Objective To investigate the role and specific molecular mechanism of action of endoplasmic reticulum transmembrane protein 166 (TMEM166) in liver lipid metabolism. Methods Tmem166^flox/flox mice were crossed with Alb-Cre mice, and the offspring Tmem166^+/+ mice were selected as group A, while the Tmem166^-/- mice were selected as group B, with 8 mice in each group. The mice were fed till 24 weeks of age, and serum and liver tissue samples were isolated. HepG2 cells were divided into group C (transfected with sh-vector), group D (transfected with sh-TMEM166), group E (transfected with sh-vector and treated with oleic acid [OA]), group F (transfected with sh-TMEM166 and treated with OA), group G (co-transfected with sh-TMEM166 and siNC and treated with OA), and group H (co-transfected with sh-TMEM166 and siCD36 and treated with OA). Western blotting was used to measure the protein expression level of TMEM166 in liver tissue of the mice in groups A and B and in cells in groups C-F; quantitative real-time PCR and Western blotting were used to measure the expression levels of CD36, FATPs, and other fatty acid uptake-related factors in cells and liver tissue; HE staining was used to observe the pathological changes of liver tissue in groups A and B, and oil red O staining was used to observe the accumulation of lipid droplets in liver tissue of groups A and B and in cells of groups E-H; corresponding kits were used to measure the serum levels of triglyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and non-esterified fatty acids (NEFA) in the mice in groups A and B, as well as the contents of TG and NEFA in the mice in groups A and B and in the cells in groups C-H. Results Animal experiments showed that compared with group A, group B had a significant reduction in the relative protein expression level of TMEM166 in liver tissue (t=6.55,P<0.01), significant increases in liver vacuolization and lipid droplets, and significant increases in the serum levels of TG, ALT, and AST and the levels of TG and NEFA in liver tissue (t=3.10-28.53,P<0.05), as well as significant increases in the mRNA and protein expression levels of CD36 and FATPs in liver tissue (t=4.48-52.27,P<0.05). Cell experiments showed that TMEM166 knockdown and OA treatment had significant influence on the levels of TG and NEFA, the protein expression level of TMEM166, and the mRNA and protein expression levels of CD36, FATPs in each group of cells (F_{TMEM166 knockdown}=23.95-185.00,F_{OA treatment}=26.99-991.80,P<0.01), with an interaction between TMEM166 knockdown and OA treatment on the level of TG and the mRNA expression level of PPARγ2 in cells (F_interaction=9.01,15.50,P<0.05), and the individual effect analysis showed that TMEM166 knockdown or OA treatment alone could cause the significant increases in all the above indicators (F=21.89-393.90,P<0.05). Oil red O staining showed that group F had a significant increase in the accumulation of lipid droplets than group E, and compared with group G, group H had significant reductions in the protein expression level of CD36 and the levels of TG and NEFA (t=7.03-12.07,P<0.01), as well as a significant reduction in the accumulation of lipid droplets. Conclusion Downregulation of TMEM166 can increase the expression of CD36 and promote fatty acid uptake and lipid droplet accumulation in liver cells, thereby affecting the lipid homeostasis in the liver, and the regulatory mechanism of TMEM166 on CD36 may be associated with the activation of PPARγ2.

