Table of Content

  

10 September 2023
Volume 61 Issue 9

Preclinical Medicine
Clinical Medicine


Preclinical Medicine

Development and characterization of π-π stacked and chemical crosslinked polymeric micelles

LYU Xiaolin, WU Yingli, YANG Yu, GONG Xujin, LIU Yanna, YAO Qingqiang

Journal of Shandong University (Health Sciences). 2023, 61(9):  1-9.  doi:10.6040/j.issn.1671-7554.0.2023.0137

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Objective To improve the stability of micelles in blood circulation and the retention of drugs in micelles by synthesizing amphiphilic block copolymer PEG-b-P(TMC-COOH)-b-P(CL-Bz). Methods PEG-b-P(TMC-Bz)block was firstly synthesized by carbonate monomers(TMC-Bz)containing benzyl side chains. Then, PEG-b-P(TMC-Bz)-b-P(CL-Bz)containing benzyl side chains was synthesized using hydroxyl end groups of PEG-b-P(TMC-Bz)as initiator. Finally, the benzyl group in P(TMC-Bz)was removed under Pd/C to obtain PEG-b-P(TMC-COOH)-b-P(CL-Bz). The synthesized monomers and polymers were characterized by nuclear magnetic resonance hydrogen spectroscopy(¹H NMR)and gel permeation chromatography(GPC). In addition, the empty and drug-loaded crosslinked micelles prepared by the resulting polymer were characterized by dynamic light scattering(DLS), and the effects of crosslinking reaction time and the amount of crosslinking agent on crosslinking degree were explored by GPC. Results PEG-b-P(TMC-COOH)-b-P(CL-Bz)polymer was successfully synthesized. The crosslinked polymer micelles prepared had a small and uniform particle size(about 80 nm), and the loading efficiency of paclitaxel in the micelles reached more than 90%. The crosslinking reaction based on the activated carboxyl group in the polymer and the amino group in the crosslinker cystamine dihydrochloride ended within 12 h, and the degree of crosslinking increased with the increase of the amount of crosslinker. Conclusion The PEG-b-P(TMC-COOH)-b-P(CL-Bz)polymeric micelles as a drug delivery system were successfully developed, which can realize the interaction of π-π stacking and chemical crosslinking.

Effects of Helicobacter pylori infection on m6A level in gastric cancer cells and its mechanism

SUN Jingguo, ZHU Wenshuai, LU Yi, MA Xiaoli, JIA Yanfei

Journal of Shandong University (Health Sciences). 2023, 61(9):  10-18.  doi:10.6040/j.issn.1671-7554.0.2022.1289

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Objective To investigate the effects of Helicobacter pylori(HP)infection on the level of m6A in gastric cancer and the mechanism. Methods After gastric cancer cell line MGC-803 was co-cultured with HP strains NCTC 11637, NCTC 26695 and PM SS1 at MOI=100 for 8 hours, the m6A level was detected with dot-blot and ELISA. The expressions of METTL3 and METTL14 were detected with qRT-PCR and Western blotting, respectively. Relevant data were downloaded from the GEO database to analyze the relationship between m6A methyltransferase and clinicopathological parameters. The difference in the expression of METTL14 in gastric cancer and normal tissues from TCGA and GTEx databases was analyzed with GEPIA. Results HP upregulated the m6A level in gastric cancer cells. HP infection promoted METTL14 expression. METTL14 expression correlated with LAUREN typing and was higher in intestinal type gastric cancer. METTL14 expression was higher in gastric cancer tissues than in normal adjacent tissues. Conclusion HP infection increases the m6A level in gastric cancer cells, which may be caused by the upregulation of METTL14.

