1.Spectrum-effect Relationship of Bupleuri Radix Processed with Trionyx sinensis Blood for Yin Deficiency Based on Saponins
Mengyu HOU ; Xia ZHAO ; Zhiyu GUO ; Ting LIU ; Yuexing MA ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):147-155
ObjectiveTo analyze the pharmacodynamic activity of Bupleuri Radix processed with Trionyx sinensis blood in the treatment of Yin deficiency and study the spectrum-effect relationship of this medicine. MethodsHigh performance liquid chromatography was employed to establish the fingerprints of 15 batches of Bupleuri Radix processed with Trionyx sinensis blood, and the similarity was evaluated according to the SOP of Similarity Evaluation System of Chromatographic Fingerprint of TCM (version 2012). A mouse model of Yin deficiency induced by thyroxine was established. The relationship between the active components and the effect on Yin deficiency was explored by grey correlation analysis and partial least squares method based on the changes in the serum levels of triiodothyronine (T3), thyroxine (T4), cyclic adenosine phosphate (cAMP), and cyclic guanosine phosphate (cGMP). The components screened out based on the spectrum-effect relationship were used for retrieval of the targets from the Traditional Chinese Medicine Systems Pharmacology and Analysis Database (TCMSP), The Encyclopedia of Traditional Chinese Medicine (ETCM), and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP). Furthermore, the Online Mendelian Inheritance in Man (OMIM), GeneCards, TTD, DisGeNET, and Drugbank were employed to establish the active component-target against Yin deficiency network of Bupleuri Radix processed with Trionyx sinensis blood. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were carried out for the core targets. Real-time PCR was conducted to verify the predicted key pathways and mechanisms. ResultsThe fingerprints of the 15 batches of Bupleuri Radix processed with Trionyx sinensis blood showed the similarities of 0.976-0.999 with the control fingerprint. Compared with the model group, the drug administration group showed elevated levels of T3 and T4 and lowered levels of cAMP, cGMP and cAMP/cGMP. The results of grey correlation analysis showed that active components in terms of the correlations followed the trend of saikosaponin B1 > saikosaponin B2 > saikosaponin C > saikosaponin D > saikosaponin A. The partial least squares analysis showed that saikosaponins A, D, B1, and B2 had higher VIP values. Network pharmacology predicted a total of 30 common targets, which were enriched in 276 GO terns and 115 KEGG pathways. The results of Real-time PCR showed that the model group had lower mRNA levels of Caspase-9, kinase insert domain receptor (KDR), and mammalian target of rapamycin (mTOR) and higher mRNA level of mouse double minute 2 homolog (MDM2) than the blank group and the drug administration group. ConclusionBupleuri Radix processed with Trionyx sinensis blood has therapeutic effect on Yin deficiency syndrome, which provides a new idea for studying Bupleuri Radix processed with Trionyx sinensis blood.
2.Diagnosis and treatment of colorectal liver metastases: Chinese expert consensus-based multidisciplinary team (2024 edition).
Wen ZHANG ; Xinyu BI ; Yongkun SUN ; Yuan TANG ; Haizhen LU ; Jun JIANG ; Haitao ZHOU ; Yue HAN ; Min YANG ; Xiao CHEN ; Zhen HUANG ; Weihua LI ; Zhiyu LI ; Yufei LU ; Kun WANG ; Xiaobo YANG ; Jianguo ZHOU ; Wenyu ZHANG ; Muxing LI ; Yefan ZHANG ; Jianjun ZHAO ; Aiping ZHOU ; Jianqiang CAI
Chinese Medical Journal 2025;138(15):1765-1768
3.CDH17-targeting CAR-NK cells synergize with CD47 blockade for potent suppression of gastrointestinal cancers.
