1.Extraction and differentiation of mature and immature dendritic cells from Lewis rat bone marrow
Liqiang LI ; Minghao LI ; Liang LI ; Wenbo LI ; Jun ZHANG ; Lingmei KONG
Chinese Journal of Tissue Engineering Research 2024;28(25):4013-4017
BACKGROUND:Dendritic cells exhibit extremely strong antigen phagocytic function in the immature stage,and they can demonstrate great advantages in immune tolerance,cancer immunotherapy,and other aspects.However,due to the extremely low content of immature dendritic cells in living organisms,its clinical and scientific applications are severely limited. OBJECTIVE:To study the extraction and identification of mature and immature dendritic cells from Lewis rat bone marrow. METHODS:Bone marrow precursor cells were isolated from the bone marrow of Lewis rats,and immature dendritic cells were induced by 20 ng/mL of granulocyte colony-stimulating factor and 10 ng/mL of interleukin-4 for 7 days,and then mature dendritic cells were induced by adding 1 μg/mL of lipopolysaccharide to immature dendritic cells for 2 days.The morphology of dendritic cells was observed using inverted fluorescence microscopy.The surface-specific molecules of mature and immature dendritic cells were identified by flow cytometry,and the secretion levels of supernatant interleukin-10,interleukin-12,and interleukin-17A in mature and immature dendritic cells were detected by ELISA.The response of mature and immature dendritic cells to T lymphocyte stimulation was measured by mixed lymphocyte reaction. RESULTS AND CONCLUSION:(1)The dendritic cells showed an obvious protrusion structure under an ordinary inverted fluorescence microscope.(2)Flow cytometry showed low expression of CD40,CD86,and other co-stimulatory molecules in immature dendritic cells.On the contrary,mature dendritic cells highly expressed the above co-stimulatory molecules.(3)The secretion of interleukin-10 and interleukin-17A in immature dendritic cells was much higher than that in mature dendritic cells(P<0.01).Interleukin-12 secretion in immature dendritic cells was much lower than that in mature dendritic cells(P<0.05).(4)Mature dendritic cells stimulated T cells significantly better than immature dendritic cells,and the stimulation ability was stronger when the ratio of mature dendritic cells to T lymphocytes reached 1:10.(5)The results indicate that Lewis rat bone marrow precursor cells can differentiate into dendritic cells and distinguish between mature and immature dendritic cells by flow cytometry identification,related factor detection,and mixed lymphocyte reaction.
2.Mechanism of Prevention and Treatment of Myocardial Ischemia-reperfusion Injury by Qi-replenishing and Blood-activating Chinese Medicines Based on Theory of Qi and Blood Interacting in Vessels
Han PENG ; Gaojie XIN ; Ce CAO ; Fan GUO ; Lingmei LI ; Jianhua FU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(23):27-34
Myocardial ischemia-reperfusion injury (MIRI) is a common injury in the treatment of ischemic heart diseases. MIRI can be categorized as chest impediment and palpitation in traditional Chinese medicine, with the pathogenesis related to Qi and blood disharmony. The simultaneous disorders of Qi and blood are the key mechanism of MIRI, and thus the differentiation of Qi and blood syndromes is the prerequisite for the treatment. The theory of Qi and blood interacting in vessels is proposed by our team based on Qi being the commander of blood and blood being the mother of Qi as well as previous pharmacological studies. Specifically, Qi marshals blood by vessels, and the blood carries Qi by vessels. Accordingly, Qi and blood interact in the vessels. MIRI is accompanied by mitochondrial dysfunction, platelet function abnormality, and vascular endothelial damage, which are correlated with Qi deficiency, blood stasis, and vessel damage, respectively. Mitochondrial, platelet, and vascular endothelial structural and functional changes triggered by their interactions are one of the mechanisms by which Qi deficiency, blood stasis, and vessel damage lead to the occurrence and development of MIRI. By exploring the correlations between Qi and mitochondria, between blood and platelets, and between vessels and blood vessels, we can explain the modern scientific content of the theory of Qi and blood interacting in vessels in traditional Chinese medicine. According to the pathogenesis of Qi and blood disharmony in vessels, we discussed the pharmacological mechanisms of Qi-replenishing medicines, blood-activating medicines, and their combinations in the prevention and treatment of MIRI. On the basis of the research achievements in the prevention and treatment of MIRI by Qi-replenishing and blood-activating Chinese medicines based on the theory of Qi and blood interacting in vessels, we analyzed the effects of these medicines on Qi, blood, and vessels. According to the theory of Qi and blood, this article reveals the theoretical basis and scientific connotations of the prevention and treatment of cardiovascular diseases, with the aim of providing new ideas and references for the clinical application of traditional Chinese medicine in the prevention and treatment of cardiovascular diseases.
