1.Analysis on Application of Ginseng Prescriptions for Generations
Journal of Zhejiang Chinese Medical University 2017;41(7):601-604
[Objective]Combing the application of ginseng in the past physicians to make readers understand the efficacy of ginseng,and to be flexible in order to better serve human health.[Methods]Through consulting the ancient books of materia medica,the understanding of the ancient physicians and the application of hanren formulae were summarized and analyzed.[Results]Although the ancient physicians of the past believed that ginseng was subject to tonify five organs,quiet spirt,set soul,stop fright,remove pathogen,in addition to bright eye,happy,educational. long taking makes people fit and live longer,but with the development of the times,the Han dynasty in to nifying Qi torelieve thirst,tonify Qi for deficiency,the Tang dynasty in tonifying heart Qi,benefiting heart spirit,the Song dynasty in complement,benefiting Qi,the Ming dynasty in benefiting qi,Qing dynasty in tonifying Qi and Yin,modern in emergency first aid application.[Conclusion]Through the analysis of the application of ginseng in past dynasties,the complement ginseng can be more widely used in clinic,and better serve human health.
2.Stability of a rat model of type 2 diabetic mellitus induced by streptozotocin combined with high-fat feeding and its eye disease characteristics
Yang YU ; Shilei ZHENG ; Xuezheng LIU
Chinese Journal of Tissue Engineering Research 2015;(27):4389-4393
BACKGROUND:Streptozotocin-induced diabetic ophthalmopathy is commonly used in animal models, but the pathological changes are local that mainly emphasize on the retina. Little evidence is found about the animal models of the pathology of diabetic ophthalmopathy. OBJECTIVE:To investigate the long-term stability of type 2 diabetic melitus rat models induced by streptozotocin combined with high-fat feeding and to observe the characteristics of eye disease. METHODS: Rats were randomly divided into control group and diabetes group. Control group was given normal feeding, while diabetes group was given intraperitoneal injection of streptozotocin combined with high-fat feeding to establish diabetic models. RESULTS AND CONCLUSION:Compared with the control group, at 1 month after modeling, the fasting blood glucose levels increased, and the insulin sensitivity index decreased in the diabetic group (P < 0.05). Evans blue staining results showed that, at 3 months after modeling, retinal cel lesions exacerbated in the diabetic group; at 5 months after modeling, retinal blood vessels traveled in circuity and disorderly, accompanied by the leakage in the diabetic group, Evans blue content in the retina increased as the time after modeling went by (P < 0.05). Under transmission electron microscopy, at 5 months after modeling, the eye lenses in the diabetes group were flocculent pieces, which were the typical characteristics of cataract. Experimental findings indicate that the rat model of type 2 diabetic melitus induced by streptozotocin combined with high-fat feeding has a long-term stability, and its eye changes are consistent with the characteristics of diabetic ophthalmopathy. Therefore, it is an ideal animal model for diabetic ophthalmopathy.
3.The role of activation of IL-6/STAT3 signaling in Th17/Tr imbalance of Kawasaki disease
Guobing WANG ; Chengrong LI ; Jun YANG ; Pengqiang WENG ; Shilei JIA
Chinese Journal of Microbiology and Immunology 2011;31(6):517-522
Objective To investigate the role of IL-6/STAT3 signaling in Th17/Tr imbalance of Kawasaki disease(KD). Methods Forty-eight children with KD and eighteen age-matched healthy children were consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of IL-17A, IL-17F, RORγt, Foxp3, SOCS1 and SOCS3 were assessed by real-time PCR. The proportion of CD4+CD25+Foxp3+ regulatory T(Tr) cells and mean fluorescence intensity(MFI) for phosphorylated-STAT3(pSTAT3) protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCS1 exon2, three potential bind sites for STAT3 in 5'-untraslated region(5'-UTR) of SOCS3 in CD4+ T cells. Results (1)Compared with healthy volunteers, plasma IL-6 concentration and MFI for pSTAT3 in CD4+ T cells were elevated significantly during acute phase of KD[IL-6:(54.02±20.58) pg/ml vs (8.72±2.06) pg/ml, P<0.05;pSTAT3 MFI:(55.41±15.08) vs (9.35±3.76), P<0.05], and the two items in KD patients with coronary artery lesion (KD-CAL+) were found to be higher than those in KD patients without coronary artery lesion (KD-CAL-)[IL-6:(84.76±29.35) pg/ml vs (38.65±13.76) pg/ml, P<0.05;pSTAT3 MFI:(72.36±16.81) vs (46.93±13.57), P<0.05]. (2)Transcription levels of IL-17A, IL-17F and RORγt in patients with KD were significantly elevated (P<0.05) while the proportion of CD4+CD25+Foxp3+ Treg and expression levels of Foxp3 were detected to be lower than those in normal controls (P<0.05). The mRNA levels of IL-17A, IL-17F and RORγt in KD-CAL+ group were higher than those in KD-CAL- group(P<0.05), as well as expression level of Foxp3 were found to be lower in KD-CAL+ group(P<0.05). (3)The mRNA levels of SOCS1 and SOCS3 in CD4+ T cells increased significantly during acute phase of KD(P<0.05), while the two items in KD-CAL+ group were lower than those in KD-CAL- group(P<0.05). Furthermore, CpG islands in SOCS1 exon2 and the third potential bind site for STAT3 in SOCS3 5'-UTR were hypomethylated in acute KD, while those in healthy controls were fully demethylated(P<0.05). Demethylation levels of SOCS1 exon2 and the third potential bind site for STAT3 in SOCS3 5'-UTR in KD-CAL+ group were lower than those in KD-CAL- group(P<0.05). CpG islands in the other two bind sites for STAT3 in SOCS3 5'-UTR were fully demethylated among all the groups(P>0.05). ConclusionAberrant activation of IL-6/STAT3 signaling caused by hypomethylation of SOCS1 and SOCS3 might be one contributing factor to unbalance of Th17/Tr in KD.
