1.The construction and evaluation of heart preservation model for empty beating donor heart based on extracorporeal membrane oxygenation technology
Shijie YIN ; Xiao YUE ; Chunhua WANG ; Wei WU ; Guanbin QIN ; Lan LUO ; Qiangxin HUANG ; Guixin HE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2026;33(05):791-798
Objective To explore the construction of heart preservation model of empty beating donor based on extracorporeal membrane oxygenation (ECMO). Methods From January 2022 to August 2023, 20 Guangxi Bama miniature pigs weighing 25-30 kg were selected, half male and half female. Under general anesthesia and heparinization, a midline thoracotomy was performed. The pericardium was cut after freeing the anterior and posterior vena cavae, and a perfusion needle was inserted near the brachiocephalic artery in the ascending aorta, connected to a blood collection bag to collect 500-600 mL of blood. The anterior and posterior vena cavae were ligated, the aorta was blocked and perfused with HTK solution to stop the heart beating. The superior and inferior vena cavae were cut off, the right pulmonary vein was decompressed, the aorta and left and right pulmonary arteries and veins were cut off, and the whole heart was removed. An ECMO device was used to continuously perfuse a cardioprotective solution mainly composed of oxygenated warm blood, maintaining the isolated pig heart beating for 8 hours, monitoring (once/hour) ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, inflammatory factors, myocardial enzymes, myoglobin, and troponin levels. Myocardial tissue was taken for hematoxylin-eosin (HE) staining to observe myocardial cell damage and evaluate the quality of heart preservation. Results Among the 20 isolated beating pig hearts, 17 successfully resumed beating, 3 experienced ventricular fibrillation, resuscitated after intracardiac electrical defibrillation, and all 20 pig hearts successfully beat for 8 hours. There was no statistical difference in ECMO perfusion parameters, blood gas indicators, perfusate electrolytes, and inflammatory factors at each time point (P>0.05). There were statistical increases in myocardial enzymes, myoglobin, and troponin levels (P<0.05). HE staining results suggested that there was no severe myocardial damage. Conclusion ECMO technology can be used for pig heart preservation with good results, and this study provides experimental evidence for improving heart preservation research in clinical heart transplantation.
2.Exploring Mechanism of Modified Danggui Yinzi in Regulating "Itch-anxiety" Cycle of Chronic Urticaria Based on STEP/NR2B Signaling Pathway
Mingyue LI ; Xinyu XIAO ; Anjing CHEN ; E LIU ; Xurui WANG ; Qin ZHOU
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(13):123-133
ObjectiveTo explore the effects and mechanism of the modified Danggui Yinzi on "itch-anxiety" model rats of chronic urticaria (CU). MethodsThe 36 SPF-grade 6-8-week-old female SD rats were randomly divided into a blank control group,a model group,a positive control group,a low-dose modified Danggui Yinzi group,a medium-dose modified Danggui Yinzi group,and a high-dose modified Danggui Yinzi group. A "itch-anxiety" model was established by intraperitoneal injection of a suspension of sodium chloride and aluminum hydroxide and ovalbumin,combined with chronic unpredictable emotional stress stimulation. After successful modeling,rats in each group were administered drugs by gavage. The positive control group was given intragastric administration of the drug solutions of cetirizine and fluoxetine (2.08 mg·kg-1·d-1 fluoxetine, 2 mg·kg-1·d-1 cetirizine), the low-,medium-,and high-dose modified Danggui Yinzi groups were administered traditional Chinese medicine at 1.44,2.88, 5.76 g·kg-1, respectively,while the blank control group and model group were given an equal volume of normal saline. All interventions lasted for 15 days. Behavioral changes were evaluated by the elevated plus-maze test (detecting the percentage of entries into the open arms (OE%),the percentage of time spent in the open arms (OT%),and the total number of entries into the open and closed arms (TNE)),the open-field test (detecting total activity,average movement speed,and latency to enter the central area),and scratching