1.Significance of human plasma and feces microRNA-92a-1 expression level in colorectal tumor screening
Jing WANG ; Jinping ZHANG ; Yinli GUI ; Jiansheng LI
Chinese Journal of Digestion 2012;(12):834-837
Objective To explore the feasibility and clinical significance of combined detection of human plasma and feces expression level of microRNA-92a-1 (miRNA-92a-1) as colorectal tumour screening marker.Methods From August to October in 2011,the feces and plasma samples of 60 colorectal cancer (CRC) patients,23 colorectal adenoma (CRA) patients and 30 healthy controls were collected.The expression level of miRNA-92a-1 was determined by quantitative reverse transcription-polymerase chain reaction.Mann-Whitney's U test was applied for the difference test between groups.Then,according to the receiver operating characteristic (ROC) curve analysis the cut-off point was determined and the sensitivity and specificity were analyzed.Results Compared with healthy controls,the expressions of miRNA-92a-1 in plasma of CRC and CRA patients increased (U=288.5 and 151.0,both P<0.01).Compared with healthy controls,the expression of miRNA-92a-1 in feces of CRC patients increased (U=627.5,P=0.0199).According to ROC curve analysis,when the cut-off point was >1.22,the sensitivities of plasma miRNA-92a-1 in CRC patients and CRA patients were 85.0% (51/60) and 73.9% (17/23) respectively.The specificity of healthy controls was 76.7% (23/30).When the cut-off point was >1.14,the sensitivities of feces miRNA-92a-1 in CRC patients and CRA patients were 31.7%(19/60) and 26.1%(6/23) respectively.The specificity of healthy controls was 90.0 % (27/30).Combined the detection results of plasma and feces,the sensitivities of miRNA-92a-1 in CRC patients and CRA patients was 88.3% (53/60) and 82.6% (19/23) respectively.The specificity in healthy controls was 73.3% (22/30).The sensitivity was higher than that of single sample testing.Conclusions The sensitivity and specificity are high in the combined detection of plasma and feces miRNA-92a-1 expression level in CRC patients and CRA patients,miRNA-92a-1 may be a potential CRC screening marker.
2.Evaluation of DNA microarray for detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis isolates
Feng WANG ; Jing GUI ; Guanglu ZHAO ; Yunyong CUI ; Xiaoli LIU
Chinese Journal of Laboratory Medicine 2012;(12):1125-1129
Objective To evaluate the performance of DNA microarray for rapid detection resistance to rifampin and isoniazid in Mycobacterium tuberculosis clinical isolates and identify suitable target sites for molecular genetic test.Methods Twenty-four clinical Mycobacterium tuberculosis isolates were collected retrospectively from Shenzhen Center for Chronic Disease Control in 2009 and 127 isolates from project on anti-tuberculosis drug resistance surveillance in Shenzhen during 2007 to 2009.Drug susceptibility to rifampin and isoniazid of the stains were determined by DNA microarray,and results were compared to that obtained with reference proportion method drug susceptibility testing for sensitivity,specificity and accuracy.The consistency of microarray and phenotypic susceptibility testing was evaluated by Kappa test.Genetic mutations in rpoB,katG,inhA,regulatory region of inhA,and regulatory region of ahpC were investigated by DNA sequencing to assess proper loci for rapid molecular diagnosis.Results Compared against results of proportion method,the sensitivity,specificity and accuracy of the DNA microarray assay for rifampin resistance were 94.4%,97.5% and 96.0% respectively,and for isoniazid resistance were 79.1%,100% and 86.8% respectively.Mutations in resistance-determining region of rpoB were observed in 97.2% (70/72) of the isolates resistant to rifampin,which contributed in the 531,526,516,511 and 533codon region.Mutations in katG315 codon,inhA-15,and ahpC regulatory region were found in 70.3% (64/91),11.0% (10/91) and 9.9% (9/90) of the isolates resistant to isoniazid,respectively.Mutations of ahpC promoter region consists of ahpC-9 (4 strains),ahpC-10 (2 strains),ahpC-6 (2 strains),ahpC-12 (1 strain),and ahpC-32 (1 strain).Conclusions DNA microarray provided a rapid method for the detection of drug-resistant Mycobacterium tuberculosis isolates,and demonstrated good performance except less sensitive for isoniazid resistance.The mutations in ahpC regulatory region might be good target loci for detection of isoniazid-resistant Mycobacterium tuberculosis,so screening the region may significantly improve the sensitivity for molecular genetic tests.
