1.Mechanism of Xuefu Zhuyutang in Intervening in Ferroptosis in Rats with Coronary Heart Disease with Blood Stasis Syndrome Based on ACSL4 Signalling Pathway
Yi LIU ; Yang YANG ; Chang SU ; Peng TIAN ; Mingyun WANG ; Ruqian ZHONG ; Xuejiao XIE ; Qing YAN ; Qinghua PENG ; Qiuyan ZHANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(6):27-38
ObjectiveTo investigate the mechanism of ferroptosis mediated by long-chain acyl-CoA synthetase 4 (ACSL4) signalling pathway in rats with coronary heart disease with blood stasis syndrome and the intervention effect of Xuefu Zhuyutang. MethodsSPF male SD rats were randomly divided into normal group, sham-operation group, model group, trimetazidine group (5.4 mg·kg-1), low-, medium-, and high-dose group (3.51, 7.02,14.04 g·kg-1) of Xuefu Zhuyutang. The coronary artery left anterior descending ligation method was used to prepare a model of coronary heart disease with blood stasis syndrome, and continuous treatment for 7 d was conducted, while the sham-operation group was only threaded and not ligated. The general macroscopic symptoms of the rats were observed, and indicators such as electrocardiogram, echocardiography, and blood rheology were detected. The pathological morphology of myocardial tissue was observed by hematoxylin-eosin (HE) staining, and the changes in mitochondria in myocardial tissue were observed by transmission electron microscopy. The level of iron deposition in myocardial tissue was observed by Prussian blue staining. The levels of 12-hydroxyeicosatetraenoic acid (12-HETE) and 15-HETE were detected in serum by enzyme-linked immunosorbent assay. A biochemical colourimetric assay was used to detect the levels of Fe2+, lipid peroxidation (LPO), glutathione (GSH), and T-GSH/glutathione disulfide (GSSG) in myocardial tissue. DCFH-DA fluorescence quantitative assay was employed to detect the levels of reactive oxygen species (ROS). Western blot and Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was adopted to detect the protein and mRNA expressions of glutathione peroxidase 4 (GPX4), ferritin heavy chain 1 (FTH1), ACSL4, and ly-sophosphatidylcholine acyltransferase3 (LPCAT3) in myocardial tissue. ResultsCompared with those in the normal group, the rats in the model group were poor in general macroscopic symptoms. The electrocardiogram showed widened QRS wave amplitude and increased voltage, bow-back elevation of the ST segments, elevated T waves, J-point elevation, and accelerated heart rate. Echocardiography showed a significant reduction in left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) (P<0.01). Blood rheology showed that the viscosity of the whole blood (low, medium, and high rate of shear) was significantly increased (P<0.01). HE staining showed an abnormal structure of myocardial tissue. There was a large area of myocardial necrosis and inflammatory cell infiltration and a large number of connective tissue between myocardial fibers. Transmission electron microscopy showed that the mitochondria were severely atrophy or swelling. The cristae were reduced or even broken, and the matrix was flocculent or even vacuolated. Prussian blue staining showed that there were a large number of iron-containing particles, and the iron deposition was obvious. The content of 12-HETE and 15-HETE in the serum was significantly increased (P<0.01). The content of Fe2+, LPO, and ROS in myocardial tissue was significantly increased (P<0.01). The content of GSH was significantly decreased (P<0.01), and T-GSH/GSSG was decreased (P<0.01). The protein and mRNA expressions of GPX4 and FTH1 in myocardial tissue were both significantly decreased (P<0.05, P<0.01), while those of ACSL4 and LPCAT3 increased significantly (P<0.01). Compared with the model group, the general macroscopic symptoms and electrocardiogram results of rats in low-, medium- and high-dose groups of Xuefu Zhuyutang were alleviated, and the differences in LVEF/LVFS ratios were all significantly increased (P<0.05, P<0.01). The differences in whole-blood viscosity (low, medium, and high rate of shear) were all significantly decreased (P<0.01). The results of HE staining and transmission electron microscopy showed that the morphology, structure, and mitochondria of cardiomyocytes were improved. The content of 12-HETE and 15-HETE in serum was reduced to different degrees in low-, medium-, and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01). The content of Fe2+, LPO, and ROS was significantly reduced in the medium- and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01), and the content of GSH and T-GSH/GSSG was significantly increased (P<0.05, P<0.01). The protein and mRNA expressions of GPX4 and FTH1 were significantly increased to varying degrees in the medium- and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01), and ACSL4 and LPCAT3 were decreased to different degrees in the low-, medium-, and high-dose groups of Xuefu Zhuyutang (P<0.05, P<0.01). ConclusionXuefu Zhuyutang can regulate iron metabolism and anti-lipid oxidation reaction to mediate ferroptosis through the ACSL4 signalling pathway, thus exerting a protective effect on rats with coronary heart disease with blood stasis syndrome.