4.Investigation of the methylation status of Foxp3 gene during acute phase of Kawasaki disease
Guobing WANG ; Chengrong LI ; Jun YANG ; Pengqiang WEN ; Shilei JIA
Chinese Journal of Microbiology and Immunology 2010;30(7):678-682
Objective To investigate the methylation status of Foxp3 gene and its roles in immunological pathogenesis of Kawasaki disease(KD). Methods Thirty children with KD and eighteen agematched healthy children consented to participate in this study. Quantitative methylation specific polymerase chain reaction(MSQP) was used to assess the methylation status of Foxp3 promoter and regulatory T cells specific demethylated region(TSDR) in CD4+ T cells. The proportion of CD4+ CD25 + Foxp3 + regulatory T (Tr) cells was analyzed by flow cytometry. Transcriptional levels of CD4+ CD25 + Foxp3 + Tr associating genes (Foxp3, CTLA4, GITR, LAG3 and CCR8 ) and Foxp3-dependent molecules (UBD and LGAIS3)were measured by real-time PCR. Results ( 1 ) Demethylation level of Foxp3 promoter in CD4 + T cells from patients with KD was lower significantly than that of health subjects( P < 0.01 ), and increased significantly after treated with intravenous gamma globulin therapy(IVIG) (P < 0.01 ). No difference of demethylation level of TSDR region was observed among all groups ( P > 0. 05 ). ( 2 ) The proportion of CD4 + CD25 + Foxp3 + Tr in peripheral blood from patients with KD , as well as mRNA levels of Foxp3 gene in CD4+ T cells, was significantly lower than those of health subjects ( P <0. 01 ), and increases significantly after IVIG therapy (P <0.01 ). Significant positive correlations between demethylation level of Foxp3 promoter and the proportion of CD4 + CD25 + Foxp3 + Tr , or expression levels of Foxp3 in CD4 + T cells, were observed during acute phase of KD (CD4+CD25+ Foxp3+ Tr: r=0.76, P<0. 01; Foxp3: r=0.89, P<0. 01). (3)Transcription level of CD4+ CD25+ Foxp3+ Tr associating factors, such as CTLA4, GITR, LAG3 and CCR8, was significantly down-regulated in acute phase of KD(P<0. 01 ), and up-regulated to some extent after treated with IVIG(P <0. 01 ). Expression levels of Foxp3-dependent molecules UBD and LGALS3 in CD4 + T cells decreased significantly during acute phase of KD (P < 0.01 ), and basically recovered to the levels of health subjects( P < 0.01 ). Conclusion Decrease of demethylation level of Fopx3 promoter is correlated with immune dysfunction in Kawasaki disease.