Liuhai ZHENG ; Youbing DING ; Xiaolong XU ; Huifang WANG ; Guangwei SHI ; Yang LI ; Yuanqiao HE ; Yue GONG ; Xiaodong ZHANG ; Jinxi WEI ; Zhiyu DONG ; Jiexuan LI ; Shanchao ZHAO ; Rui HOU ; Wei ZHANG ; Jigang WANG ; Zhijie LI
Acta Pharmaceutica Sinica B 2025;15(5):2559-2574
Gastrointestinal (GI) cancers are a leading cause of cancer morbidity and mortality worldwide. Despite advances in treatment, cancer relapse remains a significant challenge, necessitating novel therapeutic strategies. In this study, we engineered nanobody-based chimeric antigen receptor (CAR) natural killer (NK) cells targeting cadherin 17 (CDH17) for the treatment of GI tumors. In addition, to enhance the efficacy of CAR-NK cells, we also incorporated CV1, a CD47-SIRPα axis inhibitor, to evaluate the anti-tumor effect of this combination. We found that CDH17-CAR-NK cells effectively eliminated GI cancers cells in a CDH17-dependent manner. CDH17-CAR-NK cells also exhibit potent in vivo anti-tumor effects in cancer cell-derived xenograft and patient-derived xenograft mouse models. Additionally, the anti-tumor activity of CDH17-CAR-NK cells is synergistically enhanced by CD47-signal regulatory protein α (SIRPα) axis inhibitor CV1, likely through augmented macrophages activation and an increase in M1-phenotype macrophages in the tumor microenvironment. Collectively, our findings suggest that CDH17-targeting CAR-NK cells are a promising strategy for GI cancers. The combination of CDH17-CAR-NK cells with CV1 emerges as a potential combinatorial approach to overcome the limitations of CAR-NK therapy. Further investigations are warranted to speed up the clinical translation of these findings.
4.Evaluation of A Liver Yin Deficiency Mouse Model Based on Untargeted Metabolomics
Xuening HUANG ; Xia ZHAO ; Mengyu HOU ; Zhiyu GUO ; Yaohui YE
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(5):118-125
ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS), to evaluate the establishment of a mouse model of liver Yin deficiency by thyroid tablet suspension combined with 10% carbon tetrachloride(CCl4) from the perspective of non-targeted metabolomics, in order to lay the foundation for the establishment of a traditional Chinese medicine(TCM) syndrome model. MethodA total of 24 mice were randomly divided into blank group and model group. The model group was given thyroid tablet suspension(0.003 2 g·kg-1) by gavage for 14 consecutive days, and 10% CCl4(5 mL·kg-1) was intraperitoneally injected once a week to establish a liver Yin deficiency model, while the blank group was injected with an equal amount of olive oil intraperitoneally and gavaged with an equal amount of distilled water, and was fed with normal feed. After the modeling was completed, 6 mice in each group were randomly selected, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), cyclic adenosine monophosphate(cAMP), cyclic guanosine monophosphate(cGMP), interleukin(IL)-6, IL-10, tumor necrosis factor-α(TNF-α)were measured in the mice serum, and malondialdehyde(MDA), superoxide dismutase(SOD), total protein(TP), hydroxyproline(HYP) and other indicators were measured in the mice liver. Liver tissue sections were taken for hematoxylin-eosin(HE) staining and observing pathological changes. The remaining 6 mice in each group were subjected to UPLC-Q-TOF-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to screen differential metabolites in the liver Yin deficiency mouse model, Kyoto Encyclopedia of Genes and Genomes(KEGG) database was used to analyze the corresponding metabolic pathways of differential metabolites. ResultCompared with the blank group, mice in the model group showed liver Yin deficiency manifestations such as reduced body weight, fatigue and sleepiness, disheveled and lusterless hair, irritability. The levels of ALT, cAMP/cGMP, IL-6, AST, MDA, cAMP, TNF-α significantly increased(P<0.05, P<0.01), while the levels of SOD, IL-10 and cGMP significantly decreased(P<0.05, P<0.01), and the changes of HYP and TP were not statistically significant. Hepatic steatosis and distortion of the radial arrangement of the liver plate cells were seen in the section images of the model group, endogenous substances were clearly separated, and 252 differential metabolites were identified in the serum samples, which were mainly involved in the metabolic pathways of purine metabolism, steroid hormone biosynthesis and pyrimidine metabolism. A total of 229 differential metabolites were identified in the liver samples, mainly involving nucleotide metabolism, purine metabolism, steroid hormone biosynthesis, pyrimidine metabolism, antifolate resistance, insulin resistance, primary bile acid biosynthesis, prostate cancer, sulfur relay system, arachidonic acid metabolism and other metabolic pathways. ConclusionThe successful establishment of liver Yin deficiency model in mice by CCl4 combined with thyroid hormone is evaluated through the investigation of serum and liver metabolomics, combined with biochemical indicators, which provides a biological basis and experimental foundation for the Yin deficiency syndrome model of TCM.
