Journal of Shandong University (Health Sciences)

Effects of mesenchymal stem cells on ferroptosis in experimental autoimmune encephalomyelitis mice

LIU Haixia, HUANGFU Shasha, SANG Xiaoyu, CUI Dongqing, BI Jianzhong, WANG Ping

Journal of Shandong University (Health Sciences). 2024, 62(6): 1-8. doi:10.6040/j.issn.1671-7554.0.2024.0265

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Objective To investigate the effects of mesenchymal stem cells(MSCs)treatment on ferroptosis and expressions of key regulatory proteins in experimental autoimmune encephalomyelitis(EAE)mice. Methods The body weights and symptom scores of mice were evaluated daily. To assess conditions of inflammation and myelination in spinal cord, the sections were respectively stained. The contents of reduced glutathione(GSH), malondialdehyde(MDA)and total superoxide dismutase(T-SOD)were tested to detect ferroptosis. The protein expressions of transferrin receptor 1(TFR1), acyl-CoA synthetase long-chain family 4(ACSL4), glutathione peroxidase 4(GPX4)and ferroptosis suppressor protein 1(FSP1)were detected with Western blotting. Results MSCs treatment alleviated weight loss and symptoms of EAE mice. Pathologically, MSCs improved infiltration of inflammatory cells and remyelination of mice. MSCs upregulated the contents of GSH in spinal cord and brain(P<0.01, P<0.05), as well as T-SOD in spinal cord(P<0.01), and downregulated MDA concentration in brain(P<0.05). Besides, MSCs reduced the protein expressions of TFR1(P<0.05, P<0.01)and ACSL4(P<0.05, P<0.001), increased the protein expressions of FSP1(both P<0.05)in spinal cord and brain. Also, the protein expression of GPX4 in brain was promoted after MSCs treatment(P<0.05). Conclusion MSCs play a therapeutic role in inhibiting ferroptosis in EAE mice by regulating iron metabolism, lipid metabolism and promoting reactive oxygen species clearance.

Role of microfibril-associated protein 3 in regulating mesenchymal transition of glioma stem cells

GUO Shuhua, FAN Yang, TIAN Feng, WANG Chuanxin, DU Lutao, LI Peilong, GUO Xing, XU Shuo

Journal of Shandong University (Health Sciences). 2024, 62(6): 9-16. doi:10.6040/j.issn.1671-7554.0.2023.1040

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Objective To investigate the expression and clinical significance of microfibril-associated protein 3(MFAP3)in glioblastoma(GBM)and its influence on the malignant biological behavior of glioma stem cells(GSCs). Methods TCGA data and CGGA data were used to analyze the expression of MFAP3 in GBM and its correlation with patient prognosis. Western blotting was used to detect the protein expression level of MFAP3 in GSCs. The effects of MFAP3 knockdown and overexpression on the self-renewal ability and phenotype of GSCs were explored. Results MFAP3 was highly expressed in GBM and correlated with poor prognosis. The expression level of MFAP3 in mesenchymal GSCs was higher than that in preneuronal GSCs. After MFAP3 knockdown, the self-renewal ability of GSCs was weakened, and the protein levels of mesenchymal phenotype markers CD44 and YKL40 were decreased, whereas overexpression exhibited the opposite trends. Conclusion MFAP3 plays an important role in regulating the malignant biological progression of GBM and is expected to be a new clinical diagnostic marker and therapeutic intervention target for GBM.

MicroRNA-210-3p inhibits inflammatory pain in rats by regulation ten-eleven translocation 2 expression

WEI Jiacheng, YANG Baozhong, WEI Wei, XUE Yating, CUI Chenlong, FANG Jun

Journal of Shandong University (Health Sciences). 2024, 62(6): 17-29. doi:10.6040/j.issn.1671-7554.0.2024.0163

