1.Performance verification for high performance liquid chromatography detection of glycated hemoglobin
Limei LUO ; Fengxue WANG ; Guangbing ZHENG
International Journal of Laboratory Medicine 2015;(16):2341-2343
Objective To verify the detection performance of glycated hemoglobin (HbA1c)by using the BIO-RAD D-10 high performance liquid chromatography method,and to evaluate whether the detection performance can satisfy the requirements of the laboratory and clinical diagnosis and treatment.Methods The method of CLSI EP1 5A2 was used to verify the accuracy and preci-sion.The linear correlation and reference range were verified respectively by linear regression analysis and the method of the WS/T 402 2012.Results The calibration value of HbA1c calibrator level 1 was 5.3% and the validation limit of measured results was 1. 63%-8.78%.The calibration value of HbA1c calibrator Level 2 was 10.0%,and the validation limit of measured results was 4.76%-13.64%.Both calibration values were included in the 95% confidence range of measured values,so the accuracy verifica-tion passed.Laboratory variation coefficient (CV)for HbA1c with concentration of 5.24% was 1.01% and for 9.84% was 0. 54%,both less than the precision that the manufacturer declared,so the precision verification passed.The equation of linear regres-sion analysis for HbA1c linearity calibrator measured value and target value wasY =1.008X +0.023,with correlation coefficient in-dex r 2 of 0.999,so the linearity was good.The measured results of 20 healthy adults specimens were in the reference interval(4.1%-6.2%)that the manufacturer provided.Conclusion Accuracy,precision,linear correlation and reference range of HbA1c meas-ured by BIO-RAD D-10 HPLC detection method were verified.It can provide the accurate and credible measurement results for clin-ical diagnosis and treatment,and can be used in the detection of clinical specimens.
2.Study on effect of dendrobium nobile polysaccharides on expression of WT1 gene in myeloid leukemia cells
Xiaojun GE ; Limei ZHENG ; Yonglun WANG ; Yanping TANG
Chongqing Medicine 2015;(10):1305-1307,1310
Objective To explore the effect of dendrobium nobile polysaccharides on the expression of the WT 1 gene in my-eloid leukemia cells.Methods The CCK8 assay was used to detect the half maximal inhibitory concentration(IC50 )of dendrobium nobile polysaccharides in 3 kinds of leukemia cells;the each kind of leukemia cells were divided into the treatment group and the control group.The cells in the control group maintained the normal growth,while which in the treatment group were given the den-drobium nobile polysaccharides stimulation.The Hoechst33258 staining was used to detect the apoptosis situation of the cells in the two groups.The WT l gene expression level was detected by the real-time PCR and the protein expression levels of WT1,53 and BAX were detect the Western blot.Results Dendrobium nobile polysaccharides had the similar IC50 values in 3 kinds of myeloid leukemia cells,which were (110.71±6.49),(104±48.50),(96.66±5.10)mg/mL respectively,the difference among them had no statistical significance (P >0.05);the apoptosis rate of the treatment group was higher than that of the control group (P <0.05);the expression levels of WT1 gene and protein in the treatment group were decreased compared with the control group(P <0.05), while the expression of P53 and BAX protein was increased.Conclusion Dendrobium nobile polysaccharides can obviously decrease the expression level of WT1 protein,and has a certain killing effect on myeloid leukemia cells.
3.Effects of acitretin on the level of interleukin-8(IL-8)in the peripheral blood of patients with pustular psoriasis
Jing TIAN ; Yicheng ZHOU ; Shuyun ZHENG ; Limei SUN ; Xin XIE
Chinese Journal of Dermatology 2011;44(10):732-734
Objective To investigate the changes of IL-8 in peripheral blood of patients with pustular psoriasis after acitretin treatment.Methods Dual antibody sandwich enzyme-linked immunosorbent assay (ELISA)was performed to measure the IL-8 levels in the peripheral blood of 30 patients with pustular psoriasis and 30 patients with psoriasis vulgaris before and after treatment,and in 30 normal human controls.The relationship between the IL-8 level and disease severity was assessed for patients with pustular psoriasis.Results After acitretin treatment,the condition was improved greatly in patients with pustular psoriasis,together with a significant decrease in the level of IL-8 in the peripheral blood(57.07 ± 12.02 pg/ml vs.96.84 ± 14.68 pg/ml,P < 0.05).Conclusion IL-8 may be involved in the therapeutic mechanism of acitretin in pustular psoriasis.
