1.Flexible sensors and their application in healthcare
Xuesi ZHOU ; Ronghua ZHONG ; Tianhui WANG
Military Medical Sciences 2015;(11):876-880
The traditional approach to accessing healthcare information restricts the further development of healthcare services,thus unable to meet the growing needs of individual healthcare.The flexible sensor technology has emerged along with the development of new materials,machinery and manufacturing technology.As a result,textiles,accessories,human skin and even internal body organs can be integrated with various sensors.The popularization of flexible sensors provides new methods for monitoring health,improving therapeutics,investigating disease status and building the human-machine in-terface.Through a systematic investigation of literature,this paper reviews the applications of flexible sensors in health-care,discusses the key technologies,and introduces the common materials and manufacturing technology.
2.Inhibitory effects of siRNA targeting survivin on the growth of a human melanoma cell line,M14
Linyi SONG ; Jianfang SUN ; Wuqing ZHOU ; Xuesi ZENG
Chinese Journal of Dermatology 2009;42(3):189-192
Objective To study the inhibitory effects of siRNA targeting survivin on the expression of survivin,as well as the apoptosis,proliferation and invasion of a human melanoma cell line,M14.Methods Two siRNAs targeting survivin were designed,chemically synthesized,and used to construct the recombinant plasmids,pRAT-H1.1/neo-survivin-siRNA1 and pRNAT-H1.1/neo-survivin-siRNA2.Then,recombinant plasmids were transfected into M14 cells mediated by Lipofectamine 2000 reagent.Those cells untransfected or transfected with empty vector served as the control.After culture over various periods of time.cells were collected for the detection of mRNA and protein expression of survivin with RT-PCR and Western blotting,respectively,and for the examination of apoptosis and proliferation of M14 cells by flow cytometry and MTT methods,respectively.Also,Transwell assay was performed to detect the invasive capability of M14 cells.Results A statistical decrease in the mRNA and protein expressions of survivin was observed along with an increase in apoptotic rate(x2=31.55,P<0.01)in M14 cells transfectcd with siRNA-containing plasmid compared with untransfected and empty vector-transfected cells.As MTT assay indicated,on day 4 after the transfcorion,the proliferation of M14 cells was inhibited by(55.4±4.3)%,(34.5±4.3)%and(13.3±4.6)%,with pRNAT-H1.1/neo-survivin-siRNA1,pRNAT-H1.1/neo-survivin-siRNA2 and empty vector,respectively:there was a significant difference among the three groups(P<0.05).Decreased invasive capability was noticed in M14 cells transfected with siRNA-containing plasmid compared with untransfected cells(all P<0.05).Conclusions The plasmid containing siRNA against survivin can specifically inhibit the expression of sarvivin,proliferation and invasion of tumor cells,and induce cell apoptosis.The inhibition of survivin expression by siRNA may be a rational approach to the gene therapy for malignant melanoma.
3.Silence of BRAF gene in human melanoma cells by plasmid mediated shRNA
Yongzhi HAN ; Jianfang SUN ; Wuqing ZHOU ; Xuesi ZENG ; Yiqun JIANG
Chinese Journal of Dermatology 2003;0(11):-
Objective To construct the short hairpin RNA (shRNA)-expressing plasmid vectors specific for BRAF gene, and to test their effects in BRAF knockdown in human melanoma cell lines. Methods Two pairs of specific BRAF shRNA oligoes and a pair of randomly synthesized non-specific shRNA oligo were synthesized and inserted into plasmid pGenesil-1. Their fidelity was confirmed by double endonuclease digestion and sequencing. The constructed plasmids were transfected into human melanoma cell lines A375 and M14. The expression of BRAF mRNA and BRAF protein were detected by RT-PCR and Western blotting, respectively. Results The designed shRNA oligoes were precisely cloned into the plasmid pGenesil-1. The expression of BRAF mRNA and protein were down-regulated by specific plasmid braf 1 and braf 2, except to non-specific plasmid neg. The plasmid braf 1 was more effective, reducing BRAF gene expression by 90 per cent. Conclusions Plasmid mediated shRNA could successfully knockdown BRAF expression in human melanoma cells, and the suppression of the gene expression could maintain for 1 month at least.