Objective To investigate the regulatory effect of the long non-coding RNA (lncRNA) MIATNB on autop-hagy in trabecular meshwork cells and its molecular mechanism. Methods RT-qPCR was used to measure the relative expression le-vel of lncRNA MIATNB in iHTM and GTM₃ cells. The iHTM cells were divided into A1 group (transfected with siNC) and B1 group (transfected with siMIATNB), and the GTM₃ cells were divided into A2 group (transfected with siNC) and B2 group (transfected with siMIATNB); Western blotting (WB) was used to observe the regulatory effect of lncRNA MIATNB on the expression of autophagy-related proteins in the above groups of TM cells. The iHTM cells were divided into C1 group (transfected with siNC) and D1 group (transfected with siMIATNB), and the GTM₃ cells were divided into C2 group (transfected with siNC) and D2 group (transfected with siMIATNB); RT-qPCR was used to measure the relative expression level of m⁶A-related genes in cells, and RT-qPCR and WB were used to measure the relative mRNA and protein expression levels of IGF2BP1 in iHTM and GTM₃ cells. The iHTM cells were divided into E1 group (transfected with siNC) and F1 group (transfected with siIGF2BP1), and the GTM₃ cells were divided into E2 group (transfected with siNC) and F2 group (transfected with siIGF2BP1); WB was used to measure the regulatory effect of IGF2BP1 on the expression of LC3B, p62, and Beclin-1 in TM cells, and RNA co-immunoprecipitation (RNA-IP) was used to measure the specific binding capacity between the key m⁶A reader IGF2BP1 and lncRNA MIATNB. Results Compared with iHTM cells, there was a significant reduction in the relative expression level of lncRNA MIATNB in GTM₃ cells (t=4.355,P<0.05). Compared with group A1/A2, group B1/B2 had a significant increase in LC3B-Ⅱ/Ⅰ ratio (t=3.062,6.399,P<0.05), a significant increase in the relative expression level of Beclin-1 (t=5.454,4.139,P<0.05), and a significant reduction in the relative expression level of p62 protein (t=4.799,4.796,P<0.05). Compared with group C1/C2, group D1/D2 had a significant reduction in the relative expression level of IGF2BP1 in cells (t=3.003,3.832,P<0.05), and there were no significant differences in the relative expression levels of other m⁶A genes between group D1 and group C1 (P>0.05); compared with group C2, group D2 had significant reductions in the expression levels of the METTL14, FTO, ALKBH5, YTHDF1, and YTHDF3 genes (t=2.163-5.294,P<0.05) and a significant increase in the expression level of the METTL3 gene (t=3.127,P<0.05). Compared with iHTM cells, GTM₃ cells showed significant reductions in the mRNA and protein expression levels of IGF2BP1 (t=3.636,3.485,P<0.05). Compared with group E1/E2, group F1/F2 had a significant increase in LC3B-Ⅱ/Ⅰ ratio (t=10.910,6.399,P<0.05), a significant increase in the relative expression level of Beclin-1 (t=5.533,4.025,P<0.05), and a significant reduction in the relative expression level of p62 protein (t=5.035,4.790,P<0.05). The specific binding capacity between IGF2BP1 and lncRNA MIATNB in GTM₃ cells was significantly weaker than that in iHTM cells (t=3.803,P<0.05). Conclusion lncRNA MIATNB inhibits excessive autophagy in TM cells by maintaining the expression of IGF2BP1, and the absent expression of lncRNA MIATNB may be involved in the pathogenesis of glaucoma through m⁶A.

Objective To investigate the role and mechanism of action of nuclear receptor coactivator 4 (NCOA4) in acquired resistance to targeted inhibitors in pancreatic cancer with KRAS G12C mutation. Methods Human pancreatic cancer cell lines CFPAC-1, PANC-1, and MIA-PaCa-2 were cultured in vitro. MIA-PaCa-2 cells were used to establish an adagrasib-resistant cell line (Res-MIA), and a lentivirus was used to establish a drug-resistant MIA-PaCa-2 cell line with NCOA4 knockdown (Res-MIA-KD). Res-MIA cells were divided into groups a and d, while Res-MIA-KD cells were divided into groups b and c; the cells in groups c and d were treated with adagrasib (2 μmol/L), while those in groups a and b received no drug treatment. Subsequently, MIA-PaCa-2 cells were divided into groups A, B, and C, and Res-MIA cells were divided into groups D, E, and F; groups A and D were established as control groups, the cells in groups B and E were treated with sotorasib (2 μmol/L), and those in groups C and F were treated with adagrasib (2 μmol/L). The transcriptome sequencing technique was used to detect differentially expressed genes between MIA-PaCa-2 and Res-MIA cells; CCK-8 assay was used to assess cell viability in each group; Western blotting was used to measure the relative protein expression levels of NCOA4 and FTH1 in each group; Transwell assay was used to observe cell invasion and migration abilities in each group. Male BALB/c nude mice were divided into groups Ⅰ, Ⅱ, Ⅲ, and Ⅳ, and the mice in groups Ⅰ and Ⅳ received orthotopic pancreatic injection of Res-MIA cells, while those in groups Ⅱ and Ⅲ were given orthotopic injection of Res-MIA-KD cells. Since day 14, the mice in groups Ⅲ and Ⅳ were given intraperitoneal injection of adagrasib (10 mg/kg) in normal saline twice a week, while those in groups Ⅰ and Ⅱ were given intraperitoneal injection of an equal volume of normal saline, and tumors were harvested on day 28 to measure tumor volume and weight. Results CCK-8 assay showed that after treatment with adagrasib (0, 10, 50, 100, 1 000, or 2 000 nmol/L) for 72 h, there was no significant change in the viability of CFPAC-1 and PANC-1 cells (P>0.05), while the viability of MIA-PaCa-2 cells decreased significantly with the increase in the concentration of adagrasib (F=239.24,t_LSD=3.68-30.10,P<0.05). After treatment with sotorasib or adagrasib (0, 10, 50, 100, 1 000, or 2 000 nmol/L) for 72 h, there was no significant change in the viability of Res-MIA cells (P>0.05), and there was no significant difference in cell viability between groups a and d (P>0.05); groups b and c had a significantly lower cell viability than groups a and d (P<0.05), and group c had a significantly lower cell viability than group b (P<0.05). Transcriptome sequencing showed that the expression level of NCOA4 in Res-MIA cells was significantly higher than that in MIA-PaCa-2 cells (P<0.05). Western blotting showed that groups B and C had a significantly lower relative protein expression level of NCOA4 than group A, and group D had a significantly higher expression level than group A, while group E had a significantly lower expression level than group D (F=366.20,t=4.69-22.71,P<0.05); groups B and C had a significantly lower relative protein expression level of FTH1 than group A, group D had a significantly higher expression level than group A, and groups E and F had a significantly higher expression level than group D (F=702.16,t=7.36-49.15,P<0.05); the relative protein expression levels of NCOA4 and FTH1 in Res-MIA-KD cells were significantly lower than those in Res-MIA cells (t=16.27,92.26,P<0.05). Transwell assay showed that compared with Res-MIA-KD cells, Res-MIA cells had significantly lower numbers of cell migration and invasion (t=9.83,7.19,P<0.05). Animal experiments showed no significant differences in tumor volume and weight between nude mice of groups Ⅰ and Ⅳ (P>0.05), and nude mice of groups Ⅱ and Ⅲ had significantly lower tumor volume and weight than groups Ⅰ and Ⅳ (P<0.05), while nude mice of group Ⅲ had significantly lower tumor volume and weight than group Ⅱ (P<0.05). Conclusion Pancreatic cancer with KRAS G12C mutation can enhance its resistance to the targeted agent adagrasib through NCOA4-mediated ferritinophagy, and inhibition of NCOA4 expression can restore the sensitivity of pancreatic cancer cells to adagrasib.