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Objective To investigate the roles and mutual regulatory mechanisms of microRNA-210-3p(miR-210-3p)and ten-eleven translocation 2(TET2)in a rat model of inflammatory pain induced by complete Freunds adjuvant(CFA). Methods Bioinformatics and dual-luciferase reporter assays were used to analyse and identify target genes regulated by miR-210-3p in rats. The combinations of plasmid and miR-210-3p cotransfection in the experiments were grouped as follows: pmirGLO+mimics-NC group, pmirGLO+mimics-miR-210-3p group, TET2-WT-pmirGLO+mimics-NC group, TET2-WT-pmirGLO+mimics-miR-210- 3p group, TET2-MT-pmirGLO+mimics-NC group and TET2-MT-pmirGLO+mimics-miR-210-3p group; 60 rats were divided into 4 groups according to the randomised numerical table method: Normal control(CON)group(n=20), Complete Freunds adjuvant(CFA)group(n=20 ), Complete Freunds adjuvant + adeno-associated virus vector negative control(CFA + AAV NC)group(n=10 ), Complete Freunds adjuvant + adeno-associated virus miR-210-3p inhibitor(CFA + AAVi)group(n=10 ). The rat inflammatory pain model was established by subcutaneous injection of CFA into the underside of the left hind paw; the intervention model was established by tail vein injection of AAV with miR-210-3p inhibitor; the behaviour of the rats was observed and measured; the expression of miR-210-3p was detected by RT-qPCR; Western blotting and immunofluorescence staining were used to detect changes in the expression level and fluorescence intensity of TET2 protein in the spinal cord of lumbar extension segments from L4 to L6; and immunofluorescence staining was used to observe the cellular expression localisation of TET2 protein in the rat spinal cord. Results Dual-luciferase assays confirmed a negative regulatory relationship between miR-210-3p and TET2, as evidenced by a binding site. CFA injection significantly decreased the mechanical paw withdrawal mechanical threshold(PWMT)and the thermal paw thermal withdrawal latency(PTWL)(P<0.05). An increase in miR-210-3p and a concomitant decrease in TET2 protein expression were observed in the CFA group(P<0.05). Immunofluorescence showed that TET2 protein mainly colocalised with neuronal cells and its expression in the spinal cord was decreased in the CFA group(P<0.05). After AAVi treatment, PWMT and PTWL were significantly higher in the CFA+AAVi group than in the CFA+AAV NC group(P<0.05), with increased TET2 protein levels(P<0.05). Conclusion miR-210-3p downregulates TET2 protein expression; its inhibition in rats with inflammatory pain significantly alleviates pain symptoms.

Alstonia mairei total alkaloids enhance the sensitivity of colorectal cancer to 5-fluorouracil chemotherapy

WANG Xiao, KONG Wenru, CUI Weiliang, WANG Shuqi

Journal of Shandong University (Health Sciences). 2024, 62(6): 30-37. doi:10.6040/j.issn.1671-7554.0.2024.0261

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Objective To observe the sensitizing effect of Alstonia mairei total alkaloids on 5-fluorouracil(5-FU)in the treatment of colorectal cancer, and to explore the influence on the pharmacokinetics of 5-FU. Methods The cytotoxicity of Alstonia mairei total alkaloids combined with 5-FU on HCT15 human colorectal adenocarcinoma cells and CT26.WT mouse colon cancer cells was evaluated by CCK-8 kit. A mouse subcutaneous tumor model was established to observe the effects of 5-FU and Alstonia mairei total alkaloids on tumor growth when used alone or in combination. The pharmacokinetic parameters of 5-FU and its metabolites dihydrofluorouracil(FUH₂)and fluorodeoxyuridine(FUrd)in rats were determined by LC-MS/MS. Results The IC₅₀ values of Alstonia mairei total alkaloids on HCT15 and CT26.WT cells were 31.98 μg /mL and 23.71 μg /mL, respectively. The tumor inhibition rate of low dose(100 mg/kg)and high dose(200 mg/kg)Alstonia mairei total alkaloids combined with 5-FU in tumor-bearing mice increased from 50.91% to 58.56% and 62.16%, respectively. The T_1/2 of 5-FU combined with Alstonia mairei total alkaloids was significantly prolonged, and the plasma concentration of FUrd was significantly decreased. Conclusion The Alstonia mairei total alkaloids have certain cytotoxicity against colorectal cancer cells, and its anti-colorectal cancer activity can be significantly enhanced in combination with 5-FU. Alstonia mairei total alkaloids may enhance the biological activity of 5-FU by reducing the concentration of DPD to delay the catabolism of 5-FU and regulate its anabolic pathway.