2.Comparison of rat models of pulmonary fibrosis induced by one or two intratracheal bleomycin instillations
Huimeng TANG ; Qinghua SONG ; Yunyun XIE ; Xin SUN ; Fan YANG ; Shuguang YANG ; Yange TIAN
Acta Laboratorium Animalis Scientia Sinica 2024;32(9):1139-1148
Objective A rat model of pulmonary fibrosis was constructed using a single or two intratracheal drops of bleomycin(BLM)and the modeling rate and stability of the two modeling modalities were compared.Methods A total of 150 specific pathogen-free SD rats were divided randomly into blank control(control),single intratracheal drop of bleomycin(BLM-S),and two intratracheal drops of bleomycin(BLM-M)groups.Rats in the BLM-S group received a single dose of 3 mg/kg BLM by noninvasive intratracheal instillation,and rats in BLM-M group received 3 mg/kg BLM on day 1 and BLM 2 mg/kg on day 14.Rats in the control group were given intratracheal instillation of 0.9%sodium chloride(1 mL/kg).The rats were euthanized on days 28,42,56,and 84 after modelling,respectively.Deep inspiratory capacity(IC),vital capacity(VC),static lung compliance(Cchord),and dynamic lung complication(Cdyn)were measured in all rats.Pathological changes in lung tissue were observed,and the extent of alveolitis and fibrosis was graded.Collagen-Ⅲ(COL-Ⅲ)expression in rat lung tissue was detected by immunohistochemistry.Results(1)The survival rates in the control,BLM-S,and BLM-M groups were 100%,80%,and 66%,respectively.Rats in the BLM-S and BLM-M groups had significantly lower body weights on days 14~42 compared with the control group(P<0.05,P<0.01),and rats in the BLM-M group had significantly lower body weight on days 28~42 than rats in the control and BLM-S groups(P<0.05,P<0.01).(2)Regarding lung function,IC,VC,Cchord,and Cdyn were all markedly decreased in the BLM-S group compared with the control group(P<0.05,P<0.01)and IC,VC,and Cchord were significantly decreased in the BLM-M group(P<0.05,P<0.01)on day 28.IC,VC,and Cchord were significantly decreased in rats in the BLM-S group on day 42(P<0.05,P<0.01),and were also significantly decreased in rats in the BLM-M group on days 42~84(P<0.05,P<0.01).(3)In terms of lung pathology,inflammatory infiltration and fibrous cords appeared in the BLM-S group from days 28~84 and then gradually decreased(P<0.05,P<0.01),while fibrosis and alveolitis were relatively stable in the BLM-M group(P<0.05,P<0.01).(4)COL-Ⅲ expression levels in lung tissue were significantly higher in rats in the BLM-S and BLM-M groups compared with the control group(P<0.05,P<0.01),and the COL-Ⅲ content in the BLM-S group was significantly lower at 42~84 days than at 28 days(P<0.05).Conclusions Both method are capable of effectively creating pulmonary fibrosis models.The single-dose approach is straightforward,has a lower death rate,and the degree of fibrosis is clearly visible by day 28,but progressively recovers after 42 days.In contrast,the two-dose instillation model has a greater success rate and better stability,with over half the rats still exhibiting visible fibrosis on day 84.