Objective To investigate the effect of bupivacaine on the proliferation and metastasis of tumor cells in an in vitro model of perineural invasion of cholangiocarcinoma and its mechanism. Methods Human cholangiocarcinoma cell line (QBC939 cells) were divided into QBC939 group (group A), QBC939+well-differentiated pheochromocytoma cell line (PC12) group (group B), and QBC939+PC12+bupivacaine group (group C). The cells in group A were cultured in the QBC939 cell-specific medium for 48 h, those in group B were co-cultured with well-differentiated PC12 cells for 48 h in addition to the treatment in group A to establish the in vitro model of perineural invasion of cholangiocarcinoma, and those in group C were cultured with bupivacaine for 48 h in addition to the treatment in group B. CCK-8 assay, EDU staining, cell scratch assay, and Transwell assay were used to measure cell viability and assess the proliferation, migration, and invasion abilities of the three groups of cells. Well-diffe-rentiated PC12 cells were divided into PC12 group (group D) and PC12+bupivacaine group (group E), and the cells in group D were cultured in the PC12 cell-specific medium for 48 h, while those in group E were cultured with bupivacaine for 48 h in addition to the treatment in group D. Immunofluorescence staining assay was used to measure the length of cell axons, and Western blotting was used to measure the relative expression level of neuropilin-1 (NRP-1) in the two groups. Results CCK-8 assay, EDU staining, cell scratch assay, and Transwell assay showed that compared with B, group C had significantly lower cell viability and proliferation, migration, and invasion abilities of QBC939 cells (F=34.39-149.20,q=9.47-12.95,P<0.05). Immunofluorescent staining and Western blotting showed that compared with group D, group E had a significantly shorter mean length of cell axons and a significantly lower relative protein expression level of NRP-1 (t=11.75,3.09,P<0.05). Conclusion Bupivacaine may inhibit the malignant biological behaviors of cholangiocarcinoma cells such as proliferation, migration, and invasion by inhibiting the axonal growth of neural cells and the expression of NRP-1 in an in vitro model of perineural invasion of cholangiocarcinoma.

