1.Effect of sinomenine on mouse RAW264.7 macrophage cells line polarization induced by LPS or IL-4
Jinfang LUO ; Ruili ZHU ; Lang YI ; Yan DONG ; Peixun WANG
Chinese Journal of Immunology 2015;(1):56-60
Objective:To investigate sinomenine (Sinomenine,SIN) effect on RAW264.7 cells polarization to M1 or M2 phenotype induced by lipopolysaccharide (LPS) or interleukin-4 (IL-4) .Methods:RAW264.7 cells were induced to polarize to M1 by LPS ,and to M2 by IL-4.Sinomenine effects on LPS or IL-4 induced macrophages:TNF-αand IL-10 secretion induced by different condition were detected by Enzyme linked immunosorbent assay (ELISA);The expression level of mRNA of Arginase1(Arg-1),Nitric oxide synthase(iNOS),suppressor of cytokine signaling protein-2(SOCS2) and suppressor of cytokine signaling protein-3(SOCS3) of M1/M2 phenotypes were detected by real time PCR respectively.Results:Sinomenine inhibited the increase of TNF-αsecretion,iNOS and SOCS3 mRNA expression level induced by LPS.Sinomenine inhibited the increase of IL-10 secretion and Arg-1 mRNA expression level induced by IL-4,but SOCS2 mRNA expression level was not affected by Sinomenine.Conclusion: Sinomenine can inhibite the macrophage polarization to M1 and M2 induced by LPS and IL-4.Sinomenine plays a regulatory role on imbalance of M1/M2,and is conducive to maintain the dynamic balance.
2.Effect of Berberine on Mice RAW264.7 Macrophages Polarization
Ruili ZHU ; Yangyang WU ; Jinfang LUO ; Lang YI ; Yan DONG
Journal of Guangzhou University of Traditional Chinese Medicine 2014;(6):974-978
Objective To investigate the effect of berberine on the polarization of mice RAW264.7 macrophages induced separately by lipopolysaccharide (LPS) and interleukin-4 (IL-4). Methods Mice RAW 264.7 macrophages cultured in vitro were divided into model group, medication group, and blank control group. Both model group and medication group were given either LPS (in final dose of 100 ng/mL) or IL-4 (in final dose of 10 ng/mL). Additionally, the medication group was treated with berberine in final dose of 20 μmol/L. The blank control group was given the same volume of phosphate buffered saline ( PBS). Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to detect the mRNA expression of arginase-1 (Arg-1), inducible nitric oxide synthase ( iNOS) , suppressor of cytokine signaling2 ( SOCS2) and SOCS3. Enzyme-linked immunosorbent assay (ELISA) was used to determine the contents of tumor necrosis factor alpha (TNF-α) and IL-10. Results The content of TNF-αand the mRNA expression levels of iNOS and SOCS3 in macrophages induced by LPS were increased, and then were down-regulated by berberine (P<0.05 or P<0.01) . The content of IL-10 and the mRNA expression level of Arg-1 in macrophages induced by IL-4 were increased, and then were down-regulated by berberine ( P<0.05), but berberine had no effect on the mRNA expression level of SOCS2 ( P>0.05). Conclusion Berberine has an effect on inhibiting the M1 and M2 polarization of macrophages in vitro, suggesting that berberine may play a regulatory role in the dynamic balance of M1/M2.
3.Molecular genetic analysis of partial 9p trisomy in two Chinese families with mental retardation and facial anomaly.
