1.Construction of eukaryotic expression vector carrying pprI gene of Deinococcus radiodurans and its radioresistant effect
Ling WEN ; Yi SHI ; Lili REN ; Ying CONG ; Zhanshan YANG
Chinese Journal of Radiological Medicine and Protection 2014;34(8):563-568
Objective To construct the eukaryotic expression vector of pprI gene from Deinococcus radiodurans R1 and investigate its radioresistant effects in eukaryotic cells.Methods A recombinant vector pEGFP-c1-pprI was constructed by DNA recombinant technique.The empty vector pEGFP-c1 and the pEGFP-c1-pprI were transferred into human lung epithelial cells Beas-2B by LipofectamineTM 2000,respectively.Then the infected cells were screened in order to develop a cell line with stable expression of pprI gene.Cell survival rate was tested by clone-forming assay.Cell cycle distribution and apoptosis were detected by a flow cytometry.The fluorescence intensity of reactive oxygen species (ROS) was observed by a fluorescent microscope.γ-H2AX foci in the irradiated cell was detected by immunofluorescence.Results The eukaryotic expression plasmid of pprI prokaryotic gene was constructed and PprI fusion protein was expressed in human lung epithelial cells successfully,and the cell line (2BG) with a stable pprI gene expression was established.After irradiation,the cell survival fraction of 2BG cells was significantly higher than Beas-2B cells so that the value of D0 、Dq and N of the survival curve were increased.Moreover,the fluorescence intensity of ROS and the number of γ-H2AX foci in 2BG cells were also lower than those of B eas-2B cells(F =16.73,19.47,6.94,P < 0.05).Between these two cell lines,the apoptosis rate and cell cycle G2 arrest also had significant difference (F =139.73,237.92,P < 0.05).Conclusions The pprI gene from Deinococcus radiodurans RI can be stably expressed in the eukaryotic cells and it allows the transferred cells to have a radioresistant function.
2.Analysis of survey results of iodine deficiency disorders in Huai'an City Jiangsu Province in 2011
Cong-ying, JIA ; Wen-zhou, YANG ; Wei, HU
Chinese Journal of Endemiology 2013;32(4):416-418
Objective To evaluate the status of iodine deficiency disorders(IDD) in Huai'an City in 2011 and to provide a scientific basis for adopting target prevention strategies timely and adjusting the intervention policy scientifically.Methods According to The National Project of Surveillance on IDD,probability sampling method (PPS) was used to select 30 counties in Huai'an City,and one primary school was selected randomly from each county,then 40 children aged 8 to 10 were randomly selected in each school.Children's thyroids were examined by palpation and the iodine in salt samples collected in their home was tested by direct titration.At the same time,urine samples from 12 children of the 40 selected children were collected and urinary iodine was tested by As(Ⅲ)-Ce4+ catalytic spectrophotometry.Questionnaire survey of IDD was conducted among 30 students of grade 5 and 5 housewives around that school.Results A total of 1200 children aged 8 to 10 were examined,and the goiter rate was 1.3%(15/1200).All the goiters were level Ⅰ.The goiters of each age group was 0.8%(3/390),1.8%(7/390) and 1.3%(5/420),respectively.The 360 urine samples were collected from children aged 8-10,and the median of urinary iodine was 171.5 μg/L with the proportions of < 20 μg/L,< 50 μg/L and < 100 μg/L were 0(0/360),1.1% (4/360) and 5.6% (20/360),respectively.A total of 1200 salt samples were tested in children's family.The coverage rate of iodized salt was 99.4% (1193/1200) and the qualified rate of iodized salt was 97.7% (1165/1193).The consuming rate of qualified iodized salt was 97.1%(1165/1200),and non iodized salt rate was 0.6%(7/1200).The knowledge rates of IDD were 82.7% (2234/2700) and 69.8% (314/450) in 900 students and 150 housewives,respectively.Conclusions In Huai'an City,except the knowledge rate of IDD,the goiter rate and urinary iodine of children aged 8-10,iodized salt consumption maintain at the stage of the target level.In addition to strengthen the monitoring of iodized salt and to supply qualified iodized salt,it should also increase the focus on health education propaganda in the future.