3.Comparison of pulmonary fibrosis rat models induced by different dosed of bleomycin
Qinghua SONG ; Huimeng TANG ; Xin SUN ; Yang LIU ; Yunyun XIE ; Yange TIAN
Chinese Journal of Comparative Medicine 2024;34(10):28-37
Objective To compare the success rate and stability of rat pulmonary fibrosis(PF)models induced by intratracheal instillation of different doses of bleomycin(BLM).Methods One hundred and fifty Sprague Dawley rats were divided randomly into control,low-dose BLM 3 mg/kg(BL-L),and high-dose BLM 5 mg/kg(BL-H)groups.General status,mortality,and weight changes were observed,and the lung inspiratory capacity(IC),vital capacity(VC),chord compliance(Cchord),and dynamic compliance(Cdyn)were detected on days 28,42,56,and 84.Lung coefficients were recorded and pathological changes in lung tissue were observed by hematoxylin and eosin and Masson staining.The lung hydroxyproline(HYP)content was detected and collagen type Ⅲ(COL Ⅲ)was detected by immunohistochemistry.Results The mortality rates in the BL-L and BL-H groups were 20%and 28%,respectively.Body weight was significantly lower in the BL-L group compared with the control group on days 0~56,and weight recovery after day 56.Body weight was significantly lower in the BL-H group compared with the control and BL-L groups from days 0~56(P<0.01).Regarding lung function,IC,VC,Cchord,and Cdyn were significantly lower in the BL-L group compared with the control group on day 28(P<0.01,P<0.05),and IC and Cdyn were significantly lower in the BL-H group(P<0.01).IC,VC,and Cchord were significantly decreased in the BL-L group on day 42(P<0.01,P<0.05),while IC,VC,Cchord,and Cdyn were significantly decreased in the BL-H group(P<0.01,P<0.05),and IC,VC,and Cchord were significantly lower compared with in the BL-L group(P<0.01).Cchord was significantly lower in the BL-H group compared with the control and BL-L groups on day 56(P<0.01).The lung coefficients on day 28 were significantly higher in the BL-L and BL-H groups compared with the control group(P<0.01),and were significantly higher in the BL-H group from days 42~56 compared with the BL-L and control groups(P<0.01).Regarding lung histopathology and immunohistochemistry,inflammatory infiltration,fibrotic streaks,and COL Ⅲ expression were observed in the BL-L group from days 28~56,and almost complete disappearance of the fibrotic lesions on day 84.In contrast,fibrotic lesions could be observed from days 28~84 in the BL-H group,with significantly elevated COL Ⅲ expression compared with the control group(P<0.01).The HYP content was significantly higher in the BL-L group compared with the control group from days 28~42(P<0.05,P<0.01),and then gradually decreased,and the HYP content was significantly higher in the BL-H group than in the control group from days 28~84(P<0.01).Conclusions Both 3 and 5 mg/kg BLM can successfully induce PF rat models.Rats treated with 3 mg/kg BLM developed fibrosis on day 28,which lasted until day 42 and then gradually recovered.Rats treated with 5 mg/kg BLM developed fibrosis on day 28,and the degree of fibrosis was more severe with the higher compared with the lower dose,with stable fibrotic lesions up to day 56 and moderate-to-severe fibrosis still present in half of the rats until day 84.
