1.Isochlorogenic acid A alleviates reticulum stress induced by peste des petits rumi-nants virus via PERK signaling pathway
Yun MU ; Tiantian SUN ; Yongsheng KUANG ; Shuyi YUAN ; Yanfen LIU ; Shaohong CHEN ; You LIU ; Fucheng GUO
Chinese Journal of Veterinary Science 2024;44(7):1408-1417
Viral infection can induce endoplasmic reticulum stress(ERS)and unfolded protein re-sponse(UPR)in host cells,resulting in perturbation of endoplasmic reticulum homeostasis.To e-lucidate the action mechanism of isochlorogenic acid A(IAA)in regulating peste des petits rumi-nant virus(PPRV)-induced ERS and UPR,MTT assay,indirect immunofluorescence assay and Western blot were used to evaluate the anti-PPRV activity of IAA,and the effects of IAA on PPRV-induced ERS and PERK signaling pathway were studied by Western blot and quantitative real-time PCR.The results showed that the PPRV replication and virus-induced cytopathic in LDG-2 cells were significantly inhibited,and the survival rate of virus-infected cells was significantly in-creased due to IAA treatment.Compared with the virus control group,the expression levels of GRP78 and p-eIF2α,the ratios of p-PERK/PERK and p-eIF2α/eIF2α in IAA treated PPRV-infec-ted cells were significantly decreased.The expression level of GADD153 significantly decreased at 24,36 h,and significantly increased at 48,60 h.Furthermore,treatment with ERS inhibitor 4-PBA could significantly suppress the expression levels of GRP78,PPRV-N protein and GADD153 in PPRV-infected cells,and the ratios of p-eIF2α/eIF2α and p-PERK/PERK in PPRV-infected cells were also significantly decreased caused by treatment with IAA or 4-PBA and IAA combination.These findings implicated that the PPRV-induced ERS could be alleviated by inhibiting activation of the PERK-eIF2α-GADD1 53 signaling pathway,which led to restriction of PPRV replication in host cells.
2.Effect of endoplasmic reticulum stress induced by peste des petits ruminants virus on PERK signaling pathway and apoptosis in goat kidney cells
Yongsheng KUANG ; Yun MU ; Tiantian SUN ; Shuyi YUAN ; Yanfen LIU ; Shaohong CHEN ; Fucheng GUO ; You LIU
Chinese Journal of Veterinary Science 2024;44(9):1882-1891
Viral infection can induce endoplasmic reticulum stress(ERS)and unfolded protein reac-tion(UPR)in host cells.This study aims to further explore the effects of ERS induced by pest des petits ruminants virus(PPRV)infection on UPR signaling pathway,virus replication and apopto-sis of host cells.MTT assay,indirect immunofluorescence assay(IFA)and Western blot were used to observe the proliferation of PPRV in goat kidney cells(LDG-2).Western blot and real-time flu-orescence quantitative PCR(qRT-PCR)were used to observe the effects of PPRV infection on the expression levels of GRP78,PERK and its downstream signal molecules,apoptosis-related proteins Bcl-2 and Bax.The result indicated that the cell survival rate was significantly declined with evident cytopathic effect at 36 h post-infection,and the expression level of PPRV-N protein tended to be elevated,and was significantly higher than that of cell control at 30 h post-infection.Meanwhile,the expression levels of GRP78,p-eIF2α and GADD153,the ratio of p-PERK/PERK and p-eIF2α/eIF2α were significantly increased.Moreover,the expression levels of PPRV-N protein,GRP78,p-eIF2α and GADD1 53,the ratio of p-eIF2α/eIF2α and p-PERK/PERK were significantly decreased in PPRV-infected cells due to 4-PBA treatment.The expression level of apoptosis-related Bcl-2 was down-regulated,Bax was up-regulated,and the ratio of Bcl-2/Bax was significantly decreased.Therefore,the activation of PERK/eIF2α/GADD153 signaling pathway could be induced by PPRV infection resulting in alleviating of virus-induced ERS,which is beneficial to viral replication.Bloc-king PPRV-induced ERS could inhibit the activation of PERK signaling pathway and virus replica-tion.PPRV infection and prolonged ERS can induce apoptosis of LDG-2 cells.