Objective To investigate the effect of Wuzhuyu Decoction (WZYD) on alcoholic gastric ulcer and its mechanism. Methods A total of 120 female KM mice, aged 8 weeks, were randomly divided into normal control group (group A), model control group (group B), ranitidine group (group C), and low-, middle-, and high-dose WZYD groups (groups D-F), with 20 mice in each group. The mice in groups A and B were given distilled water (10 mL/kg) at room temperature by gavage, those in group C were given ranitidine (30 mg/kg) by gavage, and those in groups D-F were given WZYD by gavage at a dose of 5, 10, and 20 mL/kg, respectively. After 7 consecutive days of intragastric administration, 10 mice were selected from each group, and serum was collected under aseptic conditions for cell experiments, and the remaining mice were still given distilled water or the drug by gavage for another 21 days. Body weight and food intake of the mice in each group were recorded on days 7, 14, 21, and 28. On day 29, serum, gastric juice, and gastric tissue samples were collected; gastric morphology was observed with the naked eye, and HE staining was used to observe the histopathological changes of gastric mucosa; the volume, pH value, and total acidity of gastric juice were measured; ELISA was used to measure the serum levels of IL-6, IL-10, TNF-α, and PGE2; Western blot (WB) was used to measure the expression of PI3K/Akt/NF-κB pathway-related proteins in gastric mucosa; biochemical assays were used to measure the levels of MDA, CAT, SOD, and GSH-Px in gastric mucosa. Human gastric mucosal epithelial cells (GES-1) were divided into groups a1-e1 and a2-f2. The cells in groups a1-e1 were treated with the aseptic serum from groups A, C, D, E, and F, respectively. The cells in groups b2-f2 were treated with 0.8 mol/L ethanol, while those in group a2 were treated with an equal volume of PBS; after 4 hours of treatment, the cells in groups a2-f2 were treated with the aseptic serum from groups A-F. CCK-8 assay was used to measure cell viability in each group. Results The animal experiment showed that compared with group B, groups C-F had a significant increase in body weight at different time points, along with varying degrees of improvements in congestion, edema, punctate ulcers, and inflammatory cell infiltration of gastric mucosa, significant reductions in the volume and total acidity of gastric juice, a significant increase in the pH value of gastric juice, significant reductions in the serum levels of IL-6, TNF-α, and PGE2 and p-AKT/AKT ratio and MDA level in gastric mucosal tissue, and significant increases in the serum level of IL-10 and the levels of SOD and GSH-Px in gastric mucosa; groups C, E, and F had a significant increase in food intake on days 14, 21, and 28 and a significant reduction in the ratio of P65 in the nucleus to P65 in the cytoplasm; groups C and E had a significant increase in the level of CAT in gastric mucosal tissue; groups D-F had a significant reduction in p-PI3K/PI3K ratio in gastric mucosal tissue; group E and F had a significant increase in p-IκBα/IκBα ratio in gastric mucosal tissue (F=3.4-1 013.0,q=4.1-78.7,P<0.05). CCK-8 assay showed that there was no significant difference in cell viability between groups c1-e1 and group a1 at any time point (P>0.05), and groups c2-f2 had a significantly higher cell viability than group b2 at 12, 24, and 48 hours of culture (F=15.5-146.1,q=4.6-22.3,P<0.05). Conclusion WZYD can exert a protective effect against alcoholic gastric ulcers by modulating the PI3K/Akt/NF-κB signaling pathway, regulating inflammatory cytokines, and inhibiting oxidative stress.

Objective To investigate the status and classification of pediatric renal artery variations using the Hisense CAS computer-assisted surgery system. Methods Lower abdominal contrast-enhanced CT images were collected from the children with non-kidney disease or healthy children in The Affiliated Hospital of Qingdao University from January 2013 to December 2024, and the images were imported into the Hisense CAS system to perform three-dimensional reconstruction and construct the model of the kidney. The status and classification of pediatric renal artery variations were summarized and analyzed. Results Left renal artery variations accounted for 10.78%, while right renal artery variations accounted for 10.28%, with no significant difference between the two sides (P>0.05). There were four types of renal artery variations, i.e., prehilar renal artery branch type (type Ⅰ), accessory renal artery type (type Ⅱ), hilar accessory artery type (type Ⅲ), and mixed type (type Ⅳ), accounting for 16.79%, 5.76%, 3.01%, and 0.50%, respectively. Conclusion The model of the kidney constructed based on the Hisense CAS system can clearly show the shape of the renal artery and clarify the type of renal artery variation, which provides an anatomical basis for the preoperative planning of renal surgery and a reference for subsequent research.

Objective To investigate the expression characteristics of immune-related guanosine triphosphatase M (IRGM) in colorectal cancer (CRC) tissue and its association with clinicopathological features and prognosis. Methods Fresh tumor tissue samples and the corresponding paracancerous normal tissue samples were collected from 16 patients who underwent radical resection for CRC in The Affiliated Hospital of Qingdao University, and RT-qPCR was used to measure the mRNA expression level of IRGM. Postoperative paraffin-embedded tumor tissue samples and the corresponding paracancerous normal tissue samples were collected from 143 CRC patients who were treated in The Affiliated Hospital of Qingdao University, and immunohistochemical staining was used to measure the protein expression level of IRGM; the association of the protein expression level of IRGM with clinicopathological features and prognosis was analyzed. Results The mRNA expression level of IRGM in paracancerous normal tissue was significantly higher than that in CRC tumor tissue (t=2.836,P<0.05). The protein expression level of IRGM in paracancerous normal tissue was also significantly higher than that in CRC tumor tissue (χ²=23.738,P<0.05). The low expression of IRGM in CRC tumor tissue was significantly associated with lymph node metastasis and vascular tumor thrombus (χ²=9.827,7.690,P<0.05). The survival analysis showed that the expression level of IRGM had no significant correlation with overall survival rate or disease-free survival rate in CRC patients (P>0.05). Conclusion The expression level of IRGM is significantly downregula-ted in CRC tissue; the low expression of IRGM is closely associated with lymph node metastasis and vascular tumor thrombus, while it is not significantly associated with the overall survival rate and disease-free survival rate of patients, suggesting that IRGM may become a potential target for the treatment of CRC.

