1.Promotive effect of high expression of nerve growth factor in Schwan-like cells induced by adipose-derived stem cells on growth of rat dorsal root ganglion cell protrusion
Qinghua ZHU ; Bo YUAN ; Yilun WANG ; Miao REN ; Xiaofei LI ; Simiao WANG ; Zixuan ZHEN ; Xiumei FU
Journal of Jilin University(Medicine Edition) 2025;51(4):984-995
Objective:To discuss the promotive effect of nerve growth factor(NGF),which is highly expressed in the adipose-derived stem cell(ADSC)-induced Schwann-like cells(SCLCs),on the growth of dorsal root ganglion(DRG)cell processes in the rats,and to clarify its mechanism.Methods:The ADSCs were extracted from the epididymal adipose tissue of the SD rats,and their multidirectional differentiation potential was identified through osteogenic,adipogenic,and chondrogenic induction.The ADSCs were induced to differentiate into the SCLCs,and the expression levels of glial fibrillary acidic protein(GFAP)and S100 calcium-binding protein β(S100β)protein in the ADSCs and SCLCs were detected by immunofluorescence staining and Western blotting methods.The DRG cells were isolated and cultured,and immunofluorescence staining was used to detect the βⅢ-tubulin expression in the DRG cells for identification.The SCLCs were co-cultured with the DRG cells(co-culture group),the single-culture DRG cells were regared as DRG group and toluidine blue staining was used to observe and measure the length of DRG cell processes under the optical microscope in co-culture group and DRG group.Small interfering RNA(siRNA)transfection was used to knock down NGF,and plasmid transfection was used to over-express NGF.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the NGF mRNA expression levels in the cells in various groups;enzyme-linked immunosorbent assay(ELISA)method was used to detect the NGF protein levels in the cell supernatants.The transfected SCLCs were co-cultured with DRG cells and divided into control group,siNC/vector group,NGF knockdown group(si-NGF group),and NGF over-expression group(oe-NGF group).The lengths of DRG cell processes in various groups were observed.Results:The primary ADSCs adhered within 24 h after seeding,with a small number of lipid droplets remaining.After 3 d of culture,the cells were mostly short spindle-shaped,fusiform,or polygonal,growing rapidly in a vortex pattern.After passaging,the cells exhibited a uniform morphology,appearing as long spindles arranged in a fish-school pattern.After 14 d of adipogenic induction,the cell morphology changed from spindle-shaped to flat-round,with translucent lipid droplets forming in the cytoplasm,which were stained red by Oil Red O.After 28 d of osteogenic induction,the cells appeared sand-like with blurred morphology,and calcified nodules were observed,which were stained red by Alizarin Red and deposited in the extracellular matrix.After 28 d of chondrogenic induction in a 3D culture system,millet-sized chondrogenic spheres formed.Frozen sections of the spheres were stained with Alcian Blue,and acidic mucopolysaccharides in the cartilage tissue were stained blue under the microscope.Under the fluorescence microscope,the third-passage purified ADSCs showed positive expression of CD29[fluorescein isothiocy anate(FITC)-labeled green fluorescence]and CD44(Cy3-labeled red fluorescence).The immunofluorescence staining results showed that GFAP was labeled with FITC(green fluorescence),and S100β was labeled with Cy3(red fluorescence).The Western blotting results showed that compared with ADSCs,the expression levels of S100β and GFAP proteins in the SCLCs were increased(P<0.05).The primary DRG cells began to adhere 6 h after conventional culture,and after 3 d,the cell bodies appeared round and bright,with two linear processes extending from them.Under fluorescence microscope,the cells positively expressed the neuron-specific marker βⅢ-tubulin,confirming that the isolated cells were DRG cells.Compared with the ADSCs,the NGF protein expression level in the SCLCs was increased(P<0.05).Compared with DRG group,the length of DRG cell processes in co-culture group was the highest when DRG cells and SCLCs were co-cultured at a 1∶2 ratio(P<0.05).The RT-qPCR results showed that compared with si-NC group,the expression levels of NGF mRNA in the cell supernatant in si-NGF-1,si-NGF-2,and si-NGF-3 groups were significantly decreased(P<0.05),with si-NGF-1 showing the highest knockdown efficiency,which was selected for subsequent experiments.The ELISA results showed that compared with si-NC group,the NGF levels in the cell supernatant of si-NGF-1,si-NGF-2,and si-NGF-3 groups were decreased(P<0.05).Compared with Vector group,the expression level of NGF mRNA and NGF protein level in the supernatant in oe-NGF group were increased(P<0.05).Compared with control group and siNC/vector group,the length of DRG cell processes in si-NGF group was decreased(P<0.05),while the length of DRG cell processes in oe-NGF group was increased(P<0.05).Conclusion:ADSCs can be directionally differentiated into SCLCs,and the differentiated cells highly express NGF.Knockdown or overexpression of NGF can affect the growth of DRG cell processes.