3.An Improved Network-Enhanced Fusion Method and its Application in Papillary Renal Cell Carcinoma Subtyping using Multi-omics Data
Guojing SHI ; Lingmei LI ; Yifang WEI
Chinese Journal of Health Statistics 2024;41(3):376-381
Objective The Network Enhancement Fusion(ne-SNF)method had no denoising strategy for the networks from each omics data type,which would weaken the power of network fusion.To address this problem,we proposed an improved Network Enhancement Fusion(improved ne-SNF)model,and further applied the method to subtype identification of papillary renal cell carcinoma.Methods We conducted simulation studies to compare the performances of the improved ne-SNF method with the ne-SNF and SNF method,and applied improved ne-SNF method to integrate multi-omics data of PRCC.Cox regression model was performed to evaluate the prognostic risk of different subtypes.Differentially expressed mRNAs(DEmRNAs),miRNAs(DEmiRNAs)and differentially methylated genes(DMGs)with different subtypes were screened.KEGG pathway analysis was performed for the overlapping genes of three gene sets.Finally,the immune cell infiltration analysis was performed for patients with different subtypes.Results The improved ne-SNF method outperformed both SNF and ne-SNF approach in various simulation scenarios.In subsequent subtyping application,PRCC patients were divided into high-risk and low-risk groups,and the risk of death was 7.727 times higher in the high-risk group than in the low-risk group.A total of 3511 DEmRNAs,96 DEmiRNAs and 3426 DMGs were identified.Among them,649 overlapping genes yielded 42 KEGG pathways with statistical differences.In addition,3 immune filtrating cells showed statistical significance.Conclusion The improved ne-SNF performed better than SNF and ne-SNF,and the identified subtypes of PRCC may provide important clues and basis for treatment of PRCC patient.
4.A systematic review of two-way referral criteria for diabetes
Min XU ; Xiaoxu GE ; Rui LI ; Liling XU ; Shan HUANG ; Lingmei QIAN ; Jun MA ; Juan DU
Chinese Journal of General Practitioners 2024;23(12):1299-1306
Objective:To investigate the criteria for two-way referral of diabetes in China through systematic review.Methods:According to inclusion and exclusion criteria, articles on two-way referral criteria for diabetes in China were retrieved from databases of PubMed, Embase, Cochrane Library, CNKI and Wanfang from inception to July 15, 2023. The data were extracted, by author′s name, publish year, criteria for rupwards and downwards referral, sample size, study region, study type and main conclusions.Results:A total of 702 relavent articles were retrieved, and 15 of which were included in the analysis. Blood glucose levels were the basic criteria for two-way referral, The criteria for upward referral included fasting blood glucose, 2-h postpradial blood glucose, glycosylated hemoglobin and random blood glucose, but there were variations in measuement values. The criteria for downward referral included fasting blood glucose, 2-h postpradial blood glucose, glycosylated hemoglobin, with the variation in fasting blood glucose. Lipids and blood pressure were also included in the referral criteria in later literature, with the variation of blood pressure values. There was consensus on the upward referral criteria for patients with special types of diabetes and with newly diagnosed diabetes. In contrast, the referral criteria for diabetic complications were relatively vague, especially with the lack of severity classification for chronic complications.Conclusion:The two-way referral criteria for diabetes mellitus have been constantly developing, while the criteria for downwards referral to the community need to be further improved.
5.Lamin B1 regulates the growth of hepatocellular carcinoma cells by influencing telomerase activity.
