1.Effects of cytoplasmic transduction peptide-HBcAg18-27-Tapasin induced murine bone marrow derived dendritic cell maturation on T lymphocyte proliferation in vitro
Honghong LIU ; Xiaohua CHEN ; Liqin ZHOU ; Xueni LIU ; Yongsheng YU ; Guoqing ZANG ; Zhenghao TANG
Chinese Journal of Infectious Diseases 2012;30(10):593-597
Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) %] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )%] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)%] and medium group [(0.09±0.02)%,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.
2. Development and evaluation on the primipara social capital scale
Chi ZHOU ; Ruinan LAI ; Xueni ZHANG ; Hao CHEN ; Weijue WANG ; Baodan ZHANG
Chinese Journal of Preventive Medicine 2017;51(4):313-318
Objective:
To develop the Chinese primipara social capital scale (C-PSCS), and to evaluate its validity and reliability.
Methods:
The items of C-PSCS were developed based on Social Capital Scale by the World Bank and Social Network Scale. This scale was modified according to the characteristics of primiparas. We selected 10 experts who specialized in related field, and two rounds of seminars about content, cultural compatibility, primiparas' characteristics, and practicability. The finally C-PSCS included four dimensions: social trust, social reciprocity, social network, and social participation. Using purposive sampling to select 1 100 primiparas in their third trimesters (gestational weeks from 30 to 36 weeks). The validity analyses included content validity, construct validity and discriminant validity. The reliability analyses included Cronbach' α coefficient, and split-half reliability.
Results:
1 035 questionnaires (94.09%) were qualified and the completion time was (13.23 ± 2.53) minutes. The total score of scale was 195.38 ± 45.98, and scores for social trust, social reciprocity, social network, and social participation were 30.26 ± 4.25, 22.84 ± 4.21, 34.23 ± 7.47, and 108.05 ± 41.96, respectively. The common factor cumulative variance contribution rates of each dimension were from 52.92% to 69.37%, which achieved more than 50% of the approved standard, and all the items held factor loading >0.5 in its relevant common factor, it had good construct validity. The scale had a good content validity, the
3.Clinical value of non-coding RNA molecular markers for sepsis
Jiaojiao YUAN ; Min LI ; Xueni CHANG ; Yu CHEN ; Chenming DONG
Chinese Critical Care Medicine 2019;31(5):650-653
Sepsis and septic shock are important clinical problems in critically ill patients, accounting for the first cause of death in intensive care unit (ICU). Therefore, early diagnosis and treatment are particularly important. Recently, genome-wide expression analysis of non-coding RNA in septic patients showed that more than 80% were differentially expressed compared with healthy individuals. These molecules play important roles in biological processes, including innate immunity, mitochondrial function and apoptosis. Therefore, a class of non-coding RNAs such as microRNA (miRNA), long-chain non-coding RNA (lncRNA) and circular non-coding RNA (circRNA) are increasingly recognized as a regulator of various signaling pathways. The potential of regulatory non-coding RNA target to treat sepsis was discussed by studying non-coding RNAs that might serve as molecular markers of sepsis, and its clinical value was evaluated.
4. The situation of social support and its relationship with antenatal depression among 1 075 Zhejiang primiparas in their third trimesters
Baodan ZHANG ; Yuanchun SHAN ; Liangwen XU ; Hao CHEN ; Xueni ZHANG ; Chengyao TONG ; Yali MAO ; Chi ZHOU
Chinese Journal of Preventive Medicine 2017;51(8):740-745
Objective:
To investigate the situation of social support (SS), and explore its relationship with antenatal depression (AD) among Zhejiang primiparas in their third trimesters.
Methods:
From March to August 2016, a cross-sectional study was conducted and the questionnaire was used at the outpatient consulting room of one maternity hospital in Hangzhou. Inclusion criteria includes the primiparas over 18 years old, gestation from 30 to 36 weeks, been able to understand and complete the questionnaires independently, no family history and history of mental disorders and no use of psychotropic drugs, without serious illness and so on. Exclusion criteria was that the primiparas were unable to complete all the contents of the questionnaire and of poor compliance. 1 150 questionnaires were actually given out, and 1 075 questionnaires were valid, so the valid rate was 93.48%. AD was evaluated by the Edinburgh Postnatal Depression Scale (EPDS) and SS was evaluated by the Perceived Social Support Scale (PSSS). SS between the non-AD group and the AD group was compared. The correlation between SS and AD was analyzed. Binary logistic regression model was used to assess the relationship between SS and AD. The level of SS was divided by average scores, groups lower than the average score was defined as the low-score group, groups higher than the average score was defined as the high-score group.
