1.Identification of porcine-derived circ_PIK3C2A and its effect on PEDV replication in host cells after infection
Siqi JIA ; Rongrong LIU ; Yingjin CHAI ; Xinxin HAN ; Mingqing WEI ; Tingting WU ; Ying DING ; Shaoxiu CHEN ; Xingmei DENG ; Hui ZHANG
Chinese Journal of Veterinary Science 2025;45(10):2110-2117
Circular RNA(circRNA),as a kind of non-coding RNA,regulates a variety of biological functions.To explore the effect of circRNA on PEDV replication in the host porcine intestinal epi-thelial cells,this study screened and analyzed the differentially expressed circRNAs by bioinforma-tic software in African Green Monkey renal cells(Vero-E6 cells)infected by porcine epidemic di-arrhea virus(PEDV),the differentially expressed circRNA ssc_circ_PIK3C2A was identified and the secondary structure was analyzed.PCR was used to identify the ssc_circ_PIK3C2A circRNA structure,the model of PEDV-infected IPEC-J2 cells was constructed,the TCID50 test was used to validate the viral titer of PEDV.The expression of circ_PIK3C2A was detected by qRT-PCR in IPEC-J2 infected by PEDV.circ_PIK3C2A qRT-PCR was performed to detect the expression of N gene of PEDV when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.The results showed that ssc_circ_PIK3C2 A is a porcine circular RNA with a typical circular structure,the virus titer of PEDV reached 10-6/mL after PEDV infected IPEC-J2 cells for 48 h,the expression of circ_PIK3C2A increased extremely(P<0.01)at 6 h after PEDV-infection,with the extension of infec-tion time,its expression gradually decreased,and the expression was the lowest at 24 h,but there was no time-dependent trend.The expression of PEDV N gene decreased significantly when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.In conclusion,when PEDV infects IPEC-J2 cells,the expression of porcine circ_PIK3C2A decreases,and replication of PEDV increases signifi-cantly in IPEC-J2 cells.our result provides a basis for further study of the mechanism of circular RNA on PEDV replication and its physiological activities in host cells in the future.
2.Supramolecular Binding Behavior and Solubilization of Cationic Cyclodextrins towards Liquiritigenin
Ying-Hui DENG ; Dong-Jing ZHANG ; Hai-Kun WANG ; Jia-Xing CHEN ; Shuang SONG ; Bo YANG ; Xia-Li LIAO
Chinese Journal of Analytical Chemistry 2025;53(2):289-299,中插20-中插30
Liquiritigenin(LG)is a flavone of pharmacological importance,however,its application potential is severely limited due to its poor water solubility.LG could be disassociated slightly in water to form phenolate anion,therefore,better solubilization effect is expected by inclusion with cationic cyclodextrins(CCDs).In this work,four kinds of CCDs modified with amino groups at the primary face were synthesized,and their solid inclusion complexes with LG were successfully prepared by preparing their saturated solutions.The formation of the solid inclusion complexes was confirmed by scanning electron microscopy(SEM)and powder X-ray diffraction(PXRD),and their supramolecular binding behavior in solution was studied using multiple techniques.A 1∶1 inclusion stoichiometry of inclusion complexation was defined using Job plot by ultraviolet-visible(UV-vis)spectroscopy,and their binding stability constants(Ks)were determined as 2862.77,3494.70,6521.85 and 9599.48 L/mol using UV-vis spectroscopic titration,far more superior to that of nativeβ-CD(Ks=236.79 L/mol).This indicated that the amino side chains on CCDs could actively participate in the inclusion complexation through anion-cation interactions,significantly strengthening the host-guest binding between CCDs and LG.The inclusion modes were further elucidated based on proton and two-dimensional rotating-frame overhauser enhancement spectroscopy(2D-ROESY)nuclear magnetic resonance(NMR)experiments and molecular docking.Water solubility of LG was dramatically promoted up to 4.9 mg/mL,which was 70-fold higher than that of native LG.This study could draw inspiration for the binding and solubilization of phenols such as flavones by design of cationic macrocyclic molecules.