4.Positive pressure biological protective suit under high-risk biological contamination environment:statusquo,problems and lessons learned
Runze WANG ; Zheng WANG ; Jinhui WU ; Ying YI ; Limei HAO
Military Medical Sciences 2015;(5):390-392
People wearing positive pressure biological protective suits might face some potential safety risks when work -ing in a high-risk biological contamination environment .These risks could be effectively prevented by simulation experi-ments.This paper elaborates on the progress in risk studys on positive pressure biological protective suits in terms of dynam -ic detection methods and risk assessment , and points out the problems to be solved and the direction of future development .
5.A Comparative Study of Performances on the Open -set Monosyllable Test of Typical Children and Hearing Impaired Children at Age 3
Min SHEN ; Wenfang ZHENG ; Xiaoyue LU ; Limei YU
Journal of Audiology and Speech Pathology 2013;(5):460-464
Objective The present study analyzed the performances of hearing impaired children and normal hearing children at age 3 on the open set monosyllables test to provide reference for rehabilitation .Methods 30 typi-cal children and hearing impaired children at age 3 were randomly selected and tested using the open -set monosylla-ble test in evaluation of auditory response to speech (EARS) .Results ① There were significantly differences be-tween different test items in both groups ,the decreasing order of the score was tone> vowel>consonant>word(P<0 .01) ,and the scores on vowels ,consonants and words of hearing -impaired children were respectively signifi-cantly lower than that in normal hearing children(P<0 .001) .②There were similarities and differences between the mistakes of typical children and hearing impaired children .Blade-palatals and laterals in both groups achieved the lowest accuracy .While the accuracies on bilabials ,labiodentals ,nasals and plosives were highest in typical children , the accuracy on velar was highest in hearing impaired children .And the accuracies of consonants and nasal vowels on each place and manner of articulation in hearing -impaired children were significantly lower .Conclusion The study showed that the performances on vowels ,consomants and words of hearing -impaired children aged 3 in the open set monosyllables test were poorer than that in normal hearing children .The main errors were between the same places or methods of articulation ,especially for Blade -palatals and laterals .It is possible related to the deficiency of lan-guage input with the hearing impairment and the age with HA/CI .
6.The effect of shenfu injection on immune function in severe trauma patients
Limei LEI ; Xiang LAN ; Yan KAO ; Hao ZHENG ; Jie PAN ; Dingding HUANG
Chinese Journal of Primary Medicine and Pharmacy 2015;(19):2928-2930,2931
Objective To investigate the effect of shenfu injection on immune function in severe trauma patients.Methods 60 severe trauma patients were divided into the control group (n =30)and shefu group (n =30) by random number table.Other 30 cases were chosen as the health control group at the same phase.All patients were received conventional treatments,however,patients of the shenfu group were additionally received the shenfu injection treatment in the early stage.The CD +3 ,CD +4 ,CD +8 cell,human leukocyte antigen (HLA -DR),interleukin -1(IL -1),interleukin -6(IL -6)were detected on 3rd and 7th day by double -antibody sandwich enzyme -linked immu-nosorbent assay (ELISA).Results Compared to the health control group,the IL -1,IL -6 in the control and shenfu group were significantly higher than the health control group(f=7.128,q =9.212,10.112,all P <0.05).The IL -1and IL -6 in the control and shenfu group were significantly increased on 3rd day (t =11.126,10.013,all P <0.05)and decreased on 7th day(t =17.121,14.213,all P <0.05).The IL -1 and IL -6 in shenfu group were sig-nificantly lower than that of the control group(χ2 =4.113,10.117,all P <0.05).The CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were significantly lower than the health control group(f=11.071, q =10.229,12.032,all P <0.05).On 3rd day,the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly increased(t =10.013,P <0.05).On 7th day,CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in the control and shenfu group were both increased(t =11.126,15.932,all P <0.05).And the CD +3 ,CD +4 ,CD +8 ,HLA -DR and CD +3 /CD +8 rate in shenfu group were significantly higher than the control group(χ2 =3.771,P <0.05).Conclusion Shenfu injection can regulate immune function in severe trauma and improve clinical treatment.