3.Inflammatory response in peste des petits ruminants virus infected LDG-2 cells is mediated by TLR2/MyD88/NF-κB signaling pathway
Tiantian SUN ; Yun MU ; Yongsheng KUANG ; Shuyi YUAN ; Shaohong CHEN ; Fucheng GUO ; You LIU
Chinese Journal of Veterinary Science 2024;44(10):2204-2212,2308
MTT assay,indirect immunofluorescence assay(IFA)and real-time fluorescence quanti-tative PCR(qRT-PCR)were used to observe the proliferation of PPRV in goat kidney cells(LDG-2).Western blot and qRT-PCR were used to evaluate the effects of PPRV infection on the expres-sion levels of TLR2,MyD88,NF-κB signaling pathway-related factors and their downstream in-flammatory factors.The results indicated that the significantly decreased cell survival rate and ob-vious cytopathic effect were observed at 36 h post PPRV infection,and the mRNA expression level of PPRV-N gene was significantly up-regulated.At the same time,the expression levels of TLR2,MyD88,p-p65 and p-IκBα,the ratio of p-p65/p65 and p-IκBα/IκBα and the mRNA expression levels of downstream inflammatory factors TNFα,IL-1β,IL-4 and IL-10 were significantly increased.Mo-reover,the expression levels of PPRV-N mRNA,TLR2,MyD88,p-p65 and p-IκBα,the ratio of p-p65/p65 and p-IκBα/IκBα and the mRNA expression levels of downstream inflammatory factors IL-1β and IL-4 in PPRV-infected cells were significantly decreased in the presence of the inhibitor C29 of TLR2.These findings implied that the TLR2/MyD88/NF-κBα signaling pathway can be activated by PPRV infection,which is beneficial to the replication and spread of the virus.Blocking down the activation of TLR2 can inhibit MyD88/NF-κB signaling pathway and viral replication in PPRV-infected cells.
4.The potential targets and mechanisms of modified Baihe dihuang decoction applied in post-stroke depression
Sixing HUANG ; Shuyi WU ; Ping ZHANG ; Jinping LUO ; Min WANG ; Yanlei GUO ; Hao LI ; Li ZHANG ; Zhe QIANG
China Pharmacy 2023;34(20):2483-2489
OBJECTIVE To explore the potential targets and mechanisms of the modified Baihe dihuang decoction (MBD/ BDD) applied in post-stroke depression (PSD). METHODS Network pharmacology was used to mine the potential targets and key pathways of MBD/BDD in the treatment of PSD. PSD model rats were induced by focal cerebral ischemia surgery combined with chronic unforeseen mild stress, and then were randomly divided into PSD model group, MBD/BDD group (12.6 g/kg, by raw drug), and fluoxetine hydrochloride (FLX) group (positive control, 2.3 mg/kg); a blank control group was also set up, with 8 rats in each group. Each administration group was given a corresponding medication solution by gavage once a day for 21 consecutive days. The intervention effect of MBD/BDD on depression-like symptoms in model rats was evaluated by open field and forced swimming tests. The brain tissues of rats in each group were dissected and total RNA was extracted for transcriptome sequencing and bioinformatics analysis. The mRNA and protein expressions of genes with significant changes and common neurotrophic factors were verified based on the above results. RESULTS A total of 131 MBD/BDD antidepressant-related target genes were obtained (such as IL1B and AKT1, etc.), which were closely related to neural active ligand-receptor interactions and cyclic adenosine monophosphate signaling pathway. MBD/BDD could significantly prolong or increase the total time spent and distance traveled in the central grid of qiangzhe@cqtcm.edu.cn PSD model rats, and significantly shorten the cumulative immobility time (P<0.05). After treatment with MBD/BDD, the number of genes that changed in rat brain tissue was much higher than that in the FLX group, and there were significant differences in gene profiles among the PSD model group, MBD/BDD group, and FLX group. There were 1 351 differentially expressed genes (DEGs) between the MBD/BDD group and the PSD model group, of which 178 were significantly down-regulated and 1 173 were significantly up-regulated (P<0.05). Above 1 351 DEGs were involved in neuronal differentiation, chemical synaptic transmission regulation. They were significantly enriched in axonal guidance, cholinergic synapses and neuroactive ligand-receptor interactions. The top 30 genes in terms of up-regulation in the brain tissue of rats of MBD/BDD group were all associated with neuronal proliferation, development, differentiation, and migration. After MBD/BDD intervention, the expressions of Fezf2, Arx, Ostn, Nrgn genes, brain-derived neurotrophic factor and tyrosine kinase receptor B protein in brain tissue of rats were significantly increased (P<0.05). CONCLUSIONS The anti-PSD effect of MBD/BDD may be related to the up-regulation of the expression of genes related to neuronal proliferation, development, differentiation and migration, as well as the promotion of neural structural and functional repair.
