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.Study on protective effects of Shiyifang medicinal wine on knee osteoarthritis in rabbits based on TLR 4/MyD88/ NF-кB signaling pathway
Wentao ZHANG ; Lingmei ZHOU ; Honghai HUANG ; Huihui BAO ; Dewen MAO ; Hui TIAN ; Shuling LIU
China Pharmacy 2022;33(18):2224-2229
OBJECTIVE To study the protective effect and mechanism of Shiyifang medicinal wine (SYF) on knee osteoarthritis(KOA)of rabbit induced by papain . METHODS Thirty-five rabbits were randomly divided into blank group ,model group,positive group (Diclofenac diethylamine emulsion 200 mg/kg),SYF high -dose group (386 mg/kg)and SYF low -dose group(97 mg/kg),with 7 rabbits in each group (all had 4 males and 3 females). Except for the blank group ,the other groups ’ rabbits were injected 0.5 mL papain mixture (containing 2% papain and 0.03 mol/L L -cysteine)into the right knee cavity on day 1, 4 and 7,to replicate KOA model . Blank group was given constant volume of normal saline . From the 15th day ,drugs were applied to right hind knee joints of rabbits in each group ,twice a day for 20 days. At the same time ,the diameter of right knee joints of rabbits was measured by vernier calipers at day 0,8,14 and 35 to calculate swelling degree . After the experiment ,the levels of IL-1β,TNF-α and PGE 2 in synovial tissue were determined by enzyme -linked immunosorbent assay . Hematoxylin-eosin(HE) staining was used to prepare the sections of synovial tissue ,and the pathological changes were observed . The relative mRNA expressions of TLR 4,MyD88 and NF - кB p 65 in the TLR 4/MyD88/NF- кB signaling pathway were detected by real -time quantitative polymerase chain reaction . The relative protein expressions of TLR 4,MyD88,NF-кB p 65 and p -NF-кB p 65 were detect by Western blot . RESULTS Compared with blank group,the degree of knee swelling could be increased in model group ,the pathological damage of synovial tissue was more serious ,and the levels of IL -1β,TNF-α and PGE 2 were increased significantly in synovial tissue (P<0.05). The relative expression levels of TLR 4,MyD88,NF-кB p 65 mRNAs and TLR 4,MyD88,p-NF-кB p 65 proteins were significantly increased(P<0.05). Compared with model group ,swelling degree of right hind knee and the pathological injury degree of synovial tissue were significantly improved in each treatment group ,while the levels of IL -1β,TNF-α and PGE 2 in synovial tissue were significantly decreased (P<0.05). The relative mRNA expressions of TLR 4,MyD88 and NF -кB p 65 and relative protein expressions of TLR 4,MyD88(except for SYF low -dose group )and p -NF-кB p 65 were all significantly decreased (P<0.05). CONCLUSIONS SYF shows protective effect on KOA induced by papain ,the mechanism of which is associated with decreasing the levels of IL -1β,TNF-α and PGE 2 and down -regulating TLR 4/MyD88/NF-кB signaling pathway .
3.Application of quantitative FIT for primary colorectal cancer screening of a health check-up population in Suzhou City
Dan LIU ; Su YAN ; Daiyi ZHANG ; Wenxia MA ; Lingmei ZHAO
Chinese Journal of Health Management 2021;15(2):158-162
Objective:To explore the value of the quantitative fecal immunochemical test (FIT) for primary colorectal cancer screening in health check-up population.Methods:A total of 468 health check-up participants who underwent quantitative FIT and colonoscopy at the First Affiliated Hospital of Soochow University from January 2018 to December 2019 were enrolled into this study. The participants were divided into two groups-the negative group(FIT<100 μg/L)and the positive group(FIT≥100 μg/L) according to the results of the quantitative FIT. We compared the detection rate of colorectal polyps and colorectal advanced cancer between the two groups, and analyzed the sensitivity and specificity of the quantitative FIT for advanced colorectal cancer and the risk factors of colorectal polyps.Results:The positive rate of quantitative FIT in the healthy population was about 4.6% (95% CI: 4.27%-4.93%). The detection rate of colorectal polyps in the positive group was significantly higher than that of the negative group (51.53%, 95% CI: 45.81%-57.25%) vs (34.28%, 95% CI: 27.25%-41.31%, P<0.001). The sensitivity and specificity of the quantitative FIT for advanced colorectal cancer was 98.55% and 56.77%, respectively. The positive predictive value of the quantitative FIT for advanced colorectal cancer was 50.37%, while the negative predictive value was 98.88%. With the increase of FIT value, the detection rate of advanced cancer was increased (χ2 =20.08, P<0.001). The multivariate logistic regression of colorectal cancer suggested that the risk of colorectal polyps in patients with a history of diabetes was significantly higher ( OR=3.28, 95% CI: 1.45-7.41, P=0.004); the risk of colorectal polyps in men was also significantly higher ( OR=3.05, 95% CI: 1.40-6.65, P=0.005). Conclusions:Quantitative FIT is an ideal non-invasive examination for primary colorectal cancer screening for a health check-up population. Patients with negative FIT are less likely to develop colorectal cancer. Diabetes history, male, and age are independent risk factors for colorectal cancer.