7.Combined treatment with probiotics and fructooligosaccharides in a mouse model of nonalcoholic fatty liver disease and its possible mechanism
Fangfang YAO ; Huang HUANG ; Limei BAI ; Yirui DING ; Lu MEI ; Simeng LIU ; Pengyuan ZHENG
Chinese Journal of Microbiology and Immunology 2017;37(7):527-533
Objective To investigate the effectiveness of Lactobacillus paracasei N1115 combined with fructooligosaccharides (FOS) in the treatment of nonalcoholic fatty liver disease(NAFLD) in a mouse model and to analyze the possible mechanism.Methods Fifty male C57BL/6 mice were randomly divided into five groups and respectively given normal diet (ND group), high-fat diet (HFD group), HFD containing Lactobacillus paracasei N1115 (HFD+L) (2.2×109 CFU/ml), HFD containing FOS (HFD+FOS) (4 g/kg per day) and HFD containing Lactobacillus paracasei N1115 and FOS for 16 consecutive weeks.Levels of triglyceride (TG), total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), lipopolysaccharide (LPS) and diamine oxidase (DAO) in serum samples from each group were measured.Expression of tight-junction proteins (Claudin-1 and Occludin), p38 and phosphorylated p38 (p-p38) in intestinal tissues were analyzed.Results Compared with the HFD group, the HFD+FOS+L group showed decreased levels of TC, TG, LDL, LPS and DAO in serum samples, but increased serum HDL level (P<0.05).Moreover, combined treatment with Lactobacillus paracasei N1115 and FOS alleviated liver lipid deposition, significantly increased the expression of Claudin-1 and Occludin in intestine and inhibited the phosphorylation of p38 (P<0.05).Conclusion Lactobacillus paracasei N1115 combined with FOS may increase the expression of Claudin-1 and Occludin through inhibiting the phosphorylation of intestinal p38, which is conducive to maintaining the intestinal mucosal barrier integrity and alleviating NAFLD.
8.Effects of Dihuangyinzi on behaviors and RAGE/p38 pathway in APP/PS1 mice
Zhongyuan PIAO ; Lifen YAO ; Lin SONG ; Yichan LU ; Zhuo JIANG ; Yang ZHENG ; Zhiyong DI ; Limei AO
Chinese Journal of Behavioral Medicine and Brain Science 2017;26(7):577-581
Objective To investigate the effects of Dihuangyinzi(DHYZ) on behaviors and RAGE/p38 pathway in APP/PS1 mice.MethodTwenty APP/PS1 dementia mice were randomly divided into model group(n=10) and Chinese medicine group(n=10).The blank group was C57 BL/6 J normal mouse(n=10).The mice in Chinese medicine group were intragastric administration with DHYZ (9.75 g·kg-1·d-1).The mice in model group and blank group were treated with distilled water.After 30 days,the abilities of learning and memory of mice were detected by Morris water maze.The expression of amyloid-beta1-42(Aβ1-42) in the hippocampus and cortex was detected by immunohistochemistry.Reactive oxygen species of brain tissue were detected by DCFH-DA Methods in the brain of APP/PS1 mice.Gene expression level of receptor for advanced glycation end products(RAGE) was measured by real-time polymerase chain reaction (RT-PCR) in the cortex and hippocampus of APP/PS1 mice.The expression of phospho-mitogen-activated protein kinases (p38) was analyzed with Western blot and immunofluorescence analysis in the cortex and hippocampus of APP/PS1 mice.Results Behavioral Results showed that DHYZ significantly increased the distance((23.088±7.083)cm) and residence time((1.961±1.230)s)of effective area in Morris water maze on the fifth day(P<0.05,P<0.01)and remarkably increased the number of effective area crossings((1.607±0.405) times) and plats((0.893±0.283) times) in Morris water maze on the fifth day(P<0.01,P<0.05).DHYZ also significantly reduced the intracelluar ROS level(122.611±7.630) in the brain(P<0.01),and DHYZ could depress the expression of RAGE(1.467±0.081,7.983±0.136) and phosphorylation of p38 (0.376±0.026,0.538±0.016)in the cortex and hippocampus of APP/PS1 mice(P<0.01,P<0.05).Conclusions The Results demonstrate that DHYZ can partly improve memory impairment of APP/PS1 mice by the inhibition of RAGE/p38 pathway.