4.Anmeidan Regulates Hepatic Neurotransmitters and Circadian Rhythm in Rat Model of Insomnia via OX1R/PLCβ-1/PKCα/ERK1/2 Signaling Pathway
Bo XU ; Ping WANG ; Jing XIA ; Guangjing XIE ; Zijing YE ; Qinghua QIN ; Jing CHENG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(15):11-20
ObjectiveTo explore the effect and mechanism of the classic famous prescription Anmeidan (AMD) developed in the Qing Dynasty in regulating the hepatic neurotransmitters and circadian rhythm in the rat model of insomnia via the orexin-1 receptor (OX1R)/phosphatidylinositol-specific phospholipase Cβ-1 (PLCβ-1)/protein kinase Cα (PKCα)/extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. MethodSixty SPF-grade SD rats were randomized into blank, model, suvorexant (30 mg·kg-1·d-1), and low-, medium-, and high-dose (4.55, 9.09, 18.09 g·kg-1·d-1, respectively) AMD groups, with 10 rats in each group. The rats in other groups except the blank group were modeled by intraperitoneal injection of p-chlorophenylalanine (PCPA) and administrated with corresponding drugs by gavage, and the blank group received an equal volume of normal saline. The general condition, body mass, and 24 h autonomic activity of each group were observed. The pathological changes of the liver tissue were observed by hematoxylin-eosin(HE)staining and Masson staining. The expression of gamma-aminobutyric acid (GABA), 5-hydroxytryptamine (5-HT), epinephrine (EPI), norepinephrine (NE), and acetylcholine (ACh) in the liver tissue was detected by enzyme-linked immunosorbent assay. The glutamate (Glu) expression in the liver tissue was detected by the biochemical method. The mRNA levels of biological clock genes Per1, Per2, Cry1, Cry2, Bmal1, and Bmal2 in the liver were determined by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR). The protein and mRNA levels of factors in the OX1R/PLCβ-1/PKCα/ERK1/2 signaling pathway in the liver were determined by Western blot and Real-time PCR, respectively. ResultCompared with the blank group, the modeling decreased the body mass (P<0.05, P<0.01) and caused mania and disturbed resting rhythms (P<0.01), hepatic muscle fiber fracture, and edema with inflammatory cell infiltration. In addition, the modeling decreased the GABA, 5-HT, EPI, NE, and ACh content, increased Glu content (P<0.01), down-regulated the mRNA levels of Per1, Per2, Cry1, and Cry2 (P<0.01), up-regulated the mRNA levels of Bmal1 and Bmal2 (P<0.01), and promoted the expression of OX1R, PLCβ-1, PKCα, and ERK1/2 at both protein and mRNA levels (P<0.01). Compared with the model group, suvorexant and AMD increased the body mass (P<0.05, P<0.01), alleviated the mania, and increased the resting time and frequency (P<0.05, P<0.01). Moreover, the medications elevated the levels of GABA, 5-HT, EPI, NE, and ACh, lowered the Glu level, up-regulated the mRNA levels of Per1, Per2, Cry1, and Cry2 (P<0.05, P<0.01), down-regulated the mRNA levels of Bmal1 and Bmal2, and inhibited the expression of OX1R, PLCβ-1, PKCα, and ERK1/2 at both mRNA and protein levels (P<0.05, P<0.01). ConclusionAMD can regulate hepatic neurotransmitters and improve circadian rhythm in insomniac rats by inhibiting the OX1R/PLCβ-1/PKCα/ERK1/2 signaling pathway, and high-dose AMD demonstrated the strongest effect.
5.Effects of different fortified feeding methods on the nutrient metabolism and growth rate of very low birth weight preterm infants
Yue NING ; Liyan LUO ; Jiang DUAN ; Yunbo XIE ; Zhiye QI ; Caiying ZHANG ; Li YANG ; Qinghua ZHONG
Chinese Journal of Child Health Care 2024;32(1):84-88
【Objective】 To analyze the effects of different fortified feeding methods on nutritional metabolism and growth rate of preterm very low birth weight infants (VLBWI), in order to provide new clues for improving the prognosis of the preterm infants. 【Methods】 A total of 115 cases of premature VLBWI admitted to Department of Neonatology, The First Affiliated Hospital of Kunming Medical University from January 2019 to December 2020 were included in this study, and were divided into fortified breastfeeding group (HFM group), mixed feeding group, and premature formula feeding group (PF group) based on their feeding methods. The effects of different feeding methods on the nutritional metabolism and growth rate of premature VLBWI were analyzed. 【Results】 1) The hospitalization time of infants in the HFM group was shorter than that in PF group and mixed feeding group (t=7.185, 6.924, P<0.05). 2) The proportion of necrotizing enterocolitis (NEC) in the HFM group during hospitalization was lower than that in the PF group (P<0.05); the proportions of late onset septicemia(LOS) and extra uterine growth restriction(EUGR) in the HFM group during hospitalization were lower than those in the PF group (χ2=5.030, 4.147, P<0.05); the proportion of LOS was lower than that of the mixed feeding group(χ2=6.589, P<0.05). 3) During hospitalization, the proportions of abdominal distension, bloody stools and increased eosinophils in the HFM group were lower than those in the PF group (P<0.05), which in mixed feeding group was lower than those in PF group (Fisher exact test, P<0.05). 4) At discharge, the weight and length growth rate of the HFM group were higher than those of the mixed feeding group (t=3.722, 0.425, P<0.001) and the PF group (t =6.015, 0.496, P< 0.001). 【Conclusion】 Fortified breastfeeding can more effectively increase the growth rate of VLBWI in premature infants, improve nutritional metabolism, reduce complications and adverse feeding reactions related to premature infants, and is safer and more effective.