2.The enlightenment of artificial intelligence large-scale model on the research of intelligent eye diagnosis in traditional Chinese medicine
Yuan GAO ; Zixuan WU ; Boyang SHENG ; Fu ZHANG ; Yong CHENG ; Junfeng YAN ; Qinghua PENG
Digital Chinese Medicine 2024;7(2):101-107
Eye diagnosis is a method for inspecting systemic diseases and syndromes by observing the eyes.With the development of intelligent diagnosis in traditional Chinese medicine(TCM),artificial intelligence(AI)can improve the accuracy and efficiency of eye diagnosis.However,the research on intelligent eye diagnosis still faces many challenges,including the lack of standardized and precisely labeled data,multi-modal information analysis,and artificial in-telligence models for syndrome differentiation.The widespread application of AI models in medicine provides new insights and opportunities for the research of eye diagnosis intelli-gence.This study elaborates on the three key technologies of AI models in the intelligent ap-plication of TCM eye diagnosis,and explores the implications for the research of eye diagno-sis intelligence.First,a database concerning eye diagnosis was established based on self-su-pervised learning so as to solve the issues related to the lack of standardized and precisely la-beled data.Next,the cross-modal understanding and generation of deep neural network models to address the problem of lacking multi-modal information analysis.Last,the build-ing of data-driven models for eye diagnosis to tackle the issue of the absence of syndrome dif-ferentiation models.In summary,research on intelligent eye diagnosis has great potential to be applied the surge of AI model applications.
3.Clinicopathological characteristics and prognosis of idiopathic membranous nephropathy with or without C3 deposition
Zixuan FU ; Huifang WANG ; Jun LIU ; Chunhui JIANG ; Min LI ; Xuemei LIU
Chinese Journal of General Practitioners 2023;22(3):295-300
Objective:To investigate the clinicopathological characteristics and prognosis of idiopathic membranous nephropathy (IMN) with or without C3 deposition.Methods:Clinical and pathological data of 576 patients with IMN diagnosed in Affiliated Hospital of Qingdao University from January 2017 to January 2021 were retrospectively analyzed. The patients were divided into C3 deposition group and non-C3 deposition group according to the immunofluorescence staining of C3. The clinical and pathological characteristics were compared between the two groups. Kaplan-Meier survival curve was used to compare the prognosis of the two groups.Results:A total of 576 IMN patients (male 364 (63.20%)) were enrolled, including 400 patients (69.44%) with C3 deposition and 176 patients (30.56%) without C3 deposition. Compared with the non-C3 deposition group, the levels of total blood cholesterol ( t=0.94, P=0.002) and the proportion of phospholipase A2 receptor ( χ2=9.99, P=0.002), IgG ( χ2=10.67, P=0.001), IgM ( χ2=7.00, P=0.008), IgA ( χ2=7.87, P=0.005) and C1q ( χ2=8.28, P=0.004) depositions in renal tissues was higher in C3 deposition group, while the levels of serum C3 ( t=2.87, P=0.004), albumin ( t=3.57, P<0.001) and IgG ( Z=3.55, P<0.001) were lower in C3 deposition group. There were no significant differences in other clinicopathological indicators between the two groups. The survival analysis was performed in 460 patients who were followed for>6 months, including 319 cases (69.35%) of C3 deposition and 141 cases (30.65%) of non-C3 deposition. The end point event was defined as an eGFR decline>30% or entry into end stage renal disease (ESRD). There was no statistically significant difference in treatment method between the two groups ( P>0.05). The median follow-up time was 22 (13,32) months, 327 (71.09%) patients achieved remission, and 22 patients had renal end-point events. Compared with the non-C3 deposition group, the proportion of urinary protein remission was lower ( χ2=10.85, P<0.05), the incidence of renal end-point events was higher ( χ2=5.05, P<0.05). Kaplan-Meier survival analysis showed that patients with C3 deposition had a lower cumulative remission rate (Log-rank χ2=6.68, P=0.010), and a lower cumulative renal survival than those without C3 deposition had ( χ2=5.42, P=0.020). Conclusions:Compared with patients without C3 deposition, IMN patients with C3 deposition have more severe clinical and pathological changes, lower renal cumulative remission rate, and are more likely to have poor prognosis.