Clinical value of serum exosomal LINC02163 as a marker for distant metastasis of colorectal cancer

DONG Yaqi, WANG Xinhui, ZHAO Yinghui, WANG Chuanxin

Journal of Shandong University (Health Sciences). 2023, 61(9):  19-28.  doi:10.6040/j.issn.1671-7554.0.2023.0243

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Objective To investigate the correlation between long non-coding RNA(lncRNA)LINC02163 and distant metastasis of colorectal cancer and its biological function, and to evaluate the potential of serum exosomal LINC02163 as a marker for distant metastasis. Methods The differential lncRNAs was screened by analyzing the transcriptome data of colorectal cancer tissues. The clinicopathological parameters and demographic parameters of patients were combined to analyze the expression characteristics of LINC02163. The correlation between LINC02163 and prognosis was investigated by in situ hybridization(ISH)in colorectal cancer microarray. The effects of LINC02163 on the distant metastasis of SW620 cells in vivo was investigated by establishing a tumor metastasis model in nude mice. Serum exosomes were identified with transmission electron microscopy, nanoparticle tracking analysis and Western blotting. The stability of serum exosomal LINC02163 was analyzed with RNase A incubation assay. The receiver operating characteristic(ROC)curve was drawn to evaluate the efficacy of serum exosomal LINC02163 in the identification of distant metastasis of colorectal cancer. Results The high expression of LINC02163 in colorectal cancer was screened by five colorectal cancer tissue transcriptome data. TCGA data analysis demonstrated that the expression of LINC02163 was associated with distant metastasis of colorectal cancer(P<0.05). ISH assay showed that high expression of LINC02163 was associated with poor prognosis(P<0.05). Tumor metastasis model in nude mice revealed that LINC02163 knockdown inhibited the distant metastasis of SW620 cells in vivo(P=0.05). Experiments involving the characterization of exosomes showed that serum exosomes were effectively extracted. RNase A incubation demonstrated the high stability of serum exosomal LINC02163. ROC analysis indicated that the area under the curve of serum exosomal LINC02163 was 0.864 for distant metastatic colorectal cancer. Conclusion LINC02163 is highly expressed in colorectal cancer and is associated with distant metastasis. Serum exosomal LINC02163 may become a marker of distant metastasis.

Effects of Pulsatilla decoction on the intestinal function in mice with short bowel syndrome

CHEN Taiyu, TANG Xuegui, TANG Shiyu, ZHANG Zhibin, CHEN Simin, LI Min, PANG Jun

Journal of Shandong University (Health Sciences). 2023, 61(9):  29-37.  doi:10.6040/j.issn.1671-7554.0.2023.0309

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Objective To explore the effects of Pulsatilla decoction(BTWD)on the intestinal permeability and intestinal flora structure in mice with short bowel syndrome(SBS). Methods Kunming mice were randomly divided into the sham group, model group, high-dose group, medium-dose group, and low-dose group. Mice in the sham group underwent jejunal transection anastomosis, while in the other groups, the middle small intestine of mice were resected to establish the SBS models. In the sham and model groups, saline was instilled by gavage starting from the 2nd postoperative day. In the other groups, equal volume of BTWD with different concentrations was instilled by gavage. After 4 weeks of treatment, the mice were sacrificed. The pathological morphology of ileal tissue was observed with HE staining; the protein expressions of occludin and ZO-1 were determined with immunohistochemistry; the levels of serum tumor necrosis factor-alpha(TNF-α)and interleukin-6(IL-6)were detected with ELISA; the intestinal flora was analyzed with 16S rDNA sequencing. Results Compared with the model group, the high-, medium-, and low-dose groups showed dose-dependently reduced ileal mucosal inflammatory cell infiltration; the ileal villus height, circular muscle thickness and longitudinal muscle thickness were significantly increased; the crypt depth was significantly reduced(P<0.05); the protein expressions of ileal occludin and ZO-1 were significantly increased(P<0.01); the serum TNF-α and IL-6 levels were significantly decreased(P<0.01). Analysis of intestinal flora showed that BTWD significantly regulated the relative abundance of phylum and genus. Conclusion BTWD can improve the intestinal mucosal structure, repair the intestinal mucosal barrier, regulate the relative abundance of phyla, and increase the diversity of intestinal flora.