5.Autoimmune encephalitis with double positive anti-NMDAR and anti-GABABR secondary to herpes simplex virus encephalitis:A case report and literature review
Zhongyan ZHAO ; Zhiyu XU ; Chanji WU ; Eryi ZHAO ; Dan HUANG ; Shixiong HUANG
Journal of Jilin University(Medicine Edition) 2024;50(1):236-242
Objective:To analyze the clinical presentations and diagnostic and treatment process of one patient with autoimmune encephalitis(AE)with double positive anti-N-methyl-D-aspartate receptor(NMDAR)and anti-γ-aminobutyric acid B receptor(GABABR)secondary to herpes simplex virus encephalitis(HSVE),and to improve the clinicians'awareness of this disease.Methods:The clinical data of one AE patient with double positive anti-NMDAR and anti-GABABR secondary to HSVE were collected,the diagnostic and therapeutic processes were summarized,and the relevant literatures were reviewed.Results:The patient,a 36-year-old male,developed a headache followed by limb convulsions,and progressed to disturbed consciousness.After admission,the routine biochemistry of the cerebrospinal fluid(CSF)was abnormal,and the herpes simplex virus-1(HSV-1)IgG antibody showed positive in the CSF;both CSF and serum tests for NMDAR antibodies were positive;the head magnetic resonance imaging(MRI)results showed abnormal signals in the right occipital white matter,leading to the diagnosis of HSVE secondary to anti-NMDAR encephalitis.Several months later,the patient experienced psychiatric behavior abnormalities,cognitive dysfunction,and sleep disorders,and both the serum NMDAR and GABABR antibodies showed positive results,prompting the diagnosis of HSVE secondary anti-NMDAR encephalitis and anti-GABABR encephalitis.After treatment with steroid pulse therapy and intravenous immunoglobulin(IVIG),the patient's condition was improved and the patient was discharged.At one-year follow-up,the patient's psychiatric symptoms had completely resolved,leaving mild cognitive impairment.Conclusion:If the clinical symptoms of the patients recovering from antiviral treatment for HSVE is worsened,secondary AE should be highly suspected;it is important to complete autoimmunity antibody testing as soon as possible for the early diagnosis and treatment to improve the prognosis of the patient.
6.Predictive value of lactate/albumin ratio,interleukin-6 and CD4+T lymphocyte count in the short-term prognosis of severe pneumonia and sepsis
Zhiyu MIAO ; Lei ZHANG ; Xiaoyan LI ; Ruixia ZHANG ; Mengjiao ZHAO ; Yanqiu GAO
Journal of Xinxiang Medical College 2024;41(3):232-239
Objective To investigate the predictive value of lactate/albumin ratio(LAR),interleukin-6(IL-6)and CD4+T lymphocyte count in 28-day mortality in patients with severe pneumonia and sepsis.Methods A total of 73 patients with severe pneumonia and sepsis admitted to the Respiratory Intensive Care Unit(RICU)of Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2022 to June 2023 were enrolled and divided into the survival group(n=43)and the death group(n=30)according to their 28-day outcomes.The clinical data of the patients were collected from their electronic medical records,including age,gender,comorbidities with hypertension,diabetes,and coronary artery heart disease(CHD),as well as sequential organ failure assessment(SOFA)score,acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,mean arterial pressure(MAP),confusion,uremia,respiratory rate,blood pressure,age ≥65 years(CURB-65)score,total bilirubin(Tbil),serum creatinine(Scr),platelet count(PLT),white blood cell(WBC)count,procalcitonin(PCT),and C-reactive protein(CRP)at admission to RICU.On the 1st,3rd,and 7th day after admission to RICU,the patients'arterial blood was drawn,and the lactate level was detected by a fully automated blood gas analyzer.The peripheral venous blood was drawn,and the serum albumin and IL-6 levels were detected by enzyme-linked immunosorbent assay,and the CD4+T lymphocyte subset count was measured by flow cytometry.The LAR of patients on the 1st,3rd and 7th day was calculated.The clinical data of the patients and the LAR,IL-6 level and CD4+T lymphocyte