6.Role of endoplasmic reticulum stress in retinitis pigmentosa
Chen OU ; Wei XIE ; Qinghua PENG
International Eye Science 2024;24(12):1912-1916
Retinitis pigmentosa(RP)is a degenerative blinding disease characterized by the loss of the function of photoreceptor and retinal pigment epithelium. The activation of endoplasmic reticulum stress is a cellular defense regulatory mechanism, aimed at restoring the stability of endoplasmic reticulum function by self-regulation through a series of molecular signaling pathways. Rhodopsin mutation is a common cause of RP. Misfolding and retention of rhodopsin in endoplasmic reticulum and apoptosis of photoreceptor cells and retinal pigment epithelial cells induced by endoplasmic reticulum stress can lead to the occurrence and development of RP. This paper discusses endoplasmic reticulum stress and its role in the pathogenesis of RP and the role of endoplasmic reticulum stress inhibitors, traditional Chinese medicine and chemical drugs in regulating endoplasmic reticulum stress in RP treatment was summarized, in order to provide theoretical basis for endoplasmic reticulum stress in the clinical application of RP and provide new ideas for the research, prevention and treatment of RP.
7.The Effect of Ziyin Mingmu Formula (滋阴明目方) on Retinal Tissue Apoptosis and Endoplasmic Reticulum Stress in Retinitis Pigmentosa Model Mice
Chen OU ; Houpan SONG ; Wei XIE ; Ying WANG ; Jun PENG ; Qinghua PENG
Journal of Traditional Chinese Medicine 2024;65(17):1808-1815
ObjectiveTo study the effect of Ziyin Mingmu Formula (滋阴明目方) on retinitis pigmentosa mice and its possible mechanism. MethodsSixty rd10 mice were randomly divided into model group, vitamin A group, and Ziyin Mingmu Formula low-, medium-, and high-dose groups, 12 mice in each group; twelve C57BL/6 mice as the blank group. The mice in Ziyin Mingmu Formula low-, medium-, and high-dose groups were given 13.5, 27.0 and 54.0 g/(kg·d) of Ziyin Mingmu Formula by gavage, respectively; mice in the Vitamin A group were given 750 IU/(kg·d) of vitamin A soft capsule by gavage; mice in the blank group and model group were given 13.5 ml/(kg·d) of physiological saline by gavage. Each group was gavaged once a day for 28 days. Fundus photography and optical coherence tomography were used to detect the morphology of the fundus and retina, TUNEL method was used to detect apoptosis, microtitre digital PCR and immunofluorescence double staining were used to detect glucose-regulated protein 78 (GRP78), inositol-requiring enzyme 1 (IRE1), activated transcription factor 6 (ATF6), and C/EBP homologous protein (CHOP) mRNA and protein expression levels. ResultsIn the model group, optic discs were yellowish, fundus was atrophic with large pigmentation. The retinal blood vessels were thinned, a large number of vascular white sheaths were seen, the retinal cell nuclei disorganised, and a large number of apoptotic cells were seen; the fundus conditions of the mice in all dosages of Ziyin Mingmu Formula and the vitamin A group were improved to varying degrees compared with those of the model group, the apoptotic cells reduced, with Ziyin Mingmu Formula middle- and high-dose groups showing more significant improvement. Compared with the blank group, the retinal thickness of mice in the model group reduced, and the expression of GRP78, IRE1, ATF6, CHOP mRNA and protein elevated (P<0.01). Compared with the model group, retinal thickness increased and GRP78, IRE1, ATF6 mRNA and protein expression decreased in the vitamin A group and Ziyin Mingmu Formula medium- and high-dose groups; CHOP mRNA and protein expression decreased in low-, medium-, and high-dose groups of Ziyin Mingmu Formula (P<0.05 or P<0.01). Compared with vitamin A group and the low-dose group of Ziyin Mingmu Formula, retinal thickness increased in the high-dose group of Ziyin Mingmu Formula (P<0.01). Compared with vitamin A group, GRP78, ATF6, IRE1, and CHOP mRNA expression decreased in the medium- and high-dose groups of Ziyin Mingmu Formula, and GRP78 and CHOP protein expression decreased (P<0.05 or P<0.01). Compared with the low-dose group of Ziyin Mingmu Formula, CHOP protein expression reduced in the medium- and high-dose groups of Ziyin Mingmu Formula (P<0.01). ConclusionZiyin Mingmu Formula can reduce apoptosis of retinal tissues by inhibiting the endoplasmic reticulum stress response, thus slowing down the progression of retinopathy and protecting visual function in a dose-dependent manner.