5.The role of activation of toll-like receptors in immunological pathogenesis of Henoch-Schonlein purpura
Yuanyuan LI ; Chengrong LI ; Guobing WANG ; Jun YANG ; Shilei JIA
Chinese Journal of Rheumatology 2010;14(8):538-542
ObjectiveTo investigate the role of signal transduction of TLRs in the Henoch-Schonlein purpura (HSP). Methods Reverse-transcription PCR (RT-PCR) and real-time PCR were used to evaluate the levels of TLRs(1~10), MyD88, TRAF6, TRIF, IFN-α, IFN-β, IL-6, IL-1β, TNF-α, IP-10, RANTES,iNOS, Blys/April mRNA expression in peripheral blood mononuclear cells and their expression levels were compared using t test., while the concentration of plasma cytokines such as Blys、IFN-α、IFN-β、IL-6、IL-1、TNF-α was measured by enzyme-linked immunosorbent assay(ELISA).Expression levels of those genes were compared using t test. Results①Compared with the control group, the expression levels of TLR1, TLR2,TLR6, TLR5, TLR3, TLR7, TLR9 mRNA were up-regulated significantly(P<0.01), while no difference of TLR4 was detected (P>0.05).②Transcription levels of MyDg8(2.47±1.06) vs(0.73±0.22), TRAF6 (2.54±0.72)×10-3vs(0.70±0.20)×10-3, TRIF(3.18±0.86)×10-3vs(0.93±0.35)×10-3 were significantly up-regulated in acute phase of HSP (P<0.01).③The levels of IFN-α and IFN-β protein and mRNA were remarkable increased (P<0.01).④ The expression of cytokine/chemotactic factor such as IL-6, IL-1β, IP-10, RANTES,iNOS was higher than that of the control group(p<0.01), while TNF-αdid not change in children with HSP (P>0.05). ⑤ It was detected that the expression of Blys/April was higher than that of the control group(P<0.01). ConclusionExpressions of TLR1, TLR2, TLR6, TIR5, TLR3, TLR7, TLR9, MyD88, TRAF6, and TRIF are up-regulated during acute phase of HSP, suggesting that aberrant activation of TLRs triggered by microbes may be one of the initiating factors of immune aberrance in HSP. Over expression of cytokine/chemotactic factor or Blys/April owning to the aberrant activation of TLRs, may be correlated with immunological pathogenesis of HSP.
6.Vertebral artery hypoplasia and its clinical significance
Shuangshuang YANG ; Yan JI ; Bo SONG ; Yuming XU ; Shilei SUN
International Journal of Cerebrovascular Diseases 2015;(3):209-213
Vertebral artery hypoplasia is a congenital vessel variation. Its incidence is from 1. 9 to 26. 5% . In recent years, studies have shown that vertebral artery hypoplasia may be a potential risk factor for posterior circulation infarction, especialy when it coexists with other cerebrovascular risk factors. Vertebral artery hypoplasia may also cause regional hypoperfusion and complex neurovascular regulation, and it also has a certaln link with migralne.
7.Influence of SOCS1 and SOCS3 hypomethylation on homeostasis of Th1/Th2 in Kawasaki disease
Guobing WANG ; Chengrong LI ; Jun YANG ; Pengqiang WENG ; Shilei JIA
Chinese Journal of Rheumatology 2010;14(11):732-737
Objective To investigate the effect of SOCS1 and SOCS3 hypomethylation on homeostasis of Th1/Th2 in Kawasaki disease(KD). Methods Thirty-six children with KD and sixteen age-matched healthy children consented to participate in this study. Protein concentration of IL-6 in plasma was measured by ELISA. Transcriptional levels of SOCS1, SOCS3, T-bet, IFN-γ, GATA3 and IL-4 were assessed by realtime PCR. The proportion of Th1 and Th2 cells, and mean fluorescence intensity(MFI)for phosphorylated STAT3(pSTAT3)protein in CD4+ T cells was analyzed by flow cytometry. A quantitative methylation specific PCR based on SYBR Green was used to evaluate methylation status of CpG islands in SOCSl exon2, and three potential binding sites for STAT3 in 5'-untraslated region(5'-UTR)of SOCS3 in CD4+T cells. Comparisons between groups were performed with t-test. Results ①Compared with healthy volunteers, plasma IL-6 concentration[(51.8±16.3)pg/ml vs(8.6±2.0)pg/ml, respectively]and MFI for pSTAT3[(52±14)vs(10±4), respectively]in CD4+ T cells were elevated significantly during acute phase of KD(P<0.05), and the two items in KD patients with coronary artery lesion(KD-CAL+)were found to be higher than those in KD patients without coronary artery lesion(KD-CAL-)[IL-6:(87.2±27.4)pg/ml vs(36.2±12.8)pg/ml, P<0.05; pSTAT3 MFI:(75±15)vs(42±11), P<0.05]. ② The proportions of Th1 and Th2 cells and transcription levels of Th-associating factors(T-bet, IFN-γ, GATA3 and IL-4)in CD4+ T cells increased significantly in acute KD(P<0.05), while the rate of Thl div Th2 in KD patients was found to be lower than that in normal controls(P<0.05). In addition, the proportions of Th1 and Th2 cells and expressions levels of Th-associating factors in KD-CAL+ group were higher than those in KD-CAL-group, as well as the rate of Thl div Th2 cells in KD -CAL+ group were lower than that in KD-CAL- group(P<0.05). ③ The mRNA levels of SOCSl and SOCS3 in CD4+ T cells increased significantly during acute phase of KD(P<0.05), while the two items in KDCAL+ group were lower than those in KD-CAL- group(P<0.05). Furthermore, CpG islands in SOCSl exon2 and the third potential binding site for STAT3 in SOCS3 5'-UTR were hypomethylated in acute KD, while those in healthy volunteers were fully demethylated(P<0.05). Demethylation levels of the two items mentioned above in the KD-CAL+ group were lower than those in the KD-CAL-group(P<0.05). CpG islands in the other two binding sites for STAT3 in SOCS3 5'-UTR were fully demethylated among all the groups(P>0.05).Conclusion Relative insufficiency of SOCS1 and SOCS3 expression caused by hypomethylation may be one contributing factor for the imbalance of Th1/Th2 in KD.
