Objective Currently, there is no commercially available quantitative detection kit for the main Felis domestic allergen (Fel d 1) in China. To establish a rapid detection method for Fel d 1, this study aims to prepare monoclonal antibodies against Fel d 1 protein. Methods The codon preference of Escherichia coli was utilized to optimize and synthesize the Fel d 1 gene. The prokaryotic expression plasmid pET-28a-Fel d 1 was constructed and used to express and purify the recombinant Fel d 1 protein. Subsequently, the recombinant protein was immunized into BALB/c mice and monoclonal antibodies (mAbs) were prepared by the hybridoma technique. An indirect ELISA was established using the recombinant Fel d 1 as the coating antigen, and hybridoma cell lines were screened for positive clones. The specificity and antigenic epitopes of the mAbs were confirmed by Western blot analysis. Finally, the selected hybridoma cells were injected into the peritoneal cavities of BALB/c mice for large-scale monoclonal antibody production. Results The recombinant plasmid pET-28a-Fel d 1 was successfully constructed, and soluble Fel d 1 protein was obtained after optimizing the expression conditions. Western blot and antibody titer assays confirmed the successful isolation of two hybridoma cell lines, 7D11 and 5H4, which stably secreted mAbs specific to Fel d 1. Antibody characterization revealed that the 5H4 mAb was of the IgG2a subtype and could recognize the amino acid region 105-163 of Fel d 1, while the 7D11 mAb was the IgG1 subtype and could recognize the amino acid region 1-59. Conclusion The high-purity recombinant Fel d 1 protein produced in this study provides a promising alternative for clinical immunotherapy of cat allergies. Furthermore, the monoclonal antibody prepared in this experiment lays a material foundation for the in-depth study of the biological function of Fel d 1 and the development of ELISA detection.
Animals ; Antibodies, Monoclonal/biosynthesis* ; Mice, Inbred BALB C ; Cats ; Mice ; Allergens/genetics* ; Glycoproteins/genetics* ; Enzyme-Linked Immunosorbent Assay ; Hybridomas/immunology* ; Recombinant Proteins/genetics* ; Female ; Antibody Specificity

Recently, small nucleolar RNAs (snoRNAs) have transcended the genomic "noise" to emerge as pivotal molecular markers due to their essential roles in tumor progression. Substantial evidence indicates a strong association between snoRNAs and critical clinical features such as tumor pathology and drug resistance. Historically, snoRNA research has concentrated on two classical mechanisms: 2'-O-ribose methylation and pseudouridylation. This review specifically summarizes the novel regulatory mechanisms and functional patterns of snoRNAs in tumors, encompassing transcriptional, post-transcriptional, and post-translational regulation. We further discuss the synergistic effect between snoRNA host genes (SNHGs) and snoRNAs in tumor progression. More importantly, snoRNAs extensively contribute to the development of tumor cell resistance as oncogenes or tumor suppressor genes. Accordingly, we provide a comprehensive review of the clinical diagnosis and treatment associated with snoRNAs and explore their significant potential as novel drug targets.

Chronic obstructive pulmonary disease(COPD)has become a common chronic disease in the adult in recent years,and more attention has been gradually paid to its prevention and treatment.This paper reviewed the clinical studies about the effect of traditional Chinese medicine(TCM)exercise therapy on COPD,and indicated that TCM exercise therapy can improve the rehabilitation of COPD patients.TCM exercise therapies,such as Tai Chi,Wuqinxi,Baduanjin,and Liuzijue,have been shown to significantly improve lung function,inflammation levels,and exercise capacity in COPD patients in recent studies.Tai Chi significantly improves patient's respiratory problems although lung function indices are not changed.Liuzijue enhances the strength and endurance of respiratory muscles and limbs.Baduanjin helps to improve patient's cognitive and emotional states.Overall,TCM exercise therapy provides an effective rehabilitation option for COPD patients.However,more clinical controlled trials are needed to further confirm their effectiveness and to develop appropriate rehabilitation programs for COPD patients.

