1.Detection of microparticles in peripheral blood and its clinical significance
Changfu WANG ; Linyun LI ; Hongli WANG
Chinese Journal of Laboratory Medicine 2000;0(06):-
Microparticles in the peripheral blood are generated from the activated blood cells and vascular endothelial cells. Microparticles reflect the functional situations of their original cells directly and specifically,provide the basis for revelation of mechanisms and development trends of hematological or vascular diseases and also serve as the indications for differentiation diagnosis and evaluation of clinical treatment. Therefore it is very important to detect microparticles. This article will explain the present detection methods and the correlation of microparticles with diseases to provide basis and strategy for further study.
2.Performance Verification of XN-1000 Hematology Analyzer
Linyun LI ; Xiulin XIAO ; Changzheng WANG ; Juanan ZHANG
Journal of Modern Laboratory Medicine 2015;(3):126-128,132
Objective To verify the main index of Sysmex XN-1000 hematology analyzer and check whether performance of XN-1000 hematology analyzer meet the quality requirement.Methods The background value,carryover rate,precision,line-arity,reportable range,accuracy and comparability of different modes of sample absorbing of Sysmex XN-1000 hematology analyzer were verified and WS/T 406-2012 Analytical Quality Specifications for Routine Tests in Clinical Hematology was referred.Results The background counts,carryover rate,inter-batch precision and inter-day precision all met the require-ment.Linearity verification of WBC,RBC,HGB and PLT all met the requirement of the value of a within 1±0.05 and the correlation r≥0.975.The reportable range surpassed the range stated by the manufacture,which was accepted as the clinic reportable range.The accuracy test showed the mean and bias all met the requirement.The difference between the values de-tected by different absorbing modes all met the comparability requirement.Conclusion Performance of Sysmex XN-1000 he-matology analyzer was verified to meet the requirement.
3.Analysis on bacterial drug resistance in Hechuan District People's Hospital during 2016
Mingshuang LI ; Linyun YANG ; Yunyun TANG ; Tiantian WANG
Chongqing Medicine 2017;46(18):2521-2524
Objective To analyze the clinically isolated bacterial distribution and drug resistance characteristics in the our hospital during 2016 to provide the pathogenic drug resistance monitoring data for rational bacterial drugs use in clinic.Methods The clinicaly submitted samples were performed the pathogenic bacterial isolation according to the routine method.The isolated pathogens were identified by the Vitek2-Compact system and the drug susceptibility test was performed by adopting the MIC and KB methods.The results were statistically analyzed by adopting the WHONET5.6 software.Results A total of 2 214 non-repeat strains of bacteria were isolated in 2016,including 1 614 strains of Gram-negative bacilli,accounting for 72.9%,600 strains of Gram-positive bacteria,accounting for 27.1 %.The top five isolated bacteria were Klebsiella pneumoniae,Escherichia coli,Pseudomonas aeruginosa,Acinetobacter baumannii and Staphylococcus aureus.The detection rates of ESBLs producing Escherichia coli and Klebsiella pneumoniae were 51.8% and 27.6% respectively.The detection rates of methicillin-resistant Staphylococcus aureus (MRSA) was 26.5%.No vancomycin or linezolid resistant staphylococcal strains were found.Conclusion The main isolated pathogens in our hospital are dominated by Gram-negative bacteria.Hospital should strengthen reasonable and standardized use of antibacterial drugs to reduce the generation of drug resistant bacterial strains.