Objective To investigate the efficacy and safety of dapagliflozin combined with Bailing Capsule in the treatment of patients with diabetic kidney disease (DKD). Methods A total of 1 162 patients with early-stage DKD who were admitted to The Affiliated Hospital of Qingdao University from January 2019 to December 2023 were enrolled, and after propensity score matching was performed to balance intergroup covariates, the patients were divided into control group (dapagliflozin monotherapy) and combination group (dapagliflozin+Bailing Capsule) according to their treatment modality, with 417 patients in each group. The two groups were compared in terms of clinical outcome, blood glucose parameters, renal function parameters, safety indicators (liver function and routine blood test results), and the incidence rate of adverse events after 12 months of treatment. Results After 12 months of treatment, the combination group had a significantly better clinical outcome than the control group (Z=2.919,P<0.05); the combination group had reductions in the levels of fasting plasma glucose, 2 hour postprandial glucose, and HbA1c after treatment, with significantly greater reductions in these indicators compared with the control group (t=41.91-59.57,P<0.05). The combination group had significant reductions in serum creatinine, blood urea nitrogen, 24 hour urinary protein, and urinary albumin excretion rate after treatment, with significantly greater reductions in these indicators compared with the control group (t=12.51-154.9,P<0.05). After 12 months of treatment, liver function parameters and routine blood test results remained within normal ranges in both groups, and there were no significant differences between the two groups in the incidence rates of adverse events such as hypoglycemia and urinary tract infection (P>0.05). Conclusion Dapagliflozin combined with Bailing Capsule can significantly reduce the level of blood glucose and improve renal function parameters in patients with DKD, with a better clinical effect than dapagliflozin monotherapy and with a favorable safety profile.

Objective To investigate the association between neuroinflammation, vascular risk, and biomarkers in Alzheimer’s disease (AD). Methods A total of 1 079 individuals without AD were selected as subjects from the China Alzheimer’s Biomarker and Lifestyle study cohort, and Framingham General Cardiovascular Risk Score (FGCRS) was used to assess the vascular risk of the subjects. The multiple linear regression model was used to investigate the correlation of FGCRS with AD biomarkers in cerebrospinal fluid (CSF) (β-amyloid 42 [Aβ₄₂], β-amyloid 40 [Aβ₄₀], phosphorylated tau [P-tau], total tau [T-tau], Aβ₄₂/Aβ₄₀ ratio, T-tau/Aβ₄₂ ratio, and P-tau/Aβ₄₂ ratio], neuroinflammatory factors in CSF (soluble triggering receptor expressed on myeloid cells 2 [sTREM2] and progranulin [PGRN]), and peripheral blood inflammatory cells (white blood cell count [WBC], neutrophil count [NEU], lymphocyte count [LY], monocyte count [MONO], and neutrophil-to-lymphocyte ratio [NLR]). Mediation models were constructed with sTREM2 and PGRN as mediators to investigate the mediating effect of neuroinflammatory factors between vascular risk and AD biomarkers. Results FGCRS was positively correlated with the levels of Aβ₄₂, Aβ₄₀, P-tau, and T-tau in CSF (β=0.066-0.242,t=2.020-7.928,P<0.05), and it was also positively correlated with the levels of sTREM2 and PGRN in CSF and the levels of WBC, NEU, NLR, and MONO in peripheral blood (β=0.120-0.228,t=3.285-5.705,P<0.05). PGRN and sTREM2 mediated the association between FGCRS and AD biomarkers in CSF (Aβ₄₀, P-tau, T-tau, and Aβ₄₂), with a mediation ratio of 26.75%-81.82% for sTREM2 and 9.66%-31.03% for PGRN. Conclusion The increase in vascular risk may influence the levels of AD biomarkers in CSF by increasing the levels of neuroinflammatory factors in CSF, thereby promoting the development and progression of AD.

Objective To investigate the influencing factors for hyporesponsiveness to erythropoiesis-stimulating agents (ESAs) in patients on maintenance hemodialysis (MHD), and to construct a predictive model. Methods A total of 120 patients with end-stage chronic kidney disease (CKD) who received MHD and had hyporesponsiveness to ESAs from January 2020 to June 2023 were enrolled as hyporesponsiveness group, and 122 patients without hyporesponsiveness to ESAs who received MHD during the same period of time were enrolled as non-hyporesponsiveness group. Related data were collected for both groups, including sex, age, body height, body weight, primary diseases (chronic glomerulonephritis, chronic renal insufficiency of unknown cause, nephrotic syndrome, polycystic kidney, obstructive nephropathy, lupus nephritis, diabetes, and hypertension), duration of dialysis, and frequency of dialysis. Meanwhile, related laboratory markers were collected at 6 months of ESAs treatment, i.e., serum albumin, blood calcium, blood phosphorus, serum total cholesterol, serum triglyceride, serum parathyroid hormone, blood uric acid, hemoglobin (Hb), mean corpuscular Hb concentration, mean corpuscular volume, hematocrit, lymphocyte count, and platelet count, and platelet-to-lymphocyte ratio (PLR) was calculated. A multivariate logistic regression analysis was performed for the indicators with statistical difference in the univariate analysis, and Python 3.12 was used to plot a nomogram. The receiver operating characteristic (ROC) curve was plotted to assess the value of related indicators in predicting hyporesponsiveness to ESAs in MHD patients. Results The multivariate logistic regression analysis showed that PLR, serum albumin, and triglyceride were influencing factors for hyporesponsiveness to ESAs in MHD patients. The predictive model constructed based on the above indicators had a Youden index of 0.319 and an area under the ROC curve of 0.689. The calibration curve showed a good degree of fitting between the actual value and the predictive value, and the decision curve analysis showed a high net benefit in predicting hyporesponsiveness to ESAs in MHD patients within a range of 0.4-0.6 for threshold probability. Conclusion PLR, serum albumin, and triglyce-ride have a certain value in predicting hyporesponsiveness to ESAs in MHD patients. The risk prediction model for hyporesponsiveness to ESAs in MHD patients constructed based on PLR, triglyceride, and albumin has good predictive performance and can provide a basis for predicting hyporesponsiveness to ESAs in MHD patients.