Objective To systematically analyze the clinical pharmaceutical rehabilitation model based on data mining,so as to explore its role in improving drug treatment efficacy,reducing drug side effects,and optimizing rehabilitation experience.Methods We retrieved published literatures related to clinical pharmaceutical rehabilitation from CNKI,Wanfang,VIP,and PubMed,with a search time from inception to June 30,2022.Results Totally 11 articles were enrolled in this study.A study involving 69 randomized controlled trials(RCTs)demonstrated that the involvement of pharmacists in medication reviews significantly improved the control of hypertension,type 2 diabetes,and high cholesterol.The odds ratio(OR)for improved control was 2.71(95%confidence interval[CI]:1.03-7.01)for hypertension,3.12(95%CI:1.12-5.84)for type 2 diabetes,and 1.90(95%CI:1.05-3.35)for high cholesterol.A study involving 6 RCTs and 2 573 patients with chronic kidney disease(CKD)showed that pharmacist-led blood pressure monitoring,lifestyle,and medication regimen assessments significantly improved blood pressure control in CKD patients,with an OR of 1.51(95%CI:1.13-2.03).A Meta-analysis,including 5 RCTs and 3 observational studies,explored the impact of medication education and behavioral interventions on the treatment outcomes of gout.The results revealed that patients receiving medication education and behavioral interventions had a significantly higher rate of uric acid control and a higher proportion of patient with uric acid less than 360 μmol/L.A Meta-analysis of 14 RCTs involving 4 509 patients showed that pharmacist interventions such as medication education and counseling significantly reduced the 30-day readmission rates for hypertension and diabetes patients,with a relative risk(RR)of 0.75(95%CI:0.53-0.96).Eighteen RCTs involving 7 244 patients investigated the impact of pharmacist-led discharge counseling on the readmission rate.The findings showed that discharge counseling significantly reduced readmission rate,with a RR of 0.864(95%CI:0.763-0.997,P=0.020).Eight studies assessed the impact of pharmacist services on readmission rate,and the result showed that patients receiving pharmacy-related services had significantly reduced readmission rates,with a 32%reduction in the risk of readmission(OR=0.74,95%CI:0.62-0.89).A Meta-analysis including 123 studies investigated the effect of pharmacist interventions in patient care on the readmission rate,and the result indicated that pharmacist interventions led to a decrease in readmission rates,with the maximum decrease of 44.5%.A Meta-analysis of 23 RCTs evaluated the impact of pharmacist interventions on medication-related adverse events in elderly residents of nursing homes.The study found that pharmacist-led medication reviews and education significantly reduced adverse events.Another Meta-analysis of 14 RCTs and intervention studies explored the effect of pharmacist involvement in multidisciplinary care on medication-related adverse events.The analysis showed that pharmacist participation in multidisciplinary care was significantly associated with a reduction in the incidence of adverse events(RR=0.47,95%CI:0.28-0.77,P<0.01).A Meta-analysis of 6 RCTs involving 29 291 pediatric inpatients revealed that pharmacist interventions effectively reduced the risk of medication errors in hospitalized children(OR=0.27,95%CI:0.15-0.49).A literature analysis on the effectiveness of clinical pharmaceutical services in patient drug treatment showed that introducing a clinical pharmaceutical service model can reduce the use of antibiotics,lower medical costs,and improve medication satisfaction among patients treated with antibiotics.Conclusion In-depth exploration of clinical pharmaceutical rehabilitation model plays an important role in clinical treatment,which can improve the effectiveness of rehabilitation and reduce the incidence of adverse reactions.

