1.Protective effect of Shenfu injection against neonatal hypoxic-ischemic brain injury by inhibiting the ferroptosis
Xiaotong Zhang ; Meng Zhang ; Gang Li ; Yang Hu ; Yajing Xun ; Hui Ding ; Donglin Shen ; Ming Wu
Acta Universitatis Medicinalis Anhui 2025;60(1):31-40
Objective :
To observe the brain tissue injury during hypoxia-ischemia, as well as the pathological changes and the expression of ferroptosis-related factors after the use of Shenfu injection(SFI), and to explore the protective effect of SFI on hypoxic-ischemic brain injury(HIBD) by inhibiting ferroptosis.
Methods :
An animal model of HIBD in SD rats was constructed and intervened with SFI. Pathologic changes in brain tissue were observed by HE staining methods. Nissen staining was used to observe neuron survival. Glutathione Peroxidase 4(GPX4) and Divalent Metal Transporter 1(DMT1) expression were detected in brain tissue by Western blot, immunohistochemistry and immunofluorescence. Reduced Glutathione(GSH), Lactate Dehydrogenase(LDH), Malondialdehyde(MDA), Superoxide Dismutase(SOD) and tissue iron content were determined with the kits. BV-2 microglial cell line(BV2) cells were culturedin vitroand divided into control group(Ctrl group), oxygen-glucose deprivation group(OGD group), iron ferroptosis-inducing group(Erastin group), iron ferroptosis-inhibiting group(Fer-1 group), Shenfu injection group(SFI group), and Erastin+Shenfu injection group(Erastin+SFI group). 2′,7′-Dichlorodihydrofluorescein diacetate(DCFH-DA) reactive oxygen species(ROS) fluorescent probe was used to detect the ROS release level; Immunofluorescence was used to observe intracellular GPX4, DMT1 expression.
Results :
Compared with the Sham group, rats in the HIBD group showed significant neuronal cell damage in brain tissue, decreased GPX4 expression(P<0.01), increased DMT1 expression(P<0.01), decreased GSH and SOD levels(P<0.01), and increased LDH, MDA and tissue iron levels(P<0.05,P<0.05,P<0.01). In contrast, after the intervention of SFI, GPX4 expression was elevated(P<0.01), DMT1 expression decreased(P<0.01), GSH and SOD levels were elevated(P<0.01), and LDH, MDA, and tissue iron levels decreased(P<0.05,P<0.05,P<0.01). The cells experiments showed that compared with the Ctrl group, the OGD group had a significantly higher ROS content and a decrease in the expression of GPX4 fluorescence intensity, and an increase in the fluorescence intensity of DMT1(P<0.01), compared with the OGD group, the ROS content was reduced in the SFI group, while the expression of GPX4 was elevated and the expression of DMT1 was reduced(P<0.01).
Conclusion
Hippocampal and cortical regions are severely damaged after HIBD in neonatal rats, and their brain tissues show decreased expression of GPX4 and increased expression of DMT1. The above suggests that ferroptosis is involved in HIBD brain injury in neonatal rats. In contrast, Shenfu injection has a protective effect on HIBD experimental animal model and BV2 cell injury model by reducing iron aggregation and ROS production.