Results:
The prevalence of AD (score≥9) was 27.3% (293/1 075) among Zhejiang primiparas in their third trimesters. The scores of family support, friend support and other support, and the total score of SS among the non-AD group were 24.80±2.83, 23.40±3.00, 21.91±3.54 and 70.11±7.92, respectively, which were higher than those in the AD group (22.71±3.88, 21.45±3.59, 19.95±3.97, 64.10±10.01), (
5.Relationship between interleukin-1β and silicosis development
Junjin LIAO ; Xueni YU ; Junpeng CHI ; Luocheng SHAO ; Shizi WANG ; Zhen LIU ; Zhuo CHEN ; Huihui TAO
Chinese Journal of Primary Medicine and Pharmacy 2023;30(2):316-320
Silicosis is a diffuse pulmonary fibrosis disease caused by occupational exposure to silica, which is one of the occupational diseases with high incidence in developing countries. Up to now, there is no definite drug to relieve or reverse the lung injury caused by silicosis, so it is very important to prevent, diagnose and treat pulmonary fibrosis as soon as possible. Studies have shown that a chronic inflammatory environment contributes to pulmonary fibrosis to a certain extent. Interleukin-1β is a cytokine that increases the number of inflammatory factors in the microenvironment in the immune response and plays a key role in inflammatory reaction. Therefore, the release of interleukin-1β is of great significance in the pathogenesis of silicosis. This paper aims to systematically expound the development course of silicosis, the signal pathway of interleukin-1β production, and the relationship between them.
6.Discriminante analysis of risk factors Nomograms of myopia in children and adolescents in Yunnan Province
Chinese Journal of School Health 2023;44(9):1387-1391
Objective:
To explore the related factors of myopia among children and adolescents in Yunnan Province, and to predict and evaluate the influencing factors, so as to provide a scientific theoretical basis for the prevention and control of myopia.
Methods:
From March 9 to 14, 2023, 848 students from 6 primary and secondary schools in Dali and Lijiang of Yunnan Province were selected by multi stage stratified random cluster sampling method for visual acuity detection and questionnaire survey on myopia related factors. Multivariate Logistic regression analysis was used to establish a Nomogram prediction model for the selected influencing factors.
Results:
The overall myopia rate of the respondents was 68.3%, the myopia rate of boys (63.4%) was lower than that of girls (72.9%), and the myopia rate of primary school students (46.7%) was lower than that of junior high school students (81.1%), and the difference was statistically significant( χ 2=8.71, 108.07, P <0.05). Daily eye exercises, activities outside the teaching building during recess, having daily sleep time of 7-9 and >9 h, having both parents without myopia were negatively correlated with the occurrence of myopia in children and adolescents in Yunnan Province ( OR=0.64, 0.63, 0.56, 0.28, 0.48, P < 0.05 ). The reading and writing time after school ≥3 h per day and parents unrestricted time to play video games were positively correlated with myopia ( OR=1.94, 1.78, P <0.05). Based on the influencing factors, a Nomogram prediction model was established to quantitatively evaluate the risk of myopia. The results showed that greater risk for myopia was associated with sleep duration, parental history of myopia, and the time spent reading and writing after school every day.
Conclusion
Both genetic factors and environmental factors are related to myopia in children and adolescents. The prediction model of nomogram is beneficial for screening high risk factors of myopia and taking corresponding prevention and treatment measures.
7.Relationship of pks gene islands with virulence genes and biofilm formation in Klebsiella pneumoniae
Chenshuo LUO ; Yanshu CHEN ; Yulan LIN ; Tongzhe DENG ; Xueni HU ; Bin YANG
Chinese Journal of Microbiology and Immunology 2022;42(11):888-893
Objective:To investigate the relationship of pks gene islands with virulence genes, capsular serotypes and biofilm formation in Klebsiella pneumoniae ( Kp). Methods:A total of 113 Kp clinical isolates were collected in the First Affiliated Hospital of Fujian Medical University and divided into two groups based on the presence of pks gene islands: pks+ and pks- groups. The hypermucoviscous (HM) phenotype was detected by string test. Five virulence genes ( peg-344, rmpA, rmpA2, iucA, iroB) and six common capsular serotypes (K1, K2, K5, K20, K54, K57) were detected by polymerase chain reaction (PCR). The biofilm formation ability was measured by crystal violet staining. Results:Among the 113 Kp isolates, 46 were pks+ strains and 67 were pks- strains. The detection rate of HM phenotype was higher in the pks+ group than in the pks- group [87.0% (40/46) vs 43.3% (29/67)]. The detection rates of virulence genes ( peg-344, rmpA, rmpA2, iucA, iroB) and K1 serotype in the pks+ group were also higher than those in the pks- group ( P<0.05). The biofilm formation ability of the pks+ strains was higher than the pks- strains ( P<0.05). Conclusions:Kp strains carrying pks gene islands were likely to display a HM phenotype and mainly belonged to the K1 serotype. Most of the pks+Kp strains carried the virulence genes of peg-344, rmpA, rmpA2, iucA and iroB, and had a greater ability to form biofilms.