Aiping, FENG ; Xiaohua, DAI ; Xiaoran, WANG ; Yong, GAO ; Ruili, LUO ; Yulei, LI ; Na, ZHANG ; Jingyu, LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):570-7
Mental retardation is defined by significant limitations in intellectual function and adaptive behavior that occur before 18 years of age. Many chromosomal diseases come with mental retardation. We reported two Chinese families with partial trisomy 9p and other chromosome partial monosomy, clinical features of mental retardation and mild facial and pinkie anomalies. In the family 1, we showed that the proband carried a trisomy 9p21.3→pter and monosomy 21q22.3→qter by using fluorescence in situ hybridization analysis. Molecular genetic analysis defined the precise breakpoint on chromosome 9p between markers D9S1846 and D9S171, an interval of about 2.9 Mb on 9p21.3, and the breakpoint on chromosome 21q between markers D21S1897 and D21S1446, a region of about 1.5 Mb on 21q22.3. In the family 2, a patient with trisomy 9p21.3→pter and monosomy 5p15.33→pter, and a de novo maternal balanced translocation between chromosomes 5 and 9 was identified in his mother. Cytogenetic and molecular genetic analysis defined the precise breakpoints on chromosome 9p21.3 and chromosome 5p15.33. Further clinical investigation found that any individual had no refractoriness eczema disease except the proband in this family. These results further implicate that trisomy 9p is associated with mental retardation, and that there may be key gene duplication on chromosome 9p21.3→9pter responsible for mental retardation and mild facial anomaly. This result has been applied successfully in prenatal diagnosis of the second family.
4.Amplification,cloning,expression and protein analysis of enterovirus 71 VP1 gene
Yunfeng LIU ; Zhenwen ZHOU ; Ruili GUAN ; Qihong GUAN ; Mingyong LUO ; Shuai ZHOU
International Journal of Laboratory Medicine 2014;(16):2126-2128,2131
Objective To conduct the amplification,cloning,bioinformatics analysis,prokaryotic expression and purification of enterovirus 71 VP1 gene segment and to initially confirm the biological activity of the recombinant expression product.Methods A pair of specific primers was designed according to GenBank EV71 sequence,viral RNA as a template was extracted from the throat swab specimens in the EV71 patients.EV71 VP1 gene was amplified by RT-PCR.After enzyme digestion,the expression vector pET28a was inserted.The prokaryotic expression vector of pET28a-EV71 VP1 was constructed.Then the E.coli DH5a transforma-tion was performed.IPTG was adopted for induction expression.The expression results were analyzed by using SDS-PAGE and Western blot.The bioinformatics analysis of the sequenced results was performed by the software.Expressed protein was purified and the plates were coated,ELISA was used to test the VP1 specific IgG antibody in serum samples of EV71 positive and COX A16-positive patients.Results The BLAST alignment showed that the homology of the objective gene EV71 VP1 was 99% com-pared with other strains(JQ766207.1)in GenBank.EV71 VP1 protein was about 32×103 ,which mainly existed in the form of in-clusion body.The bioinformatics analysis showed that EV71 VP1 protein was a hydrophilic protein,without transmembrane region and N-terminal signal peptide sequence,the tertiary structure existed.The ELISA results showed that the specific IgG OD value in EV71-positive patients was(2.425±0.521),OD value in COX A16 positive patients was(1.205 ±0.314),the normal control OD value was(0.353±0.128).The sensitivity and specificity of EV71 VP1 protein detection were 84% and 88% respectively.Conclu-sion The pET28a-EV71 VP1 expression vector is successfully constructed;the preliminary analysis on the serum of the infected patients by ELISA shows that the obtained objective protein has higher sensitivity and specificity,which is initially confirmed to have biological activity and can be further used for the related study on EV71 diagnosis and vaccine.