4.Generation and Expression of Recombinant Eukaryotic Expression Plasmids of PAX3 Gene and Its Significance
Hua ZHANG ; Jiada LI ; Hunjin LUO ; Hongsheng CHEN ; Linyun MEI ; Chufeng HE ; Yong FENG
Journal of Audiology and Speech Pathology 2014;(1):67-72
Objective To study exogenous expression and subcellular localization of wild type (WT ) and mu-tant PAX3 proteins in vitro by generating their expression plasmids for further study of pathogenesis of Waarden-burg syndrome (WS) .Methods The plasmids pECE-PAX3 and pcDNA3 .0-HA were ligased after they were cut by double enzyme digestion using molecular cloning technique to generate recombinant eukaryotic expression plasmid pcDNA3 .0-PAX3-HA ,which was as a template to generate expression plasmids pcDNA 3 .0 -H80D -HA and pcDNA3 .0-H186fs-HA of novel mutations H80D and H186fs of PAX3 gene .All constructs were verified by di-rect nucleotide sequencing .NIH3T3 cells were transfected transiently with the expression plasmids of PAX3 ,H80D and H186fs respectively .The exogenous expression of WT PAX3 protein and mutant H80D ,H186fs proteins were analysed using Western blot assay ,while their subcellular distribution were observed using immunofluorescence as-say .Results The DNA sequences of expression plasmids of PAX3 and its mutant H80D ,H186fs were correct . Both WT and mutant PAX3 proteins were detected at the expected size .WT PAX3 and H80D proteins were only lo-calized in the nucleus ,whereas H186fs protein showed aberrant localization in both cytoplasm and nucleus .Conclu-sion We successfully generated the recombinant eukaryotic expression plasmids of PAX 3 gene and its mutants and drew preliminary conlusion of gene mutation having effect on subcellular distribution of WT PAX 3 proteins in vitro , which lays experimental basis for further study of the moceluar mechanism of WS caused by PAX3 gene mutations in China .
5.Study on relationship between anti-proliferation effect of resveratrol and resveratrol-induced bone morphogenetic protein 9 and its receptors in colon cancer cells
Yuhua ZENG ; Ying SHAO ; Yang LI ; Qianzhao CHEN ; Wenyan REN ; Linyun ZHOU ; Yi ZHOU ; Rongxing LIU ; Baicheng HE
Chinese Pharmacological Bulletin 2016;32(12):1705-1710,1711
Aim To study the anti-proliferation effect of resveratrol (Res)and the role of Res-induced bone morphorgenetic protein 9 (BMP9 )in this process in colon cancer cells.Methods Crystal violet staining and flow cytomtry were introduced to assay the anti-proliferation effect of Res in LoVo cells.The effect of Res on apoptosis in LoVo cells was also detected with flow cytometry.Then,RT-PCR and Western blot assay were employed to unveil the effect of Res on the ex-pression of BMP9 .The effect of BMP9 on the anti-pro-liferation of Res in LoVo cells was analyzed with crystal violet staining and flow cytometry too.Finally,the effect of Res on the expression of ALK2 and ALK3 was assayed with RT-PCR,and the inhibitor of ALK2 and ALK3 was used to figure out the possible mechanism of BMP9 on Res-induced proliferation inhibition in LoVo cells.Results Res apparently inhibited the prolifera-tion,arrested the cell cycle at S phase in LoVo and in-creased the percentage of apopotic cells in LoVo cells. Res increased the expression of mRNA and protein of BMP9 concentration dependently. Exogenous ex-pressed-BMP9 enhanced the anti-proliferation and ap-optosis inducing effects of Res in LoVo cells, but BMP9 knockdown decreased these effects of Res.Al-though Res had no apparent effect on increasing the phosphorylation of Smad1/5/8,it increased the ex-pression of ALK2 and ALK3 .Inhibition of ALK2 and ALK3 decreased the anti-proliferation effect of Res partly in LoVo cells.Conclusion Res is potent to in-hibit the proliferation of LoVo cells,Which may be mediated by up-regulating the expression of BMP9 and its receptor at least.
6.Anti-proliferation effect of resveratrol and p38 MAPK in human colon cancer cells
Qianzhao CHEN ; Yuhua ZENG ; Ying SHAO ; Yang LI ; Wenyan REN ; Linyun ZHOU ; Yi ZHOU ; Rongxing LIU ; Baicheng HE
Chinese Pharmacological Bulletin 2016;32(8):1110-1114
Aim To investigate the relationship be-tween the anti-proliferation effect of resveratrol ( Res ) and p38 MAPK in colon cancer cells .Methods Crys-tal violet staining , Western blot and flow cytometry were employed to analyze the effect of Res on the pro-liferation in LoVo cells.Western blot assay was used to detect the effect of Res on the apoptosis of LoVo cells and the phosphorylation of p 38 MAPK.Crystal violet staining and Western blot assay were used to analyze whether p38 MAPK was involved in the Res-induced proliferation inhibition and apoptosis in LoVo cells .Re-sults Res inhibited the proliferation , arrested cell cy-cle at S phase , and increased the protein level of PC-NA in LoVo cells apparently .Res increased the level of Bad in LoVo cells, but decreased the level of Bcl-2. Although Res exerted no substantial effects on total lev-el of p38 MAPK, it markedly increased the phospho-rylation level of p38 MAPK in LoVo cells.p38 MAPK inhibitor promoted the proliferation , and decreased the anti-proliferation effect of Res on LoVo cells .Moreo-ver , the effects of Res on the level of Bcl-2 and Bad were both reduced by the p 38 MAPK inhibitor .Con-clusions Res can inhibit the proliferation of LoVo cells, which may be partly mediated by promoting the phosphorylation of p38 MAPK.
