Olfactory receptors (ORs) form the largest superfamily of G protein-coupled receptors (GPCRs). Traditionally recognized for their role in the nasal olfactory epithelium, where they mediate the sense of smell, accumulating evidence has firmly established their ectopic expression in non-olfactory tissues, including the intestine, lungs, and kidneys. The intestine, as the primary site for nutrient digestion and absorption, harbors a highly complex chemical environment. To adapt to this environment, the gut employs a sophisticated network of “chemosensors” to monitor luminal contents and maintain homeostasis. Among these sensors, intestinal ORs have emerged as crucial functional components, serving as a molecular bridge that connects environmental chemical signals—such as food-derived odorants—to specific physiological responses. This discovery has significantly deepened our understanding of how dietary flavors and compounds influence intestinal physiology at the molecular level. This review systematically summarizes the expression profiles, ligand classification, and biological functions of ORs within the gastrointestinal tract. Studies indicate that intestinal ORs exhibit distinct spatial distribution patterns across different gut segments and display cell-type specificity, particularly within enterocytes and enteroendocrine cells. These receptors function as versatile sensors capable of recognizing a wide variety of ligands, including exogenous dietary components, gut microbiota metabolites such as short-chain fatty acids, and endogenous small molecules like azelaic acid. Upon activation by specific ligands, intestinal ORs trigger intracellular signaling cascades, primarily involving the AC-cAMP-PKA pathway or calcium influx channels. A major focus of this review is to elucidate the molecular mechanisms by which these receptors regulate the secretion of gut hormones. Activation of specific ORs in enteroendocrine cells has been shown to stimulate the release of hormones such as glucagon-like peptide-1 (GLP-1), peptide YY (PYY), and serotonin (5-HT), thereby modulating systemic energy metabolism, glucose homeostasis, and gastrointestinal motility. Furthermore, the review addresses the critical roles of ORs in immune regulation and pathology. Evidence suggests that specific ORs contribute to the maintenance of intestinal immune homeostasis and may offer protection against inflammation. Beyond their involvement in inflammatory responses, ORs such as Olfr78 have been shown to regulate the differentiation and function of intestinal endocrine cells. Similarly, Olfr544 has been demonstrated to alleviate intestinal inflammation by remodeling the gut microbiome and metabolome. These findings collectively suggest that specific ORs hold promise as therapeutic targets for mitigating intestinal inflammation and maintaining gut homeostasis. Additionally, the review explores the emerging role of ORs in cancer. Although OR expression is often downregulated in tumor tissues compared to normal mucosa, activation of specific ORs by certain ligands can inhibit tumor cell proliferation and migration and induce apoptosis via pathways such as MEK/ERK and p38 MAPK. Conversely, other receptors, such as OR7C1, may serve as biomarkers for cancer-initiating cells. In conclusion, intestinal ORs represent a vital component of the gut’s sensory network. The review also discusses the translational potential of these findings. By elucidating the precise pairing relationships between dietary components and specific ORs, novel therapeutic strategies could be developed. Intestinal ORs may thus emerge as promising targets for nutritional and pharmacological interventions in metabolic diseases, inflammatory bowel diseases, and malignancies.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.

Objective:To compare and observe the visual acuity and ocular anatomical outcome of different subtypes in open-globe injury (OGI) Ⅲ.Methods:A retrospective study. A total of 187 eyes of 187 patients with OGI involving zone Ⅲ who were admitted to the Department of Ophthalmology of The First Affiliated Hospital of Army Medical University from January 2020 to December 2023 were included in the study. According to the 2022 International Globe and Adnexal Trauma Epidemiology Study groups consensus, zone Ⅲ was further divided into Ⅲa zone (5-8 mm posterior to the limbus) and Ⅲb zone (>8 mm posterior to the limbus), with 58 eyes (31%, 58/187) in group Ⅲa and 129 eyes (69%, 129/187) in group Ⅲb. Best corrected visual acuity (BCVA) was examined using the international standard decimal visual acuity chart, converted into the logarithm of the minimum angle of resolution (logMAR) visual acuity when recorded. The injured zone, initial visual acuity, final visual acuity, retinal detachment (RD), uveal prolapse, and proliferative vitreoretinopathy (PVR) were collected. The follow-up time after surgery ≥ 6 months. The final visual acuity and anatomical prognosis of the two groups were observed. Silicone oil dependence, phthisis, and enucleation were defined as poor anatomical outcomes. Multiple linear regression analysis was performed to analyze the impact of zone Ⅲb of OGI on the final visual acuity.Results:At the 6-month follow-up, the logMAR BCVA of group Ⅲa and group Ⅲb was 1.49±1.0 and 2.51±0.85; there was a statistically significant difference in the logMAR BCVA between the two groups ( t=-2.736, P<0.05). Compared with group Ⅲa, the proportion with light perception in group Ⅲb was higher, and the proportions with visual acuity of hand movement, counting fingers, and >0.01 were lower, and the differences were all statistically significant ( P<0.05). Compared with group Ⅲa, RD and PVR were more likely to occur in group Ⅲb, and the differences were all statistically significant ( χ2= 16.696, 8.697; P<0.05). Among the affected eyes in group Ⅲa and group Ⅲb, there were 14 eyes (24.1%, 14/58) and 95 eyes (73.6%, 95/129) with poor final anatomical outcomes respectively; the incidence of poor final anatomical outcomes in group Ⅲb was higher, and the difference was statistically significant ( χ2= 40.332, P<0.01). The results of multiple linear regression analysis showed that initial visual acuity, RD, and uveal prolapse were independent risk factors affecting the final visual acuity (odds ratio=2.407, 4.162, 3.413; P<0.05). Conclusions:Patients with OGI in zone Ⅲb have a worse visual prognosis and a higher incidence of poor anatomical outcomes. The subclassification of zone Ⅲ is helpful for better predicting the prognosis of OGI clinically.