8.CD38 regulates macrophagic cholesterol efflux by promoting lysosome reformation via TFEB
Hao XU ; Xueni SUN ; Tianqi WU ; Jinyuan LIU ; Qianlin HUANG ; Die MO ; Jiaxin WANG ; Shenxian CHEN ; Bodan DENG ; Xiaoyang XU
Chinese Journal of Pathophysiology 2024;40(1):28-37
AIM:To explore the effects of CD38 on lysosome reformation and cholesterol efflux in macro-phages.METHODS:Bone marrow-derived macrophages from low-density lipoprotein(LDL)receptor knockout(LDLr-/-)mice were cultured as cell model.Live cell imaging system was applied to evaluate the effect of nicotinic acid adenine di-nucleotide phosphate(NAADP)on lysosome number.ELISA was conducted to measure NAADP level in macrophages.After the cells were treated with nicotinic acid(NA),RT-qPCR was conducted to detect CD38 mRNA expression,and Western blot was conducted to observe CD38 protein expression and phosphorylated transcription factor EB(TFEB)level.Laser scanning confocal microscopy was applied to evaluate the influence of CD38/NAADP signaling on lysosome number and cholesterol egression.RESULTS:NAADP remarkably increased lysosome number(P<0.05),and this effect was significantly inhibited by NAADP antagonist NED-19,Ca2+ chelator BAPTA,and calcineurin inhibitor CsA(P<0.05).CD38 markedly enhanced NAADP synthesis in macrophages(P<0.05).NAADP synthetic substrate NA prominently ele-vated the expression of CD38 mRNA and protein(P<0.05).NA significantly decreased the phosphorylated TFEB level;this effect was also attenuated by NED-19,BAPTA and CsA(P<0.05).Disrupting CD38/NAADP signaling pathway markedly inhibited NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux in macrophages(P<0.05).NA-induced enhancement of lysosome number,lysosomal free cholesterol and cytosol cholesterol ester efflux abolished in LDLr/CD38 DKO macrophages(P<0.05),whereas these effects induced by NA were recovered after CD38 gene rescue.CONCLUSION:CD38 triggers lysosome reformation via TFEB and consequently pro-motes the efflux of lysosomal free cholesterol and cytosol cholesterol ester.
9.Protective Mechanism of Wenyang Fuyuan Prescription on Nerve Injury in Rats with Cerebral Ischemia-Reperfusion Injury Based on Ferroptosis
Junjun XIANG ; Liqin LI ; Jianzheng LI ; Xueni MO ; Wei CHEN ; Yueqiang HU
Traditional Chinese Drug Research & Clinical Pharmacology 2023;34(12):1649-1657
Objective To investigate the protective mechanism of Wenyang Fuyuan Prescription on nerve injury by improving brain iron metabolism in rats with cerebral ischemia-reperfusion injury(CIRI)based on ferroptosis.Methods A total of 72 SD rats were randomly divided into sham-operation group,CIRI model group,Wenyang Fuyuan Prescription group(18.0 g·kg-1,gavage),ferroptosis inducer group(100 mg·kg-1,intraperitoneal injection),Wenyang Fuyuan Prescription(18.0 g·kg-1,gavage)+ ferroptosis inducer group(intraperitoneal injection)and ferroptosis inhibitor group(5 mg·kg-1,intraperitoneal injection),12 rats in each group.All the procedures adopted in the sham group were the same as those in the model group.But nylon thread was inserted into the internal carotid artery at a depth of 9 mm and un-plugged middle cerebral artery.The rest of the groups were used to construct middle cerebral artery occlusion/reperfusion(MCAO/R)model by thread embolism method.Ferroptosis inducer(100 mg·kg-1)and ferroptosis inhibitor(5 mg·kg-1)were administered intraperitoneally to rats according to the grouping 24 hours before modeling.Wenyang Fuyuan Prescription(18.0 g·kg-1)was administered by gavage 2 hours after anesthesia and awakening.All intervention were given once daily for 7 consecutive days.The Longa scoring standard was used to evaluate the neurological deficit on 1,3,and 7 days after MCAO/R surgery,respectively.At the end of the treatment period,brain tissues were taken to observe the morphological changes of rat neurons in each group by hematoxylin eosin staining(HE).The ultrastructural changes of neuron mitochondria in each group were observed by transmission electron microscope.The biochemical kit was used to detect the content of iron ions(Fe2+)and reduced