2.Clinical application value of intracavitary PRP infusion combined with IVF-FET in patients with chronic endometritis
Xiaotong ZHANG ; Xiaoyuan HAO ; Rui FANG ; Shuyao HU ; Linkun MA ; Yaqi ZHAO ; Wei HAN
Chinese Journal of Blood Transfusion 2025;38(3):382-387
[Objective] To evaluate the clinical application value of intrauterine perfusion with platelet-rich plasma (PRP) combined with in vitro fertilization-frozen-thawed embryo transfer (IVF-FET) in patients with chronic endometritis (CE). [Methods] A randomized controlled trial (RCT) was conducted, enrolling 60 CE patients undergoing artificial cycle frozen embryo transfer at our hospital from January 2022 to January 2024. Participants were randomly divided into three groups: Group A (routine frozen embryo transfer, n=20), Group B (routine frozen embryo transfer + one PRP intrauterine perfusion, n=20), and Group C (routine frozen embryo transfer + two PRP intrauterine perfusions, n=20). Endometrial thickness during the transformation and transplantation phases, uterine artery pulsatility index (PI), resistance index (RI), systolic peak velocity/end-diastolic velocity (S/D) ratio during transplantation, serum levels of IL-2, IL-4, IL-6, IL-10, and TNF-α during transplantation, as well as biochemical pregnancy rate, clinical pregnancy rate, live birth rate, and early miscarriage rate were compared across groups. [Results] No significant differences in endometrial thickness were observed among the three groups during the transformation phase (P>0.05). During the transplantation phase, endometrial thickness in Groups C and B was significantly higher than in Group A[9.54 (8.96-10.22) and 8.90 (8.34-9.72) vs 8.37 (7.89-8.75) mm, P<0.05], with Group C showing greater thickness than Group B (Z=3.733, P<0.05). Endometrial thickness in Groups C and B during transplantation was significantly increased compared to their respective transformation phases (Z=2.191, 2.462; P<0.05). Groups C and B exhibited lower PI, RI, and S/D values than Group A[PI:1.87 (1.77-1.97), 1.94 (1.88-2.15) vs 2.43 (2.35-2.49); RI:0.75 (0.73-0.77), 0.78 (0.75-0.81) vs 0.84 (0.83-0.86); S/D:2.61 (2.33-3.42), 3.01 (2.20-3.93) vs 3.72 (3.06-4.49); P<0.05]. Group C demonstrated lower PI and RI than Group B (P<0.05). IL-2 levels in Groups C and B were higher than in Group A[3.88 (2.71-5.01), 3.59 (2.73-4.38) vs 3.16 (2.11-3.25) ng/L, P<0.05], while IL-4, IL-6, IL-10, and TNF-α levels were significantly lower (IL-4: Z=1.428, 2.421; IL-6: Z=1.754, 2.435; IL-10: Z=1.754, 2.854; TNF-α: Z=1.961, 1.765; P<0.05). Group C had lower IL-6 levels than Group B (Z=3.976, P<0.05). Biochemical pregnancy rate, clinical pregnancy rate, and live birth rate in Group C were significantly higher than in Group A (75% vs 40%, 70% vs 35%, 60% vs 20%, P<0.05). No significant differences in early miscarriage rates were observed among the groups (χ2=3.750, P>0.05). [Conclusion] Intrauterine autologous PRP perfusion in CE patients enhances pregnancy and live birth rates, improves pregnancy outcomes post-FET, and demonstrates superior efficacy in endometrial repair and receptivity with two PRP perfusions compared to a single perfusion. This provides a safe and effective therapeutic option for optimizing outcomes in CE patients with prior implantation failure.
3.Development and validation of a prediction score for subtype diagnosis of primary aldosteronism.
Ping LIU ; Wei ZHANG ; Jiao WANG ; Hongfei JI ; Haibin WANG ; Lin ZHAO ; Jinbo HU ; Hang SHEN ; Yi LI ; Chunhua SONG ; Feng GUO ; Xiaojun MA ; Qingzhu WANG ; Zhankui JIA ; Xuepei ZHANG ; Mingwei SHAO ; Yi SONG ; Xunjie FAN ; Yuanyuan LUO ; Fangyi WEI ; Xiaotong WANG ; Yanyan ZHAO ; Guijun QIN
Chinese Medical Journal 2025;138(23):3206-3208
4.Effects of platelet isolation optimization and its activation productson on proliferation of endothelial progenitor cells
Jiajun XIAO ; Yue ZHAO ; Lu BAI ; Cheng XU ; Jinhua ZUO ; Yahui HU ; Kai XIA ; Bicheng WANG ; Xiaotong XIE ; Xiangxiang TANG
Chongqing Medicine 2025;54(10):2269-2274
Objective To optimize the platelet enrichment method,and to analyze the concentration changes of key molecules in platelet-rich plasma(PRP)before and after activation,as well as the impact of its activated products on the proliferation of rat endothelial progenitor cells.Methods The tube double-centrifu-gation method was employed to optimize platelet enrichment,and the platelet count in the enriched PRP was measured.ELISA was used to detect the concentration changes of vascular endothelial growth factor(VEGF),endostatin(ES),and P-selectin(CD62P)in PRP before and after activation.The PRP was activated by using liquid nitrogen freeze-thaw method,and the effect of its activated products on the proliferation of rat endothelial progenitor cells was evaluated by using the methyl thiazolyl tetrazolium(MTT)assay.Results The optimal enrichment coefficient of platelets achieved by the double-centrifugation method was 4.63.After low-speed,long-duration double centrifugation,the platelet count was highest in the upper layer of the buffy coat.For PRP with a platelet count of 500× 109/L obtained by machine collection,the VEGF con-centrations before and after activation were(3 418.12±488.80)pg/mL and(4 530.04±308.30)pg/mL,re-spectively,the ES concentrations were(6 168.98±253.22)pg/mL and(6 594.65±82.47)pg/mL,respec-tively,the CD62P concentrations were(6 678.23±324.15)pg/mL and(17 630.53±746.24)pg/mL,respec-tively,statistically significant differences were observed in the above indicators before and after activation(P<0.01).The activated PRP was diluted in a gradient manner by using a specialized culture medium for en-dothelial progenitor cells.MTT assay results indicated that,in the basal medium,the optimal volume fraction for promoting endothelial progenitor cell proliferation was 0.25%after 48 hours of culture;in the complete medium,the optimal volume fractions for promoting endothelial progenitor cell proliferation were 0.062 5%after 24 hours and 0.125%after 48 hours.Conclusion The concentrations of VEGF,ES,and CD62P in the optimized,enriched PRP exhibited significant changes before and after activation.The optimal volume fraction for promoting endothelial progenitor cell proliferation in the basal medium was 0.25%.