3.Application of annual progressive assessment mode in standardized training for residents in a medical laboratory base
Ying LUO ; Yuwei DI ; Yue MENG ; Qianyun DENG ; Hui LI ; Huiquan GAN ; Bing GU
Chinese Journal of Medical Education Research 2025;24(3):361-365
Objective:To explore the establishment, implementation, and application effect of the annual progressive assessment mode for standardized training of resident physicians in a medical laboratory base.Methods:Since 2020, the base has improved the annual progressive assessment system through assessment system establishment, question bank construction, routine assessment, rotation examination, and annual examination. This new assessment mode was adopted for the residency trainees enrolled in the base after 2020. The control group consisted of seven residency trainees enrolled before 2020, while the experimental group comprised 13 trainees enrolled in 2020 and later. The two groups were compared based on the rotation and annual examination scores of second- and third-year trainees, as well as their academic achievements. Additionally, teaching outcomes of the base before and after 2020 were analyzed. The t test, rank-sum test, and Fisher's exact test were performed using SPSS 27.00. Results:There were no significant differences in age, sex, and education level between the two groups. The experimental group showed significantly higher scores than the control group in theory assessment ( P<0.001 for second-year trainees and P=0.008 for third-year trainees) and skill assessment ( P<0.001 for second-year trainees and P=0.038 for third-year trainees) in rotation examination, as well as in theory assessment (both P<0.001) and skill assessment (both P<0.001) in annual examination. The improvement was particularly pronounced among second-year trainees. The trainees of the experimental group submitted 17 case reports in three years, and won awards at national conferences, provincial conferences, and institutional case speech competitions. In a national academic conference, the trainees won the second prize of "Excellent Case". Since 2020, we have been granted with our first teaching reform project and published our first teaching article. To date, we have obtained five teaching reform projects, published six teaching articles, and won multiple awards in provincial and institutional teaching competitions. Conclusions:The establishment of the annual progressive assessment mode is critical for training high-quality laboratory physicians, and puts forward higher requirements for residency training teachers.
4.Efficacy of Differential Dosage of Pueraria in Gegen Qinliantang on Acute Enteritis Model in Mice
Ruiying ZHANG ; Ping WANG ; Di ZHANG ; Hongfa CHENG ; Ying ZHANG ; Zhu DENG ; Hui FENG ; Min LIU ; Yang TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):197-204
ObjectiveTo investigate whether there are differences in the efficacy of Gegen Qinliantang with different contents of Puerariae Lobatae Radix on the acute enteritis (AE) model mice and provide a scientific basis for the interpretation of Gegen Qinliantang in the treatment of "Xie Re Li". MethodsA total of 112 male BALB/c mice were randomly divided into a blank group,model group,single Puerariae Lobatae Radix group,non-Puerariae Lobatae Radix group,regular dose Gegen Qinliantang group (regular dose group),half-dose Puerariae Lobatae Radix group,and doubled-dose Puerariae Lobatae Radix group, with 16 mice in each group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the colon tissue. Western blot was employed to detect the expression of ZO-1 (a protein in the tight junction) and Occludin in the colon tissue, as well as the changes of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). ResultsCompared with the blank group,the DAI scores of the mice in the model group were significantly higher (P<0.05),and the histopathological sections of their colon tissues showed mucosal damage,glandular atrophy,disordered arrangement,and a large number of inflammatory cells infiltration,and the expression of ZO-1 and Occludin proteins in their colon tissues was significantly down-regulated (P<0.05,P<0.01). The expression of inflammatory factors TNF-α and IL-1β was significantly up-regulated (P<0.05,P<0.01). Compared with the model group,the DAI scores of mice in all dosing groups decreased significantly (P<0.05),with the most significant effect in the regular dose group. After 7 d of drug administration,the regular dose group had the best impact on the repair of colonic mucosa in the AE mouse model. The regular dose group significantly down-regulated the expression of TNF-α (P<0.05) and significantly up-regulated the expression of ZO-1 protein (P<0.05). The doubled-dose Puerariae Lobatae Radix group significantly down-regulated the expression of IL-1β protein (P<0.01),and there was no significant difference between all dosing groups and the model group in terms of the expression of Occludin protein. After 14 d of drug administration,the best effect on the repair of colonic mucosa in the AE mouse model was observed in the doubled dose Puerariae Lobatae Radix group. All groups except the non-Puerariae Lobatae Radix group significantly down-regulated the expression of TNF-α (P<0.01). Meanwhile,the regular dose group and doubled-dose Puerariae Lobatae Radix group significantly elevated the expression level of Occludin protein (P<0.01). The doubled-dose Puerariae Lobatae Radix group also significantly inhibited the expression of IL-1β protein (P<0.05) and up-regulated ZO-1 protein expression (P<0.05). ConclusionGegen Qinliantang can reduce the pathological damage of colon tissue, protect the barrier function and structure of intestinal epithelial cells, and reduce the expression of inflammatory factors, so as to achieve the therapeutic effect of AE model mice. When comparing the therapeutic efficacy of Gegen Qinliantang containing different Gegen contents, Gegen Qinliantang with the proportion of the original formula of Zhongjing was the most effective in AE model mice.