4.Correlation between serum anti-phospholipase A2 receptor antibody combined with glomerular complement C3 deposition and clinicopathology and prognosis in patients with idiopathic membranous nephropathy
Zixuan FU ; Huifang WANG ; Chunhui JIANG ; Min LI ; Yahuan YU ; Xuemei LIU
Chinese Journal of Nephrology 2023;39(10):760-767
Objective:To investigate the correlation between serum anti-phospholipase A2 receptor antibody (SAb) combined with glomerular complement C3 (GC3) deposition and clinicopathologic features and prognosis in patients with idiopathic membranous nephropathy (IMN).Methods:It was a retrospective cohort study. The patients diagnosed with IMN in Affiliated Hospital of Qingdao University from July 1, 2019 to April 30, 2022 were enrolled, and the clinical and pathological data were collected and analyzed. The patients were divided into negative SAb and negative GC3 (SAb -/GC3 -) group, negative SAb and positive GC3 (SAb -/GC3 +) group, positive SAb and negative GC3 (SAb +/GC3 -) group and positive SAb and positive GC3 (SAb +/GC3 +) group according to the status of SAb titer and GC3 deposition. Clinical and pathological characteristics among the groups were compared. Kaplan-Meier survival curve was used to compare the cumulative renal remission rates of different groups. Cox regression analysis model was used to analyze the related factors of renal remission. Results:A total of 143 IMN patients aged (53.35±12.34) years old were included in the study, including 94 males (65.7%). There were 17 patients (11.9%) in the SAb -/GC3 - group, 30 patients (21.0%) in the SAb -/GC3 + group, 19 patients (13.3%) in the SAb +/GC3 - group, and 77 patients (53.8%) in the SAb +/GC3 + group. Compared with SAb -/GC3 - group, the level of serum albumin was lower in the SAb +/GC3 + group, and the level of 24 h urine protein, SAb titer, and the proportions of glomerular anti-phospholipase A2 receptor antigen and renal tubule atrophy were higher in the SAb +/GC3 + group (all P<0.05). After 26.0 (19.0, 36.0) months of follow-up, a total of 96 patients (67.1%) attained remission. The proportion of patients receiving immunosuppressive therapy in the SAb +/GC3 + group was higher than that in the SAb -/GC3 - group [93.5% (72/77) vs. 70.6% (12/17), fisher value=8.974, P=0.016] and the proportion of renal remission rate in the SAb +/GC3 + group was lower than that in the SAb -/GC3 - group [49.4% (38/77) vs. 100% (17/17), χ2=25.438, P<0.001]. Kaplan-Meier survival curve result showed that the cumulative renal remission rate in the SAb +/GC3 + group was significantly lower than that in the SAb -/GC3 - group (Log-rank χ2=31.538, P<0.01). Multivariate Cox regression analysis result showed that 24 h urine protein level ( HR=0.891, 95% CI 0.803-0.988, P=0.029), SAb titer ( HR=0.996, 95% CI 0.992-1.000, P=0.042) and SAb +/GC3 + (with SAb -/GC3 - group as reference, HR=0.414, 95% CI 0.204-0.827, P=0.013) were independent related factors for renal remission in patients with IMN. Conclusions:IMN patients with positive SAb and GC3 deposition have more severe clinical and pathological changes, lower renal cumulative remission rates, and are more likely to have poor prognosis. The combined assessment of SAb and GC3 deposition may be helpful for evaluating prognosis and guiding treatment in IMN patients.