5.A real-world study on the influence of Qishi Tongguan Prescription on the pregnancy outcome of patients with tubal factor infertility after interventional recanalization
Xiaoqing GAO ; Xiaole ZHANG ; Panwei HU ; Xiaotong YAN ; Cong QI
International Journal of Traditional Chinese Medicine 2024;46(2):168-174
Objective:To evaluate the effect of Qishi Tongguan Prescription on pregnancy outcomes after interventional recanalisation in patients with tubal infertility (TFI).Methods:This was a retrospective study based on real-world and propensity score matching. Totally 260 patients with TFI from January 2020 to October 2021 in Shuguang Hospital of Shanghai University of Traditional Chinese Medicine and Maternal and Child Health Hospital of Pudong New Area were selected as observation subjects, and were divided into 123 cases in the TCM combination group and 137 cases in the control group based on whether they were treated with Qishi Tongguan Prescription in combination with interventional revascularization. Propensity score matching (PSM) was used as a covariate to obtain a new sample of inter group covariate equilibrium, and confounding factors that may affect the pregnancy outcome of TFI patients undergoing interventional recanalization surgery were used as covariates. The intrauterine pregnancy rate, ectopic pregnancy rate, biochemical pregnancy rate, early abortion rate and adverse reactions of the two groups of patients within 12 months of follow-up were compared, and the influence of TFI intervention and recanalization combined with Qishi Tongguan Prescription on intrauterine pregnancy rate was evaluated.Results:Age, years of infertility, type of infertility, history of miscarriage, history of ectopic pregnancy, history of biochemical pregnancy, history of uterine surgery, history of pelvic laparotomy, and degree of tubal patency had an effect on whether intrauterine pregnancy was achieved after interventional reversal in patients with TFI ( P<0.05), with age [ OR (95% CI) was 0.843 (0.769, 0.926)], history of pelvic laparotomy [ OR (95% CI) was 0.477 (0.248, 0.920)] and the degree of tubal obstruction [ OR (95% CI) was 0.152 (0.046, 0.500)] were independent factors ( P<0.01 or P<0.05). 81 patients were seen in each of the 2 groups after PSM, of whom the intrauterine pregnancy rates in the combined herbal group at 9 and 12 months after recanalisation were 48.1% (39/81) and 58.0% (47/81) respectively, compared with 32.1% (26/81) and 35.8% (29/81) in the control group, with statistical significance between the 2 groups ( χ2 values of 4.34 and 8.03, respectively, P<0.01); there was no statistical significance in the ectopic pregnancy rate, biochemical pregnancy rate and early abortion rate between the 2 groups ( P>0.05). There were no significant adverse reactions during the treatment. Conclusion:Qishi Tongguan Prescription combined with interventional recanalization can effectively improve the intrauterine pregnancy rate and shorten the waiting time for pregnancy in patients with TFI with higher safety.
