1.Angiotensin Ⅱ activates p53/SAT1 signaling pathway to induce ferroptosis in white adipocytes
Wei DENG ; Xiyan LIU ; Liyuan GUO ; Qian XU ; Kun ZHOU ; Yuanqin ZHAO ; Zhaoyue WANG ; Xiang LI ; Xin-mei DENG ; Xinyi QIN ; Zhong REN ; Zhisheng JIANG
Chinese Journal of Arteriosclerosis 2025;33(5):385-394
Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.
2.Research progress in the regulation of host immune metabolism by Mycobacterium tuberculosis
Dan-dan ZHANG ; Jia-xu WAN ; Sha-sha FU ; Cheng-kun ZHENG ; Xiang CHEN ; Zheng-zhong XU ; Xin-an JIAO
Chinese Journal of Zoonoses 2025;41(4):358-363
Immunometabolism studies focus on the relationships between immune cell functions and cellular energy metabolism pathways.Immunometabolism plays an important regulatory role in immune-related diseases.Mycobacterium tuberculosis(M.tb),an important intracellular pathogenic bacterium,enters alveolar macrophages after infection.The confrontation between M.tb and the host is a complex and dynamic process involving multiple aspects and mechanisms,such as the immune response,granuloma formation,and immune evasion.M.tb effector proteins play key roles in maintaining bacterial virulence and regulating host cell metabolism.This article reviews the reprogramming process of glucose metabolism,lipid metabolism,and immunometabolism,as well as changes in mi-tochondrial function in M.tb-infected host cells,thereby revealing the relationship between M.tb pathogenicity and host metabolic regu-lation,which is important for understanding tuberculosis.
3.Epidemic status and control strategies of tuberculosis in sheep and goats
Jia-xing AN ; Wen-jun KONG ; Zheng-zhong XU ; Cheng-kun ZHENG ; Xiang CHEN ; Xin-an JIAO
Chinese Journal of Zoonoses 2025;41(4):346-350
Sheep and goats are important tuberculosis hosts found predominantly in plateau and mountainous regions.In recent years,the number of reported tuberculosis cases in sheep and goats has increased.The tuberculosis pathogen can spread among vari-ous animal species and even infect humans,thus further complicating disease prevention and control,and posing a serious threat to the health of both humans and livestock.This article summarizes the global prevalence of tuberculosis in sheep and goats,and specifi-cally analyzes the epidemic status in China.Frequently used tuberculosis detection methods in sheep and goats are described,and the shortcomings of each method are briefly introduced.Additionally,on the basis of frequently applied methods for monitoring,handling,and controlling tuberculosis in sheep and goats worldwide,suggestions are offered to provide a reference for tuberculosis control in sheep and goats in China.
4.Mycobacterium tuberculosis Rv3641c inhibits macrophage type Ⅰ interferon responses and promotes intracellular survival in macrophages
Wen JIN ; Min GENG ; Su-jie HU ; Xin-yang ZHANG ; Wen-qin LI ; Cheng-kun ZHENG ; Xin-an JIAO ; Xiang CHEN ; Zheng-zhong XU
Chinese Journal of Zoonoses 2025;41(4):385-391
This study was aimed at investigating the immunoregulatory function of Mycobacterium tuberculosis Rv3641c gene in modulating host type Ⅰ interferon responses.The shuttle plasmid pMV261 was used to construct Rv3641c overexpression recombinant Mycobacterium smegmatis,and the biological characteristics of the recombinant bacteria were analyzed to explore the effect of Rv3641c on the growth curve,colony morphology and stress resistance of Mycobacterium.Subsequently,RAW264.7 cells were infected with Rv3641c overexpressing Mycobacterium smegmatis,and the transcriptional expression of genes related to the inhibition of type I inter-feron pathway was determined by RT-PCR.The expression level of IFN-βprotein was determined by ELISA,and the intracellular sur-vival level was determined.As a result,the recombinant rMS::pMV261-Rv3641c was successfully constructed.The results of biologi-cal characteristics analysis showed that Rv3641c did not affect the growth of mycobacteria,but significantly changed the colony mor-phology of mycobacteria and improved its resistance to H2O2.The results of recombinant bacteria infection experiments showed that Rv3641c significantly down-regulated the transcription levels of IFN-α,IFN-βand downstream ISGs genes CXCL10,IFIT2 and IL-1β in host cells,and Rv3641c significantly down-regulated the transcription levels of IFN-α,IFN-βand downstream ISGs genes CXCL10,IFIT2 and IL-1βin host cells.The results of intracellular colonization experiments showed that the intracellular mycobacte-ria in the overexpression recombinant bacteria infection group were significantly higher than those in the empty vector group,indicat-ing that Rv3641c could promote the intracellular surviv al of mycobacteria.In summary,the Rv3641c gene of M.tuberculosis can inhibit the host type I interferon response and promote the intracellular survival of M.tuberculosis,which provides a new idea for further explor-ing the immune escape function of M.tuberculosis and the discovery of new targets for anti-tuberculosis drugs.