4.Pathway of establishing continuing education mechanism for community general practitioner in regional medical centers
Dandan SHI ; Gang YONG ; Rong ZHANG ; Jun MA ; Lingmei QIAN ; Min GE ; Kun TAO
Chinese Journal of General Practitioners 2020;19(6):532-536
Comprehensively and continuously improving the ability is the goal of continuing medical education for practicing general practitioners (GPs). It is urgent to carry out systematic planning for improvement of the training content, training methods and evaluation methods in current continuing education system. Therefore, based on the survey of the training requirements of the community GPs in Shanghai Changning District, this study proposed a pathway of establishing continuing education mechanism for community GPs in regional medical centers. This continuing education mechanism planned to build a talent evaluation system reflecting the competency of the community GPs, to develop training contents based on the community situation and the needs of the GPs, to establish the practice support and the collaboration platform of science, education and research, to promote the sustaining improvement of GPs.
5.Research progress on stomy nurse-led follow-up visit for patients with enterostomy
Lingmei XUAN ; Mengting CHEN ; Lixiu ZHANG ; Sunwen ZHUAN ; Shiliang TU ; Xuelian CAI
Chinese Journal of Modern Nursing 2019;25(17):2242-2244
This article illuminates the status quo, shortcomings and prospects of follow-up visit at home and abroad by retrieving and summarizing the related studies on follow-up visit for enterostomy at home and abroad. It reviews the nurse-led follow-up visit from the perspective of continued nursing care, in order to provide a reference for China to further standardize the follow-up content for patients with enterostomy.
6.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.
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.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
9.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.
10.Bioinformatics characteristics of lncRNA -uc.167 and its temporal and spatial expression pattern for mouse embryonic development
Lijie WU ; Guixian SONG ; Xing LI ; Yumei CHEN ; Yi FAN ; Hua LI ; Qijun ZHANG ; Lingmei QIAN
Chinese Journal of Applied Clinical Pediatrics 2016;31(24):1902-1905
Objective To explore the basic biological characteristics of lncRNA -uc.1 67,and its spatial dis-tribution,temporal expression pattern during the mouse embryonic development.Methods The UCSC genome browser of ENCODE was used to analyze preliminary bioinformatics of lncRNAs.Real -time (RT)-PCR was applied to detect the expression of uc.1 67 and neighboring genes in the embryonic mouse heart in different stages (P7.5,P1 1 .5,P1 4.5, P1 8.5).Dimethyl sulphoxide was used to induce P1 9 cell differentiation into the cardiomyocytes.RT -PCR was applied to detect the expression changes in uc.1 67 and neighboring genes on differential day 0,4,6,8 and 1 0.Results Full -length of human uc.167 was 201 bp,and human uc.167 was located in the genome 5q14.3 (chr5:88179623 -881 79824,GRCh37 /hg1 9).uc.1 67 mainly expressed in the ventricular muscle tissue.The expression of uc.1 67 was gradually decreased in the mouse embryonic heart development process(P7.5:1 .000 ±0.1 00,P1 1 .5:0.71 4 ±0.1 07, P1 4.5:0.393 ±0.043,P1 8.5:0.1 25 ±0.01 3),while the expression of its neighboring Mef2c gene was gradually in-creased(P7.5:1 .081 ±0.1 1 8,P1 1 .5:2.340 ±0.351 ,P1 4.5:3.958 ±0.542,P1 8.5:9.361 ±0.722),which showed an opposite trend.The expression of uc.1 67 during P1 9 cell differentiation into cardiomyocytes showed a an increase at first and then a decreasepattern,and the highest level expression of uc.1 67 was on differential day 4(d0:1 .071 ± 0.1 1 7,d4:4.71 4 ±0.501 ,d6:3.572 ±0.41 4,d8:2.550 ±0.31 4,d1 0:0.786 ±0.085).The expression of neigh-boring gene Mef2c was in an opposite trend(d0:1 .01 2 ±0.041 ,d4:0.353 ±0.037,d6:2.470 ±0.329,d8:6.706 ± 0.682,d1 0:7.765 ±0.705).Conclusions It is suggested that uc.1 67 may take part in the process of embryonic heart development and may play a role through negatively regulating its neighboring gene Mef2c.

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