8.Analysis of the characteristics of newly-diagnosed occupational diseases among female workers in Foshan City, 2013-2023
Zhiheng XU ; Qinghua ZHONG ; Yongshun HUANG ; Na DENG ; Yingqing XIE ; Qiang TAN
China Occupational Medicine 2024;51(3):336-340
Objective To analyze the characteristics of newly-diagnosed occupational diseases among female workers in Foshan City from 2013 to 2023. MethodsClinical data of newly-diagnosed occupational diseases among female workers in Foshan City from 2013 to 2023 were collected from the "Occupational Disease and Hazardous Detection Information System" subsystem of the "China Disease Prevention and Control Information System", and their distribution characteristics were analyzed using descriptive epidemiology. Results A total of 218 cases of newly-diagnosed occupational diseases among female workers in Foshan City were reported from 2013 to 2023, categorized into seven categories and 17 types. The median age and median length of exposure to occupational hazards among female workers were 45.1 and 7.8 years, respectively. The top three categories of newly-diagnosed cases were occupational diseases of the ear, nose, throat, and oral cavity (90 cases, all related to occupational noise-induced hearing loss), occupational chemical poisoning (61 cases), and occupational pneumoconiosis and other respiratory diseases (51 cases, including 50 cases with occupational pneumoconiosis). The newly-diagnosed cases were concentrated in Nanhai District, Shunde District, and Gaoming District, accounting for 75.2%. The newly-diagnosed cases were found in non-metallic mineral products manufacturing industry, metal products industry, and electrical machinery and equipment manufacturing industry, accounting for 50.9%. Enterprises of newly-diagnosed cases were mainly distributed in private enterprises and foreign-invested enterprises, accounting for 80.3%. The scale of enterprises of newly-diagnosed cases was mainly small- and medium-sized, accounting for 79.4%. Among the 218 cases, there were 21 cluster cases, involving 16 cases of occupational silicosis and five cases of occupational acute n-hexane poisoning, involving six manufacture industries, with 81.3% cases worked in sanitary ceramic products industry. Conclusion Occupational noise-induced deafness should be listed as a key occupational disease for female workers in Foshan City. Attention should be paid to the prevention of occupational diseases among middle-aged female workers in the non-metallic mineral products industry and small and medium-sized enterprises, and the cluster outbreaks of occupational pneumoconiosis.