3.The indirect diagnosis for lactase deficiency
cong-min, ZHAO ; wei-ming, LI ; ying, YANG
Journal of Applied Clinical Pediatrics 1986;0(01):-
0.1), there is not significant difference between them. Comparing with blood method, the sensitivity and specificity of urine method are 94 .1 % and 91. 6 % respectively.Conclusions To determine the B-gal and U-gal, O-toluidine method can be regarded as an indirect diagnostic method of LD.
4.Comparison of in vivo and in vitro Permeation Behaviors of the Ethosome Gels of Testosterone and Its Es-ters
Shu MENG ; Yang LI ; Cong ZHANG ; Jing QU ; Wei ZHANG ; Ying JIN ; Jing GUO ; Miao LI
China Pharmacy 2015;(22):3072-3074
OBJECTIVE:To compare in vivo and in vitro permeation behaviors of the ethosome gels of testosterone,testoster-one propionate and testosterone undecanoate. METHODS:The ethosome gels of testosterone,testosterone propionate and testoster-one undecanoate were prepared. With cumulative permeating amount and permeation rate as the indexes,Franz diffusion cell and HPLC were employed to compare in vitro permeation behaviors of 3 kinds of ethosome gels in mouse skin. With testosterone patch as the positive control drug, electrochemistry method was adopted to detect the concentration of testosterone in plasma 0,3,6, 9,12,24,36 and 48 h after applying such 3 kinds of ethosome gels on the back of rats,and then pharmacokinetic parameters were calculated with DAS 2.0 software. RESULTS:24 h cumulative permeating amounts of the ethosome gels of testosterone,tes-tosterone propionate and testosterone undecanoate were(234.31±13.8),(175.63±41.1)and(72.60±15.3)μg/cm2,and the per-meation rates were(10.25±1.9),(7.64±1.4)and(2.96±0.8)μg/(cm2·h),respectively. The pharmacokinetic parameters of the above-mentioned three kinds of ethosome gels and the positive control drug were respectively as follows as cmax of(20.19±2.57), (17.50±2.91),(0.23±0.04),(14.97±2.12)ng/ml,t1/2Ka of(2.80±0.45),(3.36±0.59),(4.02±0.62),(4.20±0.71)h,AUC0-48 h of(13.85±1.96),(13.93±2.13),(0.35±0.07),(11.76±2.31)ng·h/ml. CONCLUSIONS:in vivo and in vitro permeation behav-iors of the ethosome gels of testosterone and testosterone propionate are fairly good.
5.Effects of Highly Concentrated Potassium Chloride via Central Venous Catheterization on Severe Hypokalemia
Wei WEI ; Yan LI ; Ying ZHANG ; Yunxi WANG ; Su ZHANG ; Cong YANG
Journal of Kunming Medical University 2016;37(11):113-115
Objective To investigate the clinical effects and feasibility of highly concentrated Potassium Chloride via central venous catheterization by micro pump on severe hypokalemia patients in EICU.Methods A totlal of 120 severe hypokalemia patients in our department were randomly divided into experimental group (treated with highly concentrated Potassium Chloride) and control group (normal treatment group) respectively,and treated with Potassium Chloride liquid of different concentration.Potassium levels in blood were checked every hour and the time for reaching standard potassium level (4.0mmol/L) and the total volumes of infusion fluid within 24 hours in the two groups was compared.Results The mean time for reaching standard potassium level and the total volumes of infusion fluid within 24 hours in the experimental group,(12.83 ± 3.57) h and (402.56 ± 54.61) ml respectively,were significantly less than those in the control group (P <0.01),(23.18 ±4.98) h and (2875.2 ± 206.26) ml respectively.Conclusion Highly-concentrated potassium chloride injection via central venous catheterization by micro-pump is a safe,effective and feasible treatment on the patients with severe hypokalemia,especially on the patients with volume-overloaded heart and severe hypokalemia,which is worthy of further clinical research.