5.Efficacy of Differential Dosage of Pueraria in Gegen Qinliantang on Acute Enteritis Model in Mice
Ruiying ZHANG ; Ping WANG ; Di ZHANG ; Hongfa CHENG ; Ying ZHANG ; Zhu DENG ; Hui FENG ; Min LIU ; Yang TANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(21):197-204
ObjectiveTo investigate whether there are differences in the efficacy of Gegen Qinliantang with different contents of Puerariae Lobatae Radix on the acute enteritis (AE) model mice and provide a scientific basis for the interpretation of Gegen Qinliantang in the treatment of "Xie Re Li". MethodsA total of 112 male BALB/c mice were randomly divided into a blank group,model group,single Puerariae Lobatae Radix group,non-Puerariae Lobatae Radix group,regular dose Gegen Qinliantang group (regular dose group),half-dose Puerariae Lobatae Radix group,and doubled-dose Puerariae Lobatae Radix group, with 16 mice in each group. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of the colon tissue. Western blot was employed to detect the expression of ZO-1 (a protein in the tight junction) and Occludin in the colon tissue, as well as the changes of tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). ResultsCompared with the blank group,the DAI scores of the mice in the model group were significantly higher (P<0.05),and the histopathological sections of their colon tissues showed mucosal damage,glandular atrophy,disordered arrangement,and a large number of inflammatory cells infiltration,and the expression of ZO-1 and Occludin proteins in their colon tissues was significantly down-regulated (P<0.05,P<0.01). The expression of inflammatory factors TNF-α and IL-1β was significantly up-regulated (P<0.05,P<0.01). Compared with the model group,the DAI scores of mice in all dosing groups decreased significantly (P<0.05),with the most significant effect in the regular dose group. After 7 d of drug administration,the regular dose group had the best impact on the repair of colonic mucosa in the AE mouse model. The regular dose group significantly down-regulated the expression of TNF-α (P<0.05) and significantly up-regulated the expression of ZO-1 protein (P<0.05). The doubled-dose Puerariae Lobatae Radix group significantly down-regulated the expression of IL-1β protein (P<0.01),and there was no significant difference between all dosing groups and the model group in terms of the expression of Occludin protein. After 14 d of drug administration,the best effect on the repair of colonic mucosa in the AE mouse model was observed in the doubled dose Puerariae Lobatae Radix group. All groups except the non-Puerariae Lobatae Radix group significantly down-regulated the expression of TNF-α (P<0.01). Meanwhile,the regular dose group and doubled-dose Puerariae Lobatae Radix group significantly elevated the expression level of Occludin protein (P<0.01). The doubled-dose Puerariae Lobatae Radix group also significantly inhibited the expression of IL-1β protein (P<0.05) and up-regulated ZO-1 protein expression (P<0.05). ConclusionGegen Qinliantang can reduce the pathological damage of colon tissue, protect the barrier function and structure of intestinal epithelial cells, and reduce the expression of inflammatory factors, so as to achieve the therapeutic effect of AE model mice. When comparing the therapeutic efficacy of Gegen Qinliantang containing different Gegen contents, Gegen Qinliantang with the proportion of the original formula of Zhongjing was the most effective in AE model mice.
6.Effects of rice wine type and wine processing method on chemical constituents and anti-coagulation effect of Angelicae sinensis Radix
Ying WANG ; Ya-yi DENG ; Xue-qi GE ; Hui ZHU ; Yu DUAN ; Xiao-ning YAN ; Hao CAI ; Ke PEI
Chinese Traditional Patent Medicine 2025;47(5):1443-1448
AIM To investigate the effects of rice wine type and wine processing method on chemical constituents and anti-coagulation effect of Angelicae sinensis Radix.METHODS Wine-washed products and wine-stir-fried products were prepared by different types and ages of rice wine,respectively,after which HPLC was adopted in the content determination of tryptophan,chlorogenic acid,vanillic acid,phthalic acid,ferulic acid,senkyunolide I,senkyunolide H,coniferyl ferulate and ligustilide,and PT,APTT,TT were detected in rabbit plasma.RESULTS Phenolic acids and volatile constituents demonstrated lower contents in the wine-stir-fried products than those in the raw product(P<0.05),while those in the wine-washed products displayed no obvious changes(except for senkyunolide I)(P>0.05).The contents of volatile constituents in the wine-washed products were higher than those in the wine-stir-fried products(P<0.05).After being processed with dry rice wine,various constituents exhibited increased contents as compared with those after being processed with sweet rice wine(P<0.05).Compared with the raw product,prolonged PT,APTT and TT were observable in the processed products prepared by 3-year semi-dry rice wine(P<0.05).CONCLUSION The optimal rice wine type is determined to be 3-year semi-dry.Wine-washed Angelicae sinensis Radix shows high contents of ferulic acid and volatile constituents,whose activating blood and resolving stasis effect may be stronger.