5.Application of self-monitoring of blood glucose information management platform based on medical consortium in community patients with type 2 diabetes
Quanying WU ; Lixin GUO ; Chao SUN ; Qi PAN ; Qi ZHOU ; Shuyi YU
Chinese Journal of Nursing 2023;58(24):2949-2956
Objective To study the effect of self-monitoring of blood glucose information management platform based on the medical consortium on the level of blood glucose control by improving the implementation rate of self-monitoring of blood glucose prescriptions for type 2 diabetes patients in the community.Methods A convenient sampling method was used to select 121 patients with type 2 diabetes from April 2021 to April 2023 who were enrolled in a community of a tertiary hospital medical consortium in Beijing.They were randomly divided into 61 patients in an experimental group and 60 patients in a control group.The control group received routine blood glucose monitoring and guidance education,while the experimental group was managed by a self-monitoring of blood glucose information management platform based on the medical consortium with nurses as the main body.Glycated hemoglobin,glucose test,fasting blood sugar,blood glucose 2 hours after meal,and the completion of self blood glucose monitoring prescriptions were evaluated and analyzed in the first week and the twelfth week of the 2 groups.Results 61 cases in the experimental group and 58 cases in the control group completed the follow-up,and the patients who completed the follow-up were analyzed.There was no statistically significant difference in glycated hemoglobin between the 2 groups at baseline(P=0.117).After 12 weeks,the reduction standard-reaching rate in glycated hemoglobin in the experimental group was significantly different from that in the control group(P=0.014).The number of self-monitoring of fasting blood glucose and the total number of other time periods in experimental group were significantly higher than those in the control group(P<0.001);the implementation rate of self-monitoring prescription in the experimental group was better than that in the control group(P<0.001).Conclusion A self blood glucose monitoring information platform was built to manage community type 2 diabetes patients,so that patients can master and implement the personalized blood glucose monitoring prescription formulated by the medical union team with nurses as the main body,and it is conducive to improving the frequency of blood glucose monitoring and the level of blood glucose control of diabetes patients.
6.Evaluation of internal doses to workers and monitoring of 131I concentration in air in nuclear medicine workplaces
Bao LIN ; Guixiang SUN ; Chaoyu ZHANG ; Hezheng ZHAI ; Yong GUO ; Shuyi ZHANG ; Shaolong LIU ; Zixuan GUO ; Xiaojun CHENG
Chinese Journal of Radiological Medicine and Protection 2023;43(9):717-723
Objective:To asscentain the 131I activity concentration in 131I treatment workplaces and to explore the method of estimating the internal dose to workers by air sampling and to analyze its influencing factors. Methods:Air sampling method was used to collect aerosols containing radioactivity in 10 randomly selected workplaces in Zhengzhou where 131I therapy was performed. Aactivity concentration of 131I in treatment workplace was measured for gamma emitters by gamma-ray spectrometry. The internal dose due to 131I inhalation was estimated based on measurement result and field investigation result. Results:The activity concentration of 131I in air samples from 19 subpacking rooms ranged from 0.087 to 570 Bq/m 3, with an average of (51.04 ± 128.58) Bq/m 3. Those from 11 wards ranged from 0.162 to 54.6 Bq/m 3, with an average of (7.97 ± 15.89) Bq/m 3. In terms of the work hours recommended by the national standard GBZ 129-2016 Specifications for individual monitoring of occupational internal exposure, the estimated annual effective dose to radiation workers due to the inhalation of 131I ranges from 0.002 to 10 mSv, with an average of (0.61 ± 1.80) mSv, below the dose limit specified in the national standards. Conclusions:The samples with high 131I activity concentration in nuclear medicine workplaces of 10 medical institutions selected in Zhengzhou are mostly distributed in tertiary class hospitals operating large amount of radionuclide with large numbers of thyroid cancer patients adimitted. The result ing internal dose to radiation workers cannot be ignored. Estimating the internal dose based on the measurement result of air samples has a large uncertainty.However, air sampling method can promptly detect radioactive contamination in case of abnormal events or accidents, providing early warning for workers to carry out dose measurement from external exposure and internal exposure assessment.