6.Influence of hyperhomocysteinemia on signal pathway of KV1 .3-CaN-NFAT in lymphocytes of patients with acute ST-segment elevation myocardial infarction
Yuling WANG ; Shaobin JIA ; Jianjun HOU ; Guangzhi CONG ; Shaobing YANG ; Ying CUI ; Kai WANG
Journal of Xi'an Jiaotong University(Medical Sciences) 2016;37(3):412-417
Objective To observe the influence of different level of hyperhomocysteinemia on mRNA and protein expressions of KV1 .3 ,CaN,NFAT,IL-6 and TNF-αin lymphocytes of patients with acute ST-segment elevation myocardial infarction (STEMI).Methods We selected 90 STEMI patients and divided them into three groups according to the level of plasma homocysteine:the first experimental group (STEMI group,Hcy<1 5μmol/L, n=30),the second experimental group (STEMI with mild Hhcy group,Hcy 15~30μmol/L,n=30)and the third experimental group (STEMI with intermediate Hhcy group,Hcy>30 μmol/L,n=30 ).Another 30 healthy examined people were selected as control group (n=3 0 ).Peripheral lymphocytes were isolated by Ficoll density gradient centrifugation.The Hcy in the plasma was measured with the IMX assays.Real-time quantitative PCR (RT-PCR)was used to detect mRNA expressions of KV1.3,CnAα,NFAT1,IL-6 and TNF-αand Western blot technique was used to detect the expressions of KV1.3,CnAαand NFAT1.Results The mRNA and protein expression levels of KV1.3,CnAαand NFAT1 in each experimental group were significantly higher than those in control group (P<0 .0 5 or P<0 .0 1 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0.05 or P<0.01)and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.05 or P<0.01).The mRNA expression levels of IL-6 and TNF-αin each experimental group were significantly higher than those in control group (P<0.05 or P<0.01 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0 .0 5 or P<0 .0 1 )and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.01).Plasma total Hcy levels were positively correlated with mRNA and protein expressions of KV1.3 in all observed groups (r=0.503 P=0.000,r=0.726 P=0.000).Conclusion The higher level of Hcy in plasma,the higher mRNA and protein expression levels of KV1.3,CnAα,NFAT1 and the higher mRNA expression levels of IL-6,TNF-αin the lymphocyte of STEMI patients,which may be one mechanism for Hcy exacerbating the inflammatory reaction of STEMI.
7.Investigation and analysis of trace element levels in children with osteomalacia malformation from endemic fluorosis area in Shuicheng, Guizhou
Fu-cheng, LI ; Liang, QI ; Xu-zi, CONG ; Ying, YANG ; Shu-lin, YAN
Chinese Journal of Endemiology 2011;30(2):197-201
Objective To investigate the levels of trace elements such as fluorine(F), and aluminium (Al)etc. of osteomalacia malformation children and to make etiological diagnosis in reference with clinical manifestations.Methods Urine and occipitalia hairs of 14 diseased children(patient group) from endemic fluorosis area and 13 healthy children(control group) from non-endemic area were included in the study on November, 2008, and contents of 10 elements of fluorine(F), aluminum(Al), chromium(Cr), manganese(Mn), ferrum(Fe), cuprum(Cu), zinc(Zn), arsenic (As), selenium(Se), strontium(Sr), and barium(Ba) were tested. The data were analyzed with medical soft package PEMS 3.1. Results Urinary contents of F, Al, Mn, Cu, Sr, and Se(1.18 mg/L, 112.6 μg/L,6.62,29.86 mg/L, 177.5,4.23 ng/L) in patient group were significantly different from those in control group (0.48,47.1,2.04,16.61 mg/L, 55.17,15.52 ng/L, t = 4.592,2.486,4.850,2.210 2.078,2.912, all P< 0.05); Hair contents of Al, Mn, As, Sr, Ba, Fe, and Se in patient group(59.27,5.26,0.96,1.50,1.29,297.13,0.45 mg/kg)were significantly different from those of control group( 18.69,0.72,1.09,0.62,0.68,69.02,1.323 mg/kg, t = 4.583,6.318,3.309,2.704,5.606,6.294, all P < 0.05); in patient group, the correlation coefficients of urinary Fe to Al,Zn, As, and Se were all bigger tan 0.662(all P< 0.05), those of urinary Se to Mn, Ba, Cu, Zn, Sr, and As were all bigger than 0.694(all P< 0.05), those among urinary Mn, Sr, As, and Ba were bigger than 0.550(all P<0.05), those