7.Identification of porcine-derived circ_PIK3C2A and its effect on PEDV replication in host cells after infection
Siqi JIA ; Rongrong LIU ; Yingjin CHAI ; Xinxin HAN ; Mingqing WEI ; Tingting WU ; Ying DING ; Shaoxiu CHEN ; Xingmei DENG ; Hui ZHANG
Chinese Journal of Veterinary Science 2025;45(10):2110-2117
Circular RNA(circRNA),as a kind of non-coding RNA,regulates a variety of biological functions.To explore the effect of circRNA on PEDV replication in the host porcine intestinal epi-thelial cells,this study screened and analyzed the differentially expressed circRNAs by bioinforma-tic software in African Green Monkey renal cells(Vero-E6 cells)infected by porcine epidemic di-arrhea virus(PEDV),the differentially expressed circRNA ssc_circ_PIK3C2A was identified and the secondary structure was analyzed.PCR was used to identify the ssc_circ_PIK3C2A circRNA structure,the model of PEDV-infected IPEC-J2 cells was constructed,the TCID50 test was used to validate the viral titer of PEDV.The expression of circ_PIK3C2A was detected by qRT-PCR in IPEC-J2 infected by PEDV.circ_PIK3C2A qRT-PCR was performed to detect the expression of N gene of PEDV when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.The results showed that ssc_circ_PIK3C2 A is a porcine circular RNA with a typical circular structure,the virus titer of PEDV reached 10-6/mL after PEDV infected IPEC-J2 cells for 48 h,the expression of circ_PIK3C2A increased extremely(P<0.01)at 6 h after PEDV-infection,with the extension of infec-tion time,its expression gradually decreased,and the expression was the lowest at 24 h,but there was no time-dependent trend.The expression of PEDV N gene decreased significantly when ssc_circ_PIK3C2A was over-expressed in IPEC-J2 cells.In conclusion,when PEDV infects IPEC-J2 cells,the expression of porcine circ_PIK3C2A decreases,and replication of PEDV increases signifi-cantly in IPEC-J2 cells.our result provides a basis for further study of the mechanism of circular RNA on PEDV replication and its physiological activities in host cells in the future.
8.Application of annual progressive assessment mode in standardized training for residents in a medical laboratory base
Ying LUO ; Yuwei DI ; Yue MENG ; Qianyun DENG ; Hui LI ; Huiquan GAN ; Bing GU
Chinese Journal of Medical Education Research 2025;24(3):361-365
Objective:To explore the establishment, implementation, and application effect of the annual progressive assessment mode for standardized training of resident physicians in a medical laboratory base.Methods:Since 2020, the base has improved the annual progressive assessment system through assessment system establishment, question bank construction, routine assessment, rotation examination, and annual examination. This new assessment mode was adopted for the residency trainees enrolled in the base after 2020. The control group consisted of seven residency trainees enrolled before 2020, while the experimental group comprised 13 trainees enrolled in 2020 and later. The two groups were compared based on the rotation and annual examination scores of second- and third-year trainees, as well as their academic achievements. Additionally, teaching outcomes of the base before and after 2020 were analyzed. The t test, rank-sum test, and Fisher's exact test were performed using SPSS 27.00. Results:There were no significant differences in age, sex, and education level between the two groups. The experimental group showed significantly higher scores than the control group in theory assessment ( P<0.001 for second-year trainees and P=0.008 for third-year trainees) and skill assessment ( P<0.001 for second-year trainees and P=0.038 for third-year trainees) in rotation examination, as well as in theory assessment (both P<0.001) and skill assessment (both P<0.001) in annual examination. The improvement was particularly pronounced among second-year trainees. The trainees of the experimental group submitted 17 case reports in three years, and won awards at national conferences, provincial conferences, and institutional case speech competitions. In a national academic conference, the trainees won the second prize of "Excellent Case". Since 2020, we have been granted with our first teaching reform project and published our first teaching article. To date, we have obtained five teaching reform projects, published six teaching articles, and won multiple awards in provincial and institutional teaching competitions. Conclusions:The establishment of the annual progressive assessment mode is critical for training high-quality laboratory physicians, and puts forward higher requirements for residency training teachers.