Impacts of CircFAT1 on the proliferation, apoptosis and radiosensitivity of nasopharyngeal carcinoma cells by regulating miR-296-3p/MAPRE1 axis

CAO Hualin, JIA Yanzhao, QU Li, YIN Xin

Journal of Shandong University (Health Sciences). 2023, 61(9):  38-46.  doi:10.6040/j.issn.1671-7554.0.2023.0140

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Objective To explore the impacts of circular RNA fat atypical cadherin 1(CircFAT1)on the proliferation, apoptosis and radiosensitivity of nasopharyngeal carcinoma cells by regulating miR-296-3p/microtubule-associated protein RP/EB family member 1(MAPRE1)axis. Methods The expressions of CircFAT1, miR-296-3p and MAPRE1 in NP69, CNE2 and CNE2-RR cells were detected with real-time fluorescent quantitative PCR and Western blotting. CNE2 cells cultured in vitro were randomly divided into the control group, CircFAT1 knockdown group, negative control group, and CircFAT1 knockdown+miR-296-3p inhibitor group. After transfection, the expressions of CircFAT1, miR-296-3p and MAPRE1 were detected with real-time fluorescent quantitative PCR and Western blotting. The proliferation and apoptosis of CNE2 cells in each group were detected with CCK-8 assay, 5-Ethynyl-2’-deoxyuridine_(Edu)staining and flow cytometry. CNE2-RR cells cultured in vitro were randomly divided into the control group, radiation group, radiation+negative control group, radiation+CircFAT1 knockdown group, and radiation+CircFAT1 knockdown+miR-296-3p inhibitor group. After transfection, the expressions of CircFAT1, miR-296-3p and MAPRE1 were detected with real-time fluorescent quantitative PCR and Western boltting; the proliferation and apoptosis of CNE2-RR cells in each group were detected with CCK-8 assay, Edu staining and flow cytometry. The targeting regulation of CircFAT1 on miR-296-3p and miR-296-3p on MAPRE1 were detected with double luciferase reporter gene assay. Results Compared with NP69 cells, CNE2 and CNE2-RR cells had increased expression of CircFAT1 and mRNA and protein expressions of MAPRE1(P<0.05), but decreased expression of miR-296-3p(P<0.05). Compared with CNE2 cells, CNE2-RR cells had increased mRNA and protein expressions of CircFAT1 and MAPRE1(P<0.05), but decreased expression of miR-296-3p(P<0.05). Compared with the control group, the CircFAT1 knockdown group had decreased mRNA and protein expressions of CircFAT1 and MAPRE1, and reduced survival rate and proliferation rate(P<0.05), but increased expression of miR-296-3p and apoptosis rate(P<0.05). There were no significant changes in the indexes of the negative control group(P>0.05); miR-296-3p inhibitor reversed the effects of CircFAT1 knockdown on the indexes. Compared with the control group and radiation group, the radiation + CircFAT1 knockdown group had decreased mRNA and protein expressions of CircFAT1 and MAPRE1, and reduced survival rate and proliferation rate(P<0.05), but increased expression of miR-296-3p and apoptosis rate(P<0.05). There were no significant changes in the cell indexes of the radiation group and radiation + negative control group(P>0.05); miR-296-3p inhibitor reversed the effects of CircFAT1 knockdown on the indicators of CNE2-RR cells under radiation treatment. CircFAT1 targeted and downregulated the expression of miR-296-3p, and miR-296-3p targeted and downregulated the expression of MAPRE1 in CNE2-RR cells. Conclusion Knockdown of CircFAT1 can reduce the expression of MAPRE1 by upregulating miR-296-3p, thereby inhibiting the proliferation, promoting the apoptosis, and enhancing the radiosensitivity of nasopharyngeal carcinoma cells.

PM_2.5 induces inflammatory response in nasal epithelial cells through TLR4/NF-κB pathway

YANG Xiaozhe, ZHAO Yan, QING Hui, WANG Xiangdong, ZHANG Luo

Journal of Shandong University (Health Sciences). 2023, 61(9):  47-55.  doi:10.6040/j.issn.1671-7554.0.2023.0524