count on the 1st,3rd,and 7th day were compared between the two groups.The influencing factors of 28-day mortality in patients with severe pneumonia and sepsis were analyzed by logistic regression,and the predictive value of each influencing factor on the 28-day mortality in patients with severe pneumonia and sepsis was evaluated by the receiver operating characteristic(ROC)curve.Results There was no significant difference in gender,age,proportions of comorbidities with hypertension,diabetes and CHD,length of stay in RICU,and Tbil,MAP,PLT,Scr,WBC,PCT and CRP at admission to RICU(P>0.05).The APACHE Ⅱ and CURB-65 scores of the patients in the death group were significantly higher than those in the survival group(P<0.05).On the 1st,3rd and 7th day,the CD4+T lymphocyte count in the death group was significantly lower than that in the survival group,while the SOFA score was significantly higher than that in the survival group(P<0.05).On the first day,there was no significant difference in the LAR and IL-6 level be-tween the death group and the survival group(P>0.05).However,on the 3rd and 7th day,the LAR and IL-6 level in the death group were significantly higher than those in the survival group(P<0.05).The LAR,IL-6 level and SOFA score on the 3rd and 7th day in the survival group were significantly lower than those on the 1st day,and these indicators on the 7th day were sig-nificantly lower than those on the 3rd day(P<0.05);the CD4+T lymphocyte count on the 3rd and 7th day was significantly higher than that on the 1st day(P<0.05),while it showed no significant difference on the 7th and 3r day(P>0.05).The IL-6 level on the 7th day in the death group was significantly lower than that on the 1st and 3rd day(P<0.05),while there was no significant difference in IL-6 level on the 1st day compared with the 3r day(P>0.05);moreover,there was no significant difference in LAR,CD4+T lymphocyte count and SOFA score between each time point(P>0.05).Pearson correlation analy-sis showed that on the 3rd day,the LAR and IL-6 level were significantly positively correlated with the SOFA score in patients with severe pneumonia and sepsis(r=0.385,0.394;P<0.05).On the 7th day,the LAR and IL-6 level were also significantly positively correlated with the SOFA score(r=0.418,0.402;P<0.05).On the 3 rd and 7 th day,CD4+T lymphocyte count was significantly negatively correlated with the SOFA score(r=-0.451,-0.454;P<0.05).Logistic regression analysis showed that the APACHE Ⅱ score,LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day,and the IL-6 level and CD4+T lym-phocyte count on the 7th day were the influencing factors for 28-day mortality in patients with severe pneumonia and sepsis(P<0.05).The ROC curve showed that the APACHE Ⅱ score,LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day and the combination of the three,IL-6 level and CD4+T lymphocyte count on the 7th day and the combination of the two had certain predictive value for the 28-day mortality in patients with severe pneumonia and sepsis(P<0.05).The area under the ROC curve(AUC)of LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day combined to predict 28-day mortality in patients with severe pneumonia and sepsis was 0.891,and the AUC of APACHE Ⅱ score for predicting 28-day mortality in pa-tients with severe pneumonia and sepsis was 0.769.The AUC values of LAR,IL-6 level and CD4+T lymphocyte count on the 3rd day for predicting 28-day mortality in patients with severe pneumonia and sepsis were 0.795,0.757 and 0.770,respective-ly,and the AUC values of IL-6 level and CD4+T lymphocyte count on the 7th day and their combination for predicting 28-day mortality in patients with severe pneumonia and sepsis were 0.743,0.802 and 0.888,respectively.Conclusion The 3-day LAR,IL-6 level and CD4+T lymphocyte count,and the 7-day IL-6 level and CD4+T lymphocyte count after admission are re-lated to the 28-day mortality in patients with severe pneumonia and sepsis.The combined LAR,IL-6 level and CD4+T lympho-cyte count on the 3rd day can better assess the severity and prognosis of patients.
