1.Progress in preclinical studies of xenogeneic lung transplantation and single-center technical experience
Xiaoting TAO ; Xinzhong NING ; Yong LIU ; Guimei ZHANG ; He XIAO ; Shiyu LIN ; Zizi ZHOU ; Taiyun WEI ; Chunxiao HU ; Hongjiang WEI ; Kun QIAO
Organ Transplantation 2025;16(6):874-880
Lung transplantation is the ultimate therapeutic option for end-stage pulmonary diseases such as interstitial pneumonia, chronic obstructive pulmonary disease and pneumoconiosis. Currently, the shortage of allogeneic lung donors significantly limits the opportunity for end-stage lung disease patients to receive lung transplantation. In recent years, with the rapid development of biomedical engineering technologies, especially the major breakthroughs in genetic modification and cloning, xenogeneic lung transplantation has shown important potential for clinical translation. Among them, genetically modified pigs have become the most promising xenogeneic lung source due to the close similarity of organ size and physiological characteristics to humans, and the ability to perform targeted gene knockouts (such as α-Gal antigen knockout) to reduce the occurrence of hyperacute rejection. This article focuses on the research progress of porcine xenogeneic lung transplantation, systematically reviews the latest achievements and challenges in animal experiments and human trials, and introduces the technical experience accumulated by Shenzhen Third People's Hospital in the porcine-to-monkey xenogeneic lung transplantation model, in the hope of providing practical references for future research in this field.
2.Study on the machanism of Huannao Yicong Deoction targeting HAMP to regulate iron metabolism and improve cognitive impairment in AD model mice
Ning-Ning SUN ; Xiao-Ping HE ; Shan LIU ; Yan ZHAO ; Jian-Min ZHONG ; Ya-Xuan HAO ; Ye-Hua ZHANG ; Xian-Hui DONG
Chinese Pharmacological Bulletin 2024;40(7):1240-1248
Aim To explore the effects of Huannao Yicong decoction(HYD)on the learning and memory ability and brain iron metabolism in APP/PS1 mice and the correlation of HAMP knockout mice and APP/PS1 double transgenic model mice.Methods The ex-periment was divided into five groups,namely,HAMP-/-group(6-month HAMP gene knockout mice),APP/PS1 group(6-month APP/PS1-double-transgenic mice),HAMP-/-+HYD,APP/PS1+HYD,and negative control group(6-month C57BL/6J mice),with six mice in each group.The dose was ad-ministered(13.68 g·kg-1 weight),and the other groups received distilled water for gavage once a day for two months.After the administration of the drug,the mice in each group were tested for learning and memory in the Morris water maze;Biochemical detec-tion was performed to detect iron ion content in each mouse brain;Western blot and RT-qPCR were carried out to analyze hippocampal transferrin(TF),transfer-rin receptor1(TFR1),membrane iron transporter1(FPN1)divalent metal ion transporter 1(DMT1)and β-amyloid protein(Aβ)protein and mRNA expression levels in each group.Results Compared with the normal group,both HAMP-/-mice and APP/PS1 mice had reduced the learning and memory capacity,in-creased iron content in brain tissue,Aβ protein ex-pression increased in hippocampus of HAMP-/-group and APP/PS1 group mice(P<0.01),the protein and mRNA expression of TF,TFR1 and DMT1 increased in hippocampal tissues of HAMP-/-and APP/PS1 groups(P<0.01),and the FPN1 protein and mRNA expres-sion decreased(P<0.01).Compared with the HAMP-and APP/PS1 groups,respectively,HAMP-/-+HYD group and APP/PS1+HYD group had improved learning and memory ability,decreased iron content,decreased Aβ protein expression(P<0.01),decreased TF,TFR1,DMT1 protein and mR-NA expression(P<0.01),and increased expression of FPN1 protein and mRNA(P<0.01).Conclusions There is some association between HAMP-/-mice and APP/PS1 mice,HYD can improve the learning and memory ability of HAMP-/-and APP/PS1 mice and reduce the Aβ deposition.The mechanism may be related to the regulation of TF,TFR1,DMT1,FPN1 expression and improving brain iron overload.