Objective To assess the utility of real-time shear wave elastography(SWE)in diagnosing vascular erectile dysfunction(ED)and to predict the optimal timing for color Doppler flow imaging(CDFI)examination.Methods Patients diagnosed with ED who received intracavernosal injection(ICI)of vasoactive drugs were recruited and categorized based on CDFI findings into three groups:arterial ED(n=17),venous ED(n=33),and non-vascular ED(n=29).SWE technology was utilized to measure the average Young's modulus(E value)of the corpus cavernosum in these patients,both in the flaccid state prior to ICI and at four time points following ICI-induced erection.Subsequently,the differences in E values among the three groups were analyzed.Results There was no significant difference in the E value of the corpus cavernosum in the flaccid state among the arterial,venous,and non-vascular ED groups before ICI(P>0.05).However,the E value in the flaccid state for each group was significantly higher than the mean E values observed at the four time points after ICI-induced erection(P<0.01).Additionally,the mean E values at these four time points post-ICI were also statistically significant(P<0.01).ROC curve analysis revealed that the AUC for diagnosing arterial,venous,and non-vascular ED using the E value after ICI were 0.814,0.770,and 0.711,respectively,with corresponding cutoff values of 9.98,8.16 and 7.06 kPa.The combined use of CDFI and SWE cutoff values following ICI-induced erection significantly shortened the detection time for both arterial and venous ED groups(P<0.01).Conclusions SWE can accurately measure the E value of the corpus cavernosum following erection induced by the vasoactive drug ICI,thereby facilitating the differentia-tion of various types of ED.Additionally,when combined with CDFI,this technique can reduce the time required for examination.

A quadruplex RT-qPCR method was developed for rapid identification and diagnosis of transmissible gastroenteritis virus(TGEV)of swine,porcine epidemic diarrhea virus(PEDV),porcine deltacorona virus(PDCoV),and porcine rotavirus type A(PoRVA).The full-length sequences of PEDV(77 strains),TGEV(63 strains),PDCoV(17 strains)that are prevalent in China,as well as the 85 VP6 gene sequences of PoRVA,were downloaded from the NCBI database for homology analysis.Based on the relatively conserved sequences,the corresponding primers and probes for each virus were designed and used to establish the quadruplex RT-qPCR method.After optimization of the probes and the reaction conditions,the specificity,sensitivity,and repeatability were determined.Using the established method,109 clinical samples of diarrhea were tested and further compared with the results by standard method.The results showed that the quadruplex RT-qPCR method established in this experiment has good amplification effect,with the C,value linearly correlated with the copies of templates(R2＞0.99).Specificity assay demonstrated that the quadruplex RT-qPCR method can identify TGEV,PEDV,PDCoV,PRoVA strains,and do not de-tect African swine fever virus(ASFV),porcine circovirus type 2(PCV2),porcine reproductive and respiratory syndrome virus(PRRSV),classical swine fever virus(CSFV),pseudorabies virus(PRV)and other epidemic viruses.Sensitivity assay showed that the detection limits for TGEV,PEDV,PDCoV and PoRVA were 10,20,20 and 50 copies/μL,respectively.The method exhibits excellent reproducibility,with coefficients of variation(Cv)for both intra-and inter-assay repli-cates being less than 1％.Detection of 109 samples of diarrhea by this method yielded the coinci-dence rate of 100％with the industry standard,indicating high practical applicability.The devel-oped method possesses the advantages of strong strain compatibility,high sensitivity,strong speci-ficity,good repeatability and stability.It is suitable for virus diagnosis and large-scale clinical sam-ple testing,providing technical support for disease prevention and control as well as epidemiologi-cal investigation.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Colorectal cancer(CRC)is a malignant tumor formed in the colon or rectum,usually caused by uncontrolled growth and division of normal cells in the body.Cell apoptosis,pyroptosis,and necroptosis are key pathways of cell death in colorectal cancer.The comprehensive treatment strategy includes the synergistic effect of cell death inducers with chemotherapy,immunotherapy,and targeted therapy.Traditional Chinese medicine plays an important role as an adjuvant therapy in regulating cell death.The combination of traditional Chinese and western medicine has shown significant effects in precancerous lesions,improving efficacy,reducing adverse reactions,and reducing drug resistance.Although the research on the mechanism of cell death is not yet sufficient,emphasizing the unique characteristics of traditional Chinese medicine,clarifying the anti-tumor mechanism of traditional Chinese medicine,and achieving modern scientific internationalization of integrated traditional Chinese and western medicine treatment for colorectal cancer have become future research directions.This article will comprehensively review the molecular mechanisms of cell apoptosis,pyroptosis,and necroptosis from the perspective of combining traditional Chinese and Western medicine,as well as their regulatory role in the treatment of colorectal cancer.