6.Clinical characteristics and risk prediction model construction of children with Kawasaki disease complicated with pleural effusion
Hu YANG ; Jun LEI ; Li YANG ; Xiaotong PENG
Tianjin Medical Journal 2024;52(10):1089-1094
Objective To investigate the clinical features of pediatric Kawasaki disease(KD)complicated with pleural effusion(PE)and construct a clinical prediction model.Methods A retrospective review was conducted on clinical data of 462 children with KD from June 2017 to June 2023.Eleven KD children with PE were selected as the KD-PE group,and 118 patients without PE were selected as the KD control group.Using propensity score matching,the clinical data including clinical manifestation,auxiliary examination,treatment process and complications were collected and analyzed.Univariate and Lasso-Logistic regression were used to analyze influence factors of PE in children with KD.A line chart model for predicting KD-PE was constructed,and the prediction efficiency of the model was evaluated.Results The respiratory symptoms were generally mild in the KD-PE group,with 10 cases exhibiting mild PE.Two variables of C-reactive protein(CRP,OR=1.045,95%CI:1.009-1.082)and albumin(ALB,OR=0.755,95%CI:0.591-0.964)were screened out by Lasso regression and multi-factor Logistic regression to construct the prediction model.The regression equation was Logit(P)=2.221+0.044×CRP-0.281×ALB.The area under the ROC curve of the prediction model was 0.957(95%CI:0.911-1.000),and the Hosmer-Lemeshow test showed that the curve of the calibration model was well coincident with the curve of the actual model(χ2=4.320,P=0.827).The evaluation model using clinical decision curve and clinical impact curve had good clinical applicability.Conclusion KD complicated with PE in children is rare,elevated CRP is a risk factor for KD-complicated PE,and elevated ALB is a protective factor.The model has higher accuracy,discriminatory power and net benefit.
7.Construction of a stable 4T1 cell line expressing UL19 by the PiggyBac transposon system.
Xiaotong ZHAO ; Xinya WANG ; Binlei LIU ; Han HU ; Yang WANG
Chinese Journal of Biotechnology 2024;40(11):4138-4148
To investigate the mechanism of the major capsid protein VP5 (encoded by the UL19 gene) of oncolytic herpes simplex virus type Ⅱ (oHSV2) in regulating the antitumor function of immune cells, we constructed a mouse breast cancer cell line 4T1-iRFP-VP5-GFP stably expressing VP5 protein, near-infrared fluorescent protein (iRFP), and green fluorescent protein (GFP) by using the PiggyBac transposon system. Flow cytometry and Western blotting were employed to screen the monoclonal cell lines expressing both GFP and VP5 and examine the expression stability of UL19 in the constructed cell line. The results of SYBR Green I real-time PCR and Western blotting showed that the copies of UL19 and the expression level of VP5 protein in the 15th passage of 4T1-iRFP-VP5-GFP cells were significantly higher than those in the 4T1 cells transiently transfected with UL19, demonstrating the stable insertion of UL19 into the 4T1 cell genome. The real-time cell analysis (RTCA) was employed to monitor the proliferation of 4T1-iRFP-VP5-GFP cells, which showed similar proliferation activity to their parental 4T1 cells. Further studies confirmed that NK92 cells exhibited stronger cytotoxicity against 4T1-iRFP-VP5-GFP cells than against 4T1 cells. This study layed a foundation for elucidating the role of VP5 protein in regulating immune cells, including T cells and NK cells, via HLA-E in 4T1 cells to exert the anti-tumor function.
Animals
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Mice
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DNA Transposable Elements/genetics*
;
Cell Line, Tumor
;
Capsid Proteins/biosynthesis*
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Transfection
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Green Fluorescent Proteins/metabolism*
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Oncolytic Viruses/genetics*
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Female
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Simplexvirus/genetics*
8.Practice and Reflection on the Application of Tongue Image APP for Observing Tongue Image Characteristics of Epidemic Diseases
Yan YANG ; Meng WU ; Xiaotong MA ; Zhaoying ZHENG ; Xiaodong LI ; Xinhe ZUO ; Jingqing HU
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(8):2653-2659
Tongue diagnosis is an indispensable objective basis for TCM diagnosis and treatment of epidemic diseases.To understand its application in an epidemic situation and to support in the diagnosis and treatment of infectious diseases using traditional Chinese medicine,the tongue image APP was implemented in this study to monitor the tongue image features of patients with new coronavirus pneumonia.It has been discovered through practice that the tongue image APP enables medical professionals to objectively,conveniently,quickly,and flexibly collect the patient's tongue image.It has also been discovered through the analysis of the tongue image characteristic data that the tongue image APP can,to a certain extent,objectively reflect the general law of the tongue image characteristics of the new crown pneumonia.According to the tongue image data gathered by the Tongue Image APP,Xinguan pneumonia patients'tongues were typically red,their fur was typically white,yellow,or both white and yellow,and they had a greater amount of thick and greasy fur.Nevertheless,there are still several issues with the Tongue APP application that have been noted:①The consistency of tongue shape and coating was poor;for instance,the inconsistency rate between a thin and fat tongue was as high as 62.96%;②The tongue image analysis index in the APP is still mostly a qualitative index,and the degree of discriminating is insufficient.The results of this study demonstrate that the tongue image information of different ages,sexes,disease classifications,and onset times does not reflect obvious differences and certain rules.③The tongue image characteristic indexes gathered by the tongue image APP are insufficient and do not include information on glossiness of tongue image(such as dark tongue)and tongue state.To promote the adoption of the tongue image APP and better support the prevention and treatment of epidemic diseases by traditional Chinese medicine,we should fully integrate modern advanced science and technology,improve the short videos of tongue coating,quantification of qualitative indicators,comprehensive collection of tongue image characteristic indicators,etc.