3.MTHFD2 Is a Negative Regulatory Molecule for the Formation of Heterotypic Cell-in-Cell Structures
Peng-Fei FENG ; Chen-Yu LIU ; Yi-Nuo HUANG ; Zhuo-Ran SUN ; Yang-Yi ZHANG ; Hong-Yan HUANG ; Chen-Xi WANG ; Xiao-Ning WANG
Chinese Journal of Biochemistry and Molecular Biology 2024;40(6):819-826
Heterotypic cell-in-cell structures(heCICs)mediate unique non-autonomous cell death,which are widely involved in a variety of important pathological processes,such as tumorigenesis,pro-gression and clinical prognosis.Methylenetetrahydrofolata dehydrogenase 2(MTHFD2),one of the key enzymes of one-carbon metabolism,is highly expressed in a variety of tumor cells.In this study,in order to investigate the effect of MTHFD2 on the formation of heCICs,liver cancer cells and immune cells were first labeled separately by live cell dyes,and the heCIC model was established by using fluorescence mi-croscopy for cell imaging and analysis.After transiently knocking down MTHFD2 in cells by RNAi,we found that the ability of PLC/PRF/5 and Hep3B to form heCICs with immune cells was significantly in-creased(all P<0.01).MTHFD2 recombinant expression plasmid was constructed by the homologous re-combination method,and MTHFD2 overexpression cell lines were further constructed.Then,the effect of MTHFD2 overexpression on the ability to form heCICs was detected by co-culturing the overexpression cell lines with immune cells.The results showed that the rate of heCIC formation was significantly re-duced after overexpression of MTHFD2(all P<0.001).In conclusion,this study demonstrated that MTHFD2 is a negative regulator of heCIC formation,providing a research basis for targeting MTHFD2 to promote heCIC formation and enhance the in-cell killing of immune cells.
4.The Regulatory Effect of RNA m6A Methylation Modification on KDM4B Gene Expression in t(8;21)AML Cells by MeRIP-qPCR
Yu-Qing LI ; Yang-Liu SHAO ; Meng-Yue LI ; Li-Li WANG ; Xiao-Ning GAO
Journal of Experimental Hematology 2024;32(2):382-388
Objective:To confirm the direct regulatory effect of WTAP-mediated RNA m6A modification on the KDM4B gene in t(8;21)acute myeloid leukemia(AML)cells through MeRIP combined with reverse transcription real-time quantitative PCR(RT-qPCR)technology.Methods:The lentivirus-mediated shRNA target WTAP or KDM4B gene was used to transfect the t(8;21)AML cell lines:Kasumi-1 and SKNO-1,and cells transfected with randomly shuffled shRNA as the control.Using the Ultrapure RNA Extraction Kit(DNase Ⅰ)to extract RNA.The Magna MeRIPTM m6A Kit was used to enrich methylated modified fragments,and detect the m6A methylated RNA regions by RT-qPCR,and the protein and mRNA expression levels of WTAP and KDM4B in cells were detected by Western blot and reverse transcription real-time quantitative PCR(RT-qPCR).Colony formation assays were used to detect the colony ability of cells in vitro.Results:Silencing the expression of WTAP in Kasumi-1 cells,the enrichment of m6A methylation modification was significantly decreased in the 3'UTR of KDM4B mRNA(P<0.01),and the protein(P<0.001)and mRNA(Kasumi-1:P<0.001;SKNO-1:P<0.01)expression levels of KDM4B were also significantly inhibited in Kasumi-1 and SKNO-1 cells upon WTAP knockdown(all P<0.01),accompanied by a significant decrease in the colony-forming ability of both cell lines(both P<0.01).Conclusion:In t(8;21)AML cell lines,WTAP could regulate the expression of KDM4B by regulating the m6A modification of the 3'UTR of KDM4B mRNA,and silencing the expression of KDM4B could inhibit the cellular proliferation in vitro.
5.Analysis of Therapeutic Efficacy and Adverse Prognostic Factors of Secondary Central Nervous System Lymphoma
Ning WANG ; Fei-Li CHEN ; Yi-Lan HUANG ; Xin-Miao JIANG ; Xiao-Juan WEI ; Si-Chu LIU ; Yan TENG ; Lu PAN ; Ling HUANG ; Han-Guo GUO ; Zhan-Li LIANG ; Wen-Yu LI
Journal of Experimental Hematology 2024;32(5):1420-1426
Objective:To explore the therapeutic efficacy and prognostic factors of induction therapy for secondary central nervous system lymphoma(SCNSL).Methods:Clinical data of patients diagnosed with SCNSL from 2010 to 2021 at Guangdong Provincial People's Hospital were retrospectively collected.A retrospective cohort study was performed on all and grouped patients to analyze the efficacy and survival.Multivariate logistic regression analysis was used to explore the adverse prognostic factors.Results:Thirty-seven diffuse large B-cell lymphoma patients with secondary central involvement were included in the research.Their 2-year overall survival(OS)rate was 46.01%and median survival time was 18.1 months.The 2-year OS rates of HD-MTX group and TMZ group were 34.3%and 61%,median survival time were 8.7 and 38.3 months,and median progression-free survival time were 8.1 and 47 months,respectively.Multivariate logistic regression analysis showed that age,sex,IPI,Ann Arbor stage were correlated with patient survival time.The median survival time of patients with CD79B,KMT2D,CXCR4.ERBB2,TBL1XR1,BTG2,MYC,MYD88,and PIM1 mutations was 8.2 months,which was lower than the overall level.Conclusion:HD-MTX combined with TMZ as the first-line strategy may improve patient prognosis,and early application of gene sequencing is beneficial for evaluating prognosis.
