1.Construction and validation of malnutrition risk model for non-dialysis patients with stages 3 to 5 chronic kidney disease
Yuting ZHENG ; Zexuan ZHENG ; Yabo SUN ; Yafei WAN ; Zhanhong GAO ; Shuxing LI
Journal of Clinical Medicine in Practice 2025;29(11):110-117
Objective To construct and validate a risk prediction model for malnutrition in non-di-alysis patients with stages 3 to 5 chronic kidney disease(CKD)based on Logistic regression(LR)and XGBoost algorithms,and to compare the predictive performance between the two models.Methods A total of 506 CKD patients were enrolled as study subjects.According to chronological order,they were divided into training set(n=404)and test set(n=102)at the ratio of 8 to 2.The training set was divided into case group and control group based on whether they were malnourished,with 202 cases in each group.The LR and XGBoost models were established,and the model efficacy was evaluated through the area under the receiver operating characteristic(ROC)curve(AUC),sensitivity,speci-ficity,GiViTI calibration curve band and clinical decision curve.Results The LR model identified age ≥60 years,disease of stage 5,reduced appetite,hypoalbuminemia,low prealbumin,low mid-arm muscle circumference and high perceived stress as independent risk factors for malnutrition among non-dialysis CKD patients,while physical activity was identified as a protective factor(P<0.05).In the XGBoost model,the top five influential variables were serum albumin,appetite,physical activity,prealbumin and mid-arm muscle circumference.The AUC of the LR and XGBoost models in the train-ing set were 0.930 and 0.947 respectively,and those in the test set were 0.925 and 0.933.The pre-dictive ability of the latter was slightly higher(P>0.05).The GiViTI calibration curve bands all showed good calibration capability.Conclusion The XGBoost model combined with shapley additive explanation performs better in identifying malnourished patients and guiding precise care.
2.Expression of long non-coding RNA SFTA1P and its effect on biological functions in lung squamous cell carcinoma
Weiping WAN ; Weijia XIE ; Tingting XIA ; Ying XIANG ; Na WU ; Chengying LI ; Yifan SHAN ; Li BAI ; Yafei LI
Journal of Army Medical University 2024;46(11):1226-1234
Objective To investigate the expression of long non-coding RNA(lncRNA),surfactant associated 1 pseudogene(SFTA1P)in lung squamous carcinoma and its effect on the biological functions of SFTA1P in lung squamous carcinoma cell lines.Methods Based on the cancer genome atlas(TCGA)database,the differential expression of SFTA1P in tumor and normal tissues were compared in patients diagnosed with lung squamous cell carcinoma.Then,the expression of SFTA1P was detected in human normal lung epithelial cell line BEAS-2B and lung squamous cell lines SK-MES-1 and H520 with real-time quantitative polymerase chain reaction(RT-qPCR).SK-MES-1 and H520 cells with overexpression and/or knockdown of SFTA1P were constructed by transfecting the overexpression plasmids(pcDNA3.1-SFTA1P)and small interfering RNAs(si-SFTA1P-1 and si-SFTA1P-2).CCK-8 assay and Transwell assay were used to investigate the effect of SFTA1P on biological functions in lung squamous carcinoma cells.Differential gene expression analysis,correlation analysis and functional enrichment analysis were employed to explore the potential mechanism that SFTA1P may affect biological functions of lung squamous cells.Results Analysis of TCGA showed that the expression of SFTA1P was significantly lower in lung squamous cell carcinoma tissue than adjacent normal tissue(P<0.05).RT-PCR results showed that the expression of SFTA1P was obviously lower in lung squamous carcinoma cells than the human normal lung epithelial cells(P<0.05).And the expression level of SFTA1P was relatively lower in the SK-MES-1 cells than the H520 cells(P<0.05).Overexpression of SFTA1P suppressed the proliferation,migration and invasion of lung squamous carcinoma cells(P<0.05),while its knockdown promoted these abilities(P<0.05).Differential gene expression analysis,correlation analysis and functional enrichment analysis indicated that SFTA1P may inhibit MYC,G2m checkpoints and E2f signaling pathways in lung squamous cell carcinoma.Conclusion SFTA1P shows anti-cancer function in lung squamous cell carcinoma,and it may affect the biological functions of lung squamous cell carcinoma cells through down-regulating MYC,G2m checkpoints and E2f signaling pathways.