Objective To investigate the clinical features of adults with primary minimal change nephrotic syndrome (MCNS) complicated by acute kidney injury (AKI), as well as the risk factors for AKI in primary MCNS and their impact on prognosis. Methods A total of 315 adult patients who were diagnosed with primary MCNS by renal biopsy in The Affiliated Hospital of Qingdao University from October 2013 to March 2024 were enrolled, and according to the presence or absence of AKI at initial diagnosis, they were divided into MCNS group and MCNS+AKI group. The clinical features of adult patients with MCNS complicated by AKI were analyzed, and the logistic regression model was used to identify the risk factors for AKI in adult patients with primary MCNS. The impact of AKI on the prognosis of adult patients with primary MCNS was further analyzed. Results When compared with the MCNS group, the MCNS+AKI group had a significantly older age (Z=3.989,P<0.05), a significantly higher proportion of male patients (χ²=7.623,P<0.05), significantly lower levels of complement C3, lymphocyte count, and hemoglobin (t=2.248-5.619,P<0.05), and significantly higher levels of lactate dehydrogenase, blood uric acid, urinary erythrocyte count, urinary leukocyte count, D-dimer, C-reactive protein, and cystatin C (t=-3.292,-4.261,Z=2.492-5.973,P<0.05). As for renal histopathological changes, compared with the MCNS group, the MCNS+AKI group had a significantly higher proportion of patients with IgA deposition, interstitial renal lesions, or protein casts (χ²=4.567-12.013,P<0.05). The logistic regression analysis showed that age ≥60 years (OR=2.56,95%CI=1.10-5.95,P<0.05), male sex (OR=2.88,95%CI=1.36-6.11,P<0.05), a high level of lactate dehydrogenase (OR=1.00,95%CI=1.00-1.01,P<0.05), blood uric acid >385.04 μmol/L (OR=1.98,95%CI=1.03-3.78,P<0.05), and protein casts (OR=2.73,95%CI=1.20-6.22,P<0.05) were independent risk factors for AKI in adult patients with primary MCNS, while hemoglobin >139.26 g/L (OR=0.40,95%CI=0.20-0.80,P<0.05) was a protective factor against AKI. Compared with the MCNS group, the MCNS+AKI group had a significantly longer time to achieve complete remission of urinary protein (t=2.318,P<0.05), and the patients with MCNS and stage Ⅱ/Ⅲ AKI had a significantly longer time to achieve complete remission of urinary protein than those with MCNS and stage I AKI (F=3.287,t=2.080、2.048,P<0.05). Conclusion There is an increased risk of AKI in adult patients with primary MCNS who are elderly, male, have a high level of lactate dehydrogenase in blood, hyperuricemia, and the presence of protein casts found by renal pathological examination, and the clinical monitoring of renal function should be enhanced for these patients. The time to achieve complete remission of urinary protein increases with the increase in the severity of AKI, and early identification, prevention, and treatment of AKI are crucial for the prognosis of adult patients with primary MCNS.

Objective To establish an early screening model for cervical cancer based on single-cell Raman spectroscopy (SCRS) combined with machine learning algorithms, and to assess the performance of the model. Methods Cervical exfoliated cell samples were collected from 128 patients who were treated in our hospital from September 2023 to June 2024, among whom 65 had normal results of ThinPrep cytologic test (TCT), 35 had abnormal TCT results, and 28 did not receive TCT. R language was used to divide the 100 cervical exfoliated cell samples with TCT results into training set and test set at a ratio of 8∶2, and SCRS was performed for all samples. Based on the SCRS data of the training set, 7 machine learning algorithms (KNN, PLS, LDA, RF, SVM, SVMRBF, and Stack) were used to establish an early screening model for cervical cancer, which was applied in the test set to identify the optimal model. The optimal model was then used to predict the TCT results of 100 cervical exfoliated cell samples in the training and test sets, which were compared with the actual TCT results. The remaining 28 samples without prior TCT results were used as a validation set and were subjected to TCT, and the optimal model was used to predict the TCT results of these samples, which were compared with the actual TCT results. Results There were significant differences in the relative intensities of characteristic Raman peaks at 874, 935, 1 024, 1 119, 1 250, 1 328, 1 569, and 1 642 cm^-1 between the cervical exfoliated cells negative for intraepithelial lesion or malignancy, atypical squamous cells of undetermined significance, and the cervical exfoliated cells of low-grade squamous intraepithelial lesion. Among the 7 algorithms, the stacking model showed the best performance, with an AUC of 0.987, an accuracy of 99.2%, a sensitivity of 98.9%, and a specificity of 99.3%. In both training and test sets, the results predicted by the Stack model were relatively highly consistent with actual TCT results, with an accuracy of 91.0%, a sensitivity of 91.0%, a specificity of 87.4%, and an F1-score of 90.3%. In the validation set, the Stack model achieved an accuracy of 96.4%, a sensitivity of 100.0%, a specificity of 95.5%, and an F1-score of 92.3% in predicting TCT results. Conclusion The early screening model for cervical cancer based on SCRS and machine learning algorithms has a good performance and can be used as a noninvasive, efficient, and rapid tool to facilitate the early screening of cervical cancer.

