This study investigated the regulatory effects of carvacrol on lipopolysaccharide(LPS)-induced inflammatory response and endoplasmic reticulum(ER)stress in bovine mammary epithe-lial cells using in vitro cell culture techniques.Western blot analysis revealed significantly elevated expression levels of NF-κB pathway-related proteins and ER stress marker proteins in mastitis samples compared to healthy mammary tissues(P＜0.05).Cells were divided into a blank control group and carvacrol(CAV)treatment groups with varying concentrations(50,100,250,500,750,1 000 μmol/L).After 24 hours of culture,cell proliferation was assessed using the CCK-8 assay.An inflammatory model was established by stimulating MAC-T cells(a bovine mammary epithelial cell line)with LPS(10 mg/L)for 12 h,followed by measurement of the transcriptional levels of inflammatory-related genes(IL-6,IL-1β,and TNFα).MAC-T cells were pretreated with low,medi-um,and high doses of CAV for 12 h before LPS stimulation.Molecular docking analysis was per-formed to examine the interaction between CAV and GRP78,a key ER stress protein.Real-time quantitative PCR(qPCR)was used to analyze the mRNA expression levels of inflammatory cyto-kines(IL-6,IL-1β,TNFα),while Western blot was employed to assess the expression levels of NF-κB pathway proteins(p-IκB,p-NF-κB p65)and ER stress-related proteins(p-PERK,p-IRE1α,ATF6,GRP78,CHOP).The results from Western blot and qPCR demonstrated that CAV alleviated LPS-induced inflammatory response and cellular damage by inhibiting the NF-κB signa-ling pathway and ER stress.

In recent years,the intensification of dairy farming has significantly improved production efficiency but has also led to a rise in the incidence of metabolic diseases.Conditions such as keto-sis,fatty liver,and hypocalcemia pose serious threats to dairy cattle health and productivity.These diseases not only profoundly affect individual physiological function but also impose considerable economic pressures and challenges on farm management.As research advances,exosomes have e-merged as a novel intercellular signaling molecule,showing unique potential in regulating dairy cattle's nutritional metabolism.Studies suggest that exosomes hold promise as biomarkers for dis-ease and can even serve as carriers for disease detection and prevention.Acting as a crucial mediator of intercellular communication,exosomes play an important role in modulating the metabolic processes of dairy cattle.This review aims to systematically explore the role of exosomes in bovine nutritional metabolism and to provide new perspectives and theoretical support for their potential as tools for diagnosing,treating,and preventing metabolic diseases in dairy cattle,thus advancing research and practice in this field.

In recent years,the intensification of dairy farming has significantly improved production efficiency but has also led to a rise in the incidence of metabolic diseases.Conditions such as keto-sis,fatty liver,and hypocalcemia pose serious threats to dairy cattle health and productivity.These diseases not only profoundly affect individual physiological function but also impose considerable economic pressures and challenges on farm management.As research advances,exosomes have e-merged as a novel intercellular signaling molecule,showing unique potential in regulating dairy cattle's nutritional metabolism.Studies suggest that exosomes hold promise as biomarkers for dis-ease and can even serve as carriers for disease detection and prevention.Acting as a crucial mediator of intercellular communication,exosomes play an important role in modulating the metabolic processes of dairy cattle.This review aims to systematically explore the role of exosomes in bovine nutritional metabolism and to provide new perspectives and theoretical support for their potential as tools for diagnosing,treating,and preventing metabolic diseases in dairy cattle,thus advancing research and practice in this field.

This study investigated the regulatory effects of carvacrol on lipopolysaccharide(LPS)-induced inflammatory response and endoplasmic reticulum(ER)stress in bovine mammary epithe-lial cells using in vitro cell culture techniques.Western blot analysis revealed significantly elevated expression levels of NF-κB pathway-related proteins and ER stress marker proteins in mastitis samples compared to healthy mammary tissues(P＜0.05).Cells were divided into a blank control group and carvacrol(CAV)treatment groups with varying concentrations(50,100,250,500,750,1 000 μmol/L).After 24 hours of culture,cell proliferation was assessed using the CCK-8 assay.An inflammatory model was established by stimulating MAC-T cells(a bovine mammary epithelial cell line)with LPS(10 mg/L)for 12 h,followed by measurement of the transcriptional levels of inflammatory-related genes(IL-6,IL-1β,and TNFα).MAC-T cells were pretreated with low,medi-um,and high doses of CAV for 12 h before LPS stimulation.Molecular docking analysis was per-formed to examine the interaction between CAV and GRP78,a key ER stress protein.Real-time quantitative PCR(qPCR)was used to analyze the mRNA expression levels of inflammatory cyto-kines(IL-6,IL-1β,TNFα),while Western blot was employed to assess the expression levels of NF-κB pathway proteins(p-IκB,p-NF-κB p65)and ER stress-related proteins(p-PERK,p-IRE1α,ATF6,GRP78,CHOP).The results from Western blot and qPCR demonstrated that CAV alleviated LPS-induced inflammatory response and cellular damage by inhibiting the NF-κB signa-ling pathway and ER stress.

OBJECTIVE: To evaluate the efficacy and safety of hydroxychloroquine (HCQ) in the treatment of patients with moderate coronavirus disease 2019 (COVID-19). METHODS: We prospectively enrolled 30 treatment-naïve patients with confirmed COVID-19 after informed consent at Shanghai Public Health Clinical Center. The patients were randomized 1：1 to HCQ group and the control group. Patients in HCQ group were given HCQ 400 mg per day for 5 days plus conventional treatments, while those in the control group were given conventional treatment only. The primary endpoint was negative conversion rate of SARS-CoV-2 nucleic acid in respiratory pharyngeal swab on days 7 after randomization. This study has been approved by the Ethics Committee of Shanghai Public Health Clinical Center and registered online (NCT04261517). RESULTS: One patient in HCQ group developed to severe during the treatment. On day 7, nucleic acid of throat swabs was negative in 13 (86.7%) cases in the HCQ group and 14 (93.3%) cases in the control group (>0.05). The median duration from hospitalization to virus nucleic acid negative conservation was 4 (1,9) days in HCQ group, which is comparable to that in the control group [2 (1,4) days, Z=1.27, >0.05]. The median time for body temperature normalization in HCQ group was 1 (0,2) day after hospitalization, which was also comparable to that in the control group [1 (0,3) day]. Radiological progression was shown on CT images in 5 cases (33.3%) of the HCQ group and 7 cases (46.7%) of the control group, and all patients showed improvement in follow-up examinations. Four cases (26.7%) of the HCQ group and 3 cases (20%) of the control group had transient diarrhea and abnormal liver function (>0.05). CONCLUSIONS The prognosis of COVID-19 moderate patients is good. Larger sample size study are needed to investigate the effects of HCQ in the treatment of COVID-19. Subsequent research should determine better endpoint and fully consider the feasibility of experiments such as sample size.
Betacoronavirus ; isolation & purification ; China ; Coronavirus Infections ; diagnostic imaging ; drug therapy ; Humans ; Hydroxychloroquine ; therapeutic use ; Pandemics ; Pilot Projects ; Pneumonia, Viral ; diagnostic imaging ; drug therapy ; RNA, Viral ; isolation & purification ; Treatment Outcome