4.Effects of 5-aza-dc on the expression of insulin-like growth factor binding protein 7 (IGFBP7) in and proliferation of melanoma cells
Yanning XUE ; Hao CHEN ; Wuqing ZHOU ; Yali SONG ; Xuesi ZENG ; Jianfang SUN
Chinese Journal of Dermatology 2010;43(6):418-421
Objective To study the effects of 5-aza-dc on the expression of IGFBP7 in and proliferation of melanoma cell lines A375 and M14.Methods Reverse transcription-PCR and immunocytochemistry were performed to detect the mRNA and protein expression of IGFBP7 in A375 cells and M14 cells after treatment with 5-aza-dc of 10μmol/L for 48 hours,and MTT assay to measure the proliferation of both cell lines treated with 4 different concentrations (2.5,5,10,20μmol/L) of 5-aza-dc for various durations.Results The treatment with 5-aza-dc restored IGFBP7 expression at both mRNA and protein levels.The four concentrations of 5-aza-dc inhibited the proliferation of A375 and M14 cells in a dose-dependent (F=561.12,271.43,respectively,both P<0.01) and time-dependent (F=141.35,549.33,respectively,both P<0.01) manner.Conclusions DNA methylation may be involved in the modulation of aberrant IGFBP7 gene expression in melanoma,and 5-aza-dc could inhibit the proliferation of A375 and M14 cells.
5.Methylation status of CpG islands in the promoter region of IGFBP7 gene in human melanoma cell lines
Yanning XUE ; Hao CHEN ; Wuqing ZHOU ; Yuping CAO ; Yue TAO ; Xuesi ZENG ; Jianfang SUN
Chinese Journal of Dermatology 2012;45(10):714-717
Objective To assess the relationship of methylation status of CpG islands in the promoter region of insulin-like growth factor binding protein 7 (IGFBP7) gene with the expression of IGFBP7 gene in human melanoma cell lines and primary melanocytes.Methods Primary melanocytes from human forcskin tissue as well as 4 human melanoma cell lines,including A375,M14,SK-MEL-1 and MV3,were used in this study.Bisulfite sequencing PCR (BSP) was applied to detect the methylation status of 54 CpG sites in the 5'-flanking promoter region of IGFBP7 gene in all of the melanoma cell lines and primary melanocytes.Results As hierarchical cluster analysis showed,IGFBP7-positive cells (including A375,M14 and SK-MEL-1 ) differed significantly from IGFBP7-negative cells (including MV3 cells and primary melanocytes) in the methylation pattern of IGFBP7 gene promoter region.Conclusion The methylation status of CpG island in the promoter region of IGFBP7 gene may be associated with its expression in melanoma cell lines.
6.The effects of melanoma culture supernatants on production of IL-12 and expression of CD14 and CD1a by human monocytes in vitro
Ningjing SONG ; Xuesi ZENG ; Baiqing LI ; Junying WU ; Wuqing ZHOU ; Mingying ZHONG ; Qianqiu WANG
Chinese Journal of Dermatology 2003;0(12):-
85%. The concentrated MCS in different amount was added to the IFN-?(100 pg/mL) and LPS (10 ng/mL) enriched culture media. The IL-12 production by monocytes was determined by the enzyme- linked immunosorbent assay(ELISA).The expression of CD14 and CD1a was analyzed by flow cytometry 5 days after the monocytes were co-cultured with MCS. Results The production of monocytic IL-12 was down-regulated by MCS in a dose dependent manner. The amount of IL-12 from monocytes decreased along with an increased dose (25-100?L) of MCS applied in the reaction. It was also observed that the differentiation from CD14 expressing monocytes to CD1a dendritic cells was impaired by MCS. The ability of MCS to inhibit the production of IL-12 by monocytes and to suppress the differentiation of monocytes to dendritic cells in vitro could be disrupted by PD98059,an ERK specific inhibitor. Conclusions MCS appears to inhibit IL-12p40 production by monocytes and inhibit differentiation of monocytes in vitro via secretion of ERK stimulating factor. The inhibitory factors in MCS and their chemical natures need further research.