behavior observation. Pathological changes of skin tissues were observed by hematoxylin-eosin (HE) staining and toluidine blue staining,while those of amygdala tissues were observed by HE staining,Nissl staining,and immunofluorescence detection of ionized calcium-binding adapter molecule-1 (Iba-1). The content of immunoglobulin E (IgE),interleukin-33 (IL-33),histamine in serum and glutamate in the amygdala was detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the protein expression of striatal-enriched protein tyrosine phosphatase (STEP),N-methyl-D-aspartate receptor subunit 2B (NR2B), calmodulin-dependent protein kinase Ⅱ (CaMKⅡ),phosphorylated CaMKⅡ (p-CaMKⅡ),mitogen-activated protein kinase (MAPK),phosphorylated MAPK (p-MAPK),nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB),phosphorylated NF-κB (p-NF-κB),and postsynaptic density protein-95 (PSD-95) in the amygdala. ResultsCompared with the blank control group,the model group rats showed obvious anxiety-like behaviors (decreased OE%,OT%,and TNE,reduced total activity,slower average movement speed,and prolonged latency to enter the central area),increased scratching times,obvious skin inflammation and mast cell degranulation,severe amygdala tissue damage,increased glutamate content in the amygdala,and elevated levels of IgE and IL-33 in serum. The expression of STEP,NF-κB,p-NF-κB,NR2B,MAPK,p-MAPK,CaMKⅡ,and p-CaMKⅡ proteins in the amygdala increased,while the expression of PSD-95 protein decreased (P<0.05). Compared with the model group,the modified Danggui Yinzi group of each dose had increased OE%,OT%,TNE,total activity,and average movement speed,shortened latency to enter the central area, reduced scratching times,alleviated skin inflammation and mast cell degranulation,relieved amygdala tissue damage,decreased glutamate content in the amygdala,and reduced levels of IgE and IL-33 in serum. Moreover,compared with the model group,the low -,medium-,and high-dose modified Danggui Yinzi groups showed decreased expression levels of STEP,NF-κB,p-NF-κB,NR2B,MAPK,p-MAPK,CaMKⅡ,and p-CaMKⅡ proteins in the amygdala,and increased expression of PSD-95 protein. There was a significant dose-effect relationship,with the high-dose group showing the most significant regulatory effect (P<0.05). ConclusionThe modified Danggui Yinzi has a therapeutic effect on "itch-anxiety" model rats of CU. Its mechanism may be related to regulating glutamate metabolism in the amygdala,modulating the STEP/NR2B/CaMKⅡ/MAPK/NF-κB pathway,and regulating the expression of PSD-95.
3.Gene Mutation Analysis of an Individual with a RHD Variant RHD*DV.5 and RHD Negative Gene Heterozygote
Shi-Shi WU ; Juan PENG ; Li-Bo WU ; Hong-Xiao CHEN ; Dong-Mei ZHAN ; Yue-Mei DONG ; Wan-Qin WANG ; Liang WU
Journal of Experimental Hematology 2025;33(6):1758-1764
Objective:To perform RHD gene detection on a blood sample with serological weak D phenotype.Methods:A specimen received by the People's Hospital of Zhijin County was serologically identified by the microcolumn gel method and saline method.RHD gene detection was conducted by the PCR-SSP method,and the full sequence determination of the 10 exons amplified was performed.The sequencing results were compared with the ISBT database to determine the genotype.Bioinformatics tool was used to predict the functional damage of mutant proteins,and Alphafold-3 was used for tertiary structural modeling of wild-type and mutant RhD proteins,and the structures of the two proteins were compared and analyzed to explore the reasons why mutations lead to weak serological manifestations.Results:The patient's genotype was identified as RHD*DV.5/RHD*01N.01 heterozygote,with the complete deletion of RHD genes on one chromosome,unable to express the D antigen.On the other chromosome,a G>A mutation occurred at the 697th base of the 5th exon,resulting in a partial D phenotype.This mutation causes internal hydrogen bond changes at the 233 position of RhD protein,resulting in a change in the conformation of the protein,affecting binding to the corresponding antibody.Conclusion:The patient is a heterozygous mutant individual with RHD*DV.5/RHD*01N.01,exhibiting a partial D phenotype serologically.This variation is extremely rare and has been scarcely reported globally.