7.Global fatality rate and transmission factors of Nipah virus disease:A meta-analysis
Hongli LI ; Qixun CUI ; Weiwen YAN ; Yifei LIU ; Yuan LIANG ; Zhiyu WANG ; Dongx-Ia MENG ; Yanping ZHAO
Chinese Journal of Veterinary Science 2024;44(6):1229-1238,1247
Nipah virus(NipahVirus,NiV)is a zoonotic virus that can cause encephalitis and severe respiratory symptoms in humans and animals,and is at risk of being introduced into China.At present,there are many factors related to the transmission of Nipah virus disease(NVD),but the actual effects of these factors are controversial.Therefore,with the help of meta-analysis,this paper aims to determine the current mortality and the main means of transmission of NVD,so as to pro-vide reference for the input of controlling Nipah virus disease and making emergency plans for pre-vention and control in our country.By searching the articles published in Pubmed,Embase,Web of knowledge,CNKI,VIP Chinese Sci-tech Journals Database and Wanfang Database up to December 31,2022,the literature of cross-sectional,cohort and case-control studies with NiV encephalitis fa-tality rate(CFR)and risk factors were selected.Results showed that a total of 25 literatures were included in the meta-analysis after retrieval and screening,and the fatality rate of NiV encephalitis was 64.6%(95%CI,60.8-68.2;I2=96.8%).Subgroup analysis showed that tree climbing(OR=1.43;95%CI:1.05-1.96),bats were seen near their nighttime residence(OR=2.38;95%CI:1.74-3.25)and direct contact with date palm SAP(OR=6.01;95%CI:4.07-8.89)Bananas(OR=2.25;95%CI:1.31-3.85)OR porcine(OR=11.87;95%CI:1.15-122.23)was significantly associated with NiV encephalitis.The results of this study suggest that NiV encephalitis has a high mortality rate,and tree climbing,nocturnal exposure to bats,and exposure to pig,banana,and date palm SAP increase the risk of NiV encephalitis.In order to prevent NiV transmission,epidemic surveillance,education and publicity should be strengthened and protective measures should be taken.
8.Association of copy number variation in X chromosome-linked PNPLA4 with heterotaxy and congenital heart disease
Han GAO ; Xianghui HUANG ; Weicheng CHEN ; Zhiyu FENG ; Zhengshan ZHAO ; Ping LI ; Chaozhong TAN ; Jinxin WANG ; Quannan ZHUANG ; Yuan GAO ; Shaojie MIN ; Qinyu YAO ; Maoxiang QIAN ; Xiaojing MA ; Feizhen WU ; Weili YAN ; Wei SHENG ; Guoying HUANG
Chinese Medical Journal 2024;137(15):1823-1834
Background::Heterotaxy (HTX) is a thoracoabdominal organ anomaly syndrome and commonly accompanied by congenital heart disease (CHD). The aim of this study was to analyze rare copy number variations (CNVs) in a HTX/CHD cohort and to examine the potential mechanisms contributing to HTX/CHD.Methods::Chromosome microarray analysis was used to identify rare CNVs in a cohort of 120 unrelated HTX/CHD patients, and available samples from parents were used to confirm the inheritance pattern. Potential candidate genes in CNVs region were prioritized via the DECIPHER database, and PNPLA4 was identified as the leading candidate gene. To validate, we generated PNPLA4-overexpressing human induced pluripotent stem cell lines as well as pnpla4-overexpressing zebrafish model, followed by a series of transcriptomic, biochemical and cellular analyses. Results::Seventeen rare CNVs were identified in 15 of the 120 HTX/CHD patients (12.5%). Xp22.31 duplication was one of the inherited CNVs identified in this HTX/CHD cohort, and PNPLA4 in the Xp22.31 was a candidate gene associated with HTX/CHD. PNPLA4 is expressed in the lateral plate mesoderm, which is known to be critical for left/right embryonic patterning as well as cardiomyocyte differentiation, and in the neural crest cell lineage. Through a series of in vivo and in vitro analyses at the molecular and cellular levels, we revealed that the biological function of PNPLA4 is importantly involved in the primary cilia formation and function via its regulation of energy metabolism and mitochondria-mediated ATP production. Conclusions::Our findings demonstrated a significant association between CNVs and HTX/CHD. Our data strongly suggested that an increased genetic dose of PNPLA4 due to Xp22.31 duplication is a disease-causing risk factor for HTX/CHD.
