1.The establishment of a high resolution melting analysis based rapid screening system for UGT1A1 gene mutation
Hui YANG ; Liye YANG ; Lei ZHENG ; Zhen CAI
Chinese Journal of Laboratory Medicine 2017;40(2):101-104
Objective To develop a high-resolution melting ( HRM ) assay for rapidly screening Gilbert syndrome ( GS) and Crigler-Najjar syndrome ( CNS) associated with UGT1A1 defects.Method Methodology was developed .Then, we applied the established method to analyze 61 clinical samples from neonatal patients with severe unexplained unconjugated hyperbilirubinemia .Neonates with known risk factors for developing hyperbilirubinemia , such as ABO hemolysis, G6PD deficiency, sepsis, hypoxic ischemic encephalopathy were excluded .Five pairs of PCR primers were designed to detect the five common mutations (G211A, C686A, C1091T, C1352T and T1456G) in Asia population.PCR and HRM Assay conditions were optimized.UGT1A1 genotyping in clinical samples was performed by using the established HRM analysis , and all results were subsequently confirmed by direct DNA sequencing .Results The mutants were readily differentiated by using HRM analysis .In this study, 42 neonates were identified with UGT1A1 mutation, and 4 different known variants were detected .Conclusion HRM analysis in this study was economical, convenient, rapid, effective for screening UGT1A1 gene mutations, which can serve as an reliable method for the clinical diagnosis of GS and CNS and the large-scale molecular epidemiological research of UGT1A1 gene-related diseases.
2.Studies on macroscopic and microscopic characteristics of Ophiocordyceps xuefengensis.
Hao LIU ; Lan-ping ZHEN ; Ru-cai ZHU ; Shui-han ZHANG ; Hui-yong HUANG
China Journal of Chinese Materia Medica 2015;40(14):2820-2824
The macroscopic characteristics, tissue, caterpillar body wall and powder of Ophiocordyceps xuefengensis in different batch numbers were observed and researched by the macroscopic and microscopic identification methods. The result shows that the morphology, size, abdominal annulations of caterpillar, etc. of 0. xuefengensis are the macroscopic identification characteristics, the caterpillar body surface mycelium, body wall sculpture and crochets on abdominal legs are the microscopic identification characteristics. These characters are stable and regular discriminant features, which are proved to be the identification basis of O. xuefengensis. In addition, The characters such as crochets on abdominal legs arrange in two parallel ellipse rings, the inner crochets are long strip, and the external toes are unciform, are specific.
Animals
;
Hypocreales
;
cytology
;
Moths
;
anatomy & histology
;
cytology
;
microbiology
3.Expression of CXCR4 in Acute Leukemic Cells of Children and Its Signific ance
ri-ling, CHEN ; kang-rong, CAI ; ming-zhen, CHEN ; hui-qiong, ZHANG ; xi-min, FANG
Journal of Applied Clinical Pediatrics 2004;0(09):-
Objective To analyze the chemokine receptor CXCR4 expression in acute leukemic cells of children and its relationship with extramedullary infiltration.Methods The immunotypes of cases of acute leukemia in children and the expression of CXCR4 in marrow leukemic cells were studied by flow cytometry respectively. The relationship between CXCR4 expression and extramedullary infiltration of leukemic cells were analyzed by statistical method.Results The expression rates of CXCR4 in ALL children were higher than those in NALL children(P
4.Inhibitory effect of platelet-derived growth factor-α receptor silencing on the proliferation of human lens epithelial cell
Xiao-hui, LIU ; Yan-yi, PENG ; Cai-wen, FAN ; Lan-zhen, HUANG
Chinese Journal of Experimental Ophthalmology 2013;31(8):749-753
