Acta Universitatis Medicinalis Anhui

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Anhui Provincial Education Department

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Anhui Medical University

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Weng Jianping

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Editorial Board of Acta Universitatis Medi-cinalis Anhui Meishan Road , Hefei 230032

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ISSN 1000 - 1492

CN 34 - 1065/R

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Construction and validation of a prognostic risk assessment model for lung adenocarcinoma based on miR-34 family target genes

Gu Lingyu1, Ang Gelema2,Yang Dan2,Wang Huifeng3,Wang Lixin1, Dong Hui4

DOI: 专辑：医药卫生科技

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The role of DPP7 protein in hepatocellular carcinoma and its underlying mechanisms

Wang Bijun1, Cui Haodong2,3, Wu Wenyong2,3

DOI: 专辑：医药卫生科技

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Mechanism of miR-21 targeting inhibition of the PTEN/AKT/mTOR pathway in ameliorating chronic renal fibrosis in mice

Qi Jiao, Xu Shanshan, Qi Qige, Meng Yan, Zhao Jianrong, Zhang Liying

DOI: 专辑：医药卫生科技

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abstract:
To investigate the mechanism through which miR‑21 improves chronic renal fibrosis in mice via targeted modulation of the phosphatase and tensin homolog (PTEN)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway. Methods Thirty‑two chronic kidney disease model mice were randomly divided into four groups (n=8 each group): model group, miR‑21 overexpression group, miR‑21 inhibition group, and miR‑21 inhibition + MK‑2206 group. Eight healthy mice were included as the control group. The miR‑21 overexpression, miR‑21 inhibition, and miR‑21 inhibition + MK‑2206 groups received tail‑vein injections of lentivirus (50 μL, 1×10⁸ TU per mouse) once weekly for three weeks. The control and model groups were injected with an equal volume of empty vector (LV‑NC). The miR‑21 inhibition + MK‑2206 group additionally received gavage of the AKT/mTOR pathway inhibitor MK‑2206 (480 mg/kg) once weekly for three weeks. The expressions of miR‑21, 24‑h urinary protein, serum creatinine (Scr), blood urea nitrogen (BUN), and renal tissue levels of collagen I, collagen III, α‑smooth muscle actin (α‑SMA), and PTEN protein, as well as p‑AKT/AKT and p‑mTOR/mTOR ratios, were compared among groups. HE staining was used to observe pathological changes in renal tissue, and Masson staining was used to observe the degree of renal fibrosis. A dual‑luciferase assay was performed to verify the targeting relationship between miR‑21 and PTEN. Results Compared with the model group, miR‑21 expression in renal tissue increased in the miR‑21 overexpression group (P<0.05) and decreased in the miR‑21 inhibition group (P<0.05). Compared with the model group, the miR‑21 overexpression group showed increased 24‑h urinary protein, Scr, BUN, and renal tissue expression of collagen I, collagen III, and α‑SMA (all P<0.05), while these indicators decreased in the miR‑21 inhibition group (all P<0.05). Compared with the miR‑21 inhibition group, the miR‑21 inhibition + MK‑2206 group exhibited lower 24‑h urinary protein, Scr, BUN, and renal tissue expression of collagen I, collagen III, and α‑SMA (all P<0.05). Compared with the model group, the miR‑21 overexpression group showed decreased PTEN protein expression (P<0.05) and increased p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05), while the miR‑21 inhibition group showed increased PTEN expression (P<0.05) and decreased p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05). Compared with the miR‑21 inhibition group, the miR‑21 inhibition + MK‑2206 group had lower p‑AKT/AKT and p‑mTOR/mTOR ratios (P<0.05), with no significant difference in PTEN protein expression (P>0.05). HE and Masson staining showed normal kidney structure and almost no fibrosis in the control group. The model group exhibited glomerular enlargement, capillary loop adhesion, and focal fibrosis. The miR-21 overexpression group showed severe destruction of glomerular structure, accompanied by extensive fibrosis and renal tubular atrophy. The pathological changes and degree of fibrosis were alleviated in the miR-21 inhibition group. The miR-21 inhibition + MK-2206 group showed only mild pathological changes and mild fibrosis, with the interstitium being largely normal. Compared with PTEN-WT + NC mimics 1, the relative luciferase activity in the PTEN-WT + miR-21 mimics group decreased (P<0.001). There was no statistically significant difference in relative luciferase activity between PTEN-WT + NC mimics group and PTEN-MUT + miR-21 mimics group (P>0.05). Conclusion miR‑21 may improve renal function indicators and alleviate renal fibrosis in chronic kidney disease mice via targeted modulation of PTEN and subsequently inhibiting theAKT/mTOR pathway.