3.Effects of icariin on adhesion and cytoskeleton of osteoblasts in response to the extreme mechanical environment of hypergravity
Licheng SONG ; Huafeng ZHANG ; Wei CHENG ; Ya LI ; Dong LI ; Yafei QIN ; Xin WAN ; Ruixin LI ; Hui LI ; Xizheng ZHANG
Chinese Journal of Orthopaedics 2020;40(6):362-371
Objective:To establish a hypergravity loading model with a high-acceleration centrifugal loading device and to investigate the effects of different hypergravity loading and icariin on osteoblast adhesion and cytoskeleton.Methods:MC3T3-E1 cells were seeded in the dishes of cell culture at a density of 2×10 5/cm 2. And the experiment was divided into 6 groups: control group (without icariin and loading); simple administration group (only icariin); 10 G loading group (only loading); 10 G administration group (with icariin and loading); 40 G loading group (only loading); 40 G administration group (with icariin and loading). The experimental loading group was loaded with MC3T3-E1 cells using a high-acceleration centrifugal loader. And continuous loading for 3 d, 30 min per d. The control group and the simple administration group were exposed to normal gravity, and the remaining conditions were not different from the experimental group. Icariin was used at a concentration of 10 -7 mol/L in all administration groups, and the experiments were carried out according to the method of preventive administration. At the same time, the related molecular biological techniques such as alizarin red staining, alkaline phosphatase (ALP) activity measurement, CCK-8 cell proliferation experiment, cytoskeleton phalloidin staining, qPCR and Western Blot were used to detect the effects of icariin on osteoblasts adhesion protein integrin α5 and integrin β1 and cytoskeleton protein F-actin under hypergravity extreme mechanical environment. Results:All models were successfully prepared. The alizarin red staining: The icariin could significantly promote the formation of osteoblastic calcified nodules. And the 10 G loading could also promote the mineralization of osteoblasts and increase the number of mineralized nodules, while the mineralization and the number of mineralized nodules of osteoblasts are significantly reduced in 40 G loading. ALP activity test: The OD values of simple administration group, 10 G loading group and 40 G loading group were 0.246, 0.331 and 0.163, respectively. Compared with 0.207 in the control group, the differences were statistically significant ( P<0.05). The 10 G administration group and the 40 G administration group were 0.373 and 0.180, and the differences were statistically significant ( P<0.05). The results of CCK-8 proliferation experiments: The OD value of simple administration group were 0.650, which was statistically significant compared with 0.551 of control group ( P=0.031). The 10 G loading group and 40 G loading group were 1.193 and 0.245, and their differences with the control group were both statistically significant ( P<0.05). The OD value of 10G administration group and the 40 G administration group were 1.300 and 0.310, which were significantly different from the respective loading groups ( P<0.05). Phalloidin staining: 10 G loading significantly increased the number of cells, but the changes in cells morphology and skeleton were not obvious. 40 G loading significantly inhibited the increase of the number of cells, meanwhile, made the pseudopods of cells more shorter and even disappeared. 40 G loading made the seriously damage of the cytoskeleton and even cause the cells to death. Icariin had no effect on the cells morphology, but it did has a certain repair effect after the cells loading. The results of qPCR and Western Blot experiments all confirmed that the expressions of integrin α5, integrin β1 and F-actin were up-regulated after icariin treatment. 10 G loading could promote the expression of integrin α5, integrin β1 and F-actin, and 40 G loading significantly inhibited the expression of the mRNA and proteins. Conclusion:Both 10 G condition and icariin can promote the development, cell adhesion and the cytoskeleton's stability of osteoblasts, while 40 G has a significant inhibitory effect.