9.Inhibitory effect of miR-30c-5p on proliferation,migration,and invasion of prostate cancer cells and its mechanism
Bin ZHAO ; Jinye YANG ; Zhiyao LI ; Chengwei BI ; Libo YANG ; Zhiyu SHI ; Xin LI ; Jianpeng ZHANG ; Yuanlong SHI ; Yong YANG ; Guoying ZHANG
Journal of Jilin University(Medicine Edition) 2024;50(6):1632-1643
Objective:To discuss the effect of microRNA(miR)-30c-5p on the proliferation,migration,and invasion of the human prostate cancer cells(LNCap),and to clarify its possible mechanism.Methods:The LNCap cells were divided into LNCap group(without plasmid transfection),miR-30c-5p mimic group(transfected with miR-30c-5p mimic),mimic NC group(transfected with miR-30c-5p mimic NC),sh-DNA damage inducible transcript 4(DDIT4)group(transfected with sh-DDIT4),sh-NC group(transfected with sh-DDIT4 NC),miR-30c-5p mimic+pc-DNA3.1-NC group(co-transfected with miR-30c-5p mimic and pc-DNA3.1 empty vector),and miR-30c-5p mimic+pc-DNA3.1-DDIT4 group(co-transfected with miR-30c-5p mimic and pc-DNA3.1-DDIT4 over-expression plasmid).The RWPE-1 cells were cultured normally.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of miR-30c-5p and DDIT4 mRNA in the cells in various groups;Western blotting method was used to detect the expression levels of DDIT4 protein in the cells in various groups;CCK-8 method was used to detect the proliferation rates of the LNCap cells in various groups;Transwell assay was used to detect the numbers of the invasion LNCap cells in various groups;Scratch assay was used to detect the scratch healing rates of LNCap cells in various groups;dual-luciferase reporter assay was used to detect the targeting relationship between miR-30c-5p and DDIT4.In the in vivo tumor formation experiment,18 male BALB/c nude mice were divided randomly into blank group,agomiR-NC group(transfected with agomiR-30c-5p NC),and agomiR-30c-5p group(transfected with agomiR-30c-5p);there were six mice in each group.The mice in agomiR-NC group and agomiR-30c-5p group were subcutaneously injected with LNCap cells,while the mice in blank group were given an equal volume of physiological saline.The volumes of tumor of the mice in various groups were detected.HE staining was used to observe the morphology of prostate cancer tissue the mice of in various groups;RT-qPCR method and immunofluorescence staining were used to detect the expression levels of miR-30c-5p and DDIT4 mRNA and the fluorescence intensities of DDIT4 protein in prostate cancer tissue of the mice in various groups.Results:The In vitro prostate cancer cell experiment results showed that compared with RWPE-1 cells,the expression level of miR-30c-5p in the prostate cancer LNCap cells was decreased(P<0.01),and the expression levels of DDIT4 mRNA and protein were increased(P<0.05 or P<0.01).After 48 of transfection,compared with LNCap group and mimic NC group,the expression level of miR-30c-5p in the LNCap cells in miR-30c-5p mimic group was increased(P<0.01).Compared with LNCap group and sh-NC group,the expression level of DDIT4 mRNA in the LNCap cells in sh-DDIT4 group was decreased(P<0.01).Compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of miR-30c-5p in The LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 mRNA in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the expression level of DDIT4 protein in the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.05).The CCK-8 method results showed that compared with LNCap group and mimic NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the proliferation rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the proliferation rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The Transwell assay results showed that compared with LNCap group and mimic NC group,the number of the invasion LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the number of invasion LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the number of the invasion LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The scratch assay results showed that compared with LNCap group and mimic NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic group was