HK-2 cells with AQP1 knockdown. The results indicated that SQW alleviated kidney injury and renal collagen deposition in the kidneys of mice induced by adenine, increased the protein expression of E-cadherin and AQP1 expression, and decreased the expression of vimentin and α-smooth muscle actin (α-SMA). Similarly, treatmement with SQW-containing serum significantly halted EMT process in TGF-β1 stimulated HK-2 cells. The expression of snail and slug was significantly upregulated in HK-2 cells after knockdown of AQP1. AQP1 knockdown also increased the mRNA expression of vimentin and α-SMA, and decreased the expression of E-cadherin. The protein expression of vimentin increased, while the expression of E-cadherin and CK-18 significantly decreased after AQP1 knockdown in HK-2 cells. These results revealed that AQP1 knockdown promoted EMT. Furthermore, AQP1 knockdown abolished the protective effect of SQW-containing serum on EMT in HK-2 cells. In sum, SQW attentuates EMT process in RIF through upregulation of the expression of AQP1.
Drugs, Chinese Herbal/pharmacology* ; Humans ; Animals ; Mice ; Male ; Cell Line ; Rats ; Kidney/physiology* ; Fibrosis/drug therapy* ; Renal Insufficiency, Chronic/drug therapy* ; Adenine ; Epithelial-Mesenchymal Transition ; Aquaporin 1/metabolism*