Objective To investigate the value of three-dimensional speckle-tracking imaging (3D-STI) in assessing left atrial function in patients with cirrhotic cardiomyopathy (CCM). Methods A total of 165 patients with liver cirrhosis who were treated in The Affiliated Hospital of Qingdao University from January 2021 to May 2024 were enrolled as cirrhosis group, and according to the presence or absence of cardiomyopathy, the patients were divided into CCM(+) group with 57 patients and CCM(-) group with 108 patients. In addition, 51 healthy individuals who underwent physical examination during the same period of time were enrolled as control group. Conventional echocardiography and 3D-STI were used to assess the left atrial structure and function of the subjects in each group, and the results were compared. Meanwhile, a multivariate logistic regression model was used to analyze the association between 3D-STI parameters and the onset of CCM in both CCM (+) and CCM (-) groups. Results Compared with the control group, the cirrhosis group had significant increases in LAD, late diastolic peak flow velocity (A), and mitral valve early diastolic peak flow velocity (E)/early diastolic mitral annular velocity (e') and significant reductions in e' and E/A (t=1.999-8.079,P<0.05), and there were no significant differences in these parameters between the CCM(+) group and the CCM(-) group (P>0.05). Compared with the control group, the liver cirrhosis group had significant reductions in LA-GLS, LA-GRS, LA-GCS, and LAEF and significant increases in LAV_max, LAVI_max, LAV_min, and LAVI_min (t=4.666-19.286,P<0.05). Compared with the CCM(-) group, the CCM(+) group had significant reductions in LA-GLS, LA-GRS, LA-GCS, and LAEF and significant increases in LAV_max, LAVI_max, LAV_min, and LAVI_min (t=2.637-16.307,P<0.05). The multivariate logistic regression analysis showed that LA-GLS and LA-GRS were significantly associated with the onset of CCM (χ²=4.771,5.601,P<0.05). Conclusion Compared with conventional echocardiography, 3D-STI can provide more comprehensive data for left atrial volume measurement and functional assessment and help to achieve early detection of occult abnormal changes in the left atrium. The reductions in LA-GLS and LA-GRS may suggest potential abnormalities in left atrial structure and function and thus assist in the prediction and early diagnosis of CCM in clinical practice.

Objective To investigate the risk factors for recurrence in patients with acute ischemic stroke (AIS). Methods A total of 4 051 AIS patients reported to the Shandong Province Chronic Disease Monitoring Information Management System by Yanzhou People’s Hospital of Jining from January 2017 to December 2020 were enrolled, and according to the number of episodes, they were divided into initial onset group (1 episode) and recurrence group (>1 episode at an interval of >28 days). The two groups were matched at a ratio of 1∶1 according to sex, age (with a difference of ≤±2 years), and onset time (the same month in the same year). General information and laboratory markers were collected and compared between the two groups, and the logistic regression model was used to investigate the risk factors for recurrence in AIS patients. Results A total of 814 patients were enrolled in this study, with 407 patients in each group. There were significant differences between the two groups in the history of hypertension, coronary heart disease, and diabetes mellitus and the levels of diastolic pressure, serum aspartate ami-notransferase, serum high-density lipoprotein cholesterol, serum glucose, blood sodium, and blood chlorine on admission (W=68 110-90 655,χ²=21.491-46.964,P<0.05). The multivariate logistic regression analysis showed that the history of hypertension, coronary heart disease, and diabetes mellitus, a high diastolic pressure, and a short plasma thrombin time on admission were risk factors for recurrence in AIS patients (P<0.05). Conclusion The history of hypertension, coronary heart disease, and diabetes mellitus, a high diastolic pressure, and a short plasma thrombin time on admission are risk factors for recurrence in AIS patients. Health management should be enhanced for the population with the above factors, in order to prevent disease recurrence and improve the quality of life of patients.