Incidence and determinants of posttraumatic stress disorder at three months following a road traffic accident

Yang Luodong1, Li Haohao2, Meng Yao2, Jiang Liang1, Hu Min1 , Zhang Guiqing1

DOI: 专辑：医药卫生科技

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abstract:
To investigate the incidence and influencing factors of posttraumatic stress disorder (PTSD) three months after a traffic accident, and to explore the role of social support and coping strategies. Methods A total of 117 individuals exposed to trauma following road traffic accidents were recruited. General demographic and clinical information was collected within one week, and the hamilton anxiety rating scale (HAMA), the hamilton depression rating scale-24 (HAMD-24), the social support rating scale (SSRS), and the simplified coping style questionnaire (SCSQ) were administered. A 3-month follow-up was subsequently conducted, during which PTSD symptoms were assessed using the post-traumatic stress disorder checklist for DSM-5 (PCL-5). Participants were divided into a PTSD group and a non-PTSD group according to whether PTSD occurred. Between-group comparisons were performed using the Mann-Whitney U non-parametric test or the χ2 test, as appropriate. Spearman’s correlation analysis was used to examine the associations between general characteristics and PCL-5 scores. Binary logistic regression was applied to identify factors influencing PTSD, and receiver operating characteristic (ROC) curve analysis was conducted to evaluate the diagnostic value of the SCSQ and SSRS. Results During the 3-month follow-up of the 117 trauma-exposed individuals, 17 cases developed PTSD, with a higher proportion of females (70.59%). Between-group comparisons showed that, compared with the PTSD group, the non-PTSD group had higher scores for positive coping, objective support, and subjective support (P<0.05), and lower scores for negative coping, HAMA, HAMD, and PCL-5 (P<0.05). Correlation analysis indicated that female gender, negative coping, and higher HAMA and HAMD scores were associated with greater PTSD severity. Logistic regression analysis demonstrated that educational level (OR=1.715, 95% CI: 1.020-2.883, P=0.042) and negative coping (OR=1.590, 95% CI: 1.003-2.522, P=0.048) were risk factors for PTSD, whereas objective support (OR=0.646, 95% CI: 0.451-0.925, P=0.017) was a protective factor. The ROC analysis showed that the total SCSQ score and its negative and positive coping dimensions, the total SSRS score and its subjective and objective support dimensions, as well as their combined use, all demonstrated good discriminative ability in distinguishing between the PTSD and non-PTSD groups. Conclusion The results suggest that individuals who are female, with higher HAMA and HAMD scores after a motor vehicle accident, and those with lower social support and negative coping strategies, should be given particular attention. Early interventions for these individuals may reduce the incidence of PTSD.

B7-H3 molecule inhibits apoptosis of non-small cell lung cancer cells via the SIRT1/p53 signaling pathway

Zheng Lin1, Zhong Jianxin2, Niu Ke1, Xu Qing1, Ling Huijuan1, Zhu Yayu1, Chen Bing1,3, Chen Liwen1,4

DOI: 专辑：医药卫生科技

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abstract:
To explore the role of the histone deacetylase Sirtuin-1 (SIRT1)/p53 signaling pathway in promoting apoptosis of non-small cell lung cancer cells (NSCLC) induced by the co- stimulatory molecule B7-H3. Methods The GEPIA 2 platform was used for survival analysis of NSCLC patients based on B7-H3 gene expression levels. The Gene Enrichment Analysis (GSEA) method was used to analyze the enrichment characteristics of B7-H3 molecules in the gene set of cell apoptosis. In the non-small cell lung cancer A549 cell line, B7-H3 was knocked down, and the protein expression levels of SIRT1 and p53 were detected by Western blot. B7-H3 was overexpressed in A549 cells and the apoptosis rate was analyzed by flow cytometry after Annexin V/PI double staining. Overexpression of B7-H3 and knockdown ofSIRT1 were performed inA549 cell line. The expression levels of p53 and apoptosis-related proteins B-cell lymphoma/leukemia-2 (Bcl-2) and Bcl-2-associated X protein (Bax) were detected respectively by Western blot. Cell apoptosis rate was analyzed by flow cytometry after Annexin V/PI double staining. Results The overall survival of the B7-H3 high-expression group was significantly lower than that of the low- expression group (P<0.01). B7-H3 was significantly enriched in the cell apoptosis signaling pathway and the p53 signaling pathway (P<0.05). Compared with the control group, the expression of SIRT1 was significantly downregulated, and p53 was significantly upregulated in the B7-H3 knockdown group (both P<0.001). Overexpression of B7-H3 significantly up-regulated SIRT1 protein expression (P<0.05), down-regulated p53 expression (P<0.01), and markedly increased the Bcl-2/Bax ratio of apoptosis-related proteins (P<0.001). The results of Annexin V/PI double staining showed that the apoptosis rate of A549 cells with overexpressed B7-H3 decreased (the apoptosis rate of the control group was 26.72%±4.13%, while that of the B7-H3 overexpression group was 13.87%±0.82%; P<0.01). In B7-H3-overexpressing cell lines, SIRT1 knockdown significantly reversed apoptosis (P<0.05), up-regulated p53 protein expression (P<0.001), and markedly reduced the Bcl-2/Bax ratio (P<0.001). Conclusion B7-H3 molecule inhibits the apoptosis of non-small cell lung cancer cells via the SIRT1/p53 signaling pathway.