9.Ziyin Mingmu Formula improve retinitis pigmentosa through the GRP78/IRE1/ATF6 pathway in vitro study
Chen OU ; Houpan SONG ; Wei XIE ; Jun PENG ; Meiyan ZENG ; Qinghua PENG
Journal of Beijing University of Traditional Chinese Medicine 2024;47(6):773-781
Objective We aimed to explore the mechanism of Ziyin Mingmu Formula in improving retinitis pigmentosa.Methods Twenty SD rats were randomly divided into the blank serum group and the Ziyin Mingmu Formula containing serum group using a random number table method,with 10 rats in each group.The rats in the Ziyin Mingmu Formula containing serum group were given Ziyin Mingmu Formula(46.875 g/kg),while the rats in the blank serum group were given distilled water.The Ziyin Mingmu Formula containing serum and blank serum were prepared.Analysis of chemical components in Ziyin Mingmu Formula containing serum using liquid chromatography-mass spectrometry technology.Adult retinal pigment epithelial cell line-19(ARPE-19)cells were treated with tunicamycin to induce endoplasmic reticulum stress injury model.The optimal volume fraction of Ziyin Mingmu Formula containing serum was screened by CCK-8.ARPE-19 cells were divided into the blank group,the model group,the blank serum group,the Ziyin Mingmu Formula containing serum group and the tauroursodeoxycholic acid group,each group is intervened separately.After 24 hours of intervention,the morphological observation of cells was performed using a multi-time dynamic cell function analysis system,cell survival rate was detected by CCK-8,cell apoptosis rate was detected by Annexin V-FITC/PI,and protein and mRNA expressions of 78 kDa glucose regulated protein(GRP78),inositol-requiring enzyme 1(IRE1)and activating transcription factor 6(ATF6)were detected by Western blotting and ddPCR.Results The main chemical components in Ziyin Mingmu Formula containing serum were obtained by liquid chromatography-mass spectrometry analysis,such as arbutin,salicylic acid,luteolin,Salvianolic acid A,erysovine,taurine,quercetin,maltol,baicalin,and danshensu.Compared with the model group,the number of cells in Ziyin Mingmu Formula containing serum group increased,the growth was more uniform,and the floating dead ARPE-19 cells and fragments decreased.The cell survival rate of Ziyin Mingmu Formula containing serum group and taursodeoxycholic acid group increased,and the cell apoptosis rate decreased(P<0.05).The protein expressions of GRP78,IRE1,and ATF6 in Ziyin Mingmu Formula containing serum group were decreased,and IRE1 and ATF6 in taursodeoxycholic acid group were decreased(P<0.05).The mRNA expressions of GRP78,IRE1,and ATF6 in Ziyin Mingmu Formula containing serum group and tauroursodeoxycholic acid group were decreased(P<0.05).Conclusion Ziyin Mingmu Formula can reduce the cell apoptosis of ARPE-19 cells induced by endoplasmic reticulum stress,and its molecular mechanism is related to down-regulating the expression of GRP78,IRE1,ATF6 and inhibiting the endoplasmic reticulum stress response.
10.Factors influencing success of external cephalic version: analysis of 118 cases
Genxia LI ; Panpan ZHAO ; Chunhua CHENG ; Mingkun XIE ; Xianrong MENG ; Qinghua XU ; Jiao LI
Chinese Journal of Perinatal Medicine 2024;27(3):220-225
Objective:To investigate the factors influencing the success of external cephalic version.Methods:Pregnant women who underwent an external cephalic version due to breech or transverse presentation by the same operator in the Third Affiliated Hospital of Zhengzhou University from July 2015 to July 2021 were selected as the study objects. Univariate analysis and logistic regression analysis were used to explore the clinical factors influencing the success of the external cephalic version. The receiver operating characteristic (ROC) curve was used to analyze the best cut-off value of gestational week and amniotic fluid index at the time of operation and to evaluate the predictive value of the influencing factors on the success of the external cephalic version.Results:(1) A total of 118 cases finally entered this study. Among the 118 cases,77 cases (65.3%) succeeded in the external cephalic version, among which the success rate was 49.1% (27/55) for primipara and 79.4% (50/63) for multipara. The vaginal delivery rate was 56.8% (67/118). (2) Complications occurred in 19 (16.1%) of the 118 cases. The main complications were abnormal fetal heart rate (13 cases, 11.0%), umbilical cord presentation, and fetal position reversion (two cases and 1.7% in each), and the serious complications were intrauterine fetal death and placental abruption (one case and 0.8% in each).The complication rate of patients with successful external cephalic version was 7.8% (6/77), which was lower than that of those who failed the external cephalic version [31.7%(13/41)] ( χ 2=11.33, P=0.001). (3) Multivariate analysis showed that gestational week at surgery before 38, amniotic fluid index >11.10 cm, and multipara were the factors affecting the success of the external cephalic version [ OR(95% CI)=0.561(0.351-0.897), 1.173(1.018-1.351) and 4.201(1.547-11.404), all P<0.05]. (4) The area under the ROC curve of the combination of the gestational week at surgery, amniotic fluid index, and parity was 0.744 (95% CI: 0.640-0.848, P<0.001), and the Youden index was 0.518, with a sensitivity of 70.0% and a specificity of 81.8%. Conclusion:Gestational weeks, amniotic fluid index, and multipara are related to the success of the external cephalic version, and the combination of the three has certain predictive power for the success of the surgery.

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