Ruiguan WANG ; Si CHEN ; Zhijia SUN ; Shikun WANG ; Jie WANG ; Lingmei QIN ; Jiangbo LI
Chinese Journal of Biotechnology 2023;39(4):1609-1620
Lamin B1 (LMNB1) is highly expressed in liver cancer tissues, and its influence and mechanism on the proliferation of hepatocellular carcinoma cells were explored by knocking down the expression of the protein. In liver cancer cells, siRNAs were used to knock down LMNB1. Knockdown effects were detected by Western blotting. Changes in telomerase activity were detected by telomeric repeat amplification protocol assay (TRAP) experiments. Telomere length changes were detected by quantitative real-time polymerase chain reaction (qPCR). CCK8, cloning formation, transwell and wound healing were performed to detect changes in its growth, invasion and migration capabilities. The lentiviral system was used to construct HepG2 cells that steadily knocked down LMNB1. Then the changes of telomere length and telomerase activity were detected, and the cell aging status was detected by SA-β-gal senescence staining. The effects of tumorigenesis were detected by nude mouse subcutaneous tumorigenesis experiments, subsequent histification staining of tumors, SA-β-gal senescence staining, fluorescence in situ hybridization (FISH) for telomere analysis and other experiments. Finally, the method of biogenesis analysis was used to find the expression of LMNB1 in clinical liver cancer tissues, and its relationship with clinical stages and patient survival. Knockdown of LMNB1 in HepG2 and Hep3B cells significantly reduced telomerase activity, cell proliferation, migration and invasion abilities. Experiments in cells and tumor formation in nude mice had demonstrated that stable knockdown of LMNB1 reduced telomerase activity, shortened telomere length, senesced cells, reduced cell tumorigenicity and KI-67 expression. Bioinformatics analysis showed that LMNB1 was highly expressed in liver cancer tissues and correlated with tumor stage and patient survival. In conclusion, LMNB1 is overexpressed in liver cancer cells, and it is expected to become an indicator for evaluating the clinical prognosis of liver cancer patients and a target for precise treatment.
Animals
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Mice
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Telomerase/metabolism*
;
Carcinoma, Hepatocellular/genetics*
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Liver Neoplasms/genetics*
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Telomere Shortening
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In Situ Hybridization, Fluorescence
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Mice, Nude
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Telomere/pathology*
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Carcinogenesis
6.Establishment and Evaluation of Rat Model of Acute Coronary Syndrome with Combined Blood Stasis and Poison
Yue YOU ; Gaojie XIN ; Zixin LIU ; Lingmei LI ; Hongxu MENG ; Yue SHI ; Yanlei MA ; Ziyan WANG ; Jianxun LIU ; Lei LI ; Jianhua FU
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(19):49-60
ObjectiveTo compare the feasibility of establishing the rat model of acute coronary syndrome with combined blood stasis and poison by lipopolysacharide (LPS) injection, ligation of coronary artery and different combinations of the two methods. MethodA total of 225 male SD rats were randomly divided into sham operation group, simple coronary artery ligation group, first injected LPS group [LPS(5 mg·kg)injection 24 h before coronary artery ligation] and follow injected LPS group [LPS(5 mg·kg)injection 10 min after coronary artery ligation]. The indexes of each group were detected at 3, 24, 72 h after modeling, and the model was comprehensively evaluated. The general state and macroscopic evaluation indexes of traditional Chinese medicine (TCM) syndrome (tongue and pulse) of rats in each group were observed. ECG and echocardiography were used to evaluate cardiac function, and the myocardial ischemia and infarction areas were measured by Evans blue/2,3,5-triphenyltetrazolium chloride (TTC) staining. The content of creatine kinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), creatine kinase (CK), and troponin T (cTnT) in serum as well as interleukin-1 β (IL-1β) and IL-6 changes were determined by biochemical method or enzyme-linked immunosorbent assay (ELISA). Hematology analyzer was adopted to determine the white blood cell (WBC) count, and the four coagulation indexes, platelet aggregation rate, hemorheology and other coagulation evaluation indexes were also detected. The myocardial tissue was observed by hematoxylin-eosin(HE)staining. ResultAfter 3 h of modeling, compared with the conditions in sham operation group, the R, G and B values of tongue of rats (P<0.01), pulse amplitude (P<0.01), and cardiac function in simple coronary artery ligation group were decreased, and the color of hypoglossal veins became purple(P<0.01). The content of CK, LDH, cTnT, IL-1β and IL-6 in serum(P<0.05), myocardial infarction area(P<0.01), and total number of WBCs (P<0.05)were increased. Compared with simple coronary artery ligation group, first injected LPS group and follow injected LPS group had increased hypoglossal veins, decreased R value of tongue and elevated cTnT content (P<0.01), while follow injected LPS group had reduced B value of tongue, decreased cardiac