5.Meta-analysis on causes of ulcerative colitis
Ruili LUO ; Lijuan HUO ; Jie ZHANG ; Qiannan ZHANG
Chinese Journal of Epidemiology 2015;36(12):1419-1423
Objective To analyze the main influencing factor of ulcerative colitis (UC).Methods Literature retrieval was conducted by using English databases (PubMed,Cochrane and Embase) and Chinese databases (CNKI,Wanfang,SinoMed and VIP) to collect the studies on the influencing factors of UC published both at home and abroad from January 2000 to October 2014.According to the inclusion and exclusion criteria,data were extracted and methodological quality was assessed.Then,a Meta-analysis was performed with Stata 12.0 software.Results A total of 24 casecontrol studies were included,involving 5 653 patients and 20 218 controls.The results of Meta-analysis showed that the influencing factors of UC would include family history of inflammatory bowel disease,ex-smoker,gastrointestinal infections,regular consumption of milk,fat diet,appendectomy,smoking and high educational level,with the pooled OR values as 4.68 (95% CI:3.59-6.11),1.81 (95%CI:1.58-2.09),5.10 (95%CI:2.38-10.92),2.26 (95%CI:1.65-3.09),2.21 (95% CI:1.49-3.27),0.40 (95% CI:0.32-0.51),0.44 (95% CI:0.32-0.60) and 0.50 (95% CI:0.36-0.69),respectively.Conclusion Current evidence showed that the risk factors influencing the incidence of UC were family history of inflammatory bowel disease,ex-smoker,gastrointestinal infections,regular consumption of milk and fat diet,whereas appendectomy,smoking and high educational level were protective factors for UC.
6.Inhibition of Long Noncoding RNA SNHG15 Ameliorates Hypoxia/Ischemia-Induced Neuronal Damage by Regulating miR-302a-3p/STAT1/NF-κB Axis
Chunting HU ; Chen LI ; Qiaoya MA ; Ruili WANG ; Ya HE ; Hui WANG ; Guogang LUO
Yonsei Medical Journal 2021;62(4):325-337
Purpose:
Ischemic brain injury results in high mortality and serious neurologic morbidity. Here, we explored the role of SNHG15 in modulating neuronal damage and microglial inflammation after ischemia stroke.
Materials and Methods:
The hypoxia/ischemia models were induced by middle cerebral artery occlusion in mice and oxygenglucose deprivation/reoxygenation (OGD/R) in vitro. Quantitative real-time PCR (qRT-PCR) and Western blot were conducted to determine the levels of SNHG15, miR-302a-3p, and STAT1/NF-κB. Moreover, gain- or loss-of functional assays of SNHG15 and miR-302a-3p were conducted. MTT assay was used to evaluate the viability of HT22 cells, and the apoptotic level was determined by flow cytometry. Furthermore, enzyme-linked immunosorbent assay was performed to detect oxidative stress and inflammatory mediators in the ischemia cortex and OGD/R-treated BV2 microglia.
Results:
The SNHG15 and STAT1/NF-κB pathways were both distinctly up-regulated, while miR-302a-3p was notably down-regulated in the ischemia cortex. Additionally, overexpressing SNHG15 dramatically enhanced OGD/R-mediated neuronal apoptosis as well as the expression of oxidative stress and inflammation factors from microglia. In contrast, knocking down SNHG15 or overexpressing miR-302a-3p relieved OGD/R-mediated neuronal apoptosis and microglial activation. Moreover, the rescue experiment testified that overexpressing miR-302a-3p also attenuated SNHG15 up-regulation-induced effects. In terms of the mechanisms, SNHG15 sponged miR-302a-3p and activated STAT1/NF-κB as a competitive endogenous RNA, while miR-302a-3p targeted STAT1 and negatively regulated the STAT1/NF-κB pathway.
Conclusion
SNHG15 was up-regulated in the hypoxia/ischemia mouse or cell model. The inhibition of SNHG15 ameliorates ischemia/hypoxia-induced neuronal damage and microglial inflammation by regulating the miR-302a-3p/STAT1/NF-κB pathway.