Research on the diagnostic value of coagulation parameters and proteinuria in preeclampsia and its severity

Feng Xiaoqian, Di Ping, Li Ruibing, Li Mianyang

DOI: 专辑：医药卫生科技

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abstract:
To investigate the diagnostic value of coagulation function indicators and proteinuria in preeclampsia and its severity assessment. Methods A total of 239 pregnant women who visited our hospital from January 2022 to December 2023 were selected as research subjects. They were divided into healthy control group (n=50), non-gestational hypertension group (n=39), and gestational hypertension group (n=150) based on clinical diagnosis. The gestational hypertension group was further subdivided into three subgroups: simple hypertension group (n=50), mild preeclampsia group (n=50), and severe preeclampsia group (n=50). The study compared differences in plasma protein S (PS), plasma protein C (PC), fibrin degradation products (FDP), fibrin monomer (FM), and proteinuria levels among different groups. The correlation between proteinuria and coagulation function indicators in patients with gestational hypertension was analyzed, and ROC curves were used to analyze the diagnostic efficacy of coagulation indicators and proteinuria for gestational hypertension and different stages of preeclampsia. Results The gestational hypertension group showed lower PS and PC values compared to the non- gestational hypertension group and healthy control group, while FDP, FM values, and proteinuria levels were higher than those in the non-gestational hypertension group and healthy control group (P<0.05). Significant differences in coagulation indicators and proteinuria levels were observed among pregnant women with different degrees of gestational hypertension (P<0.05). The AUC values for diagnosing gestational hypertension using PS, PC, FDP, FM, and proteinuria were 0.927, 0.957, 0.967, 0.947, and 0.932 respectively (P<0.000 1). For diagnosing simple hypertension and mild preeclampsia, the AUC values were 0.875, 0.777, 0.830, 0.678, and 0.935 respectively (P<0.000 1). For diagnosing mild and severe preeclampsia, the AUC values were 0.900, 0.776, 0.779, 0.806, and 0.847 respectively (P<0.000 1). Conclusion Coagulation function indicators and proteinuria show significant differences between healthy pregnant women and those with gestational hypertension. PS, PC, FDP, FM, and proteinuria levels vary among pregnant women with different stages of preeclampsia. The aforementioned indicators exhibit certain diagnostic efficacy for gestational hypertension and preeclampsia.

Culture and identification of rat corpus cavernosum smooth muscle cells by modified tissue block adherence method

Zhang Tao1,2, Yu Maobin1,2, Liu Meijun2, Ma Ziyang1,2, Zhang Peihai1,2

DOI: 专辑：医药卫生科技

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Curcumin-loaded exosomes from hypoxia-treated mesenchymal stem cells alleviate microglial inflammatory response in a combined therapy approach

Huang Xiaobin1,2, Li Qianqian2, Zhang Peng2,3, Zhou Yanhua2,3*, Fan Anran2,3

DOI: 专辑：医药卫生科技

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abstract:
To investigate the effects of hypoxia-treated mesenchymal stem cell (MSCs) exosomes (Exo) and their loading with curcumin on microglial inflammatory responses, and to explore the enhancing effect of hypoxia treatment on the function of MSCs Exo. Methods The supernatants of hUC-MSCs cultured under normal and hypoxic conditions were collected, and Exo were isolated using ultracentrifugation. After identification by transmission electron microscopy and Western blot, curcumin was loaded using the co-incubation method. The lipopolysaccharide (LPS)-induced microglial inflammation model was treated with dimethyl sulfoxide (DMSO), curcumin, normoxia Exo, hypoxia Exo, normoxic Exo loaded with curcumin, and hypoxic Exo loaded with curcumin, respectively. The expression of the M1-type marker inducible nitric oxide synthase (iNOS) in BV2 cells was detected by immunofluorescence (IF). Western blot and enzyme-linked immunosorbent assay (ELISA) were used to measure the expression and secretion levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in the cells and their culture supernatants. Results Normoxia Exo, hypoxia Exo, normoxic Exo loaded with curcumin, and hypoxic Exo loaded with curcumin exhibited a "saucer-like" shape with a diameter ranging from 30~150 nm, and the expression of exosomal markers CD9, CD81, and TSG101 were positive. After treating the BV2 cell inflammation model, IF results showed that, compared with the normoxia Exo group, treatment with hypoxic Exo significantly reduced the expression of iNOS. Moreover, when compared with the curcumin group and the normoxic Exo loaded with curcumin group, the expression level of iNOS significantly decreased after treatment with hypoxic Exo loaded with curcumin. The results of Western blot and ELISA indicated that, in comparison with the normoxia Exo group, treatment with hypoxic Exo significantly reduced the expression and secretion of the inflammatory cytokines TNF-α, IL-1β, and IL-6. Additionally, when compared with the curcumin group and the normoxic Exo loaded with curcumin group, both the expression and secretion of TNF-α, IL-1β, and IL-6 significantly decreased after treatment with hypoxic Exo loaded with curcumin. Conclusion Hypoxia preconditioning can enhance the ability of hUC-MSCs-Exo in the inhibition of microglial polarization and inflammatory factors, secretion. Additionally, using Hypoxia-MSCs-Exo as a drug-delivery carrier of curcumin can improve its solubility and stability, facilitating its absorption by cells and exerting the therapeutic effect of combination therapy.

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