YBX3 regulates thermogenesis and energy expenditure by regulating UCP1 in brown adipose tissue

WANG Wenqin, GAO Xianlong, GONG Yongfeng, FENG Ke

Journal of Shandong University (Health Sciences). 2024, 62(6): 38-47. doi:10.6040/j.issn.1671-7554.0.2024.0227

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Objective To observe the effects and mechanisms of Y-box protein 3(YBX3)deletion in brown adipose tissue(BAT)on thermogenesis and energy consumption in mice. Methods The mice were placed in a 4 ℃ environment as a cold stimulation method. Lentiviral infection assay was used to construct a human stromal vascular fraction cell line that overexpresses YBX3 or inhibits YBX3 expression. Oxygen consumption rate was used to assess mitochondrial function. Mouse BAT was injected with YBX3 siRNA(si-Ybx3)to inhibit YBX3 expression in BAT. The oxygen consumption rate, CO₂ release rate and energy consumption rate of mice were detected to evaluate the energy metabolism of mice. qRT-PCR, Western blotting, and immunofluorescence staining assays were used to detect peroxisome proliferator-activated receptor gamma coactivator 1α(PGC-1α), uncoupling protein 1(UCP1)and YBX3 expression. HE staining was used to detect the size of lipid droplets of adipocytes in BAT. RNA-binding protein immunoprecipitation was used to extract the mRNA bindsing specifically to YBX3. Next-generation sequencing assay was used to identify mRNA. Dual-luciferase reporter gene systems were used to detect the binding of YBX3 to the target sequence. Results YBX3 expression in BAT of wild-type mice was induced by cold stimulation(P<0.05). Overexpressing YBX3 promoted the expression of the thermogenic gene PGC-1α and UCP1 after human stromal vascular fraction cell line cells being induced into mature brown adipocytes(P<0.05). Silencing YBX3 in human stromal vascular fraction cell line cells inhibited the expression of PGC-1α and UCP1(P<0.05)and mitochondrial oxidative phosphorylation after being induced into mature brown adipocytes(P<0.05). Compared with the control group, mice injected with si-Ybx3 had significantly lower energy consumption(P<0.05), lower body temperature in a cold environment(P<0.05), and lower expression levels of thermogenic genes(P<0.05). The size of lipid droplets in adipocytes increased significantly(P<0.05). Compared with at 25 ℃, the expression of YBX3 in the nucleus of brown adipocytes significantly increased from mice kept at 4 ℃(P<0.05). YBX3 specifically bound to the target sequence in the 3' untranslated region of PGC-1α mRNA(P<0.05). YBX3 specifically bound to PGC-1α and UCP1 promoter regions(P<0.05). Conclusion YBX3 in BAT can affect thermogenesis and energy metabolism by regulating the expressions of PGC-1α and UCP1.

The predictive value of systemic immune-inflammation index for immune checkpoint inhibitor treatment-related adverse reactions in patients with primary liver cancer

TIAN Lijun, SANG Yujie, SUN Yujing, HAN Bing, QIN Chengyong, QI Jianni

Journal of Shandong University (Health Sciences). 2024, 62(6): 48-53. doi:10.6040/j.issn.1671-7554.0.2024.0267

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Prediction of lateral lymph node metastasis in papillary thyroid carcinoma larger than 10 mm

LIN Yuyang, WANG Bei, LI Fei

Journal of Shandong University (Health Sciences). 2024, 62(6): 54-64. doi:10.6040/j.issn.1671-7554.0.2024.0074

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Objective To predict the occurrence of lateral lymph node metastasis(LLNM)in papillary thyroid carcinoma(PTC)larger than 10 mm in diameter(non-T1A stage)and analyze the metastatic pattern. Methods A retrospective analysis was conducted on 166 PTC patients with the foci diameters larger than 10 mm who underwent thyroidectomy combined with lateral lymph node dissection. Clinicopathological characteristics were evaluated, and the occurrence and pattern of LLNM were analyzed. Univariate and multivariate Logistic regression analyses were used to assess the factors associated with LLNM. A nomogram model was established based on the results, and the receiver operating characteristic(ROC)curve was utilized to evaluate the models accuracy. Results LLNM was most commonly observed in area-Ⅲ(38.1%), and the two-segment metastasis pattern was the most prevalent(42.2%). There was no significant difference in the pattern of LLNM when the lesions were located in different areas of the gland(P>0.05). Multivariate Logistic regression analysis revealed that the location of the lesion, levels of free triiodothyronine(FT3), thyroid-stimulating hormone(TSH), thyroid peroxidase antibodies(TPOAB), and the number of central lymph node metastasis(CLNM)were independent risk factors for LLNM. The area under the curve of the nomogram prediction model is 0.851, indicating a high level of diagnostic accuracy for the established model. Conclusion The nomogram model, incorporating clinicopathological and ultrasonic characteristics, and laboratory results(the number of CLNMs, location of the lesion, FT3, TSH, TPOAB levels), proves to be effective in predicting LLNM in PTCs larger than 10 mm and holds promise for clinical application.