5.Diagnosis and Treatment Reasoning of Integrated Traditional Chinese and Western Medicine Against Acute Abdomen Based on Knowledge Graph
Zixuan FU ; Peng ZHOU ; Haiyan REN ; Chuhao SHANG ; Jingjing LUO ; Yi GUO ; Ximo WANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(11):190-199
ObjectiveIn view of the standardization of clinical diagnosis and treatment of the acute abdomen and the inheritance of diagnosis and treatment experience of prestigious veteran traditional Chinese medicine(TCM) doctors, a diagnosis and treatment reasoning algorithm based on association rule mining under incomplete evidence(AMIE)+ random walk was proposed to provide information services and technical support for primary doctors by recommending personalized diagnosis and treatment plans based on medical records. MethodThe experience of diagnosis and treatment of acute abdomen of prestigious veteran TCM doctors and the text data of clinical diagnosis and treatment guidelines of integrated TCM and western medicine were collected to complete the task of knowledge extraction and construct acute abdomen knowledge graph based on Neo4j. On the basis of ontology-supported rule-based reasoning, the rule reasoning based on similar syndromes was used to expand the syndrome combinations whose Jaccard similarity was greater than the threshold in the syndrome recommendation results. The semantic path coverage algorithm was used to calculate the semantic similarity between the symptom nodes. The symptom nodes were divided into 10 categories, and the symptom nodes in the same category were extended. The random walk algorithm was used to search the symptom nodes connected with the syndrome, and the connection rules between the syndrome and symptom nodes were extended to realize the knowledge reasoning of AMIE+ random walk. ResultThe acute abdomen knowledge graph included 1 320 nodes and 2 464 relationships. According to the link prediction evaluation index of knowledge reasoning, the reasoning results of the three algorithms in the auxiliary diagnosis and treatment of acute abdomen were compared. The AMIE+ random walk algorithm complemented the knowledge graph by extending the similar syndrome connection rules and the syndrome-symptom connection rules. Compared with the knowledge reasoning algorithm based on ontology rules, the area under the curve (AUC) was 15.18% higher and the accuracy was 30.36% higher, which achieved more accurate and effective knowledge inference. ConclusionThis study used knowledge graph technology to visualize the diagnosis and treatment of acute abdomen with TCM and western medicine, assisting primary clinicians in intuitively viewing the diagnosis and treatment process and data relationship. The proposed diagnosis and treatment reasoning algorithm can realize the personalized diagnosis and treatment plan recommendation at the level of "disease-syndrome-diagnosis-treatment-prescription", which can assist primary doctors in disease diagnosis and treatment and clinical decision-making, contribute to the knowledge sharing and application of diagnosis and treatment experience and clinical guidelines of prestigious veteran TCM doctors, improve the level of primary clinical diagnosis and treatment, and promote the normalization and standardization of the diagnosis and treatment process of acute abdomen with integrated TCM and western medicine.
6.Clinical characteristics and risk factors of antineutrophil cytoplasmic antibody-associated vasculitis complicated with infection
Chunhui JIANG ; Huifang WANG ; Dandan GUO ; Zixuan FU ; Min LI ; Xuemei LIU
Chinese Journal of Nephrology 2022;38(9):811-819
Objective:To investigate the characteristics and risk factors of infection in newly diagnosed patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV).Methods:The clinical data of AAV patients (followed up for at least 6 months) in Affiliated Hospital of Qingdao University from September 2012 to September 2020 were retrospectively collected. According to whether infection occurred during follow-up, the patients were divided into infection group and non-infection group. The clinical characteristics and infection status of the two groups were analyzed, and the Cox regression analysis model was used to explore the influencing factors of infection.Results:A total of 236 AAV patients were enrolled in this study, including 128 females (54.2%) and 108 males (45.8%), with a median age of 66.00 (59.76, 71.99) years. There were 202 patients (85.6%) with positive myeloperoxidase (MPO)-ANCA and 34 patients (14.4%) with positive protease 3 (PR3) -ANCA. There were 77 cases in the infection group and 159 cases in the non-infection group. A total of 121 infections occurred in 77 patients, and 54 infections (44.6%) occurred within 6 months after initial diagnosis. In the infection group the proportion of patients with hypertension history, pulmonary underlying diseases and patients who received hormone pulse therapy or plasma exchange, the incidence of lung, kidney, heart and gastrointestinal involvement, the level of serum creatinine and five factors score (FFS) at initial diagnosis were significantly higher than those in the non-infection group (all P<0.05), while the estimated glomerular filtration rate (eGFR) was significantly lower ( P<0.05). Lung (73.6%) was the main infection organ of AAV patients. The most common pathogenic microorganisms were bacteria (64.0%), mainly Pseudomonas aeruginosa and Staphylococcus aureus, followed by fungi (33.7%, mainly Candida albicans). Multivariate Cox regression analysis showed that lung involvement ( HR=1.682, 95% CI 1.034-2.734, P=0.036) and gastrointestinal involvement ( HR=2.976, 95% CI 1.219-7.267, P=0.017) were the independent influencing factors for infection in AAV patients. Conclusions:AAV patients have a higher incidence of infection within 6 months after initial diagnosis. The most common organ of infection in AAV patients is the lung, and the common pathogens are Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. Lung involvement and gastrointestinal involvement are the independent risk factors for infection in AAV patients.