4.A study on the effect of annexinⅤpositive microparticles on hypercoagulability in patients with hip fracture
Xin WAN ; Yafei QIN ; Chunlei XU ; Licheng SONG ; Na LI ; Huafeng ZHANG ; Hui LI
Chinese Journal of Orthopaedics 2020;40(9):584-592
Objective:To study the correlation between hypercoagulant status of patients with hip fracture and the level of microparticles (MPs) in their peripheral blood, and to explore the feasibility of removing MPs to correct hypercoagulant status in patients with hip fracture.Methods:Sixty-five patients from December 2018 to September 2019 with hip fracture were included. There were 24 males and 41 females with the average age of 75.6±9.8 years old (range 58-95 years). Among them, 27patients (43.1%) were femoral neck fracture and38 patients (56.9%) were intertrochanteric fracture.All patients were diagnosed with X-ray and CT. Meanwhile, about 20 healthy people in the physical examination center included as controls in our study. There were 8 males and 12 females with the average age of 72.3±6.5 years old (range 56-81years). 2 ml of anticoagulant whole blood was taken on an empty body in the morning, and purified microparticlesby whole-blood density gradient centrifugation in whole blood were identified by electron microscope and nanoparticle tracking analyzer. After cell free plasma (CFP) was obtained by whole-blood density gradient centrifugation, the number of whole annexin V (AV) positived MPs and these MPs which from platelet (PMPs) was determined by flow cytometry. The activated clotting time (ACT) was determined by coagulation and platelet function analyzer to evaluate the degree of hypercoagulability. Then, Logistic analysis was performed on risk factors associated with hypercoagulability to determine whether the level of MPs was an independent risk factor for hypercoagulability, and the correlation between ACT value and MPs level was analyzed. Finally, the four coagulation items of each sample CFP before and after MPs removal were determined by automatic coagulation analyzer.Results:Under electron microscopy, MPs presented vesicular appearance,with a complete double-layer membrane structure, the size was in the range of 100-1 000 nm, and the morphology was not uniform. there were irregular vesicular and circular vesicular general shapes. The average size of MPs in peripheral blood of patients with hip fractures was 239.7±4.0 nm. The mean size of MPs distribution in the control group was 247.7±3.3 nm, and there was no statistically significant difference in MPs diameter between the two groups. The average level of circulating AV +MPs in patients with hip fracture was 564±171/μl, and the average level of PMPs was 326±104/μl. In the control group, the average level of AV +MPs was 252±82/μl, the average level of PMPs was 192±41/μl, the difference between AV +MPs and PMPs was statistically significant ( P<0.05). The average ACT level of patients with hip fracture was 324±94 s, while the average ACT level of the normal population was 535±76 s, and the difference between the two was statistically significant ( P<0.05). Single factor logistic regression analysis showed that the levels of APTT, PMPs and AV +MPs may be risk factors for hypercoagulability, and multivariate logistic regression analysis showed that AV +MPs is an independent risk factor for hypercoagulability.It has a highly negative correlation with ACT ( r=-0.822, P<0.05). The results of four coagulation items determined by CFP were PT 10.8±0.46 s, APTT 30.6±1.56 s, Fib 3.08±0.36 g/L, INR 0.98±0.04 and TT 19.3±0.62 s. After the removal of MPs, the coagulation function was PT 10.8±0.52 s, APTT 32.4±3.0 s, Fib 2.90±0.33 g/L, INR 0.99±0.05 s, and TT 19.9±0.63 s. There was no statistically significant difference before and after coagulation function. Conclusion:There is a hypercoagulable state in patients with hip fracture, moreover, the level of AV +MPs is an independent risk factor for hypercoagulability, which is highly correlated with ACT, and MPs has no significant effect on the classic four factors of coagulation.

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