glutathione(GSH)in brain tissue.The protein and mRNA expressions of transferrin receptor 1(TFR1),iron regulatory protein 1(IRP1)and ferroportin(FPN)were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and Western Blot.Results① Compared with sham group,the neurological deficit scores of rats in model group increased at each time point(P<0.01).HE staining showed neurons were sparse and disordered,the nuclei underwent pyknosis,and vacuoles appeared at the edges.Under electron microscopy,it was observed that the number of neuronal mitochondria decreased,the density of mitochondrial membranes increased,massive numbers of mitochondrial membranes ruptured and dissolved,and mitochondrial cristae disappeared.The content of Fe2+,both mRNA and protein expressions of TFR1 were significantly increased(P<0.01),while GSH content,as well as expressions of mRNA and protein for IRP1 and FPN were significantly decreased(P<0.05,P<0.01).② Compared with the model group,the neurological deficit scores of rats in the Wenyang Fuyuan Prescription group decreased at various time points(P<0.05).The number of neurons increased,their arrangement was relatively neat,the morphology of the nucleus is complete and clear,the mitochondrial structure of neurons was relatively complete,the mitochondrial membrane was relatively intact,and the mitochondrial cristae were clear.The content of Fe2+,both mRNA and protein expressions of TFR1 were decreased(P<0.05,P<0.01),while GSH content,as well as expressions of mRNA and protein for IRP1 and FPN increased(P<0.05,P<0.01).③ Compared with the Wenyang Fuyuan Prescription group,the neurological deficit scores of rats in ferroptosis inducer group and the Wenyang Fuyuan Prescription + ferroptosis inducer group increased at all time points(P<0.05).Distribution of neurons was in disorder,the nucleus shrinked,and vacuoles appeared at the edges.The density of mitochondrial membranes increased,some ruptured and dissolved mitochondrial membranes were found.The number of mitochondria decreased and mitochondrial cristae disappeared.The content of Fe2+,both TFR1 mRNA and protein expression increased(P<0.05,P<0.01),while the content of GSH,as well as expressions of mRNA and protein for IRP1 and FPN decreased(P<0.05,P<0.01).However,there was no statistically significant difference in all observed indicators between the ferroptosis inhibitor group and the Wenyang Fuyuan Prescription group(P>0.05).Conclusion Wenyang Fuyuan Prescription can improve the neurological function and pathological damage of CIRI rats.Its mechanism may be related to regulating the expression of IRP1 protein,improving the brain iron metabolism pathway,and inhibiting ferroptosis.
10.Role of Akt during Islet-1 inducing C3H10T1/2 cells to differentiate specifically into cardiomyocytes
yu Xing LAI ; Jing ZHU ; Jie TIAN ; Qin YI ; Xueni CHEN
Basic & Clinical Medicine 2018;38(1):13-19
Objective To study the effect and mechanism of Akt during Islet -1 inducing the mesenchymal stem cells of C3H10T 1/2 cells to differentiate specifically into cardiomyocytes.Methods Developing a model of Islet-1 over-expression cells, Lentivirus infection efficiency of cells was detected by flow cytometry detection tech-nology.Cell proliferation was tested by CCK-8.The protein expression of Islet-1, cTnT and p-Akt/T-Akt was measured by Western blot.The mRNA expression of GATA4,Nkx2.5 and Mef2c were tested by real-time PCR. Results With the increasing of MK-2206 concentration, the inhibition rate of cell proliferation increased ( P<0.05) ,and the best inhibition concentration of Akt was 8nmol /L; With prolongation of Islet-1 inducing time,the protein expression of p-Akt/TA-kt reduced ( P<0.05 );The gene expression of myocardial specific transcription factor GATA4, Nkx2.5 andMe f2c increased( P<0.05); treated with MK-2206, the geneexpres-sion of GATA4, Nkx2.5 and Mef2cincreased significantly at the first week and then reduced ( P<0.05) . Conclu isons Akt plays different effects in different differentiation stages during the Islet-1 inducing the cells into cardiacs-pecific differentiation process .