Background Platelet-derived growth facto(PDGF) affectthe proliferation of human lenepithelial cell(LECs),and human LECexpresPDGF-α recepto(PDGFR-α) throughoutheilifetime.The binding of activated PDGF-α receptowith PDGF promotethe synthesiof DNA.Othestudiedemonstrated thasilencing of PDGFR-α by antisense oligodeoxynucleotide(ASODN) inhibitthe growth of RPE cellin proliferative vitreoretinopathy (PVR),buwhethethitechnique ifeasible foLECiunclear.Objective Thistudy wato investigate the effecof the knockdown of the PDGFR-α on the proliferation of human LECin vitro,and to offean experimental basifothe gene therapy of posteriocapsule opacification.MethodHuman LECstrain SRA01/ 04 wacultured in α-MEM containing fetal bovine serum.The cellwere incubated in 6-well platea5 × 104 cells/ well and transfection of ASODN-containing liposome waperformed.The cellwere divided into the blank control group (with blank liposome),PDGFR-α missense oligodeoxynucleotide(MSODN) group (with PDGFR-α MSODN + liposome),0.5 μmol/L PDGFR-α ASODN group (with 0.5 μmol/L PDGFR-α ASODN+liposome) and 1.0 μmol/L PDGFR-α ASODN group (with 1.0 μ mol/L PDGFR-α ASODN+liposome).The morphology of LECwaexamined undean inverse microscope 24 houraftetransfection.The expression of PDGFR-α mRNin the cellwadetected by reverse transcription-PC(RT-PCR).The rate of proliferation (A490) of the cellwaassayed using Mtand the inhibitory rate of PDGFR-α ASODN on proliferation wameasured.The percentage of LECin G1 phase waanalyzed by flow cytometer.ResultThe LECgrew well and exhibited polygonal shape in the blank control group and PDGFR-α MSODN group 24 houraftetransfection.Buin the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups,the cellappeared round in shape and the numberof cellwere obviously decreased.The expression of PDGFR-α mRNdetected by RT-Pcdemonstrated highelevel in the blank control group and PDGFR-α MSODN group;however,the PDGFR-α mRNexpression waobviously lowein the 0.5 μmol/L and 1.0 μmol/L PDGFR-α ASODN groups.The A490 value wa0.661 ± 0.036,0.655 ± 0.016,0.529 ± 0.030 and 0.441 ± 0.039 in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,respectively,showing significandecline in the 0.5 μmol/L PDGFR-α ASODN group and 1.0 μ mol/L PDGFR-α ASODN group in comparison with the blank control group (F=34.08,P<0.01).The percentageof LECin G1 phase were (47.73±1.18)%,(49.48±1.09)%,(53.31±1.30)% and (59.98±0.95) % in the blank control group,PDGFR-α MSODN group,0.5 μmol/L PDGFR-α ASODN group and 1.0 μmol/L PDGFR-α ASODN group,showing significandifference among them (F =68.41,P<0.01),and thain the 0.5 μmol/L PDGFR-α ASODN group o1.0 μmol/L PDGFR-α ASODN group showed significantly increase in comparison with the blank control group (P<0.05).ConclusionPDGFR-α silencing could inhibithe proliferation of human LECin vitro.
5.Molluscicidal effect of suspension concentrate of niclosamide ethanolamine salt
Yi YUAN ; Shunxiang CAI ; Zhengwen HE ; Bo LI ; Youbin WANG ; Zhen TU ; Zhaogang XU ; Hui HE ; Bo XIONG
Chinese Journal of Schistosomiasis Control 2017;29(4):416-419,435
Objective To evaluate the molluscicidal effect of suspension concentrate of niclosamide ethanolamine salt(SC-NE)against Oncomelania hupensis snails in laboratory and field. Methods The experiment of SCNE against the snails by using the immersing and spraying methods was performed in laboratory and field,with control groups of wettable powder of ni-closamide ethanolamine salt(WPN). Results In the laboratory,LC50(s) of SCNE for 24,48 h and 72 h by using the immersion method were 0.0926,0.0629 mg/L and 0.0549 mg/L,respectively. The mortality rates of snails for 24,48 h and 72 h by using the immersion method were all 100% with the concentrations of 0.25 mg/L. The mortality rates of snails were all 100% while spraying SCNE for 3 d in the laboratory with the concentrations of 0.25 g/m2. In Jiangling County,except 0.5 g/m3 SCNE immers-ing the snails for 24 h,the mortality rates of snails by using SCNE with the immersing method were all 100%. While the concen-tration of SCNE was 0.5 g/m3 or above,the mortality rates were all 100%after the use of it with the immersion method for 2 d in Gong'an County. In Jiangling County,the mortality rates of snails by using SCNE 0.5 g/m3 for 1 d,3 d,and 7 d with the spray-ing method were 87.5%,92.82%and 97.40%respectively. While the concentration of SCNE was 0.5 g/m3,the mortality rates were 85.94%,86.78%and 94.21%respectively after the use of it with the spraying method for 1 d,3 d,7 d in Gong'an Coun-ty,and the molluscicidal effect of SCNE(1.0 g/m2)was higher than that of WPN. Conclusion SCNE has a high molluscicidal effect in the laboratory and field,and it is a novel and simple formulation of niclosamide.