4.Inhibition of excessive inflammatory response of macrophages by Ebselen against acute Escherichia coli infection
Xiao-wen LIU ; Xiao-qin MOU ; Chuang CHENG ; Shuang-shuang GONG ; Hao-ran ZHANG ; Jing HE ; Xi ZHENG ; Jun WANG ; Yue-qing WANG ; Li-li ZOU
Chinese Pharmacological Bulletin 2025;41(7):1346-1353
Aim To investigate the pharmacological mechanism of Ebselenin(Ebselen,EbSe)in the treat-ment of Escherichia coli(E.coli)infection,which had no significant inhibitory effect on Gram-negative bacte-ria,based on previous studies.Methods After EbSe intervention in E.coli infected Raw264.7 cells,the via-bility of Raw264.7 cells was determined by CCK-8 method,the morphology and structure of Raw264.7 cells were observed by electron microscope,and the in-tracellular bacterial load of Raw264.7 cells was calcu-lated by coated plate method.Polarization status of peritoneal macrophages,Raw264.7 intracellular NO and ROS content and intracellular HO-1 expression in Raw264.7 and E.coli acutely infected mice after E.co-li infection by flow cytometry.qPCR was used to detect the expression of related mRNAs in Raw264.7 cells.qPCR was used to detect the intracellular GSH content in Raw264.7 cells by spectrophotometric assay,and the state of cytoskeletal proteins was observed by immuno-fluorescence.Western blot assay was performed to de-tect the intracellular Txnrd1 expression level.Results Microtiter method,CCK-8,and electron microscopy observations showed that EbSe had no effect on the growth of E.coli and Raw264.7 cells in vitro.The re-sults of smear plate counting showed that EbSe reduced the intracellular bacterial load of Raw264.7 in the in-fected group.Flow cytometry results showed that EbSe upregulated the number of M2-type macrophages.The EbSe-treated infected group had reduced intracellular NO and ROS levels and increased GSH levels.The qPCR results showed that the expression of IL-6,IL-1β,and iNOS was decreased,and the expression of HO-1,Txnrd1,and Glut1 was increased in DHB4-in-fected Raw264.7 cells after EbSe treatment.Cytoskel-etal staining showed that the morphology of the EbSe-treated infected cells was similar to that of oxPAPC-in-duced cells.Western blot results showed the expres-sion of Txnrd1 protein in EbSe-treated infected cells in-creased.Conclusion EbSe exerts anti-E.coli acute infection effect by regulating macrophage polarization and inhibiting macrophage excessive inflammatory state.
5.Mechanism of G9a histone methyltransferase inhibitor BIX01294 inhibiting the proliferation of vascular smooth muscle cell
Ze-lan WANG ; Wen-jian LUO ; Jun-yong ZHAO ; Xiao-lin LUO ; Zhe-xue QIN
Journal of Regional Anatomy and Operative Surgery 2025;34(4):279-283
Objective To investigate the effect of G9a histone methyltransferase inhibitor BIX01294 on the proliferation of vascular smooth muscle cell(VSMC)and its underlying mechanism.Methods Twelve SD rats were divided into the control group and the treatment group,with 6 rats in each group.The rats were anesthetized by intraperitoneal injection with sodium pentobarbital,then exposed the common carotid artery and the internal and external carotid artery after disinfection and skin preparation,ligated the distal end of the internal and external jugular veins,clamped the proximal end of the common carotid artery,cut the external carotid artery,inserted the balloon catheter,blocked the blood flow by compression,and ligated the proximal end of the external carotid artery after withdrawing the balloon.The common carotid artery clamping state of rats in the treatment group was maintained after withdrawing the balloon,then restored the blood flow after perfusion with 100 μmol/L BIX01294 solution for 30 seconds;rats in the control group were restored the blood flow after perfusion with PBS for 30 seconds.VSMC were divided into the normal group and the BIX01294 groups.Cells in the normal group were cultured in low glucose medium containing 2%fetal bovine serum,and cells in the BIX01294 groups were co-treated with 2.5,5.0,7.5 and 10.0 μmol/L BIX01294 on the basis of the normal group,respectively.CCK-8 assay and EDU assay were used to detect the cell activity and proliferative ability respectively,to screen appropriate concentration of BIX01294.The cell apoptosis was detected by TUNEL assay between the normal group and the BIX01294 group,and the control group and the treatment group.Western blot was used to detect the expression levels of autophagy-and apoptosis-related proteins in the normal group and BIX01294 group.Results Compared with 0 μmol/L,the activity and proliferative ability of VSMC decreased in a concentration-dependent manner after treatment with BIX01294 at different concentrations(P<0.05).Compared with the normal group,the apoptosis level of VSMC in the BIX01294 group was increased(P<0.05),the expression of VSMC autophagy-and apoptosis-related proteins of LC3Ⅰ/Ⅱ and Bax were up-regulated(P<0.05),and the expression of Bcl-2 and p62 proteins were down-regulated(P<0.05).In vivo test results showed that BIX01294 local perfusion aggravated the apoptosis of carotid VSMC.Conclusion BIX01294 activates VSMC autophagy and apoptosis and inhibits its proliferation.