9.Research Progress of Tai Chi's Influence on Brain Structure and Function of the Elderly Based on Magnetic Resonance Imaging Technology
An WU ; Boran ZHANG ; Xiaotong WANG ; Chang HUANG ; Boyi HU ; Yunpeng YANG ; Tingchao WU ; Feng ZHANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2023;25(12):3961-3968
Magnetic Resonance Imaging(MRI)technology can directly show the changes of brain network and explain the central mechanism of Tai Chi remodeling of brain structure and function.In this paper,we collected the domestic and foreign research on the influence of Tai Chi movement on the brain network by using MRI technology,and combed it from the perspective of brain structure and function changes.The results revealed that Tai Chi may promote memory function,cognitive flexibility,inhibitory control,and working memory capacity by remodeling the structure and function of the medial temporal lobe and prefrontal cortex in older adults,which may be a potential central mechanism for Tai Chi to improve memory and cognitive control in the elderly.However,there are some problems in the current research,such as small sample size,insufficient long-term follow-up,and difficult evaluation of exercise intensity.It is necessary to carry out large-sample and long-term detailed research to further verify the current research results.
10.Potential impact of outdoor intervention on the prevalence of myopia: a simulation study
Yanxian CHEN ; Jian ZHANG ; Xiaotong HAN ; Yin HU ; Lili WANG ; Mingguang HE
Chinese Journal of Experimental Ophthalmology 2023;41(12):1203-1207
Objective:To evaluate the long-term impact of three different outdoor activity intervention strategies on the prevalence of myopia among school-aged children in China.Methods:Cross-sectional data from the Refractive Error Study in Children (RESC) on the baseline prevalence of myopia and 10-year longitudinal data from the Guangzhou Twin Eye Study (GTES) on the baseline incidence of myopia were utilized.The effect size of outdoor activity intervention was derived from longitudinal data of the Guangzhou Outdoor Activity Longitudinal Study (GOALS). Three intervention strategies were proposed: intervention for children in grades 1 to 6 (Strategy A), intervention for children in grades 1 to 3 (Strategy B), and intervention for children from the last year of kindergarten to grade 3 (Strategy C). Outdoor activity interventions across all strategies included an extra 40 minutes of daily outdoor activity time.Simulated analyses assumed that the prevalence of myopia would decrease with declining incidence and increasing school grade, and the degree of myopia among highly myopic children would decrease with delayed myopia onset.This study was approved by the Ethics Committee of Zhongshan Ophthalmic Center (No.2006-5).Results:With prolonged intervention time, the overall prevalence of myopia in children showed a significant declining trend.After 5 years of implementation, Strategy A resulted in a decrease in myopia prevalence from 34.67% to 25.71%, which decreased to 24.89% after 10 years, with an average annual reduction of 0.98%.Strategies B and C had similar long-term effects, with myopia prevalence after 10 years being 26.00% and 26.20% respectively, with an average annual reductions of 0.87% and 0.85%.Strategy A yielded the greatest reduction in myopia severity, with a decrease of -5.94 D after 5 years and -5.88 D after 10 years in the initial 15-year-old highly myopic individuals.Strategy B followed with -6.08 D and -6.02 D after 5 and 10 years, respectively.Conclusions:Implementing intensified outdoor activity intervention among children in grades 1 to 6 may be the recommended strategy to reduce the overall prevalence of myopia in regions with high rates of myopia among school-aged children.


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