6.Simultaneou determination of twenty-eight constituents in Dayuan Drink by UPLC-MS/MS
Yu-Jie HOU ; Xin-Jun ZHANG ; Ming SU ; Xin-Rui LI ; Yue-Cheng LIU ; Yu-Qing WANG ; Dan-Dan SUN ; Hui ZHANG ; Kang-Ning XIAO ; Long-Yun DUAN ; Lei CAO ; Zhen-Yu XUAN ; Shan-Xin LIU
Chinese Traditional Patent Medicine 2024;46(11):3545-3552
AIM To establish a UPLC-MS/MS method for the simultaneous content determination of gallic acid,protocatechuic acid,neomangiferin,catechin,caffeic acid,mangiferin,isomangiferin,albiflorin,paeoniflorin,vitexin,liquiritin,scutellarin,baicalin,liquiritigenin,timosaponin BⅡ,quercetin,wogonoside,benzoylpaeoniflorin,isoliquiritigenin,honokiol,magnolol,norarecaidine,arecaidine,arecoline,epicatechin,baicalein,glycyrrhizinate and wogonin in Dayuan Drink.METHODS The analysis was performed on a 35℃thermostatic Syncronis C18 column(100 mm×2.1 mm,1.7 μm),with the mobile phase comprising of 0.1%formic acid-acetonitrile flowing at 0.3 mL/min in a gradient elution manner,and electron spray inoization source was adopted in positive and negative ion scanning with select reaction monitoring mode.RESULTS Twenty-eight constituents showed good linear relationships within their own ranges(R2≥0.991 0),whose average recoveries were 95.60%-103.53%with the RSDs of 0.60%-5.45%.CONCLUSION This rapid,simple,selective,accurate and reliable method can be used for the quality control of Dayuan Drink.
7.Research progress of fluorescent probes in uric acid detection
Di-Di XING ; Ruo-Jin LIU ; Jia-Yu QI ; Ning MA ; Ya-Kun JI ; Jia-Xin ZHOU ; Yu-Shan XING ; Xiao-Lan ZHEN
Chinese Medical Equipment Journal 2024;45(6):93-104
The advantages of fluorescence detection of uric acid were introduced compared to the traditional detection methods.The preparation process,detection principle and performance of organic,inorganic and organic-inorganic hybrid fluorescent probes were reviewed.The advantages and disadvantages of kinds of fluorescent probes were analyzed when used for uric acid detection,and the futural directions were pointed out for related research.[Chinese Medical Equipment Journal,2024,45(6):93-104]
8.Diagnostic value of magnetic resonance T2*mapping sequence combined with texture feature indicator for postmenopausal osteoporosis
Xiao-Sheng YANG ; Mao-Ting LI ; Yan-Ning LI ; Song-He LIU
Chinese Medical Equipment Journal 2024;45(8):68-72
Objective To explore the application value of T2*mapping sequence combined with texture feature indicatoror in the diagnosis of postmenopausal osteoporosis(PMOP)in women.Methods The clinical data of 50 PMOP patients admitted to some hospital from March 2019 to April 2021(enrolled into an osteoporosis group)were analyzed retrospectively,and another two groups were established including a normal bone mineral density(BMD)group composed of 50 postmenopausal physical examiners with normal BMD(BMD>120 mg/cm3)and a reduced BMD group consisting of 50 postmenopausal women with decreased BMD(80 mg/cm3<BMD<120 mg/cm3).All the 3 groups underwent magnetic resonance lumbar spine conventional sequence and T2(mapping scanning and image texture feature extraction,which were compared in terms of lumbar T2*value and texture feature indicators including energy,contrast,correlation,deficit distance and entropy.Spearman correlation analysis was used to analyze the correlation between lumbar spine T2*values,texture feature indicators and the occurrence of POMP in women,independent predictors of the occurrence of PMOP in women were determined by multifactorial logistic regression analysis,the diagnostic efficacy of T2*mapping sequence combined with texture feature indicatoror for PMOP was analyzed using ROC curves,and a column-line diagram prediction model for POMP occurrence was constructed and validated.Results There were significant differences in T2*value,energy,contrast,correlation,deficit distance and entropy among the groups(P<0.05).Spearman correlation analysis showed that with the occurrence of POMP lumbar spine T2*value,contrast and entropy were significantly positively correlated,while energy,correlation and deficit distance were significantly negatively correlated(P<0.05).T2*value,energy,contrast,correlation,deficit distance and entropy were all independent predictors of the occurrence of POMP(P<0.05).T2*mapping sequence combined with texture feature indicatoror behaved better than any single indicator in AUC,sensitivity and specificity.The column-line diagram prediction model predicted a 75.00%probability of POMP in women,which was validated with advantages ind1iscrimination,calibration and net benefit rate.Conclusion Magnetic resonance T2*mapping sequence combined with texture feature indicator behaves well in diagnosing POMP of women,and T2*value,energy,contrast,correlation,deficit distance and entropy can be used as sensitive indicators for clinical screening of people at high risk for POMP.[Chinese Medical Equipment Journal,2024,45(8):68-72]