3.Relationship between ABO hemolytic disease of newborn and maternal antibody
Qin GUI ; Yu WANG ; Jing CHEN ; Liqiang WU
Journal of Third Military Medical University 2003;0(07):-
Objective To study the relationship between ABO hemolytic disease of newborn and maternal antibody. Methods The titer of blood group antibody in 122 mothers of O blood group during prenatal diagnosis and blood group serology, bilirubin and hemoglobin level of newborn infants were tested with routine methods. The relationship between ABO hemolytic disease of newborn and the titer of blood group antibody was studied. Results The titer of blood group antibody was remarkably related with ABO hemolytic disease of newborn (P
4.Genetic and phenotypic characterization of drug-resistant Mycobacterium tuberculosis isolates in Shenzhen of China
Jing GUI ; Feng WANG ; Jinli LI ; Daoquan LUO ; Litai YAO
Chinese Journal of Microbiology and Immunology 2010;30(5):466-471
Objective To characterize the relationship between such phenotypes and the patterns of genetic mutations in the corresponding resistance genes in drug-resistant Mycobacterium tuberculosis (MTB)isolates in Shenzhen of China during 2007-2008.Methods According to standard of WHO,International Union Against Tuberculosis and Lung Disease(IUATLD),136 strains of MTB were collected by performing drug sensitivity test(DST)to isoniazid,rifampicin,streptomycin,ofloxacin and kanamycin on Lowenstein-Jensen in 1%proportion method.Genetic mutations in the corresponding resistance genes (rpoB,katG,rpsL,rrs1,gyrAB,rrs2)in these MTB isolates were identified by PCR,followed by DNA sequencing of the purified PCR products.The minimal inhibitory concentration(MIC)values of the aforementioned anti-tuberculosis drugs were determined for these MTB clinical isolates by two-fold dilution method in vitro.Results A total of 123 isolates were collected,73 isolates were drug resistant.50 isolates were drug susceptible.Among the isolates that were resistant to isoniazid,rifampicin,streptomycin.ofloxacin and kanamycin,the proportion of isolates that harboured mutations in the respective genes was 84.6%,93.6%,65.9%,100%,61.1%.For katG gene,the mutation detected were S315T or S315N.For rpoB,the most frequently found changes were S531L(30/44,68.2%)and H526D(9/44,20.5%)or H526R(1/44,2.3%).The reported mutations that K43R and KS8Q were founded in the rpsL locus and 491C→T and 513A→C were founded in the rrs1 gene related with streptomycin-resistant strains.For gyrA,all gyrA mutations were clustered in codons 90,91,and 94 apart from the S95T that was natural polymorphism.accounted for 81.1% of the ofloxacin-resistant isolates,and condon 91 was the most frequently mutated.No mutation were found in gyrB.The most frequent substitution were 1400 A→G(9/11,81.8%)and 1483 G→T(2/11,18.2%)in a specific region of the rrs2 gene related with kanamycin-resistant strains.No mutations except S95T of gyrA detected in the drug-susceptible isolates.The MIC values of clinical drug-resistant strains that the same drug-resistant group contains a different resistance phenotype are basically the same with the relevant resistance genes in the same mutation.Associated resistance mutations in different sites varied significantly with their MIC values.Conclusion The mutation characterization of drug-resistant and drug-suscep-tible isolates of MTB have been shown to vary according to geographic region,phenotypic characteristics exist difference in resistance levels due to different muntants of drug-resistant gene.
5.Relationship between pathology of concomitant exotropia medial rectus and surgical results
Zun-Jing, WANG ; Qing-Lan, KONG ; Gui-Qiu, ZHAO
International Eye Science 2009;9(5):828-830
AIM: To evaluate the relationship between the medial rectus cells counts in concomitant exotropia and surgical results. METHODS: A total of 32 pieces of medial rectus muscle were collected for HE staining in this study, of which 18 pieces were from patients with concomitant exotropia and 14 pieces were from healthy individuals. A method of strabismus score was used to assess the operative effect.RESULTS: The difference of strabismus score before and after the operation in the intermittent exotropia group was significantly higher than that in constant exotropic group (P<0.01). Under light microscope, the loosen muscle fibers and the increased stromal components in the cross sectional area of medial rectus were observed in strabismic group. The muscle cells counts was obviously lower in strabismic group than in control group (P<0.01), which was related to the difference of strabismus score before and after the operation (P<0.05).CONCLUSION: The decreased medial rectus cells counts induce concomitant exotropia directly. It is the crucial causes of the bad surgical results.