Abnormalities of morphology and function of meibomian gland and their effects on tear film in patients with systemic lupus erythematosus

KANG Suifang, WANG Yingxue, ZHANG Qichen, WANG Zhilun, YIN Zhe, XIAO Juan, CHEN Guoling

Journal of Shandong University (Health Sciences). 2024, 62(6): 65-70. doi:10.6040/j.issn.1671-7554.0.2024.0207

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Objective To observe the abnormalities of morphology and function of meibomian gland and their effects on tear film in systemic lupus erythematosus(SLE)patients. Methods Eighty-two eyes of 41 SLE patients were recruited as SLE group and 52 eyes of 26 healthy volunteers were recruited as control group. The grade of meibomian gland dropout was evaluated by Keratograph 5M ocular surface analyzer, the tear film lipid layer thickness(TFLLT)was measured by LipiView interferometer, the dynamic tear film objective scatter index(OSI)was measured by the double-pass Optical Quality Analysis System(OQAS II). The eyelid margin, meibomian gland secretion, fluorescein breakup time(FBUT)and fluorescein cornea staining(FCS)were observed under slit lamp microscope. Schirmer Ⅰ test(SⅠt)was conducted using standard Schirmer paper strip. The correlations between meibomian gland parameters and tear film function were assessed by Spearman rank correlation analysis. Results Compared with the control group, the meibomian gland dropout score(MGDS), meibomian gland secretion score(MGSS), eyelid margin score(EMS), dynamic tear film OSI and FCS were higher, while FBUT and TFLLT were lower in SLE group(all P<0.05). There was no significant difference in the SⅠt between SLE group and control group(P>0.05). MGDS, MGSS, and EMS were negatively correlated with FBUT(r_MGDS=-0.335, r_MGSS=-0.424, r_EMS=-0.318, P<0.05), while MGDS was positively correlated with dynamic tear film OSI(r=0.254, P<0.05). Conclusion Abnormalities in the morphology and function of meibomian glands can be caused by SLE, leading to decreased TFLLT and tear film stability.

Gargle-bubble method for the treatment of pharyngeal fishbone impaction

WANG Xinyue, WANG Xinglei, LU Meiqi, ZHAO Jie, WANG Xiaochuan, JIAO Ya, JIANG Duyin, XU Anting

Journal of Shandong University (Health Sciences). 2024, 62(6): 71-75. doi:10.6040/j.issn.1671-7554.0.2024.0133

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Objective To explore the feasibility of using the gargle-bubble method(GBM)to treat laryngopharyngeal fishbone impaction. Methods A total of 531 patients with laryngopharyngeal fishbone impaction from January 2019 to December 2022 were retrospectively analysed. Patients in group A(n=263)swallowed vinegar or steamed buns by themselves, and patients in group B were treated directly with GBM(n=268). The effective rates of the two groups were compared. Results In group A, the effective rate of swallowing vinegar or steamed buns was 15.2%(n=40), and 84.8%(n=223)of patients failed and received further medical treatment. In group B, the effective rate of GBM was 98.1%(n=263), 0.8%(n=2)of patients refused to undergo GBM and 1.1%(n=3)of patients failed to treatment. Conclusion GBM can be considered as a simple and effective self-treatment for laryngopharyngeal fish bone impaction, which is worthy of popularisation and application.

Clinical features and genetic analysis of three cases of Wiedemann-Steiner syndrome with KMT2A gene variants

WANG Mengqin, ZHANG Yaodong, CHEN Jiajia, ZHANG Zixia, HU Jiaqian, WANG Xi, ZHAO Yixuan, ZHANG Yingxian, WEI Haiyan, CHEN Yongxing

Journal of Shandong University (Health Sciences). 2024, 62(6): 76-81. doi:10.6040/j.issn.1671-7554.0.2023.1067