7.Effect of T-cell immunoglobulin and mucin domain-3 on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells
Yalin LYU ; Xiaowei ZHOU ; Bin HU ; Qiong WU ; Xuesi ZENG ; Yi LIU ; Jianfang SUN
Chinese Journal of Dermatology 2016;49(2):82-87
Objective To evaluate the effect ofT-cell immunoglobulin and mucin domain-3 (TIM-3) on TRP-2180-188 peptide-stimulated murine spleen lymphocytes co-cultured with B16F10 murine melanoma cells.Methods A recombinant plasmid pFUSE-TIM-3-mIgG2Aae1-Fc2 encoding TIM-3 was constructed.Then,the recombinant plasmid and an empty plasmid pFUSE-mIgG2Aae1-Fc2 were transfected into human 293T epithelial cells followed by 48-hour culture for the preparation of supernatants containing TIM-3 and Ig-tail respectively.C57BL/6 mice were immunized with the TRP-2180-188 peptide vaccine for 4 sessions.One week after the last vaccination,C57BL/6 mice were sacrificed,and spleen lymphocytes were collected and then cultured with the TRP-21180-188 peptide and interleukin-2 (IL-2) for 5 days,with lymphocytes untreated with the TRP-2180-188 peptide or IL-2 serving as the control group.Mitomycin-treated B16F10 murine melanoma cells and TRP-2180-188 peptide-stimulated lymphocytes were co-cultured with the presence of supernatants of 293T cells that had been cultured for 48 hours (blank control group),TIM-3-containing supernatants (TIM-3 group) and Ig-tail-containing supernatants (negative control group) separately.After 24 and 48 hours of co-culture,cell counting kit-8 (CCK-8) assay was performed to estimate the proliferative activity of lymphocytes,enzyme-linked immunosorbent assay (ELISA) to determine the supernatant levels of interferon (INF)-γ and tumor necrosis factor (TNF)-α,flow cytometry to determine the percentage of CD8 + T cells in the co-culture system.Results Enzyme digestion and sequence analysis showed that the TIM-3 gene was successfully inserted into the eukaryotic expression plasmid.After 48-hour culture,TIM-3 and Ig-tail expressions were detected in the supernatants of 293T cells transfected with the recombinant plasmid and empty plasmid respectively.As CCK-8 assay showed,the proliferative activity of lymphocytes was significantly lower in the TIM-3 group than in the blank control group and negative control group after 24-and 48-hour culture (78.06% ± 6.37% vs.100.00% ± 10.42% and 108.70% ± 9.90% at 24 hours,42.93% ± 5.93% vs.100.00% ± 6.24% and 168.00% ± 2.98%at 48 hours,all P < 0.05),so was the ratio of cellular proliferative activity at 48 hours to that at 24 hours (all P < 0.05).Compared with the blank control group and negative control group,the TIM-3 group showed significantly decreased supernatant levels of IFN-γ and TNF-α after 24-hour (IFN-γ:192.96 γ 5.05 ng/L vs.216.44 ± 7.85 ng/L and 223.67 ±7.79 ng/L,both P< 0.05;TNF-α:58.43 ± 0.26 ng/L vs.26.43 ± 0.01 ng/L and 86.85 ± 1.12 ng/L,both P< 0.05) and 48-hour culture (IFN-γ:54.95 ± 0.57 ng/L vs.230.06 ± 4.23 ng/L and 167.24 ± 3.33 ng/L,both P < 0.05;TNF-α:30.23 ±0.26 ng/L vs.26.84 ± 0.20 ng/L and 45.34 ± 0.22 ng/L,both P < 0.05).In addition,the median percentage of CD8+ T cells was significantly increased in the TIM-3 group compared with the blank control group and negative control group after 24-and 48-hour culture (3.30% vs.0.421% and 2.22% at 24 hours,4.06% vs.0.577% and 0.691% at 48 hours,all P< 0.05).Conclusion TIM-3 in vitro can suppress the proliferative activity of and secretion of IFN-γand TNF-α by lymphocytes,but increase the percentage of CD8 + T cells in the co-culture system of TRP-2180-188 peptide-stimulated lymphocytes and B16F10 cells.