Recently,small nucleolar RNAs(snoRNAs)have transcended the genomic"noise"to emerge as pivotal molecular markers due to their essential roles in tumor progression.Substantial evidence indicates a strong association between snoRNAs and critical clinical features such as tumor pathology and drug resistance.Historically,snoRNA research has concentrated on two classical mechanisms:2'-O-ribose methylation and pseudouridylation.This review specifically summarizes the novel regulatory mecha-nisms and functional patterns of snoRNAs in tumors,encompassing transcriptional,post-transcriptional,and post-translational regulation.We further discuss the synergistic effect between snoRNA host genes(SNHGs)and snoRNAs in tumor progression.More importantly,snoRNAs extensively contribute to the development of tumor cell resistance as oncogenes or tumor suppressor genes.Accordingly,we provide a comprehensive review of the clinical diagnosis and treatment associated with snoRNAs and explore their significant potential as novel drug targets.

An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism. Hepatic fibrosis is characterized by activated hepatic stellate cells (aHSCs) with an excessive production of extracellular matrix. Although promoted activation of HSCs by M2 macrophages has been demonstrated, the molecular mechanism involved remains ambiguous. Herein, we propose that the vitamin D receptor (VDR) involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes. We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation. The exosomes derived from M2 macrophages can promote HSC activation, while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes. Smooth muscle cell-associated protein 5 (SMAP-5) was found to be the key effector protein in promoting HSC activation by regulating autophagy flux. Building on these results, we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect. In this study, we aim to elucidate the association between VDR and macrophages in HSC activation. The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis, and provide potential therapeutic targets for its treatment.
Humans ; Hepatic Stellate Cells/pathology* ; Receptors, Calcitriol ; Liver Cirrhosis/pathology* ; Macrophages/metabolism*

ObjectiveTo investigate the therapeutic effect of the frozen and fresh preparations of human umbilical cord mesenchymal stem cells （hUC-MSC） on a rat model of liver cirrhosis after transplantation via the portal vein or the caudal vein. MethodsA total of 70 specific pathogen-free healthy male Sprague-Dawley rats were randomly divided into normal group （13 rats fed with ordinary tap water and rat food） and liver cirrhosis model group （57 rats given subcutaneous multi-point injection of mixed carbon tetrachloride/olive oil solution）. At week 8， the growth of rats was observed for both groups， and 3 rats were selected from each group for histopathological examination to confirm the formation of liver cirrhosis. A total of 50 rats were selected from the liver cirrhosis model and were divided into model group， portal vein group+fresh cell preparation group， portal vein+frozen cell preparation group， caudal vein+fresh cell preparation group， and caudal vein+frozen cell preparation group using a random number table， with 10 rats in each group. Fresh or frozen hUC-MSC were transplanted via the portal vein or the caudal vein， and after 4 weeks of administration， the different groups were compared in terms of the changes in liver function parameters and liver fibrosis degree. Continuous data were expressed as mean±standard deviation， and the independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsAt week 8 of modeling， the model group showed the formation of pseudolobules of different sizes in the liver and met the diagnostic criteria for liver cirrhosis， with significant increases in the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， total bilirubin （TBil）， and alkaline phosphatase （ALP） compared with the normal group （all P<0.001）， suggesting that the rat model of liver cirrhosis was established successfully. There were significant differences in the levels of ALT， AST， TBil， and ALP between the five groups （F=232.00， 177.10， 112.30， 121.70， all P<0.001）. Further comparison between two groups showed that the model group had significantly higher levels of ALT， AST， TBil， and ALP than the normal group （all P<0.01）， and the portal vein group+fresh cell preparation group， the portal vein+frozen cell preparation group， the caudal vein+fresh cell preparation group， and the caudal vein+frozen cell preparation group had significantly lower levels of ALT， AST， TBil， and ALP than the model group （all P<0.01）. ConclusionThere are significant improvements in liver function and liver fibrosis degree in a rat model of liver cirrhosis at week 4 after the transplantation of hUC-MSC， and frozen or fresh cell preparation and different transplantation approaches have no significant influence on treatment outcome.