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Objective To explore the clinical characteristics and genetic etiology of 3 children with Wiedemann-Steiner syndrome(WSS)and to improve the understanding of the disease. Methods A retrospective analysis was conducted on the general data, clinical characteristics, laboratory tests such as growth hormone stimulation test, and imaging examinations such as pituitary magnetic resonance imaging of three children with WSS who visited Chidrens Hospital Affiliated to Zhengzhou University from August 2016 to July 2022. Recombinant human growth hormone(rhGH)treatment was performed, and genetic testing was conducted on the proband and family. Results Among the 3 patients, 2 were males and 1 was female, aged 4.8 to 11.3 years. All patients were diagnosed with short stature, accompanied by intellectual disability, special facial features, and hirsutism. Growth hormone stimulation test results showed that the case 3 had growth hormone deficiency with significantly delayed bone age. All case showed normal pituitary magnetic resonance imaging, then received rhGH treatment for 1 to 5 years. The height increased by 0.9 SD, 1 SD, and 2.2 SD, respectively, with case 3 followed up to a lifetime height of 157.9 cm. Genetic testing results showed heterozygous mutations in the KMT2A gene. The results showed heterozygous deletions of 2.5 Kb fragments in the chr11:118353970-118356519 region, c.11081delp.K3694Sfs*3, and c.5803-3T>G. All three gene mutations were newly reported. The final diagnoses were WSS. Conclusion For children with delayed growth and development, intellectual disability, special facial features and hirsutism, WSS should be considered. This study broadens the variant spectrum and clinical phenotypic spectrum of the KMT2A gene.

Two family reports and literature review of autosomal dominant osteopetrosis type II

ZHANG Di, NIE Chenyu, LIU Jidong, HOU Xinguo, CHEN Li

Journal of Shandong University (Health Sciences). 2024, 62(6): 82-90. doi:10.6040/j.issn.1671-7554.0.2024.0064

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Objective To analyze the clinical features of two patients with autosomal dominant osteopetrosis(ADO), and to explore the mutations in the causative genes in the probands and their family lines. Methods The clinical characteristics and laboratory examination data of the two ADO cases were collected and analyzed, and relevant literature was reviewed to summarize the diagnosis and treatment of the disease. Results Genetic testing revealed a new missense mutation in exon 24 of the chloride channel protein 7(CLCN7)gene, p.Gly765Cys, in proband 1, and a known missense mutation, p.Arg286Trp, in exon 10 of the CLCN7 gene in proband 2. Both probands manifested abnormally high bone mass and "sandwich" vertebral body changes, but both had normal blood calcium, phosphorus and alkaline phosphatase levels and elevated lactate dehydrogenase and creatine kinase levels. Conclusion Patients with ADO mainly exhibit abnormally high bone density, increased bone fragility and susceptibility to fracture. Currently, the disease is mostly treated symptomatically. In severe cases, anemia, thrombocytopenia with hemorrhage, frequent infections, and liver and spleen enlargement may occur. Further literature review will better summarize the clinical presentation, diagnosis and therapeutic features of ADO.

Transjugular vein bypass for the treatment of hemodialysis arteriovenous fistula dysfunction caused by subclavian vein occlusion

HE Lanlan, YANG Ke, REN Wanjun, WANG Xiaoping

Journal of Shandong University (Health Sciences). 2024, 62(6): 91-95. doi:10.6040/j.issn.1671-7554.0.2024.0011

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Objective To investigate the clinical effect of transjugular vein bypass for the treatment of hemodialysis arteriovenous fistula dysfunction caused by subclavian vein occlusion. Methods Clinical characteristics, key points of transjugular venous bypass, and postoperative fistula function of three cases with hemodialysis arteriovenous fistula dysfunction caused by subclavian vein occlusion were retrospectively analyzed. Results Transjugular venous bypass was successful in all three patients, and none of them had any related postoperative complications. Patient 1 was followed up for 9 months postoperatively, with a patent transposition vessel and good arteriovenous fistula function. Patient 2 was followed for 33 months postoperatively, with PTA twice for anastomotic stenosis; the artificial blood vessel was patency and the arteriovenous fistula function was good. Patient 3 was followed up for 15 months postoperatively, with a patent artificial blood vessel and good arteriovenous fistula function. Conclusion Transjugular venous bypass can be an option when endovascular treatment for subclavian vein occlusion is unsuccessful or risky.