5.Clinicopathological and molecular genetic analysis of 10 cases of nephrogenic ade-noma/metaplasia in bladder
Wenwu LUO ; Mei LI ; Kun ZHONG ; Yu YIN
Chinese Journal of Clinical and Experimental Pathology 2025;41(5):584-590
Purpose To investigate the clinicopathological characteristics of nephrogenic adenoma/metaplasia(NA/M)of the bladder and analyze key points for diagnosis and differential diagnosis.Methods A retrospective a-nalysis was conducted on 10 cases of NA/M of the bladder,including clinical data,cystoscopic findings,microscopic morphology,and immunohistochemistry(IHC)results.Subsequently,a whole-exome sequencing(WES)was per-formed and a comprehensive literature review was supplemented.Results The cohort included 6 cases female and 4 cases male,aged 20-72 years(median:46.5 years).Three patients had a history of hydronephrosis(2 cases with nephrolithiasis and hydronephrosis,1 case with acute leukemia and bone marrow transplantation),2 cases had renal tuberculosis,1 case had erectile dysfunction,3 cases had a history of invasive high-grade urothelial carcinoma of the bladder,and 1 case had bladder endometriosis.All patients underwent cystoscopic biopsy,with pathological confirma-tion of NA/M.IHC showed that all 10 cases expressed PAX-8 and CK7 but were negative for GATA-3,p63/CK5/6,and PSA.The Ki67 proliferation index ranged from 1%to 10%.WES analysis of 7 cases revealed somatic single nu-cleotide variants(SNVs)involving multiple genes,with the highest mutation frequencies in FAM186A(86%),GOL-GA6L2(86%),and AQP7(86%).Conclusion NA/M is a benign but recurrent lesion,with rare malignant trans-formation after multiple recurrences.Comprehensive evaluation of histomorphological atypia and IHC results is recom-mended for accurate diagnosis and differential diagnosis to avoid misdiagnosis.Long-term follow-up is advised.WES findings suggest associations between NA/M and mutations in FAM186A,GOLGA6L2,and AQP7,providing potential directions for future research.
6.Retrospective analysis of quality results of mid-trimester serum screening in China from 2019 to 2023
Jinming ZHANG ; Xingtong CHEN ; Wei WANG ; Kun ZHONG ; Shuai YUAN ; Yuxuan DU ; Chuanbao ZHANG ; Falin HE
Chinese Journal of Preventive Medicine 2025;59(5):628-633
Objective:To analyze the quality level of maternal mid-trimester serum prenatal screening in China from 2019 to 2023.Methods:A questionnaire survey was conducted to collect information from laboratories participating in the National Health Commission′s Clinical Laboratory Center inter-laboratory quality assessment program for prenatal screening from 2019 to 2023. The collected data included screening protocols, detection methods, testing systems, and monthly laboratory screening quality indicators. The Chi-square test was used to compare the initial screening positive rates among different screening protocols, provinces, detection methods, and testing systems. A log-transformed linear regression analysis was performed to evaluate the relationship between the average annual sample size and the out-of-control rate of various median multiple of the median (mMoM) values.Results:This study included 806 laboratories from 29 provinces (including autonomous regions and municipalities). The five-year average out-of-control rates for mMoM values across indicators ranged from 15.8% to 31.3%. The initial positive rates of dual, triple, and quadruple screening protocols were statistically different ( χ2=760.2, P<0.001). The initial positive rates across different provinces ranged from 4.7% to 10.3%, with statistically significant differences ( χ2=35 388.0, P<0.001). There were also statistically significant differences in initial positive rates between different testing systems ( χ2=2 493.2, P<0.001). Testing systems using chemiluminescence methods had significantly higher initial positive rates compared to systems using time-resolved fluorescence methods (7.2% vs. 6.5%, χ2=533.6, P<0.001). Log-linear regression analysis showed that testing systems with larger annual average sample sizes had lower out-of-control rates for mMoM values (AFP: β=-0.100, P=0.005; hCG: β=-0.123, P=0.008; uE3: β=-0.139, P=0.007). Conclusion:There are significant differences in the quality of maternal mid-trimester serum screening across different detection methods, testing systems and provinces in China.