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Objective To investigate the mechanism of TLR4/NF-κB signaling pathway in the inflammatory response of nasal mucosal epithelial cells induced by fine particulate matter(PM_2.5). Methods After healthy human nasal epithelial cells(HNEpCs)were exposed to PM_2.5, the RNA was extracted for transcriptome analysis by Illumina sequencer. The expressions of TLR4, NFKB1 and NFKB1A were detected with quantitative real-time PCR(qRT-PCR). Results PM_2.5 caused cell toxicity and abnormal regulation of transcriptome of HNEpCs, and abnormal expressions of TLR4, NFKBIA and IL1B. KEGG pathway analysis indicated that TLR4/NF-κB pathway was significantly enriched. The qRT-PCR validation experiment revealed that after PM_2.5 exposure, TLR4 was up-regulated while NFKB1 was down-regulated, and NFKBIA,which inhibited NF-κB, was significantly down-regulated, thus leading to abnormal regulation of the TLR4/NF-κB pathway. Conclusion The abnormal regulation of TLR4/NF-κB in HNEpCs may play a crucial role in the nasal mucosa inflammatory response induced by PM_2.5 exposure.

Bioinformatics identification of the Hub genes and mechanism of hypoxia-induced mitochondrial damage in mouse kidney

GAO Yujie, LONG Qifu, HU Ying, XU Yuzhen, WANG Ru, YONG Sheng

Journal of Shandong University (Health Sciences). 2023, 61(9):  57-68.  doi:10.6040/j.issn.1671-7554.0.2023.0427

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Objective To explore the Hub genes of hypoxia-induced mitochondrial damage by bioinformatics analysis, and to analyze the correlation between hub genes and apoptosis of mouse kidney tissue, so as to elucidate the molecular mechanism of hypoxia-induced mitochondrial damage in mice at high altitude. Methods After animal models of 30 d normoxicity group(n=5)and hypoxia group(n=5)were established, blood gas and renal indexes were determined. Histopathological changes of renal tissues were observed with H&E staining, and transcriptomics sequencing was performed. The differentially expressed genes(DEGs)were correlated with mitochondrial database and apoptosis-related gene database to identify DEGs related to mitochondrial function(DE-MFRGs)and DEGs related to apoptosis(DE-ARGs), and to screen Hub genes for protein-protein interaction(PPI). KEGG enrichment analysis was conducted on DEGs, DE-MFRGs and DE-ARGs. Hub genes and DE-ARGs were verified with RT-qPCR and Western blotting. Results Compared with the normoxicity group, the hypoxia group had significantly decreased kidney indexes. H&E staining results showed that the glomeruli in the hypoxia group were significantly atrophied; the renal tubule epithelial cells were swollen, ruptured, or infiltrated by inflammatory cells; the volume of some renal tubule epithelial cells was increased; the cytoplasm was loose and light stained, with cavitation degeneration. A total of 3,007 DEGs were identified under hypoxic exposure, among which 464 were related to mitochondrial function and were defined as DE-MFRGs. KEGG enrichment analysis of DE-MFRGs showed that they were involved with Parkinsons disease signaling pathway, oxidative phosphorylation signaling pathway, thermogenesis signaling pathway, Alzheimers disease signaling pathway, tricarboxylic acid cycling signaling pathway, NAFLD signaling pathway, metabolic signaling pathway and other pathways related to mitochondrial function. Three Hub genes(SOD2, TUFM and MRPL12)were identified through the construction of PPI network. The mRNA and protein expressions of SOD2, TUFM and MRPL12 were down-regulated. Furthermore, Hub genes were positively or negatively correlated with 33 ARGs genes, and the mRNA expression of CASPASE3 and BAK were up-regulated, while that of BCL2 was down-regulated. Conclusion The high altitude hypoxia environment can regulate the mRNA expressions of SOD2, TUFM and MRPL12, mediate the mitochondrial dysfunction of mouse kidney tissue, and induce damage and apoptosis.

Clinical Medicine

Screening of prognosis-related RNA editing sites in clear cell renal cell carcinoma

SONG Zhaolu, DONG Zhengxuan, PENG Chuanzhen, HUANG Caina, HU Keqing, HUANG Yongsheng, YAN Lei

Journal of Shandong University (Health Sciences). 2023, 61(9):  69-78.  doi:10.6040/j.issn.1671-7554.0.2023.0150