decreased(P<0.01);compared with LNCap group and sh-NC group,the scratch healing rate of the LNCap cells in sh-DDIT4 group was decreased(P<0.01);compared with miR-30c-5p mimic group and miR-30c-5p mimic+pcDNA3.1 NC group,the scratch healing rate of the LNCap cells in miR-30c-5p mimic+pc-DNA3.1-DDIT4 group was increased(P<0.01).The dual-luciferase reporter assay results showed that compared with the LNCap cells co-transfected with WT-DDIT4 and mimic NC,the luciferase activity of the LNCap cells co-transfected with WT-DDIT4 and miR-30c-5p mimic was decreased(P<0.01).The in vivo nude mouse tumor formation experiment results showed that on the 3 rd,6 th,9 th,12 th,and 15th days after cell injection,compared with blank group and agomiR-NC group,the tumor volumes of the nude mice in agomiR-30c-5p group were decreased(P<0.05).The HE staining results showed that in prostate cancer tissue of the mice in blank group and agomiR-NC group,the cell nuclei were enlarged,and nucleoli were prominent and deformed.In the mice in agomiR-30c-5p group,some regions of prostate cancer tissues results showed neatly arranged cells with normally shaped nuclei.The RT-qPCR and immunofluorescence staining showed that compared with agomiR-NC group,the expression level of miR-30c-5p in prostate cancer tissue of the mice in agomiR-30c-5p group was increased(P<0.01).Compared with blank group and agomiR-NC group,the expression level of DDIT4 mRNA in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).DDIT4 protein was mainly expressed in the cytoplasm.Compared with blank group and agomiR-NC group,the fluorescence intensity of DDIT4 protein in prostate cancer tissue of the mice in agomiR-30c-5p group was decreased(P<0.01).Conclusion:The expression level of miR-30c-5p in the prostate cancer LNCap cells is decreased,and it inhibits the proliferation,migration,and invasion of the prostate cancer cells by targeting downregulation of DDIT4,thereby participating in the occurrence and development of prostate cancer.
10.Construction and evaluation of a risk prediction model for linezolid-related neurological adverse reactions in patients with multidrug-resistant tuberculosis
Haojie TANG ; Zilong YANG ; Zhaoxian YU ; Zhiyu FENG ; Haiping DONG ; Xiang LI ; Wei ZHAO ; Haobin KUANG
The Journal of Practical Medicine 2024;40(19):2690-2695
Objective To investigate the determinants of linezolid-associated neurological adverse reactions in patients with multidrug-resistant tuberculosis and develop a risk prediction model for such adverse events.Methods A prospective cohort study design was employed to select 120 patients with drug-resistant pulmonary tuberculosis who received a chemotherapy regimen containing linezolid at Guangzhou Chest Hospital from April 2023 to January 2024 as the study population.Clinical data,adverse reactions,and plasma concentration of linezolid were collected during fasting and at 2 hours post-medication.Univariate analysis and multivariate logistic regression were conducted to identify factors influencing linezolid-related neurological adverse reactions.Furthermore,a prediction model for such adverse reactions was developed,and its predictive efficacy and calibration ability were evaluated using ROC analysis.Results Re-treatment(OR=2.540,P=0.028),coexistence of cavities(OR=4.092,P=0.021),anemia(OR=10.921,P=0.005),and Cmin≥0.7665 mg/L(OR=6.813,P<0.001)are independent risk factors for the occurrence of linezolid-related neurological adverse reactions.The prediction model,based on these four factors,exhibits an AUC of 0.851(95%CI:0.774~0.929),accompanied by a Youden index of 0.590,a sensi-tivity of 66.7%,and a specificity of 92.3%.Moreover,the prediction model demonstrates excellent calibration ability.(Hosmer-lemeshow χ2=8.719,P=0.273).Conclusion In MDR/RR-TB patients,the presence of cavita-tion,retreatment,and anemia may confer a heightened risk of linezolid-related neurological adverse reactions.A risk prediction model incorporating these four indicators demonstrates significant predictive value for the occurrence of such adverse events.

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