6.Esophageal carcinoma with ductal differentiation of esophageal gland:clinicopath-ological characteristics and whole exome sequencing analyses
Zhu ZHU ; Xiao HU ; Zhengyang WANG ; Jiajing LI ; Feng WANG ; Hui QIN ; Xiangyu JIAN ; Wencai LI ; Yihui MA
Chinese Journal of Clinical and Experimental Pathology 2025;41(3):291-297
Purpose To summarize the clinical pathological and immunohistochemical characteristics of esophage-al carcinoma with ductal differentiation of esophageal gland,and analyze the somatic mutation characteristics,key driv-ing mutation genes,and significantly mutated genes based on whole exome sequencing.Methods The clinicopatho-logical features of 9 cases of esophageal carcinoma with esophageal duct differentiation were retrospectively analyzed,and the immunohistochemistry EnVision two-step method was used to stain them,and 3 of the samples were subjected to whole exome sequencing and data analysis.Results Among the 9 patients,6 were males and 3 were females.The average age was 68.3 years old(61-80 years old).All 9 cases were located in the middle-lower segment of the e-sophagus.The diameter of the lesion was from 1.5 cm to 3.5 cm.Most areas of the tumor had a double-layer epithelial structure,including the inner layer of luminal epithelium and the outer layer of basal epithelium.Focal areas could be seen with keratinization and mucinous cells.Immunohistochemistry showed that CK7 was positive in the inner epitheli-um,while p63 was positive in the outer basal epithelium.S-100,SOX10 and c-myb were all negative,and p53 was mutated(diffuse strongly positive).The results of whole exome sequencing analysis showed somatic mutation character-istics(796 SNV,37 InDel,482 CNV),key driving mutation genes(12),and significantly mutated genes(TP53).No intraepithelial neoplasia was observed on the surface squamous epithelium of all cases,and no Barrett's esophagus or ectopic gastric mucosa was observed.The average follow-up time was 21.9 months(8 days-51 months),with 8 ca-ses surviving and 1 case dying of severe pulmonary infection 8 days after surgery.Conclusion Esophageal carcinoma with ductal differentiation of esophageal gland is a rare epithelial derived malignant tumor of the esophagus,character-ized by unique morphological,immunohistochemical,and molecular changes.