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Objective To explore RNA editing sites associated with survival and prognosis in clear cell renal cell carcinoma(ccRCC). Methods After the expression data of RNA editing sites in the Cancer Genome Atlas(TCGA)were downloaded, the minimum absolute contraction, selection operator(LASSO)and Cox regression were used to identify RNA editing sites closely associated with ccRCC prognosis. A prognostic model and nomogram were constructed after the risk scores were calculated based on the multivariate Cox results. The accuracy of the prognostic model and nomogram was evaluated with the receiver operating characteristic(ROC)curve, and the relevant functional analysis was performed. Results A total of 25 RNA editing sites associated with ccRCC prognosis were screened. Patients were divided into the high-risk group(n=222)and low-risk group(n=226)based on the median risk score. Kaplan-Meier survival analysis showed that the overall survival(OS)and progression-free survival(PFS)were in shorter in the high-risk group than in the low-risk group(P<0.001). The area under the ROC curve(AUC)of 1-year, 3-year and 5-year survival predicted by the prognosis model were 0.801, 0.824 and 0.806, and those predicted by the nomogram were 0.858, 0.833 and 0.821, respectively. There were differences in biological function and drug sensitivity between the two groups. Conclusion The 25 RNA editing sites are expected to become new prognostic markers and drug therapeutic targets for patients with ccRCC.

Efficacy of Fuyueshu plant drink in the treatment of primary dysmenorrhea

ZHAO Tong, SU Xing, WANG Kai

Journal of Shandong University (Health Sciences). 2023, 61(9):  79-83.  doi:10.6040/j.issn.1671-7554.0.2023.0400

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Objective To explore the effects of Fuyueshu plant drink on primary dysmenorrhea. Methods A total of 32 patients with dysmenorrhea treated during Aug. 26 and Oct. 3, 2022 were included. The patients received ibuprofen sustained-release capsules in the first menstrual cycle and Fuyueshu in the second menstrual cycle. The pain, breast pain, chest and hypochondriac pain, chills, cold sweat, nausea, vomiting, diarrhea, dizziness, fatigue, irritability and serological indicators were compared between the two cycles. Results The total response rate of Fuyueshu was higher than that of ibuprofen(81.25% vs 43.75%). Fuyueshu was significantly better than ibuprofen in improving chills, cold sweat, diarrhea, dizziness, headache, fatigue and irritability(P<0.05). The two had comparable effects on the improvement of breast pain, nausea and vomiting, serum prostaglandin E2(PGE2), prostaglandin 2α(PGF2α)and β-endorphin(β-EP). Fuyueshu was superior to ibuprofen in increasing the level of tumor necrosis factor-α(TNF-α)(P<0.001). Conclusion Fuyueshu can effectively improve the clinical symptoms and serological indexes of primary dysmenorrhea.

Protective effects of crocin on intervertebral disc degeneration

LIU Jinbo, LIU Kaiwen, XIANG Chongxin, CHENG Lei

Journal of Shandong University (Health Sciences). 2023, 61(9):  84-93.  doi:10.6040/j.issn.1671-7554.0.2023.0460