of hair Al to Mn, Cr, Fe, and Cu were bigger than 0.732(all P< 0.05), those of hair Ba to Mn,Cr, Fe, and Sr, and of hair Mn to Cr and Fe, and those between Cr and As, between Cu and Sr were all bigger than 0.686 (all P < 0.05). In control group, the correlation coefficients of urinary Cu to Zn, Se, and Ba, those of Zn to Se and Ba, and those of Cr to Mn and Ba were all bigger than 0.516(all P < 0.05), those of hair Al to Mn,Fe, Cu, As, and Se, and those of hair Se to Fe, Cu, and As, those of hair Fe to Mn, Cu, and As, those of hair Cu to Zn and As, and that between Zn and As were bigger than 0.739(all P < 0.05). The correlation coefficient of urinary F to Se in patient group(0.762) was significantly different from that in control group( - 0.469, u = 2.079,P < 0.05). Conclusions The burden of F and Al of osteomalacia malformation children in endemic fluorosis area of Shuicheng county is too high. The contents of multi-elements in urine and hairs and their correlation are coincident with high levels of Al and F and they cause network increase of multi-element content changes and their correlation. According to bone X-ray features combining with the living environment, the diagnosis of endemic Al-F fluorosis can be made. The biological significance of reducing urinary and hair Se levels and the correlations of F and Al need to be further studied.
8.Review considerations on the design and evaluation of the oral sensory features in pediatric medications
Yan-zhe SUN ; Duan-duan CONG ; Ying GENG ; Zhi-min YANG
Acta Pharmaceutica Sinica 2023;58(11):3160-3164
Recent years, China has released a series of policies to encourage drug research and development in pediatric populations, aiming to meet pediatric populations' medical needs. Because of the physical and psychological developmental characteristics, tolerance of aversive feelings when taking medications are different between pediatric population and adults. So pediatric populations are at a relatively higher risk of not taking medications as prescribed when the medication tasted unpleasant. Therefore, sound design and evaluation of oral sensory features have important clinical significance and value in developing pediatric medications. "Technical guidance for the design and evaluation of the oral sensory features of pediatric drugs (trial version) " was released in November 2022, by Centre for Drug Evaluation, National Medical Products Administration of China. Based on the guidance, this article will introduce the drafting background and review considerations, hoping to provide reference for the design and evaluation of oral sensory features, and promote drug developing in pediatric population.
9.Study on identification of Sarcandra glabra and Chloranthus spicatus's leaves by PCR amplification of specific alleles.
Yi-cong WEI ; Ying CHEN ; Lin-quan LUO ; Qun-xiong YANG ; Yi-Juan CHEN ; Yi-chi LIANG ; Su-Rong CHEN
China Journal of Chinese Materia Medica 2014;39(17):3259-3262
The paper is aimed to identify SNP in Sarcandra glabra and Chloranthus spicatus, and authenticate S. glabra from Ch. spicatus and the mixture by using PCR amplification of specific alleles. SNPs in the ITS sequences of S. glabra and Ch. spicatus were found by ClustulX 2. 1 program and Bioedit software. Primers for authentic S. glabra and Ch. spicatus was designed according to the SNP site, and ITS sequence universal primers plus to the authentic primer to construct a multi-PCR reaction system, and then optimized the PCR reaction system. Five hundred and eighty band special for S. glabra and 470 bp band special for Ch. spicatus were found by using multi-PCR reaction. The multi-PCR reaction system could be applied to identify S. glabra and Ch. spicatus's leaves.
DNA, Plant
;
analysis
;
genetics
;
DNA, Ribosomal
;
genetics
;
DNA, Ribosomal Spacer
;
analysis
;
genetics
;
Magnoliopsida
;
classification
;
genetics
;
Plant Leaves
;
genetics
;
Polymerase Chain Reaction
;
Polymorphism, Single Nucleotide
;
RNA, Ribosomal
;
genetics
;
RNA, Ribosomal, 18S
;
genetics
;
RNA, Ribosomal, 5.8S
;
genetics
;
Species Specificity