8.In vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a malignant melanoma cell line,A375
Jia CHEN ; Wuqing ZHOU ; Hao CHEN ; Yali SONG ; Limin CAI ; Chonmei XIN ; Xuebao SHAO ; Yi LIU ; Xuesi ZENG ; Jianfang SUN
Chinese Journal of Dermatology 2009;42(4):255-258
Objective To investigate the in vitro anti-proliferation effect of a histone deacetylase inhibitor,chidamide,on a cutaneous malignant melanoma cell line,A375.Methods Cultured A375 cells were treated with different concentrations of chidamide(5,10,50,100,500 μmol/L)and aichostatin A (TSA)(0.1,0.25,0.5,1.0 μmol/L),respectively,for various durations(24,48,72,96,120 hours).Subsequently,cell proliferation,apoptosis and cell cycle were detected by MTT assay,annexin Vfluorescein isothiocyanate and propidium iodide double staining,and DNA ploid analysis,respectively.Results The proliferation of A375 cells was inhibited in a dose-dependent manner by chidamide of 5-500μmol/L and TSA of 0.1-1 μmol/L,and in a time-dependent manner from 0 to 120 hours after the beginning of trealment with ehidamide of 5-500μmol/L and TSA of 0.25-1μmol/L.The 48-hour 50% growth inhibition concentration(IC50)of ehidamide and TSA on A375 cells was about 250 μmol/L and 0.7μmol/L,respectively.After 48-hour treatment,the apoptosis mte was 80.27%±3.06%,79.53%±5.70%,83.13%±6.90%in A375 cells treated with chidamide of 62.5,125,250 μmol/L,respectively,16.27%±2.46%,28.83%±2.55%,83.40%±8.65%in those treated with TSA of 0.175,0.35,0.7 μmol/L,respectively,10.43%±0.96%in ontreated cells;a statistical increase was noticed in chidamide-treated cells and TSA-treated cells vs.untreated cells(all P<0.001).A positive correlation was observed between the apoptosis rate and concentrations of TSA(r=0.955,P=0.000).Cell cycle analysis indicated that treatment with chidamide induced cell cycle arrest in G0/G1 phase,with the cell proportion in G0/G1 phase being 76.30%±6.06%,82.79%±0.74%,88.91%±5.29%in A375 cells treated with chidamide of 62.5,125,250μmol/L,respectively,versus 38.73%±3.36%in untreated cells.While after 48-hour treatment with TSA of 0.35 and 0.7 μmol/L,the proportion of cells in G2/M phases was 25.15%±2.71%and 58.71%±3.45%,respectively,compared to 15.73%±0.23%in untreated cells(P<0.01).Conclusion Chidamide and TSA could induce cell cycle arrest and apoptosis,as well as inhibit the growth of A375 ceils in vitro.