7.Molecular Genetic Analysis of Partial 9p Trisomy in Two Chinese Families with Mental Retardation and Facial Anomaly
FENG AIPING ; DAI XIAOHUA ; WANG XIAORAN ; GAO YONG ; LUO RUILI ; LI YULEI ; ZHANG NA ; LIU JINGYU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(4):570-577
Mental retardation is defined by significant limitations in intellectual function and adaptive behavior that occur before 18 years of age.Many chromosomal diseases come with mental retardation.We reported two Chinese families with partial trisomy 9p and other chromosome partial monosomy,clinical features of mental retardation and mild facial and pinkie anomalies.In the family 1,we showed that the proband carried a trisomy 9p21.3→pter and monosomy 21q22.3→qter by using fluorescence in situ hybridization analysis.Molecular genetic analysis defined the precise breakpoint on chromosome 9p between markers D9S1846 and D9S171,an interval of about 2.9 Mb on 9p21.3,and the breakpoint on chromosome 21q between markers D21S1897 and D21S1446,a region of about 1.5 Mb on 21q22.3.In the family 2,a patient with trisomy 9p21.3→pter and monosomy 5p15.33→pter,and a de novo maternal balanced translocation between chromosomes 5 and 9 was identified in his mother.Cytogenetic and molecular genetic analysis defined the precise breakpoints on chromosome 9p21.3 and chromosome 5p15.33.Further clinical investigation found that any individual had no refractoriness eczema disease except the proband in this family.These results further implicate that trisomy 9p is associated with mental retardation,and that there may be key gene duplication on chromosome 9p21.3→9pter responsible for mental retardation and mild facial anomaly.This result has been applied successfully in prenatal diagnosis of the second family.
8.Effect of Lactobacillus coryniformis FZU63 on the flavor quality of black tea beverage.
Ruili LI ; Yifeng LIU ; Weibo LUO ; Huilin HUANG ; Meiting HUANG ; Chi CHEN ; Ronghui XIAO ; Jinzhi HAN ; Xucong LÜ
Chinese Journal of Biotechnology 2022;38(12):4731-4743
The tea beverages will be endowed with distinct aroma and taste, as well as various biologically active compounds including probiotic factors, when fermented with lactic acid bacteria (LAB). However, at present, few studies on the dynamics of flavors in tea soup at different fermentation stages were conducted. In this study, the composition of monosaccharides, aromatic components, free amino acids, and organic acids were measured, when the black tea beverages were fermented with Lactobacillus coryniformis FZU63 which was isolated from Chinese traditional kimchi. The results indicated that monosaccharides including glucose, fructose, mannose and xylose in black tea beverages are the main carbon sources for fermentation. In addition, the abundance of aromatic compounds in black tea soup are increased significantly at different fermentation stages, which endow the fermented black tea soup with fruit aroma on the basis of flowery and nutty aroma. Moreover, some bitter amino acids are reduced, whereas the content of sweet and tasty amino acids is elevated. Furthermore, the levels of lactic acid, malic acid, citric acid and other organic acids are accumulated during the fermentation. Additionally, sensory evaluation displays that black tea beverage is acquired with comprehensive high-quality after being fermented for 48 h. This study provides a theoretical basis to steer and control the flavor formation and quality of the fermented tea beverages during LAB fermentation.
Tea/chemistry*
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Beverages/microbiology*
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Camellia sinensis
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Fermentation
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Acids
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Amino Acids
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Glucose
9.COVID-19 Vaccination for Cancer Patients: Progress and Preliminary Recommendations.
Luo WANG ; Yan XU ; Lu ZHANG ; Junping FAN ; Ruili PAN ; Jinglan WANG ; Mengzhao WANG
Chinese Journal of Lung Cancer 2021;24(6):377-383
The pandemic of coronavirus disease 2019 (COVID-19) has had a serious impact on global health. COVID-19 vaccines may be one of the most effective measure to end the pandemic. High infection risk and higher serious incident and mortality rates have been shown in cancer patients with COVID-19. Therefore, cancer patients should be the priority group for COVID-19 prevention. Until now, data of COVID-19 vaccination for cancer patients is lacking. We review the interim data of safety and immune-efficacy of COVID-19 vaccination in cancer patients based on the latest studies. Due to the complicated immune systems of cancer patients caused by the malignancy and anticancer treatments, we proposed preliminary specific COVID-19 vaccination recommendations for cancer patients with different anticancer treatments and at different stages of the disease. Preventing COVID-19 with vaccinations for cancer patients is crucial, and we call for more large-scale clinical trials and real-world studies, for further COVID-19 vaccination recommendations development.
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