6.Iodine nutrition level of children aged 8 - 10 in low-coverage area of iodized salt of Yushu Qinghai province in 2009: an analysis of surveillance results
Lan-sheng, HU ; Hui-zhen, YU ; Li-lin, CHEN ; Ya-nan, LI ; Pei-zhen, YANG ; Sheng-hua, CAI ; Shu-bang, LI ; Fa-rong, ZHANG
Chinese Journal of Endemiology 2011;30(3):316-318
Objective An analysis was conducted to investigate the iodine nutrition level of children aged 8 - 10 in low-coverage area of iodized salt of Yushu Qinghai province for providing a scientific basis for the development of effective preventive measures. Methods Yushu, Chengduo, Nangqian and Zaduo counties with higher non-iodized salt coverage rate in Yushu Qinghai province were chosen as survey counties in 2009. Three townships were selected in each county, and 2 primary schools were selected in each township and 40 urine samples of children aged 8-10 were collected randomly in each primary school. The content of urinary iodine was analyzed by As-Ce catalytic spectrophotometery. Results Median urinary iodine of children aged 8 - 10 in Nangqian and Zaduo was < 100 μg/L. The percentage of median urinary iodine < 50 μg/L in Yushu was over 20%. Median urinary iodine of children aged 10 in Zaduo was 81.5 μg/L, the percentage of median urinary iodine < 50 μg/L of children aged 9 and 10 was over 20%. The percentage of median urinary iodine < 50 μg/L in children aged 9 and 10 of Yushu was over 20%. Median urinary iodine of girls in Zaduo was 87.1 μg/L, the percentage of median urinary iodine < 50 μg/L of boys in Zaduo was over 20%. The percentage of median urinary iodine < 50 μg/L of girls in Yushu was over 20%. Conclusions The iodine nutrition level of children aged 8 - 10 in Nangqian, Zaduo and Yushu counties were very low due to the impact of non-iodized salt. We propose salt market in the region to strengthen management and improve the coverage and consumption rates of iodized salt to improve the level of iodine nutrition for effective prevention of iodine deficiency disorders.
7.Insulin-like growth factor 1 protects dopaminergic neurons from L-dopa-induced toxicity via JAK/STAT pathway
Rong-Ni HE ; Hui-Fang XIE ; Zhen-Hua LIU ; Zhen-Xin YAN ; Wei-Wei CAI ; Kong XIN
Chinese Journal of Neuromedicine 2011;10(5):475-479
Objective To explore the protective effect of insulin-like growth factor 1 (IGF-1) on apoptosis of dopaminergic neurons induced by L-dopa via JAK/STAT signaling pathway. Methods PC12 cells were induced to differentiate into dopaminergic neurons with 100 μg/L β-NGF; MTT assay was employed to identify the changes in the viability of PC12 cells following L-dopa treatment at 0, 10,20, 50, 100, 150 and 200 μmol/L, and the different concentrations of IGF-1 at 0, 10, 25, 50 and 100 nmol/L with the same concentration of L-dopa (150 μmol/L); Western blotting was used to detect the levels of P-JAK2/P-STAT3 in PC12 cells treated with PBS (controls), L-dopa, L-dopa+IGF-1 and L-dopa+IGF-1+AG490 for 24 h, and then the apoptosis rate was assessed by flow cytometry and Hchest33258 staining. Results Western blotting showed that the expressions of P-JAK2 and P-STAT3 were detected in the L-dopa+IGF-1 and L-dopa+IGF-1+AG490 treatment groups but not in the control group or L-dopa treatment group; the expression of P-STAT3 in the L-dopa+IGF-1+AG490 treatment group was obviously lower than that in the L-dopa+IGF-1 treatment group (P<0.05). Hchest33258 staining indicated that L-dopa treatment group had the most obvious karyopyknosis and karyorrhexis,much more apoptotic bodies than the L-dopa+IGF-1 and L-dopa+IGF-1+AG490 treatment groups. Flow cytometry showed that the apoptosis rate was significantly different among the 4 groups (F=180.991,P=0.000): as compared with the control group, the other 3 groups had a higher apoptosis rate (P<0.05);L-dopa treatment group (38.13 ±2.54 %) enjoyed the highest level, followed by L-dopa+IGF-1 +AG490treatment group (25.60±1.30 %) and L-dopa+IGF-1 treatment group (20.17±1.54 %). Conclusion L-dopa has toxic effect on PC12 cells; IGF-1 could protect the PC12 cells from the neurotoxic effect of L-dopa and JAK2/STAT3 signaling pathway is activated in this procedure.
8.Concordance between hepatitis C virus serotype and genotype in chronic hepatitis C patients in China.
Hui ZHAO ; Ming-hui LI ; Yao XIE ; Shao-cai DU ; Dao-zhen XU
Chinese Journal of Experimental and Clinical Virology 2007;21(4):355-357
OBJECTIVETo investigate the relationship of hepatitis C virus (HCV) serotype with genotype.
METHODSThe serotypes of HCV in the serum of 104 patients with chronic hepatitis C from 14 cities in China for which HCV genotypes were available, were determined by ELISA using Murex HCV Serotyping 1-6 Assay.