Objective To assess the utility of real-time shear wave elastography(SWE)in diagnosing vascular erectile dysfunction(ED)and to predict the optimal timing for color Doppler flow imaging(CDFI)examination.Methods Patients diagnosed with ED who received intracavernosal injection(ICI)of vasoactive drugs were recruited and categorized based on CDFI findings into three groups:arterial ED(n=17),venous ED(n=33),and non-vascular ED(n=29).SWE technology was utilized to measure the average Young's modulus(E value)of the corpus cavernosum in these patients,both in the flaccid state prior to ICI and at four time points following ICI-induced erection.Subsequently,the differences in E values among the three groups were analyzed.Results There was no significant difference in the E value of the corpus cavernosum in the flaccid state among the arterial,venous,and non-vascular ED groups before ICI(P>0.05).However,the E value in the flaccid state for each group was significantly higher than the mean E values observed at the four time points after ICI-induced erection(P<0.01).Additionally,the mean E values at these four time points post-ICI were also statistically significant(P<0.01).ROC curve analysis revealed that the AUC for diagnosing arterial,venous,and non-vascular ED using the E value after ICI were 0.814,0.770,and 0.711,respectively,with corresponding cutoff values of 9.98,8.16 and 7.06 kPa.The combined use of CDFI and SWE cutoff values following ICI-induced erection significantly shortened the detection time for both arterial and venous ED groups(P<0.01).Conclusions SWE can accurately measure the E value of the corpus cavernosum following erection induced by the vasoactive drug ICI,thereby facilitating the differentia-tion of various types of ED.Additionally,when combined with CDFI,this technique can reduce the time required for examination.

Colorectal cancer(CRC)is a malignant tumor formed in the colon or rectum,usually caused by uncontrolled growth and division of normal cells in the body.Cell apoptosis,pyroptosis,and necroptosis are key pathways of cell death in colorectal cancer.The comprehensive treatment strategy includes the synergistic effect of cell death inducers with chemotherapy,immunotherapy,and targeted therapy.Traditional Chinese medicine plays an important role as an adjuvant therapy in regulating cell death.The combination of traditional Chinese and western medicine has shown significant effects in precancerous lesions,improving efficacy,reducing adverse reactions,and reducing drug resistance.Although the research on the mechanism of cell death is not yet sufficient,emphasizing the unique characteristics of traditional Chinese medicine,clarifying the anti-tumor mechanism of traditional Chinese medicine,and achieving modern scientific internationalization of integrated traditional Chinese and western medicine treatment for colorectal cancer have become future research directions.This article will comprehensively review the molecular mechanisms of cell apoptosis,pyroptosis,and necroptosis from the perspective of combining traditional Chinese and Western medicine,as well as their regulatory role in the treatment of colorectal cancer.

The amino-terminal pro-C-type natriuretic peptide(NT-proCNP)is a diagnostic inflam-matory marker clinically used for diagnosing bacterial infections.This study aims to establish a phage display library of single-chain variable fragment(scFv)antibodies against canine NT-proC-NP and to screen for scFvs with high binding affinity to NT-proCNP.Initially,NT-proCNP was prepared using prokaryotic expression system and was used to immunize New Zealand White rab-bits.Upon achieving the desired serum titer,total RNA was extracted from the splenocytes of rab-bits and reverse transcribed into cDNA.Using this cDNA as a template,degenerate primers were employed to amplify the genes of the rabbit antibody light chain variable region(VL)and heavy chain variable region(VH).The VL and VH regions were spliced together to form a complete scFv fragment via overlap extension PCR.The scFv was then ligated into the phagemid pComb3XSS and electroporated into competent E.coli TG1 cells to construct a rabbit-derived anti-NT-proCNP scFv immunological library.This library underwent four rounds of enrichment and screening to isolate specific single-chain antibodies.The selected antibody was subsequently ex-pressed in a soluble form within a prokaryotic system,and its immunological activity was evalua-ted.Using phage display technology,this study successfully identified a single-chain antibody scFv-1-CNP with strong antigen-binding activity and genetic sequence characteristics of scFvs,providing a research direction for further exploration of scFv applications in the detection of NT-proCNP.