9.Study on the prognosis of drug-eluting stent implantation in the elderly patients with coronary heart disease
Qiang ZHANG ; Changsheng MA ; Shaoping NIE ; Qiang LV ; Junping KANG ; Xin DU ; Yin ZHANG ; Changqi JIA ; Rong HU ; Xinmin LIU ; Xiaohui LIU ; Jianzeng DONG ; Fang CHEN ; Yujie ZHOU ; Shuzheng LV ; Xuesi WU
Chinese Journal of Geriatrics 2009;28(1):37-41
Objective To evaluate the prognosis of drug-during stent (DES) implantation in elderly patients versus non-elderly patients, and to determine the clinical outcome of complete revascularization strategy versus incomplete revascularization strategy in elderly patients. Methods Patients who were treated with at least 1 DES in our hospital were enrolled in the study. They were divided into 3 groups: the elderly group (aged 75~89 years), the presenium group (age 60~74years) and the non-elderly group (aged 40~59 years). The patients aged 60~89 years were further divided into complete revascularization group and incomplete revascularization group according the Percutaneous interventional the rapy (PCI) strategy. Clinical characteristics, angiographical and interventional data were collected. Results The success rate of PCI procedure was 99.3% in elderly group(n=137), 98.7% in presenium group(n= 1006), and 99.3% in non-elderly group(n= 1031).There were no significant differences among the three groups(P>0.05). The in-hospital mortality was highest in the elderly group among the three groups (1.5%, 0.4%, 0.1%, P<0.05), but the in-hospital rates of re-infarction, repeated revascularization and stroke had no significant differences among the three groups (P>0.05). During follow-up, the rates of death and stroke were highest in the elderly group(3.1%, 2.3%, 0.7%, P<0. 01;1.5% , 1.3%, 0.3%, P<0.05, respectively),but the rates of re-infarction and repeated revascularization among the three groups had no significant differences (all P>0.05). By Cox regression analysis, serum creatinine (OR= 2.961,95%CI=1. 643~5.338,P<0.01), gender (OR=2.661,95%C1=1.376~5.145 ,P<0.01), age(OR=2.687,95%CI=1.329~5.434, P<0.01), multi-vessel disease(OR= 1.735,95 %CI= 1.132~2.661, P<0.05), and old myocardial infarction (OR = 2.041 ; 95% CI = 1.026~4.061; P<0.05) were the independent predictors for all-cause death in patients aged 60~74 years. The in-hospital mortality was higher in the incomplete revascularization group than in complete revascularization group in patients aged 60~74 years (1.4% vs. 0.2%, P<0.05). Multiple logistic regression analysis revealed that the incomplete revascularization strategy was not the independent predictor of in-hospital death (OR=0.307; 95%CI=0.011~8.467; P>0.05). Conclusions Although DES implanting is successfully procedured in presenium and elderly patients, it is associated with higher in-hospital mortality, especially in patients aged ≥75 years . Presenium and elderly patients are to be more benefit from complete revascularization strategy, but the incomplete revascularization strategy does not influence the long-term outcomes.
10.Discussion on the acupuncture and moxibustion thoughts of diagnosis and treatment for secondary dysmenorrhea of adenomyosis on the basis of disease location and pathogenesis.
Xuesi HOU ; Qingchen ZHOU ; Lei WANG ; Lu CHENG ; Jiping ZHAO
Chinese Acupuncture & Moxibustion 2017;37(10):1108-1112
To discuss the acupuncture and moxibustion thoughts of diagnosis and treatment for secondary dysmenorrhea of adenomyosis on the basis of disease location and pathogenesis. In clinic, we take the "principle, method, prescription, acupoint and technic" as the outline, paying attention to identify disease location and establishing the method of "promoting blood to remove stasis, regulating thoroughfare vessel and conception vessel" on the basis of the pathogenesis of "stasis obstructing uterus, disharmony of thoroughfare vessel and conception vessel". The prescription combines "dredging" with "conditioning", and the emphasis should be different in different periods. In menstrual period, we put emphasis on activating the circulation ofand blood as well as clearing meridians to relieve pain and choose the acupoints on the spleen meridian of foot-and experimental points, such as Diji (SP 8), Sanyinjiao (SP 6), Ciliao (BL 32), Shiqizhui (EX-B 8). In the intermenstrual period, we regulate theand blood of thoroughfare vessel and conception vessel, and the function of viscera. The acupoints for the disease root are mainly at spleen meridian of foot-and conception vessel, such as Sanyinjiao (SP 6), Guanyuan (CV 4), Zigong (EX-CA 1), Zusanli (ST 36). And the reinforcing and reducing technic are applied accordingly.