9.Effect of demethylase FTO knockout on 5-HT-induced abnormalities of coronary smooth muscle contractile function in mice with diabetes mellitus
Zi-fan WANG ; Liu-xiang JIANG ; Mei-ying LIANG ; Meng-yun LIU ; Mei-jiang CHEN ; Hui YANG ; Fang RAO ; Chun-yu DENG
Chinese Pharmacological Bulletin 2025;41(2):315-322
Aim To explore the influence of demethy-lase fat mass and obesity-related genes(FTO)on the abnormal contractile function of diabetic coronary smooth muscle.Methods Smooth muscle specific FTO knockout mice(FTOSMKO)were prepared by Cre-loxP recombinant technology.They were divided into four groups:control(WT)group,diabetes model group(DM),FTO knockout group(FTOSMKO),and FTOSMKO diabetic group(FTOSMKO-DM),with 15 mice in each group.Diabetic mice were prepared by intraperitoneal injection of streptozotocin(STZ);the remaining mice were injected with an equal amount of citric acid-sodi-um citrate buffer.The effects of 5-HTon the contractile response of coronary artery smooth muscle in the four groups of mice were observed by the technique of small-vessel ring tensiometry.Western blot and Dot blot were used to detect the changes of FTO protein and N6-methyladenine(m6A)methylation modification levels in mouse vascular tissues.Results Compared with the WT group,the DM group had significantly higher blood glucose(P<0.01)and lower body weight(P<0.05);the level of FTO protein in aorta of DM group increased(P<0.01),and the level of m6A methylation modification decreased(P<0.01).The 5-HT-induced contractile response significantly de-creased in the DM group compared with the WT group(P<0.01),whereas the contractile response signifi-cantly increased in the FTOSMKO-DM group compared with the DM group(P<0.01);the non-L-type calci-um channel-mediated vascular smooth muscle contrac-tile response was enhanced in the FTOSMKO-DM group,among which,the contractions induced by 1,4,5-triphosphate inositol receptor(IP3R)and caffeine-ac-tivated ranine receptor(RyR)mediated by sarcoplas-mic reticulum calcium release both significantly in-creased(P<0.05).Conclusions Specific knock-down of smooth muscle FTO improves coronary artery responsiveness to the vasoconstrictor 5-HT in diabetic mice,which may be related to abnormalities in the FTO-mediated 5-HT receptor signaling pathway.
10.Effect of demethylase FTO knockout on 5-HT-induced abnormalities of coronary smooth muscle contractile function in mice with diabetes mellitus
Zi-fan WANG ; Liu-xiang JIANG ; Mei-ying LIANG ; Meng-yun LIU ; Mei-jiang CHEN ; Hui YANG ; Fang RAO ; Chun-yu DENG
Chinese Pharmacological Bulletin 2025;41(2):315-322
Aim To explore the influence of demethy-lase fat mass and obesity-related genes(FTO)on the abnormal contractile function of diabetic coronary smooth muscle.Methods Smooth muscle specific FTO knockout mice(FTOSMKO)were prepared by Cre-loxP recombinant technology.They were divided into four groups:control(WT)group,diabetes model group(DM),FTO knockout group(FTOSMKO),and FTOSMKO diabetic group(FTOSMKO-DM),with 15 mice in each group.Diabetic mice were prepared by intraperitoneal injection of streptozotocin(STZ);the remaining mice were injected with an equal amount of citric acid-sodi-um citrate buffer.The effects of 5-HTon the contractile response of coronary artery smooth muscle in the four groups of mice were observed by the technique of small-vessel ring tensiometry.Western blot and Dot blot were used to detect the changes of FTO protein and N6-methyladenine(m6A)methylation modification levels in mouse vascular tissues.Results Compared with the WT group,the DM group had significantly higher blood glucose(P<0.01)and lower body weight(P<0.05);the level of FTO protein in aorta of DM group increased(P<0.01),and the level of m6A methylation modification decreased(P<0.01).The 5-HT-induced contractile response significantly de-creased in the DM group compared with the WT group(P<0.01),whereas the contractile response signifi-cantly increased in the FTOSMKO-DM group compared with the DM group(P<0.01);the non-L-type calci-um channel-mediated vascular smooth muscle contrac-tile response was enhanced in the FTOSMKO-DM group,among which,the contractions induced by 1,4,5-triphosphate inositol receptor(IP3R)and caffeine-ac-tivated ranine receptor(RyR)mediated by sarcoplas-mic reticulum calcium release both significantly in-creased(P<0.05).Conclusions Specific knock-down of smooth muscle FTO improves coronary artery responsiveness to the vasoconstrictor 5-HT in diabetic mice,which may be related to abnormalities in the FTO-mediated 5-HT receptor signaling pathway.

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