7.Epidemiological characteristics of COVID-19 patients in Shaanxi Province
Zixuan LI ; Guotao FU ; Xiaowei LI
Shanghai Journal of Preventive Medicine 2022;34(10):973-976
ObjectiveTo describe the epidemiological characteristics of COVID-19 patients in Shaanxi Province from December 2021 to January 2022. MethodsAll COVID-19 patients’ information was obtained from the Health Committee of Shaanxi Province. SPSS 26.0 and Stata MP 16.0 were used to analyze the distribution of Time, Population and Region. Descriptive statistical method was used to investigate the correlation between age, gender and clinical syndrome types of patients. ResultsThe duration of this epidemic was 43 days, and2 080 confirmed cases in total, which distributed in cities of Xi’an (2 053 cases), Xianyang (13 cases) , Yan’an (13 cases) and Weinan(1 cases). The mean age of the patients was 35.91±17.72 years old, the number of male patients was higher than that of female, and 93.7% of the patients had mild symptoms. The age and gender of the patients were statistically correlated with the symptom type (P<0.001). ConclusionPreventing the imported and the spread of domestic cases are currently the main measures to prevent COVID-19 in China. People should abide by the requirements and duties of epidemic prevention and control. Health management and strict quarantine should apply for keynote areas and populations. Meanwhile, the discovery of asymptomatic patients is important to prevent the potential epidemic.
8.The clinical features of patients with lymphoplasmacytic diseases harboring MyD88 L265P mutation
Yuan REN ; Biqi ZHOU ; Yang XU ; Chengcheng FU ; Hongjie SHEN ; Zixuan DING ; Depei WU
Chinese Journal of Hematology 2016;37(12):1054-1059
Objective To explore the clinical features oflymphoplasmacytic diseases with MyD88 L265P mutation.Methods To analyze the distribution of MYD88 L265P mutation in patients with lymphoplasmacytic diseases by using of ARMS PCR-CE.Results There were 25(30.9%) MyD88 L265P mutated patients in 81 patients.The mutation was frequently observed in 14 patients with WM (77.8%,14/ 18),2 patients with lymphoplasmacytic lymphoma (66.7%,2/3),1 acute lymphocytic leukemia patient (50.0%,1/2),3 multiple myeloma patients (30.0%,3/10),1 patient with monoclonal gammopathy of undetermined significance (25%,1/4),3 patients with chronic lymphocytic leukemia (13.0%,3/23) and 1 lymphoma patient (4.8%,1/21).20 (80%,20/25) patients were identified with IgM subtype.Compared with wild-type group of 56 cases,mutated patients were older (median age:67 years vs 55 years,P< 0.001),with lower WBC count (median count:5.23 × 109/L vs 10.80× 109/L,P=0.001),lower HGB level (median count:85 g/L vs 119 g/L,P<0.001).Conclusion MyD88 L265P mutation was mainly observed in patients with IgM subtype lymphoplasmacytic diseases,and Waldenstrom's macroglobulinemia was the most common disease.Compared with the wild-type group,patients with MyD88 L265P mutation were older and had lower WBC count,lower level of HGB.However,further studies were needed to test the prognostic value of MyD88 L265P mutation.

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