6.Comparative measurements of exodeviations in the three types of intermittent exotropia
Hong, WANG ; Gui-Xiang, LIU ; Xiao-Jing, PAN ; Wei, HUANG
International Eye Science 2014;(12):2162-2164
AlM: To compare the results of 4 methods for measuring angle of exodeviation in the three types of intermittent exotropia, including when looking at indoor distance target of 6m, looking at indoor distance target of 30m, looking at outdoor far distance target, after 1h diagnostic occlusion test.
METHODS: Prospective case series study. Sixty-five patients with intermittent exotropia between June 2013 and June 2014 were enrolled in the Department of Ophthalmology, Affiliated Hospital to Qingdao University, included 37 males and 28 females with average age ( 12. 5 ± 6. 2 ) years. All the patients were measured when looking at indoor distance target of 6m, looking at indoor distance target of 30m, looking at outdoor far distance target, after 1h diagnostic occlusion test. lntermittent exotropia was divided into basic type, convergence insufficiency type and divergence excess type, which was based on the different result of between the distance and near measurements. The One-way test was applied to analyze the four methods of measuring angle of exodeviation in the three types of intermittent exotropia. LSD - t test was applied to compare the differences between each two methods in each type.
RESULTS: The distance exodeviations tested with looking at indoor distance target of 6m, looking at indoor distance target of 30m , looking at outdoor far distance target, after 1h diagnostic occlusion test were basic type (45. 4 ± 21. 0, 55. 0 ± 15. 0, 64. 68 ± 17. 7, 68. 75 ± 16. 6PD), convergence insufficiency type (33. 3 ± 14. 0, 44. 9 ± 12. 9, 43. 6±11. 8, 54. 6±11. 2PD), divergence excess type (55. 6± 17.4, 66.3±18.8, 76.9±16.4, 78.1±15.6PD). There were obviously differences between each two methods in each type ( basic type F = 9. 649, P = 0. 00; convergence insufficiency type F=6. 886, P=0. 001; divergence excess type F = 7. 989, P = 0. 00 ). Compared with looking at indoor distance target of 30m, looking at outdoor far distance target ( basic type P=0. 044, divergence excess type P = 0. 048 ) and after 1h diagnostic occlusion test (basic type P=0. 04, divergence excess type P=0. 027) had the statistical difference in the basic type and divergence excess type, and there was no obviously difference between looking at outdoor far distance target and after 1h diagnostic occlusion test ( basic type P=0. 353, divergence excess type P=0. 815). Compared with the other three measurements, 1h diagnostic occlusion test can elicit larger angle of deviation in the convergence insufficiency type.
CONCLUSlON: Both measurement with looking at outdoor far distance target and after 1h diagnostic occlusion test can elicit the larger angle of deviation in the basic type and divergence excess type; The measurement with after 1 hour diagnostic occlusion test can elicit the larger angle of deviation in the convergence insufficiency type.