output (CO)(P<0.05), increased IL-1β content, and thinned left ventricular anterior walls at end-systole (LVAWs)(P<0.01). After 24 h of modeling, compared with sham operation group, simple coronary artery ligation group presented significantly decreased R, G and B values of tongue, lengthened purplish dark hypoglossal veins (P<0.01), reduced pulse amplitude(P<0.01) and cardiac function, enlarged myocardial infarction area(P<0.01), increased whole blood viscosity, platelet aggregation rate, fibrinogen (FIB), shortened prothrombin time (PT) and thrombin time (TT)(P<0.01), and elevated total number of WBCs (P<0.01)and content of CK, LDH, cTnT and IL-6 in serum(P<0.05). Compared with the conditions in simple coronary artery ligation group, the pulse amplitude, R, G and B values of tongue (P<0.01), and ejection fraction (EF) and fractional shortening (FS) scores (P<0.05)dropped, and hypoglossal veins were deepened and lengthened(P<0.05), and cTnT content was increased(P<0.01)in first injected LPS group and follow injected LPS group. However, follow injected LPS group had thinned LVPWs, increased LDH content, platelet aggregation rate(P<0.05), myocardial infarction area, and total number of WBC, level of IL-1β(P<0.05), and shortened TT(P<0.01). Additionally, 72 h after modeling, compared with sham operation group, simple coronary artery ligation group showed significantly reduced pulse amplitude, lowered R, G and B values of tongue, thickened and lengthened hypoglossal veins(P<0.01), decreased cardiac function, and increased content of cTnT, FIB, whole blood viscosity(P<0.01),platelet aggregation rate, level of IL-6 and IL-1β(P<0.05). Compared with the conditions in simple coronary artery ligation group, the hypoglossal veins of the first injected LPS group and the follow injected LPS group were more purple, and the content of cTnT was boosted(P<0.01), whereas follow injected LPS group had decreased pulse amplitude, R, G and B values of tongue, EF and FS scores (P<0.05), and enlarged myocardial infarction area(P<0.01). ConclusionCompared with the other modeling methods and models at different modeling time, the established model by LPS injection 10 min after coronary artery ligation for 24 h was more consistent with the clinical characteristics of acute coronary syndrome with combined blood stasis and poison.
7.The influencing factors of cognitive function in coal workers
Yuliang ZHANG ; Lei ZHANG ; Wenyou MA ; Lingmei YUE ; Congzhe WANG ; Qian FENG ; Ke JIANG ; Chao ZHANG ; Li ZHANG ; Tianbang QIN
Chinese Journal of Behavioral Medicine and Brain Science 2018;27(5):455-458
Objective To explore the influencing factors of coal workers''''cognitive function. Meth-ods There was a physical examination on 3205 workers in the coal mine enterprise.The physical examina-tion included height,weight,blood pressure,blood routine examination and routine urine test,and Mini Men-tal State Examination (MMSE) was used to evaluate the cognitive function.The coal workers were divided in-to cognitive dysfunction group (84 cases) and cognitive function normal group (3121 cases) according to MMSE scores.Logistic regression analysis was used to explore the influencing factors of cognitive function. Results The prevalence of cognitive dysfunction was 2. 62% in coal workers. The age of the cognitive dys-function group(47.27±8.24) was significantly higher than that in the cognitive function normal group (41.39 ±8.65)(P<0.05),and the degree of culture in cognitive dysfunction group was significantly lower than those in the cognitive function normal group (χ2= 46. 610, P<0. 01 ) . The low density lipoprotein cholesterol ((2.72±0.65)mmol/L),urine pH(5.54±1.51) in the cognitive dysfunction group were significantly lower than those in the cognitive function normal group((2.89±0.73)mmol/L,(5.92±1.28))(P<0.05). The lo-gistic regression analysis showed that age, culture level, BMI, low density lipoprotein cholesterol, urine pH were the impacting factors of cognitive dysfunction. Conclusion Cognitive dysfunction is influenced by age, culture level,BMI and low density lipoprotein cholesterol. The higher of the age and the lower of the culture level,BMI,low density lipoprotein cholesterol,urine pH,the more vulnerable to cognitive dysfunction.
8.The long non-coding RNA uc.4 influences cell differentiation through the TGF-beta signaling pathway
Zijie CHENG ; Qijun ZHANG ; Anwen YIN ; Mengwen FENG ; Hua LI ; Hailang LIU ; Yun LI ; Lingmei QIAN
Experimental & Molecular Medicine 2018;50(2):e447-
In a previous study, we screened thousands of long non-coding RNAs (lncRNAs) to assess their potential relationship with congenital heart disease (CHD). In this study, uc.4 attracted our attention because of its high level of evolutionary conservation and its antisense orientation to the CASZ1 gene, which is vital for heart development. We explored the function of uc.4 in cells and in zebrafish, and describe a potential mechanism of action. P19 cells were used to investigate the function of uc.4. We studied the effect of uc.4 overexpression on heart development in zebrafish. The overexpression of uc.4 influenced cell differentiation by inhibiting the TGF-beta signaling pathway and suppressed heart development in zebrafish, resulting in cardiac malformation. Taken together, our findings show that uc.4 is involved in heart development, thus providing a potential therapeutic target for CHD.