Objective To investigate the impact of RAD51 protein on the sensitivity of ovarian cancer to neoadjuvant chemotherapy (NACT). Methods Tumor biopsy tissue samples were collected before NACT from 47 patients with advanced high-grade serous ovarian cancer (HGSOC) who underwent NACT and intermediate tumor cell reduction surgery in Department of Gynecology, The Affiliated Hospital of Qingdao University, from June 2017 to June 2019. Immunohistochemistry was used to measure the expression level of RAD51 protein in tumor tissue, and the correlation between its expression level and the sensitivity to NACT was analyzed. SKOV3 cells were divided into groups a-f, and cisplatin (DDP)-resistant SKOV3 cells were divided into groups A-F; the cells in groups b-f or groups B-F were transfected with NC-siRNA, siRNA-RAD51-264, siRNA-RAD51-416, siRNA-RAD51-608, and siRNA-RAD51-807, respectively, and then quantitative real-time PCR was used to measure the mRNA expression levels of RAD51 in each group of SKOV3 cells and DDP-resistant SKOV3 cells. The DDP-resistant SKOV3 cells were divided into SKOV3-Control group (control group), SKOV3-DDP-Control group (DDP control group), SKOV3-DDP-NC-siRNA group (transfected with NC-siRNA), and SKOV3-DDP-siRNA-RAD51 group (transfected with siRNA-RAD51), and Western blotting was used to measure the expression level of RAD51 protein in each group of cells. The DDP-resistant SKOV3 cells were divided into NC-siRNA group (transfected with NC-siRNA) and siRNA-RAD51 group (transfected with siRNA-RAD51-264), and DDP at various concentrations (0.39, 0.78, 1.56, 3.12, 6.25, 12.50, and 25.00 mg/L) was added for culture for 48 h; CCK-8 assay was used to measure the viability of DDP-resistant SKOV3 cells after transfection. The DDP-resistant SKOV3 cells were divided into Control group, NC-siRNA group (transfected with NC-siRNA), siRNA-RAD51 group (transfected with siRNA-RAD51), and siRNA-RAD51+DDP group (transfected with siRNA-RAD51 and subsequently treated with DDP), and flow cytometry was used to observe cell cycle alterations and apoptosis rate in DDP-resistant SKOV3 cells. Results Immunohistochemistry showed that there were no significant differences in age, FIGO stage (stage Ⅲ or Ⅳ), lymph node metastasis, ascites, and initial serum CA125 level between the patients with high RAD51 expression and those with low expression (P>0.05), and there was a significant difference in the expression level of RAD51 protein in tumor tissue between the chemotherapy-sensitive group and the insensitive group (χ²=10.85,P<0.05). RT-qPCR showed that the relative mRNA expression level of RAD51 in SKOV3-DDP cells was significantly higher than that in SKOV3 cells (t=6.73,P<0.05), and the mRNA expression level of RAD51 in group c/C was significantly lower than that in group a/A (F=7.81,6.58,P<0.05); there was no significant difference in the mRNA expression level of RAD51 between groups d/D, e/E, f/F and group a/A (P>0.05). Western blotting showed that compared with group A, group B had a significant increase in the protein expression level of RAD51 in cells, and compared with groups A and B, group D had a significant reduction in the protein expression level of RAD51 (F=10.56,P<0.05). CCK-8 assay showed that compared with the NC-siRNA group, the siRNA-RAD51-264 group had a significant reduction in cell viability at DDP concentrations of 3.12, 6.25, 12.50, and 25.00 mg/L (t=3.72-63.90,P<0.05). Flow cytometry showed that compared with the Control group and the NC-siRNA group, the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G₀/G₁ phase, and compared with the siRNA-RAD51 group, the siRNA-RAD51+DDP group had a significant increase in the proportion of cells in the G₀/G₁ phase (F=23.29,P<0.05); compared with the Control group and the NC-siRNA group, the siRNA-RAD51+DDP group had a significant reduction in the proportion of cells in the S phase (F=12.95,P<0.05). Compared with the Control group and the NC-siRNA group, both the siRNA-RAD51 group and the siRNA-RAD51+DDP group had a significant increase in apoptosis rate (F=16.34,P<0.05). Conclusion Patients with a high expression level of RAD51 protein in ovarian cancer tissue tend to have poorer sensitivity to NACT, and knocking down the expression of RAD51 gene in DDP-resistant SKOV3 cells can effectively reverse the resistance of these cells to DDP and enhance the sensitivity to chemotherapy.

As a rare pulp lesion, internal tooth resorption often has no obvio clinical symptoms and signs and is only accidentally identified during medical imaging examination, and its diagnosis and treatment have always been a difficult issue for dentists. Misdiagnosis or missed diagnosis of this disease can lead to the failure in root canal therapy and even root fracture and premature tooth loss. This article reviews the etiology, diagnosis, and treatment of internal tooth resorption and summarizes the latest research advances, in order to provide theoretical support and treatment strategies for dentists in the diagnosis and treatment of internal tooth resorption in clinical practice and further improve the success rate of diagnosis and treatment.