RESULTSThe serotypes of 86 (82.69 percent) of the 104 serum specimens were determined, and HCV serotypes were determined for 91 strains. Overall the concordance between hepatitis C virus serotype and genotype was 62.1 percent, and the concordance of serotype, with genotypes 1, 2 and 3 were 69.4 percent, 51.2 percent and 70.0 percent, respectively. The false-negative rate and concordance of genotype 2b was lower (54.5 percent).
CONCLUSIONThe specificity of HCV serotyping was affected by HCV strains' genotype and sometimes HCV serotype was not in concordance with genotype.
Genotype ; Hepacivirus ; classification ; Hepatitis C, Chronic ; virology ; Humans ; Serotyping
9.Cell membrane phospholipid variation and protein kinase C expression effects on hepatic metastasis of large intestinal carcinoma.
Shi-yong LI ; Bo YU ; Ping AN ; Zhen-jia LIANG ; Shu-jun YUAN ; Hui-yun CAI
Chinese Journal of Surgery 2004;42(2):84-87
OBJECTIVETo study cell membrane phospholipid variation and protein kinase C (PKC) isoenzyme expression and their effects on hepatic metastasis of large intestinal carcinoma.
METHODSHigh function liquid chromatography was used to separate and detect cell membrane phospholipids of phosphatidylinosital (PI), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylcholine (PC) in primary foci, paratumor intestine mucosa and hepatic metastasis of large intestinal carcinomas. And mRNA expression levels of PKC-alpha, -beta II, -delta, -epsilon, -lambda, -zeta isoenzymes were detected using QRT-PCR technique.
RESULTSFifty-eight cases of colorectal cancer were examined.
CONTENTSof PI, PC and PE in primary foci and hepatic metastasis were higher than those in paratumor mucosa. PE content in hepatic metastasis was much higher than that in primary foci (t = 98.88, P < 0.01). But PI and PC contents had no significant differences between primary and hepatic metastasis (t = 1.73, 1.36, P > 0.05). PKC-beta II, -delta, -epsilon, -lambda, -zeta expression were enhanced in primary foci and hepatic metastasis, but PKC-alpha level decreased in comparison with paratumor mucosa. And PKC-delta, -epsilon, -lambda, -zeta levels in hepatic metastasis were higher than those in primary foci (t = 4.31, P < 0.05). PI and PC had positive correlations with PKC-beta II expression. PE had positive correlations with PKC-delta, -epsilon, -lambda, -zeta, but a negative correlation with PKC-alpha.
CONCLUSIONSThe increases of PI and PC and PKC-alpha/PKC-beta II ratio change are related with colorectal cancer genesis. High content of PE and enhanced expression of PKC-delta, -epsilon, -lambda, -zeta isoenzymes and decreased PKC-alpha level improved hepatic metastasis of large intestinal carcinoma.
Adult ; Aged ; Aged, 80 and over ; Chromatography, Liquid ; Female ; Humans ; Intestinal Neoplasms ; pathology ; Isoenzymes ; genetics ; Liver Neoplasms ; secondary ; Male ; Membrane Lipids ; analysis ; Middle Aged ; Phosphatidylcholines ; analysis ; Phosphatidylethanolamines ; analysis ; Phosphatidylinositols ; analysis ; Phosphatidylserines ; analysis ; Protein Kinase C ; genetics ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
10.Sonographic features of lymphoma in the abdominal lymph nodes.
Ya-hong WANG ; Sheng CAI ; Lei WANG ; Xiao-dong ZHANG ; Zhen-hong QI ; Zhong-hui XU ; Jian-chu LI
Acta Academiae Medicinae Sinicae 2013;35(2):140-144
OBJECTIVETo summarize the sonographic features of the lymphoma in the abdominal lymph nodes.
METHODThe clinical data and sonographic findings of 41 lymphoma patients with original appearance of abdominal mass or lymphadenectasis were retrospectively analyzed.
RESULTSAmong these 41 cases, the retroperitoneal and mesenteric lymph nodes were most commonly involved.These patients were divided into two types according to sonographic appearances:solitary mass(n=14) and multiple nodules(n=27) .Solitary hypoechoic mass in abdomen was found in the former type, mostly with irregular or lobular shape, and the inner echo was often heterogenous.The typical appearances of the latter type were multiple enlarged lymph nodes, mostly round or oval, with homogeneous inner echo and clear margins.Some other sonographic characteristics were also helpful for the diagnosis of lymphoma, such as cobblestone sign, intranodular reticulation, vessels-embedded sign, and hilar vascularity.
CONCLUSIONUltrasonography can provide useful information in the diagnosis of the lymphoma in the abdominal lymph nodes.
Abdominal Neoplasms ; diagnostic imaging ; Adult ; Aged ; Female ; Humans ; Lymph Nodes ; diagnostic imaging ; Lymphoma ; diagnostic imaging ; Male ; Middle Aged ; Retrospective Studies ; Ultrasonography ; Young Adult