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Objective To investigate the therapeutic effects and possible mechanism of crocin on intervertebral disc degeneration(IVDD). Methods Human nucleus pulposus tissue Pfirrmann II samples were divided into two parts. One part was used to extract nucleus pulposus cells. According to the composition of the culture medium, the cells were divided into 4 groups, including blank control group, tumor necrosis factor α(TNF-α)stimulation group, low-concentration of crocin treatment group(10 μg/mL)and high-concentration of crocin treatment group(40 μg/mL). After 24 h of incubation, the mRNA expressions of inflammatory factors and metabolic markers in the 4 groups were measured with real-time quantitative PCR. After 0 min, 15 min, 30 min and 24 h of incubation, the protein expressions of inflammatory factors, metabolic markers, apoptotic markers and p-P65, a key protein of the nuclear factor-activated B-cell κ light chain enhancer(NF-κB)signaling pathway, were measured with Western blotting. The effects of crocin on the NF-κB signaling pathway was determined with dual luciferase reporter gene assay. Another part of the samples was cultured in vitro and divided into blank control group, TNF-α stimulation group and crocin treatment group(40 μg/mL). The protein expressions of inflammatory factors and cellular metabolic markers were detected with immunohistochemistry. Results The results of real-time quantitative PCR and Western blotting showed that the inflammatory response and extracellular matrix degradation in the TNF-α stimulation group intensified, and the gene transcription and protein expression levels of inducible nitric oxide synthase(iNOS), cyclo-oxygenase-2, a disintegrin and metalloproteinase with thrombospondin motifs 5(ADAMTs-5), and matrix metalloproteinase 13(MMP-13)were elevated compared with those in the blank control group(P<0.05), while the gene transcription and protein expressions of aggrecan and collagen-2 were significantly decreased(P<0.05). Western blotting also showed that the protein expression of anti-apoptotic factor B lymphoblastoma-2 gene(Bcl-2)in the TNF-α stimulation group was significantly decreased(P<0.001)compared to the blank control group, while the protein expressions of Bcl-2-related X protein(P=0.004)and cleaved-caspase 3 were significantly higher(P=0.005). In contrast, the above changes induced by TNF-α were significantly suppressed in both the 10 μg/mL and 40 μg/mL crocin treatment groups(P<0.05). In addition, the protein expression of p-P65 was significantly increased in the TNF-α stimulation group compared with the blank control group(P<0.05), whereas it was significantly decreased in the 10 μg/mL and 40 μg/mL crocin treatment groups(P<0.05), and significantly suppressed in the 15 min and 30 min groups(P<0.05). The dual luciferase reporter gene assay showed that the fluorescence intensity increased in the TNF-α stimulation group compared with the blank group(P<0.001), and significantly decreased after crocin treatment(P=0.006). Conclusion Crocin can inhibit the inflammatory response, extracellular matrix degradation and apoptosis of nucleus pulposus cells, and the NF-κB signaling pathway plays an important role in the inhibition of IVDD by crocin.

Therapeutic effects of SpyGlass DS system combined with endoscopic retrograde cholangiopancreatography(ERCP)on patients with choledocholithiasis

HAN Zhuoxing, LIN Hui, DUAN Shigang, YE Zhirong, CHEN Xuanting, LIN Kangqiang

Journal of Shandong University (Health Sciences). 2023, 61(9):  94-100.  doi:10.6040/j.issn.1671-7554.0.2023.0351

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Objective To investigate the effects of SpyGlass DS system combined with endoscopic retrograde cholangiopancreatography(ERCP)on patients with choledocholithiasis. Methods Clinical data of patients who received SpyGlass DS combined with ERCP(n=23, experimental group)and patients who received ERCP only(n=51, control group)were collected. The operation indicators and outcomes, Oddi sphincter base pressure, peak systolic blood pressure and systolic frequency, as well as differences in the expression levels of serum alanine aminotransferase(ALT), aspartate aminotransferase(AST)and total bilirubin(TBil)were analyzed. Results Compared with the control group, the experimental group had longer operation time, higher hospitalization expenses, shorter hospital stay(P<0.05), higher stone clearance rate(95.65% vs 72.55%, P<0.05), and lower incidence of postoperative pancreatitis(8.70% vs 21.57%, P>0.05). The indicators of sphincter and hepatobiliary function in experimental group and control group were decreased after surgery, and the sphincter base pressure, peak systolic blood pressure and systolic frequency of experimental group were higher than those of control group(P<0.05). There were no significant differences in the expression levels of ALT, AST and TBil between the two groups(P>0.05). Conclusion SpyGlass DS system has a good application effect in ERCP lithotomy. It may be used in clinical practice as an effective surgical method for the treatment of choledocholithiasis.

Comprehensive bioinformatics analysis to identify differentially expressed genes for aberrant methylation modification in HBV-associated HCC

CHEN Yingjun, LIU Tonggang

Journal of Shandong University (Health Sciences). 2023, 61(9):  101-117.  doi:10.6040/j.issn.1671-7554.0.2023.0468