9.Investigation of the change of CD4~+ T cell subset from children with type 1 diabetes
Shilei JIA ; Chengrong LI ; Guobing WANG ; Xia LIU ; Jun YANG ; Ying ZU ; Dan FU
Chinese Journal of Microbiology and Immunology 2010;30(4):309-314
Objective To investigate the changes of CD4~+ T cell subset in the role of immuno-pathogenesis of type 1 diabetes mellitus(T1DM). Methods Real-time PCR was used to evaluate the ex-pression levels of transcriptional factors (T-bet, GATA-3, Foxp3, ROR-γt), cytokines (IFN-γ, IL-4, IL-10, IL-17A) and negative regulatory factors (CTLA-4, GITR) mRNA from CD4~+ T cells. The proportions of Th1, Th2, Tr and Th17 cells in peripheral blood were detected by flow cytometric analysis. Plasma cyto-kine (IFN-γ, IL-4, TGF-β and IL-6) concentrations were measured by enzyme-linked immunosorbent assay (ELISA). Results (1) The proportions of Th1 cells in peripheral blood from children with T1DM were siguificanfly increased than that of healthy controls, and proportions of Th2 were decreased (P < 0.01). There were no significant differences between diabetic patients and healthy controls regarding the proportions of Tr cells and Th17 cells(P >0.05). (2) Transcription levels of T-bet and IFN-γ mRNA were significantly up-regulated, while GATA3 and IL-4 were significantly down-regulated in children with T1DM. The mRNA expression levels of Tr negativity regulatory factors such as IL-10, CTLA-4 and GITR were lower in CD4~+ T cells from children with TIDM compared with the controls(P <0.01). There were no statistically differences to be observed in mRNA expression levels of ROR-γt and IL-17A genes between two groups(P > 0.05).(3) In comparison with controls, serum concentrations of IFN-γ or IL-4 were remarkable increased or de-creased respectively (P < 0. 01), while TGF-β and IL-6 did not change in children with T1DM (P > 0.05). Conclusion The Th1/Th2 imbalance might be play an important role in immunopathogenesis of T1DM. Functional deficiency of Tr cell might further exacerbate Th1/Th2 imbalance and lead to disturbance of cellu-lar immune response.
10.Effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats
Jun FU ; Yi ZHANG ; Chengsheng ZHANG ; Changchun YANG ; Shilei WANG ; Qishui LIN ; Haixing XUAN ; Dai ZHOU
Chinese Journal of Geriatrics 2008;27(10):775-779
ObjectiveTo observe the effects of human IL-10 gene transfection on the mRNA and protein expressions of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion injury in rats and to investigate its neuroprotective mechanism. MethodsRats were divided into four groups: normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group. The mRNA and protein expressions of IL-1β and TNF-α in the penumbra area were detected by fluorescence real-time quantitative PCR and ELISA respectively. ResultsIn normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfected group, the levels of protein expression of TNF-α in penumbra area were(0.66±0. 04) ,(1.16±0.26),(1. 155±0. 26)ng/g and(0. 84±0. 05)ng/g, and the levels of protein expression of IL-1βin penumbra area were(0.37±0.05), (1.25±0.39), (1.21±0.57) ng/g and(0.62+0.05)ng/g, respectively. Compared with normal control group, the levels of protein expression of TNF-α and 1L-1β were significantly higher in other three groups(all P<0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P<0. 01). In normal control group, ischemic control group, empty plasmid group and human IL-10 gene transfectedgroup, the levels of mRNA expression of TNF-α in penumbra area were 1.00 ±0.53,9.42±1.83,9.69±1.96 and 3.53±1.09, and the levels of mRNA expression of IL-1β in penumbra area were 1.00 ±0.51,27. 81±4.84,23.96 ± 4.90 and 13.55± 4.45, respectively. Compared with normal control group, the levels of mRNA expression of TNF-α and IL-1β were significantly higher in other three groups(all P<0. 01), and lower in human IL-10 gene transfected group than in ischemic control group and empty plasmid group(all P<0. 01). ConclusionsThe human IL-10 gene transfection may play an protective effect on cerebral ischemia through inhibiting mRNA and protein expression of IL-1β and TNF-α in the penumbra area following focal cerebral ischemia-reperfusion in rats.