9.Effects of cholesterol-lowering probiotics on the metabolism of bile acid in a rat model of non-alcoholic fatty liver disease and the possible mechanism
Limei BAI ; Pengyuan ZHENG ; Jun ZHANG ; Fuguang LI ; Lu MEI ; Huang HUANG ; Yirui DING ; Simeng LIU
Chinese Journal of Microbiology and Immunology 2016;36(2):110-116
Objective To investigate the effects of two cholesterol-lowering probiotics, DM9054 (Lac-tobacillus Rhamnosus GG, LGG) in combination with 86066 (Lactobacillus plantarum WCFS1, LP), on the metabolism of bile acid via a rat model of non-alcoholic fatty liver disease (NAFLD) and the possible mecha-nism. Methods Twenty-one SD male rats were randomly divided into three groups including control group, NAFLD model group and probiotics intervention group. Rats in the control group received normal diet. The rat model of NAFLD was established by feeding rats with chronic high fat diet (45% of calories derived from fat di-et) for 20 weeks. Rats in the probiotics intervention group were given high fat diet together with cholesterol-low-ering probiotics through oral gavage. General indexes of each group including body weight and the levels of tri-glyceride, cholesterol and CK18-M30 in serums samples were detected. The expression of cholesterol 7-alpha hydroxy-lase (CYP7A1), fibroblast growth factor receptor 4 (FGFR4), farnesoid X receptor (FXR), fibroblast grwoth factor 15 (FGF15) and apical sodium-dependent bile acid transparter(ASBT) at mRNA level were de-tected by using real-time polymerase chain reaction (real-time PCR). Western blot assay was used to detect the protein expression of CYP7A1, FXR in liver tissues and ASBT in ileum tissues. The expression of FXR in liver and ileum tissues were analyzed by immunohistochemistry. Results Rats with NAFLD showed loss of body weight and decreased levels of the serological markers after treating with the probiotics (P<0. 05). Compared with the rats in model group, enhanced expression of CYP7A1 and inhibited expression of FXR in liver tissues, activated FXR-FGF15 pathway in ileum tissues as well as down-regulated expression of ASBT in ileum tissues were detected in rats receiving probiotics intervention (P<0. 05). No significant difference in the expression of FGFR4 at mRNA level was observed between NAFLD rats with or without probiotics intervention (P>0. 05). Conclusion Probiotics intervention might up-regulate the expression of CYP7A1 by suppressing the FXR path-way in liver tissues and inhibiting the expression of ASBT in ileum tissues. Treating NAFLD rats with cholester-ol-lowering probiotics could activate the FXR-FGF15 pathway in ileum tissues and enhance the metabolism of bile acid, which contributed to the alleviation of NAFLD.
10.Effects of lactitol and Bifidobacterium infantis on AQP3 and ICC in a rat model of constipation
Yirui DING ; Pengyuan ZHENG ; Fuguang LI ; Lu MEI ; Huang HUANG ; Limei BAI ; Simeng LIU
Chinese Journal of Microbiology and Immunology 2015;35(12):890-895
Objective To analyze the roles and mechanisms of lactitol and Bifidobacterium infantis in the treatment of rat constipation and to investigate their effects on aquaporin3 (AQP3) and interstitial cells of Cajal (ICC) in colon tissues.Methods Thirty SD male rats were recruited in this study,6 of which were randomly selected as the control and the rest were given 4 mg/kg.d of loperamide for 5 consecutive days to construct the rat model of constipation.The rats with constipation were randomly divided into four groups including model group,lactitol treatment group,Bifidobacterium infantis treatment group and lactitol in combination with Bifidobacterium infantis treatment group.General indexes including food intake,water intake,body weight,fecal water content and intestinal transit rate of each rat were measured after receiving corresponding treatments for 7 consecutive days.The levels of substance P (SP) and vasoactive intestinal peptide (VIP) in serums samples were detected by ELISA.The expression of protein kinase A (PKA) and neurokinin-1 receptor (NK-1) at mRNA level in colon tissues were detected by real-time polymerase chain reaction (real-time PCR).Western blot assay and real-time PCR analysis were used to detect the expression of AQP3 and c-kit at protein and mRNA levels,respectively.Results Compared with the rats in model group,the levels of fecal water content and intestinal transit rate,the concentrations of SP and VIP in serums samples,the expression of PKA and NK-1 at mRNA level and the expression of AQP3 and c-kit at mRNA and protein levels were significantly increased in rats from the three treatment groups (P<0.05).The most effective treatment was lactitol in combination with Bifidobacterium infantis,followed by the lactitol treatment and then the Bifidobacterium infantis treatment.Conclusion The combination therapy with lactitol and Bifidobacterium infantis increased the serum levels of SP and VIP in rats with constipation.SP could enhance the contraction of gastrointestinal smooth muscles and improve the intestinal motility by binding to the NK-1 receptor on the membrane of ICC.VIP could promote the absorption of water in intestinal tracts,soften stools and alleviate constipation by upregulating the expression of AQP3 at both protein and mRNA levels via the cyclic adenosine monophosphate-PKA (cAMP-PKA) signaling pathway.