9.Application research of R language-based autoregressive integrated moving average model for predicting short-term consumption of medical consumables
Ze-Hua LIU ; Hong-Tao LU ; Wei LI ; Fei WEI ; Si-Si WANG ; Xiao-Ning FU ; Xin-Ming DONG
Chinese Medical Equipment Journal 2024;45(10):84-87
Objective To explore the effect of a R language-based autoregressive integrated moving average(ARIMA)model for predicting the consumption of medical consumables.Methods The monthly consumption data of a certain type of pre-filled flush syringe from July 2018 to June 2023 was selected as the sample data,which underwent smoothness test and difference operation with R language.An ARIMA model was established and the optimal model was determined according to the Akaike and Bayesian information criteria.The corresponding data of the third quarter of 2023 was used as the validation set to predict the consumption,and the prediction result was compared with the actual values to evaluate the prediction effect of the ARIMA model.Results The ARIMA model with the best fitting was ARIMA(0,1,1)(1,0,0)12,all the predicted data were within 95%confidence interval,and its mean absolute percentage error MAPE was 9.92%.P-value proved to be higher than 0.05 when the residual series were tested using the Ljung-Box statistics,which meant the prediction result was satisfactory.Conclusion The R language-based ARIMA model behaves well in predicting the consumption of medical consumables,and provides references for demand planning,budgeting,purchasing and management of medical consumables.[Chinese Medical Equipment Journal,2024,45(10):84-87]
10.Melatonin alleviates autophagy in cortical neurons of neonatal rats with hypoxic-ischemic brain damage via the PI3K/AKT pathway
Chen-Meng LIU ; Xiao-Tian GAO ; Hai-Mo ZHANG ; Hui-Ning BI ; Chen LIANG ; Jing-Ying JIANG ; Pei-Lun XIAO ; Xiao-He YU ; Xiao-Li WANG
Chinese Journal of Contemporary Pediatrics 2024;26(6):631-638
Objective To observe the effects of melatonin on autophagy in cortical neurons of neonatal rats with hypoxic-ischemic brain damage(HIBD)and to explore its mechanisms via the PI3K/AKT signaling pathway,aiming to provide a basis for the clinical application of melatonin.Methods Seven-day-old Sprague-Dawley neonatal rats were randomly divided into a sham operation group,an HIBD group,and a melatonin group(n=9 each).The neonatal rat HIBD model was established using the classic Rice-Vannucci method.Neuronal morphology in the neonatal rat cerebral cortex was observed with hematoxylin-eosin staining and Nissl staining.Autophagy-related protein levels of microtubule-associated protein 1 light chain 3(LC3)and Beclin-1 were detected by immunofluorescence staining and Western blot analysis.Phosphorylated phosphoinositide 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)protein expression levels were measured by immunohistochemistry and Western blot.The correlation between autophagy and the PI3K pathway in the melatonin group and the HIBD group was analyzed using Pearson correlation analysis.Results Twenty-four hours post-modeling,neurons in the sham operation group displayed normal size and orderly arrangement.In contrast,neurons in the HIBD group showed swelling and disorderly arrangement,while those in the melatonin group had relatively normal morphology and more orderly arrangement.Nissl bodies were normal in the sham operation group but distorted in the HIBD group;however,they remained relatively intact in the melatonin group.The average fluorescence intensity of LC3 and Beclin-1 was higher in the HIBD group compared to the sham operation group,but was reduced in the melatonin group compared to the HIBD group(P<0.05).The number of p-PI3K+and p-AKT+cells decreased in the HIBD group compared to the sham operation group but increased in the melatonin group compared to the HIBD group(P<0.05).LC3 and Beclin-1 protein expression levels were higher,and p-PI3K and p-AKT levels were lower in the HIBD group compared to the sham operation group(P<0.05);however,in the melatonin group,LC3 and Beclin-1 levels decreased,and p-PI3K and p-AKT increased compared to the HIBD group(P<0.05).The correlation analysis results showed that the difference of the mean fluorescence intensity of LC3 and Beclin-1 protein in the injured cerebral cortex between the melatonin and HIBD groups was negatively correlated with the difference of the number of p-PI3K+and p-AKT+cells between the two groups(P<0.05).Conclusions Melatonin can inhibit excessive autophagy in cortical neurons of neonatal rats with HIBD,thereby alleviating HIBD.This mechanism is associated with the PI3K/AKT pathway.

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