7.Globular adiponectin-mediated vascular remodeling by affecting the secretion of adventitial-derived tumor necrosis factor-α induced by urotensin II.
Jun LI ; Limin LUO ; Yonggang ZHANG ; Xiao DONG ; Shuyi DANG ; Xiaogang GUO ; Wenhui DING
Journal of Zhejiang University. Science. B 2022;23(12):1014-1027
OBJECTIVES:
In this study, we explored how adiponectin mediated urotensin II (UII)-induced tumor necrosis factor-α (TNF-α) and α-smooth muscle actin (α-SMA) expression and ensuing intracellular signaling pathways in adventitial fibroblasts (AFs).
METHODS:
Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UII and inhibitors of signal transduction pathways for 1‒24 h. The cells were then harvested for TNF-α receptor (TNF-α-R) messenger RNA (mRNA) and TNF-α protein expression determination by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. Adiponectin and adiponectin receptor (adipoR) expression was measured by RT-PCR, quantitative real-time PCR (qPCR), immunohistochemical analysis, and cell counting kit-8 (CCK-8) cell proliferation experiments. We then quantified TNF-α and α-SMA mRNA and protein expression levels by qPCR and immunofluorescence (IF) staining. RNA interference (RNAi) was used to explore the function of the adipoR genes. To investigate the signaling pathway, we applied western blotting (WB) to examine phosphorylation of adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK). In vivo, an adiponectin (APN)-knockout (APN-KO) mouse model mimicking adventitial inflammation was generated to measure TNF-α and α-SMA expression by application of qPCR and IF, with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling.
RESULTS:
In both cells and tissues, UII promoted TNF-α protein and TNF-α-R secretion in a dose- and time-dependent manner via Rho/protein kinase C (PKC) pathway. We detected marked expression of adipoR1, T-cadherin, and calreticulin as well as a moderate presence of adipoR2 in AFs, while no adiponectin was observed. Globular adiponectin (gAd) fostered the growth of AFs, and acted in concert with UII to induce α-SMA and TNF-α through the adipoR1/T-cadherin/calreticulin/AMPK pathway. In AFs, gAd and UII synergistically induced AMPK phosphorylation. In the adventitial inflammation model, APN deficiency up-regulated the expression of α-SMA, UII receptor (UT), and UII while inhibiting TNF-α expression.
CONCLUSIONS
From the results of our study, we can speculate that UII induces TNF-α protein and TNF-α-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways. Thus, it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.
Mice
;
Rats
;
Animals
;
Adventitia/metabolism*
;
Tumor Necrosis Factor-alpha/metabolism*
;
Calreticulin/metabolism*
;
Vascular Remodeling
;
AMP-Activated Protein Kinases/metabolism*
;
Cells, Cultured
;
RNA, Messenger/genetics*
;
Inflammation
8.Family sex education and associated factors among preschoolers in Wuhu
WEI Liangchen, ZHOU Shu, GUO Zhiyuan, XIE Shuyi, LIANG Yali, HUANG Yueer
Chinese Journal of School Health 2021;42(5):675-678
Objective:
To understand the status of family sex education and associated factors among preschoolers in Wuhu, providing scientific basis for child family sex education.