OBJECTIVE To compare the efficacy， safety and economy of tacrolimus （TAC）， cyclosporin A （CsA）， cyclophosphamide （CTX） and rituximab （RTX） in the treatment of membranous nephropathy （MN）. METHODS Retrieved from Pubmed， the Cochrane Library， Wanfang data， CNKI and health technology assessment （HTA） official website， HTA reports， systematic reviews/meta-analysis and pharmacoeconomic studies about TAC， CsA， CTX and RTX combined with glucocorticoid in the treatment of MN were collected during the inception and Mar. 2022. After data extraction and quality evaluation， descriptive analysis was performed on the results of the included studies. RESULTS A total of 15 articles were included， involving 13 systematic reviews/meta-analysis and 2 pharmacoeconomic studies. In terms of efficacy， TAC and CsA showed significant advantages in increasing the response rate， and could improve the levels of urine protein， serum albumin， serum creatinine and serum total cholesterol. In terms of safety， the incidence of adverse reaction induced by TAC， CsA and RTX was low and the symptoms were mild. In terms of economics， CTX cost lower but caused severe adverse reaction； TAC cost higher but showed higher remission rate and good safety. CONCLUSIONS TAC combined with glucocorticoid may be the recommended scheme for MN.

Gaining a better understanding of autoprotection against drug-induced liver injury(DILI)may provide new strategies for its prevention and therapy.However,little is known about the underlying mechanisms of this phenomenon.We used single-cell RNA sequencing to characterize the dynamics and functions of hepatic non-parenchymal cells(NPCs)in autoprotection against DILI,using acetaminophen(APAP)as a model drug.Autoprotection was modeled through pretreatment with a mildly hepatotoxic dose of APAP in mice,followed by a higher dose in a secondary challenge.NPC subsets and dynamic changes were identified in the APAP(hepatotoxicity-sensitive)and APAP-resistant(hepatotoxicity-resistant)groups.A chemokine(C-C motif)ligand 2+endothelial cell subset almost disappeared in the APAP-resistant group,and an R-spondin 3+endothelial cell subset promoted hepatocyte proliferation and played an important role in APAP autoprotection.Moreover,the dendritic cell subset DC-3 may protect the liver from APAP hepatotoxicity by inducing low reactivity and suppressing the autoimmune response and occurrence of inflammation.DC-3 cells also promoted angiogenesis through crosstalk with endothelial cells via vascular endothelial growth factor-associated ligand-receptor pairs and facilitated liver tissue repair in the APAP-resistant group.In addition,the natural killer cell subsets NK-3 and NK-4 and the Sca-1-CD62L+natural killer T cell subset may promote autoprotection through interferon-y-dependent pathways.Furthermore,macrophage and neutrophil subpopulations with anti-inflammatory phenotypes promoted tolerance to APAP hepatotoxicity.Overall,this study reveals the dynamics of NPCs in the resistance to APAP hepatotoxicity and provides novel insights into the mechanism of autoprotection against DILI at a high resolution.

Objective:To construct the post competency evaluation index system for Da Vinci robot specialist nurses based on onion model, so as to provide references for the training and evaluation of Da Vinci robot specialist nurses in China.Methods:From August to September 2022, using the "onion model" as the theoretical framework, a preliminary draft of the index system was developed using domestic and foreign literature review, semi-structured interviews and group discussions. The Delphi method was applied to 22 nursing experts in the relevant fields to conduct two rounds of expert correspondence, screen and modify the items, and calculate the weight using the analytic hierarchy process. Finally, the post competency index system for Da Vinci robot specialist nurses based on the onion model was formed.Results:Two rounds of expert correspondence were conducted. The positive coefficient of experts was 90.91% (20/22) in the first round and 100.00% (20/20) in the second round, the coefficient of expert authority was 0.840 and 0.873, and the Kendall's coordination coefficient was 0.181 and 0.324 ( P<0.01), respectively. The final post competency evaluation index system for Da Vinci robot specialist nurses consisted of 4 first-level indicators, 14 second-level indicators and 58 third-level indicators. Conclusions:This study constructs a comprehensive post competency evaluation index system for Da Vinci robot specialist nurses based on onion model, with high expert enthusiasm and authority. It can provide references for the training and evaluation of Da Vinci robot specialist nurses in China.