7. Determination of ethylenediamine in the air of workplace by Ion chromatography
Changmei ZHOU ; Hao ZHANG ; Jian WU ; Linyun ZHANG ; Xiaojuan LI
Chinese Journal of Industrial Hygiene and Occupational Diseases 2018;36(10):776-778
Objective:
To establish a method to detect the concentrations of ethylenediamine in the air of workplace by Ion chromatography.
Methods:
Ethylenediamine in the air of workplace adsorpted by Silicone tube, then ultrasonic desorped by Sulfuric acid solution (0.1 mol/L) , the desorption solution qualitative and quantitative determination by Ion chromatography.
Results:
For ethylenediamine, the calibration curves were liner in the range of 0.1-100.0 μg/ml, the method detection limit is 0.1 μg/ml, the minimum detectable concentration is 0.13 mg/m3 (in terms of sampling 7.5 L) . The desorption efficiency is more than 90% on spiked levels of 2.0, 4.0 and 8.0 μg/ml, the average desorption efficiency of the method was 96.1%, the precision was 1.8%-2.1%. The recovery rate of the added standard is 98.5%-104.2%.
Conclusion
The method simplifies the preprocessing steps and improves the sensitivity. All the indexes are in conformity with the requirements of the method formulation guide. It is suitable for the determination of ethylenediamine in the air of the workplace.
8.Effects of photodynamic therapy alone or in combination with antifungal agents on the apoptosis of planktonic and biofilm cells of Exophiala dermatitidis
Yuting XU ; Wenqian ZHENG ; Lujuan GAO ; Yi SUN ; Linyun LI ; Ming LI ; Tongxiang ZENG
Chinese Journal of Dermatology 2018;51(7):515-518
Objective To evaluate the in vitro effects of photodynamic therapy alone or in combination with antifungal agents on the apoptosis of planktonic and biofilm cells of Exophiala dermatitidis (E.dermatitidis).Methods The planktonic suspensions of E.dermatitidis were prepared,and the biofilms of E.dermatitidis were prepared via a modified 96-well plate-based methods.Planktonic and biofilm cells of E.dermatitidis were separately divided into several groups:antifungal agent groups treated with antifungal agents alone,photodynamic therapy group receiving photodynamic therapy alone,combination groups receiving photodynamic therapy followed by the treatment with antifungal agents,and blank control group receiving no treatment.These antifungal agents included amphotericin B,posaconazole,voriconazole and itraconazole.The concentrations of these antifungal agents were all 1 mg/L,and the treatment with antifungal agents lasted 2 hours.Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to detect the apoptosis of planktonic and biofilm cells of E.dermatitidis in all the groups.Results The antifungal agents and photodynamic therapy both affected the apoptosis of planktonic (both P < 0.001) and biofilm cells (beth P < 0.05) of E.dermatitidis.The apoptosis rates of E.dermatitidis planktonic cells in the control group,amphotericin B group,posaconazole group,voriconazole group and itraconazole group were 11.67% ± 0.21%,13.30% ± 1.78%,14.30% ± 3.61%,14.51% ± 1.91%and 36.17% ± 4.00% respectively.The apoptosis rate of E.dermatitidis planktonic cells was significantly higher in the itraconazole group than in the control group (P < 0.05),but no significant differences were observed between the other 3 antifungal agent groups and control group (all P > 0.05).The photodynamic therapy group also showed a significantly higher apoptosis rate of E.dermatitidis planktonic cells (41.37% ±7.80%) compared with the control group (P < 0.05).After the treatment with photodynamic therapy combined with amphotericin B,posaconazole,voriconazole or itraconazole,the apoptosis rates of E.dermatitidis planktonic cells were 29.23% ± 6.71%,37.23% ± 10.86%,43.57% ± 6.42% and 69.87% ± 3.53% respectively.Moreover,the photodynamic therapy + voriconazole group and photodynamic therapy + itraconazole group both showed significantly higher apoptosis rates compared with the voriconazole group and itraconazole group respectively (both P < 0.05).The apoptosis rate of E.dermatitidis biofilm cells was significantly higher in the photodynamic therapy group than in the control group (32.00% ± 0.43% vs.25.30% ± 1.31%,P < 0.05),as well as in the photodynamic therapy + amphotericin B than in the amphotericin B group (P < 0.05).Conclusion Photodynamic therapy combined with antifungal agents can markedly promote the apoptosis of planktonic and biofilm cells of E.dermatitidis.