Methods:
From July to August 2020, random cluster sampling was used to select 481 children from 5 kindergartens in Wuhu. A questionnaire survey among parents of these preschoolers regarding family sex education status and its influencing factors.
Results:
A total of 285(59.25%) parents reported family sex education for children, and 196(40.75%) did not practice family sex education for their children. Multivariate Logistic regression analysis showed parental awareness of sex education content(OR=3.06, 95%CI=1.95-4.78), parental anxiety for child sexual assault (OR=1.82, 95%CI=1.11-2.99) were associated with higher rate of family sex education.
Conclusion
Family sex education among preschoolers in Wuhu should be further promoted. Sex education training towards parents might help improve children s family sex education.
9.Evaluation of the effects of mindfulness training on sleep, anxiety and depression in college students
YUAN Xinyun,JIA Shuyi,FU Shirui,GUO Wei,PENG Yan
Chinese Journal of School Health 2021;42(11):1655-1659
Objective:
To investigate the effect of online and offline mindfulness training on improving anxiety and depression and sleep quality of college students,and to provide a reference for mental health promotion among college students.
Methods:
From October 2020, a total of 1 203 university students from North China University of Technology were screened with the Self rating Anxiety Scale (SAS), Self rating Depression Scale (SDS) and Pittsburg Sleep Quality Index (PSQI) using the whole group radom cluster sampling method. Totally 103 students who met the inclusion criteria were randomly divided into 64 online and 39 offline groups. The degree of improvement in anxiety, depression and sleep quality was assessed after the intervention.
Results:
The SAS, SDS and PSQI scores of college students after the online and offline the mindfulness training intervention significantly decreased compared with score before the intervention( t =5.57, 5.31, 3.99; 4.88,5.02, 5.88, P <0.01). The difference in the degree of improvement in sleep quality between the two interventions, online and offline, was statistically significant ( t =-2.55, P <0.05). The less the three symptoms of anxiety, depression and sleep were combined in university students, the higher the symptom remission rate of the positive mindfulness training (25% remission rate for all three symptoms together, 40% remission rate for two symptoms together and 100% remission rate for only one symptom).
Conclusion
Both online and mindfulness training can be used as an effective intervention for sleep, anxiety and depression; offline mindfulness training is more effective than online in improving sleep quality in university students; mindfulness training is more effective in relieving single symptoms.
10.Multivariate analysis of peripheral blood mononuclear cells from children donors
Yixin GAO ; Shuyi MO ; Gang WANG ; Suying HE ; Shaowen LI ; Chuxia GUO ; Zhigang LU ; Yongzheng PENG
Chinese Journal of Blood Transfusion 2021;34(7):688-691
【Objective】 To analyze the characteristics of peripheral blood mononuclear cells (PBMC) collection in healthy children, and explore the factors affecting collection efficiency (CE). 【Methods】 The PBMC data, involving 70 episodes of apheresis from 42 children during January 2017 and June 2020 were retrospectively analyzed All children were collected in Zhujiang Hospital of Southern Medical University. 【Results】 Multiple linear regression analysis showed that mononuclear cells (MNC) in donor collections from healthy children were positively correlated with anticoagulant dosage, lymphocyte count and monocyte count (P<0.05), meanwhile, negatively correlated with age and platelet count. The PBMC CE was negatively correlated with age, platelet count, and processed whole blood volume (P<0.05). CD34+ cells (×107 /kg)was negatively correlated with age, meanwhile, positively correlated with numbers of collection and processed whole blood volume(r=-0.79). No statistical differences in red blood cell count, platelet count, lymphocyte count, monocyte count of healthy child donors were notable before versus after apheresis. 【Conclusion】 MNC can be collected effectively in children of different ages. The PBMC collection efficiency was related to age. Meanwhile, the higher the lymphocytes and monocytes were before apheresis, the more MNC were collected. The efficiency of MNC collection would decrease when the apheresis volume of the children exceeded their total blood volume twice. However, the absolute value of CD34+ cells in the final yields would increase.


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