Influencing factors on the incidence of hand, foot and mouth disease in Xian based on MGWR model

LIANG Kemeng, LI Shufen, NI Zhisong, SONG Sihao, XI Rui, CHENG Chuanlong, ZUO Hui, DUAN Yuqi, LIU Kun, BAI Yao, LI Xiujun

Journal of Shandong University (Health Sciences). 2024, 62(6): 96-101. doi:10.6040/j.issn.1671-7554.0.2024.0082

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Objective To investigate the relationship between the incidence of hand, foot and mouth disease(HFMD)and factors related to environment and socioeconomic in Xian at a spatial grid scale of 5 km, and provide a basis for the development of regional control and prevention measures. Methods The 2019 HFMD report data in Xian were collected and analyzed by spatial autocorrelation to characterize the spatial distribution. The role of natural environmental factors and socioeconomic factors on the incidence of HFMD was analyzed based on a multiscale geographically weighted regression(MGWR)model and compared with those of the ordinary least square(OLS)regression model and the geographically weighted regression(GWR)model. Results The annual reported incidence rate of HFMD in Xian in 2019 was 157.99/100,000, with a positive correlation in spatial distribution(global Morans I=0.349, P<0.001). The MGWR model fit was better than the GWR model and the OLS model(MGWR: R2=0.530; GWR: R2=0.473; OLS: R2=0.327). On the scale of influence, the GDP, land urbanization level, and average temperature had larger scale effects, while normalized difference vegetation index(NDVI)had smaller scale influence. GDP was significantly negatively correlated with the reported incidence of HFMD, land urbanization level and average temperature were significantly positively correlated with the reported incidence, and NDVI had a significant negative effect on HFMD incidence in parts of Xian. Conclusion The influence of environmental and socioeconomic factors on the incidence of HFMD is significant and there are spatial differences in the role of each influencing factor. The results are helpful for the formulation of regional prevention and control strategies for HFMD.

Trajectory analysis of post-craniotomy headache in brain tumor patients based on growth mixture modeling

ZHANG Xinrui, SUN Peng, LI Feng

Journal of Shandong University (Health Sciences). 2024, 62(6): 102-107. doi:10.6040/j.issn.1671-7554.0.2024.0178

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Objective To determine the trajectory of post-craniotomy headache symptoms in patients with brain tumors treated with craniotomy. Methods A longitudinal study design was used to assess postoperative headache symptoms using the visual analogue scale(VAS)score at the 7th day, the first month, the third month postoperatively in patients who were diagnosed with brain tumors and hospitalized in the Shandong Cancer Hospital Affiliated to Shandong First Medical Univerisity from March to June 2023. The higher VAS scores indicated more severe headache symptoms. Changes in VAS scores over time and possible underlying categories were determined using Latent Growth Curve Models as well as Growth Mixture Modeling. Results The VAS scores of 304 patients with brain tumors who completed a complete 3-month follow-up were analyzed and found to be decreased during the three follow-up measurements and the trend existed in three different categories. The plus group and the stable group accounted for 1.3% and 19.0%, and the remission group accounted for 79.6%. Conclusion There are individual differences in the development of post-craniotomy headache in patients with brain tumors and there are three types of headache symptom trajectories. Understanding the developmental trajectory of post-craniotomy headache symptoms can help clinics to regulate post-craniotomy headache more scientifically.

Influencing factors on grandparenting training demands of urban middle-aged and older adults based on Anderson model

SONG Dongyu, LYU Jingran, LI Yuli, LIU Yuzhou

Journal of Shandong University (Health Sciences). 2024, 62(6): 108-115. doi:10.6040/j.issn.1671-7554.0.2024.0070

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Objective To explore the status and influencing factors of grandparenting training demands among urban middle-aged and older adults. Methods Participants were recruited from community in Jinan, Shandong Province, China, using convenience sampling. Based on the Anderson model, χ2 test and binary Logistic regression analysis were used to explore the effects of various factors on the grandparenting training demands among middle-aged and older adults. Results Seventy point seven percent of the participants had demands for training in grandparenting, in which a propensity factor of previous residence in town(OR=2.403, 95%CI: 1.016-5.682), an enabling factor of previous frequent availability of grandparenting resources in the community(OR=2.112, 95%CI: 1.215-3.671), and demand factors of occasional conflict with children due to grandparenting(OR=6.250, 95%CI: 1.543-17.866), and without depressive symptoms(OR=0.557, 95%CI: 0.319-0.972)were domimnt. Conclusion Propensity factors, enabling factors and demand factors all have different degrees of influence on the demands of grandparenting training for the middle-aged and older adults. It is of great social value and significance to actively carry out grandparenting training. In addition to paying attention to the middle-aged and older adults with grandparenting training demands, society should also pay attention to the mental health status of those without grandparenting training demands.

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