9.Protective effect of low salt diet on vascular remodeling induced by high-fructose
Lingyu ZHANG ; Siqi LI ; Dong WU ; Haojia CHEN ; Lijin WANG ; Linyun ZHENG
Chongqing Medicine 2018;47(7):875-878,882
Objective To investigate the effect of low salt diet on vascular remodeling of rat induced by high fructose(HF).Methods Wistar male rats weighed 180-200 g were fed for 8 weeks and randomly divided into 6 groups:(1) control group was given the normal fodder and distilled water;(2) high fructose group(HF) was given normal fodder (0.5 % NaCl,w/w) and fructose water(10 %,w/v);(3) high-salt group (HNa) was given high salt fodder (7 % NaCl,w/w) and distilled water;(4) high fructose combined with high salt diet group(HFNa) was simultaneously given high salt fodder and 10 % fructose water;(5)high fructose combined low salt group(HFLNa) was simultaneously given low salt fodder and 10% fructose water;(6) high fructose combined with spirotaclone group(HFE) was given 10% fructose water for 4 weeks and then added with spirotaelone(50 mg · kg-1 · d-1 by tube feeding) for continuous 4 weeks.The changes of arterial blood pressure,vascular wall histological evaluation and expression of α-SMA and fibronectin in vascular wall were detected in each group.Results (1) Compared with the blood pressure[(111.03 ±9.17) mm Hg] in the control group,the blood pressure in the HF and HNa groups were (133.94± 5.86) mm Hg and (128.09±7.56) mm Hg respectively,which were significantly increased(P<0.05);(2) HF mainly caused the hyperplasia of vascular wall middle layer smooth muscle.The a-SMA expression results in the HF group was (0.006 3 ±0.000 21),which in the control group was (0.004 6 ± 0.000 31),the difference was statistically significant(P<0.05),moreover which promoted the elastic fibers increase;while HNa mainly stimulated the elastic fibers to thicken and extracellular matrix deposition,the fibronectin expression was 0.002 6 ± 0.000 2 in the HNa group and (0.004 7±0.000 2)in the HF group,compared with(0.001 3±0.000 1)in the normal group,which were significantly increased(P<0.001);(3) the blood pressure was (106.04±9.59) mm Hg in the HFLNa group,(103.99±7.12) mm Hg in the HFE group,compared with(133.94±5.86) mm Hg in the HF group,showing that the blood pressure in the HFLNa group and HFE group was significantly decreased compared with the HF group (P<0.05);moreover the vascular remodeling in the HFLNa group(0.006 8±0.000 2) and HFE group (0.004 2±0.000 4) was improved,and compared with the HF group(0.006 3±0.000 2),α-SMA expression was significantly decreased (P<0.05).Conclusion Low salt diet can effectively improve vascular remodeling induced by HEF.
10.The Effect of Hydroxyapatite Ultrofine Powder on the Immunity Function of Tumor-bearing Mice
Qinghua XIA ; Haiyang NIE ; Daoda CHEN ; Yuhua YAN ; Linyun FENG ; Shipu LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2001;21(2):143-144
The inhibitory effect of hydroxyapatite ultrofine powder (HAUFP) on tumor and the effect on the immunity function of body were investigated. The levels of IL-2 in the spleen cells and serum TNF levels in the tumor-bearing mice at the 7th day and 14th after peritoneal injection of HAUFP were detected by using the methods of colorimetric analysis of MTT and crystal purple decoration, respectively. The disappearance of the ascites of the mice was observed. The results showed that the levels of IL-2 and TNF in the tumor-bearing mice were higher obviously in the drug-treated group than in the control group (P<0.01), the ascites growth was inhibited. It was suggested that HAUFP could increase the levels of IL-2 and TNF of the tumor-bearing mice and improve the immune function of body.