7.Efficacy analysis of plasma exchange treatment for thymoma-associated myasthenia gravis
Miao HONG ; Dongdong CAI ; Caihui WEI ; Bing HU ; Kun XIAO ; Fangming RUAN ; Piaoping HU ; Aiping LE ; Zhanglin ZHANG ; Chang ZHONG
Chinese Journal of Blood Transfusion 2025;38(9):1188-1194
Objective: To evaluate the efficacy and safety of plasma exchange (PE) in thymoma-associated myasthenia gravis (MG), thereby to provide theoretical support for its application in the treatment of thymoma-associated MG. Methods: A total of 133 patients with thymoma-associated MG admitted from January 2018 to September 2024 were retrospectively analyzed. Patients were matched using propensity score to reduce selection bias, yielding 22 matched pairs for both PE group (n=22) and non-PE group (n=22). Patient characteristics including gender, age of disease onset, course of disease, history of thymoma resection, clinical absolute scores [clinical absolute scores (CAS) and clinical relative scores (CRS)], and synchronized immunotherapy regimen of the two groups were analyzed. The CAS scores before and after treatment were compared between the two groups, and the CRS was used to assess the treatment efficiency. Safety of the two treatment regimens were also compared. Continuous variables were compared using the t-test or ANOVA, while categorical data were compared by the chi-square test. Results: A total of 133 patients were included and divided into two groups according to whether they underwent plasma exchange treatment: the PE group (n=22) and the non-PE group (n=111). To exclude bias caused by large difference in the number of cases between the two groups, we performed propensity score matching. After matching, the number of cases in both groups was 22. There was no significant difference in baseline clinical characteristics between the two groups (P>0.05), including gender, age of onset, duration of disease course, history of thymectomy and baseline CAS score before treatment. Compared to the non-PE group, patients in the PE group showed more significant improvement in CAS score (5.09±1.95 vs 3.59±1.50, P<0.05) and a higher CRS score (75.00% vs 50.00%, P<0.001). Compared to the non-PE group, PE group had significantly longer ICU stay, longer hospital stay and higher hospitalization cost (P<0.05). There was no statistically significant difference in adverse events between the two groups during treatment (P>0.05). During long-term follow-up, both the PE and non-PE groups showed relatively low 1-, 3-, and 5-year recurrence rate, with no significant difference between the two groups (P>0.05). Conclusion: This study indicates that plasma exchange has clear value in the treatment of patients with thymoma-associated myasthenia gravis. It can not only significantly improve patients' muscle strength to alleviate motor dysfunction and enhance quality of life, but also does not significantly increase the incidence of adverse reactions. Therefore, it can be regarded as one of the preferred treatment options that achieve a "balance between efficacy and safety" for such patients, and provides an important basis for optimizing treatment strategies, improving prognosis, and promoting the application of subsequent treatment regimens.
8.The protective effect of Qingjie Huagong decoction on intestinal mucosal barrier of SAP rats based on regulation of mitochondrial autophagy by PGAM5/Drp1/PINK1/Parkin axis
Xiao-dong ZHU ; Kun-rong LIU ; Min-chao FENG ; Ying BAN ; Xi-ping TANG ; Guo-zhong CHEN
Chinese Pharmacological Bulletin 2025;41(3):561-567
Aim To investigate the mechanism of in-testinal mucosal barrier protective effect of Qingjie Huagong decoction(QJHGD)on rats with severe acute pancreatitis(SAP).Methods The SAP rat model was constructed,and the sham-operation group,the model group,the group administered with different dosages of QJHGD,and the positive control group were set up respectively.HE staining was used to observe the histopathological changes.ELISA was employed to detect the serum levels of diamine oxidase(DAO)and D-lactic acid(D-LA)in rats.Transmission electron microscopy was utilized to observe the mitochondria of ileal tissues.qRT-PCR and Western blot were applied to detect the mRNA and protein expression of PGAM5,Drp1,PINK1,Parkin,LC3B in ileal tissues of rats.Results Compared with the sham-operated group,the pancreas and ileum tissues of rats in the model group showed obvious pathological changes,with abnormal mitochondrial structure and reduced number of autoph-agic vesicles in the ileum tissues.The levels of DAO and D-LA in serum increased(P<0.01),and the mRNA and protein expression of PGAM 5,Drp 1,PINK1,Parkin,and LC3B in the ileum tissues de-creased significantly.Compared with the model group,pancreatic and ileal pathology were improved,mito-chondrial damage in the ileum was reduced,and the number of autophagic vesicles increased in the QJHGD group.The serum levels of DAO and D-LA were re-duced,and the expression of PGAM5,Drp1,PINK1,Parkin,and LC3B mRNA and protein in the ileal tis-sues increased significantly.Conclusions QJHGD may exert a protective effect on the SAP intestinal mu-cosal barrier by regulating the PGAM5/Drp1/PINK1/Parkin axis in order to elevate the level of mitochondri-al autophagy in the intestinal epithelial cells,thereby improving the level of repair of the intestinal epithelial cells.