9.Influence of CYP2C19 gene polymorphisms on the efficacy of clopidogrel treatment for the prevention of ischemic stroke following coronary stent implantation.
Guode LI ; Jingjuan CHEN ; Zuohang XU ; Yukai WANG ; Nan DING ; Lingmei PENG ; Chengguo ZHANG ; Yan SHAO
Chinese Journal of Medical Genetics 2017;34(6):839-843
OBJECTIVETo assess the association of CYP2C19 gene polymorphisms with the incidence of ischemic stroke among patients receiving clopidogrel therapy following coronary stenting for coronary artery disease.
METHODSClinical data of patients receiving clopidogrel therapy after coronary stenting were retrospectively studied. For a case-control study, 137 patients with acute cerebral infarction and 122 non-stroke patients were selected. Based on the variants of the CYP2C19 gene detected by a DNA microarray assay, the patients were further divided into the wild-type group(CYP2C19*1/*1) and mutant group(defined by the presence of at least one loss-of-function allele, including CYP2C19*1/*2, CYP2C19*1/*3, CYP2C19*2/*2, CYP2C19*2/*3 and CYP2C19*3/*3). The incidences of ischemic stroke in the two groups were compared through a chi-square analysis. The influence of CYP2C19 gene polymorphisms and clopidogrel therapy on the incidence of ischemic stroke was analyzed through multivariable logistic regression.
RESULTSA total of 259 patients were enrolled. The case and control groups showed no difference in terms of gender and age. There were 123 cases (47.5%) in the CYP2C19 wild-type group and 136 cases (52.5%) in the mutant group. The incidence of ischemic stroke of mutant group was significantly higher than that of wild-type group (59.9% vs. 44.3%, X2=6.398, P=0.042). Multivariate analysis revealed that loss-of-function polymorphisms of the CYP2C19 gene carried a 1.13 times greater risk for ischemic stroke compared to wild-type genotype (OR=2.13, 95%CI: 1.23-3.71).
CONCLUSIONThe efficacy of clopidogrel for the prevention of ischemic stroke in post-coronary stent patients may be reduced by the insufficiency of the CYP2C19 gene. The dosage of clopidogrel therapy should be adjusted based on its polymorphisms.
Brain Ischemia ; prevention & control ; Cytochrome P-450 CYP2C19 ; genetics ; Genotype ; Humans ; Percutaneous Coronary Intervention ; adverse effects ; Platelet Aggregation Inhibitors ; therapeutic use ; Polymorphism, Genetic ; Stents ; adverse effects ; Stroke ; prevention & control ; Ticlopidine ; analogs & derivatives ; therapeutic use
10.Case control study on the mental health, perceived stress,and coping styles of executives and underground workers in coal enterprise
Lingmei YUE ; Congzhe WANG ; Qian FENG ; Ke JIANG ; Chao ZHANG ; Li ZHANG ; Wenyou MA
Chinese Journal of Modern Nursing 2017;23(4):538-541
Objective To carry out a control study on the mental health,perceived stress,and coping styles of executives and underground workers in coal enterprise.Methods Totally 238 executives of state-owned coal enterprise and 240 underground workers were tested by symptom self-assessment scale (SCL-90), perceived stress scale and simple coping style scale. Then the test results were analyzed.Results according to the SCL-90 results,the total score of coal enterprise executives was (123.670±27.850) and it was lower than that of national norm (t=-3.151,P<0.01). The total score of underground workers was (138.610±49.720) and it was higher than that of national norm (t=2.882,P<0.01). The difference between them was significant (t=-4.05,P<0.01);the score of perceived stress of enterprise executives (15.70±7.37) was lower than that of underground workers (t=-9.553,P<0.01);the scores of positive coping of executives were (2.04±0.43),and they were significantly higher than those of the underground workers (t= 2.886,P<0.01);the scores of negative coping of executives were (1.06±0.49),and they were significantly lower than those of the underground workers (t=-9.225,P<0.01).Conclusions Executives of coal enterprise get a better psychological health level than underground workers;the executives are not sensitive to perceived stress;and they use more positive coping styles to deal with difficulties and setbacks. The underground workers are sensitive to perceived stress,and they use more negative coping styles to deal with difficulties and setbacks.

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