7.Design of integrated device for cleaning,sterilizing and drying glass graduated pipette
Xiao-hong ZHU ; Xiao-qin HA ; Cheng-yu WANG
Chinese Medical Equipment Journal 2025;46(4):108-112
Objective To design an integrated device for cleaning,sterilizing and drying the glass graduated pipette with considerations on its characteristics.Methods The integrated device had its shell made of 304 stainless steel and hardware part composed of three modules for control,cleaning and drying.The control module consisted of a human machine interface(HMI)touch screen,a programmable logic controller(PLC),solenoid valves and etc;the cleaning module was made up of an ultrasonic release disk,a heat preservation cylinder,a bracket and etc;the drying module comprised a blower,a heating plate,a heat preservation cylinder and connected pipes.The HMI was developed with the programming language of EasyBuilder,and the PLC control logic was written with SCL high-level programming language.Results The device developed could be used for cleaning,sterilizing and drying the glass graduated pipette,and the pipette could be stored in the device after drying.Conclusion The integrated device developed eliminates manual operation and enhances the efficiency of the cleaning,sterilization and drying of the glass graduated pipette.[Chinese Medical Equipment Journal,2025,46(4):108-112]
8.Clinical effects of Supplemented Jiao'ai Decoction combined with warm acupuncture and moxibustion on patients with endometriosis due to Congealing Cold with Blood Stasis
Jing-fen ZHAN ; Jie CHEN ; Shu-qin SHEN ; Xiao-hong WANG ; Xi WU
Chinese Traditional Patent Medicine 2025;47(4):1157-1161
AIM To investigate the clinical effects of Supplemented Jiao'ai Decoction combined with warm acupuncture and moxibustion on patients with endometriosis due to Congealing Cold with Blood Stasis.METHODS Eighty-six patients were randomly assigned into control group(43 cases)for 3-menstrual cycle intervention of warm acupuncture and moxibustion,and observation group(43 cases)for 3-menstrual cycle intervention of both Supplemented Jiao'ai Decoction and warm acupuncture and moxibustion.The changes in clinical effects,TCM syndrome scores,dysmenorrhea degree indices(VAS score,PGE2,PGF2α),hemodynamic indices(RI,PI),sexhormones(E2,FSH,LH),inflammatory factors(IL-1β,IL-6,TNF-α)and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P<0.05).After the treatment,the two groups displayed decreased TCM syndrome scores,dysmenorrhea degree indices,hemodynamic indices,sexhormones and inflammatory factors(P<0.05),especially for the observation group(P<0.05).No significant difference in incidence of adverse reactions was found between the two groups(P>0.05).CONCLUSION For the patients with endometriosis due to Congealing Cold with Blood Stasis,Supplemented Jiao'ai Decoction combined with warm acupuncture and moxibustion can safely and effectively regulate sexhormone levels,endometrial hemodynamics,inhibit inflammatory responses,and alleviate dysmenorrhea symptoms.
9.GBP3 negatively regulates HTLV-1 replication in a GTPase-dependent manner
Yanzi LIU ; Xin ZHAO ; Xiao QIN ; Yulu HUANG ; Xi YANG ; Qingqing FAN ; Bo YANG ; Jie WANG
Chinese Journal of Immunology 2025;41(3):535-539
Objective:To investigate the effect of GBP3 on replication of adult T-lymphocytic leukemia virus type 1(HTLV-1).Methods:Expression of GBP3 in HTLV-1-infected HeLa cell and THP1 cell was detected by Western blot.The knock-down efficiency of siRNAs targeting GBP3 in HeLa and THP1 cells was evaluated by Western blot.Effects of GBP3 overexpression or knockdown on expression of HTLV-1 proviral transcripts Tax,px,HBZ,Gag,ENV,5'UTR,and viral proteins Tax,p19 were investigated by RT-qPCR and Western blot.GTPase-defective mutant of GBP3,GBP3K51A was constructed to explore whether the effects of GBP3 on HTLV-1 infection were dependent on its GTPase activity.Results:GBP3 expression was upregulated in HTLV-1 infected HeLa and THP1 cells.GBP3 overexpression decreased expression of HTLV-1 proviral transcripts and viral proteins,whereas the knockdown of GBP3 has the opposite effects.Overexpression of GBP3K51A increased expression of HTLV-1 proviral transcripts and viral proteins.Conclusion:HTLV-1 virus infection can induce expression of GBP3;overexpression of GBP3 inhibits virus replication and may depend on GTPase.
10.Present situation of sensors applied to monitoring of spinal morphology and motion
Shi-yu ZHOU ; Ya-qin LI ; Yang-xi HUANG ; Xiao CHEN ; Jing WANG ; Zhi-min LIANG ; Yu-chen GUO ; Xue YANG ; Ling-li LI
Chinese Medical Equipment Journal 2025;46(6):105-110
The application of sensors to the monitoring of spinal morphology and motion was reviewed in terms of the research object and monitoring index.The present situation of the application of sensors was introduced,such as inertial sensor,stretchable strain sensor and electromagnetic sensor.The deficiencies of sensors applied to the monitoring of spinal morphology and motion were analyzed,and the future directions of the application were pointed out.[Chinese Medical Equipment Journal,2025,46(6):105-110]

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