9.Effect of hydrogen sulfide on calcification during macrophage phagocytosis of lipids and its mechanisms
Yanwen LIU ; Qian XU ; Kun ZHOU ; Ziyi LI ; Shuiqing LIU ; Jun ZHANG ; Zhong REN ; Zhisheng JIANG
Chinese Journal of Arteriosclerosis 2025;33(4):317-325
Aim To investigate the effect of hydrogen sulfide on macrophage calcification and its underlying mo-lecular mechanisms.Methods Oil red O staining was used to observe intracellular lipid accumulation,and von Kossa staining and atomic absorption spectroscopy were used for morphological and quantitative analysis of calcium deposition and intracellular calcium content in a mononuclear macrophage calcification model.Western blot and RT-PCR were used to detect the mRNA and protein expression of osteopontin(OPN)at different doses and treatment times of hydrogen sulfide.At the same time,Western blot was used to detect the expression changes of early growth response factor 1(EGR1),endo-plasmic reticulum stress-related markers C/EBP homologous protein(CHOP)and glucose-regulated protein 78(GRP78).Reactive oxygen species levels were evaluated by fluorescence probe staining,and the effect of hydrogen sulfide on macro-phage calcification was evaluated by combining von Kossa staining and calcium ion fluorescence probe staining.The mo-lecular mechanisms of hydrogen sulfide affecting macrophage calcification were explored by interfering with EGR1 expression and using endoplasmic reticulum stress inhibitor 4-phenylbutyric acid(4-PBA).Results Compared with oxidized low density lipoprotein(ox-LDL)group,β-glycerophosphate(β-GP)+40 g/L ox-LDL group showed a significant increase in intracellular lipid accumulation,while hydrogen sulfide significantly inhibited macrophage calcification in a con-centration-and time-dependent manner.Compared with the β-GP+ox LDL group,the most significant effect was observed after incubation with 100 μmol/L NaHS for 4 days.The hydrogen sulfide group showed a 66%decrease in intracellular calcium content(P<0.01),a 71%decrease in intercellular calcium deposition(P<0.01),and a 50%and 48%decrease in OPN mRNA and protein expression,respectively(P<0.05).Hydrogen sulfide treatment upregulated the ex-pression of EGR1 by 21%,while downregulating the expression of CHOP and GRP78 by 58%and 59%,respectively(P<0.01).The endoplasmic reticulum stress inhibitor 4-PBA could downregulate OPN expression by 73%(P<0.01),while interfering with EGR1 expression completely counteracts the inhibitory effect of hydrogen sulfide on OPN expression and calcium deposition(P<0.01).Conclusion Hydrogen sulfide significantly inhibits macrophage calcification by upregulating EGR1 expression and suppressing endoplasmic reticulum stress.
10.Design and application of positive and negative pressure sequential irrigation suction device
Kun-feng SANG ; Xian-jin ZHONG ; Jia-li HUANG
Chinese Medical Equipment Journal 2025;46(1):114-117
Objective To design a positive and negative pressure sequential irrigation suction device and explore its efficacy when applied to unblocking nasoenteric tubes.Methods The device consisted of a positive-pressure irrigation component,a negative-pressure suction component,a three-way adapter and a precise unblocker.The positive-pressure irrigation compo-nent was composed of a positive-pressure irrigation unit,a positive pressure regulator,a pressure sensor,a pressure bag and etc;the negative-pressure suction component was made up of a negative-pressure suction unit,a negative pressure regulator,a negative-pressure gauge,a negative-pressure bottle and etc.The pressure bag was hollow cylinder-shaped,and made of ordinary polyvinyl chloride for its inner layer and acrylonitrile-styrene-butadiene copolymer for its outer layer.Sixty patients with nasoenteric tube blockage were selected and randomly divided into a control group and an observation group,with 30 cases in each group.Irrigation suction was carried out with a 10 mL syringe for the control group and with the device for the observation group,and then unblocking was performed with lukewarm water between 38 and 40 degrees.The two groups were compared in terms of the unblocking efficacy,time required for unblocking and the days and cost of nutritional support.SPSS 22.0 software was used for statistical analysis.Results The observation group gained advantages over the control group in terms of the total effective rate(93.3%vs 73.4%),unblocking efficacy,time required for unblocking((7.63±4.97)min vs(10.73±4.57)min),days of nutritional support((10.07±3.07)d vs(13.03±4.05)d)and cost of nutritional support((3 180±260)yuan vs(4 550±660)yuan).Conclusion The device developed effectively enhances the efficacy for unblocking nasoenteric tubes,shortens the time required and decreases the days and cost of nutritional support,and thus is worthy promoting practi-cally.[Chinese Medical Equipment Journal,2025,46(1):114-117]

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