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Objective To explore the differentially expressed genes(DEGs)and molecular mechanism of abnormal methylation modification associated with the development of hepatitis B virus(HBV)-associated hepatocellular carcinoma(HCC)for the early diagnosis of this disease. Methods After the expression profile chips GSE121248, GSE107170 and DNA methylation chip GSE136319 were downloaded from the Gene Expression Database(GEO), the DEGs and differentially methylated genes(DMGs)between HBV-associated HCC tissues and adjacent tissues were screened with R language, and the visual volcano map was drawn. Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analysis of the methylated-differentially expressed genes(MDEGs)were performed to construct protein-protein interaction(PPI)networks. The molecular complex detection(MCODE)was conducted with Cytoscap, and key genes were screened with cytoHubba plugin. The mRNA expression levels of key genes were verified with the Cancer Genome Atlas(TCGA). The relationship between methylation and gene expression of key genes in HCC was determined with Pearson correlation coefficient. HPA database, Cox proportional hazard regression model, Kaplan Meier-plotter database and receiver operating characteristic(ROC)curve were used to verify the protein expressions of key genes, survival analysis and prediction accuracy. The correlation between the expressions of key genes and clinical indicators(tumor size, pathological stage)were analyzed. Results A total of 921 and 1,172 DEGs were screened from the GSE121248 and GSE107170 datasets, respectively, with 570 and 714 down-regulated and 351 and 458 upregulated genes, respectively. After differential analysis of GSE136319 data, 7 952 genes were hyptrmethylated and 2 630 genes were hypomethylated. A comprehensive analysis of DEGs and DMGs yielded 33 genes upregulated under hypomethylation modification and 158 genes downregulated under hypermethylation modification. GO enrichment analysis showed that the DEGs with abnormal methylation modification were mainly involved in organic acid catabolic process, carboxylic acid catabolic process and heme binding; KEGG pathways were mainly involved in chemical carcinogenesis, complements, coagulation cascades and PPAR signaling pathway. STRING and Cytoscape screened out 12 key genes related to methylation, including FTCD, HRG, C8A, FOXM1, FGA, KLKB1, MBL2, FETUB, TTK, AURKA, PRC1and MAD2L1. After clinical verification, FTCD, HRG, C8A, FOXM1, AURKA, PRC1, TTK and MAD2L1 were confirmed to be differentially expressed in HBV-related HCC and were associated with poor prognosis. The expression levels of FTCD, HRG, FOXM1, TTK, AURKA, PRC1 and MAD2L1 were correlated with tumor size and pathological stage. Conclusion FTCD, HRG, C8A, FOXM1, TTK, AURKA, PRC1 and MAD2L1 may play important roles in the pathogenesis of HBV-related HCC, which may serve as potential diagnostic markers and therapeutic targets.

Autoimmune enteropathy in children: a case report and literature review

FU Xiaoli, WEI Xuxia, XU Junjie, XUE Ning, ZHANG Le, CHEN Hongling

Journal of Shandong University (Health Sciences). 2023, 61(9):  118-124.  doi:10.6040/j.issn.1671-7554.0.2023.0217

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Objective To investigate the clinical and histopathological characteristics, diagnosis, treatment and prognosis of autoimmune enteropathy(AIE)in children. Methods The clinical data of a child with AIE treated at Department of Digestion, Childrens Hospital Affiliated to Shandong University in Dec. 2021 were retrospectively analyzed. With "childrens autoimmune enteropathy" "autoimmune enteropathy in children" "autoimmune enteropathy" as the key words, relevant literature in CNKI, Wanfang Data Knowledge Service Platform and PubMed database were retrieved from the inception to March 2023 to summarize the clinicopathological characteristics, diagnosis and treatment and prognosis of AIE. Results The girl, aged 3 years and 10 months, was admitted due to diarrhea for 18 days. According to the clinical symptoms and typical pathological changes, including refractory watery diarrhea, electrolyte disorder, hypoproteinemia and malnutrition, the diagnosis was AIE, after excluding infectious diarrhea, chylous diarrhea, extremely early inflammatory bowel disease, congenital genetic metabolic disease, and immunodeficiency disease. The child received methylprednisolone for more than 8 months, relapsed after the medication stopped, and then received tacrolimus. After tacrolimus use for more than six months, up to March 2023, the childs condition was improved. Literature retrieval collected 66 eligible cases of AIE in foreign children and 5 in Chinese children. Conclusion The incidence of AIE in children is low, and the main clinical manifestation is refractory diarrhea. Conventional treatment and avoidance diet are ineffective. Characteristic pathological histological changes provide an important basis for a clear diagnosis, and early diagnosis and timely treatment can help improve the prognosis.