7.Protective effect of mesenteric lymph duct ligation on organ functions in MODS rats
Zigang ZHAO ; Chunyu NIU ; Jing ZHANG ; Gui FAN ; Shuqiang WANG ; Yuzhen WANG
Chinese Journal of Pathophysiology 1986;0(02):-
AIM: To study the protective effect of mese nt eric lymph duct ligation on the functions of liver, kidney and heart, and morpho logy in multiple organ dysfunction syndrome (MODS) rats subjected to two-hit. METHODS: Male Wistar rats were divided into three groups: the me senteric lymph duct ligation group, the non-ligation group and sham group. The M ODS model of two-hit was established by bleeding and LPS administration in both ligation group and non-ligation group. After 24 h, all rats were cannulated to f acilitate blood withdrawal for serum sample, then all rats were killed and organ s including kidney, liver, lung and heart were collected for making microscopic sectio ns. The biochemical indexes of hepatic and renal functions and myocardial enzyme in s erum were determined before and after experiment. RESULTS: After two-hit, the serum contents of AST, ALT, TBA, BUN , Cr and LDH-1 in both non-ligation group and ligation group, and UA content in non-ligation group were obviously increased than those in pre-experiment and sh am group (P
9.Analysis of test results of ABO hemolytic disease of newborn in Chizhou area
Conggang WANG ; Jing CHEN ; Jianguo BAO ; Shuqi ZHU ; Fazhi RONG ; Xiufang GUI ; Yusheng MAO
International Journal of Laboratory Medicine 2015;(5):615-616
Objective To understand the laboratory testing current situation of ABO hemolytic disease of the newborn(ABO-HDN)in Chizhou area,and to analyze the test results of serological three indexes tests in order to provide the basis for clinical diag-nosis.Methods The ABO blood group identification and serological three indexes tests(direct antiglobulin test,free antibody test, antibody release test)were performed by using microcolumn gel method.Results A,B,O and AB blood groups were 29.13%, 31.09%,37.82% and 1.96%;the total positive rate of ABO-HDN was 22.41%(80/357),the positive rates of ABO-HDN in A and B blood groups were 38.46% (40/104)and 36.04% (40/111 )respectively;the occurrence rate of ABO-HDN had no statistical difference between blood group A and B (P >0.05);the positive rates of the direct antiglobulin test,free antibody test and antibody release test were 1.96%(7/357),4.76%(17/357)and 22.41%(80/357)respectively.Conclusion The serological three indexes tests are the main basis for the diagnosis of ABO-HDN,the antibody release test shows the highest positive rate.If clinically consid-ering HDN,the newborns should conduct the ABO-HDN screening as early as possible for clarifying the diagnosis and performing the early treatment.
10.Preparation and evaluation of intra-articular injectable sinomenine hydrochloride-loaded in situ liquid crystals.
Yu-lin CHEN ; Shuang-ying GUI ; Xin LIANG ; Sheng-mei WANG ; Xiao-jing JIANG
Acta Pharmaceutica Sinica 2016;51(1):132-139
Phytantriol (PT), ethanol (ET) and water were used to prepare in situ cubic liquid crystal (ISV2). The pseudo-ternary phase diagram of PT-ET-water was constructed and isotropic solution formulations were chosen for further optimization. The physicochemical properties of isotropic solution formulations were evaluated to optimize the composition of ISV2. In situ hexagonal liquid crystals (ISH2) were prepared based on the composition of ISV2 with the addition of vitamin E acetate (VitEA) and the amount of VitEA was optimized by in vitro release behavior. The phase structures of liquid crystalline gels formed by ISV2 and ISH2 in excess water were confirmed by crossed polarized light microscopy and small angle X-ray scattering, respectively. Rheological properties of ISV2 and ISH2 were studied by a DHR-2 rheometer. In vitro drug release studies were conducted by using a dialysis membrane diffusion method. Pharmacokinetics was investigated by determination of sinomenine hydrochloride (SMH) concentration in synovial membrane after intra-articular injection of SMH-loaded ISH2 in adjuvant-induced arthritis rats. The optimal ISV2 (PT/ET/water, 64 : 16 : 20, w/w/w) loaded with 6 mg x g(-1) of SMH showed a suitable pH, injectable and formed a cubic liquid crystalline gel in situ with minimum water absorption in the shortest time. The optimal ISV2 was able to sustain the drug release for 144 h. The optimal ISH2 system was prepared by addition of 5% VitEA into PT in the optimal ISV2 system. This ISH2 (PT/VitEA/ET/water, 60.8 : 3.2 : 16 : 20, w/w/w/w) was an injectable isotropic solution with suitable pH. The new ISH2 was able to sustain the drug release for more than 240 h. Local pharmacokinetics study indicated that the retention time and AUC(0-∞) of ISH2 group were increased significantly compared with that of SMH solution group and the AUC(0-∞) of ISH2 group was 6.01 times higher than that of SMH solution group. The developed ISH2 was suitable for intra-articular injection that may apply to patients in the treatment of rheumatoid arthritis.
Animals
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Chemistry, Pharmaceutical
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Diffusion
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Ethanol
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Fatty Alcohols
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Gels
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Injections, Intra-Articular
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Liquid Crystals
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Morphinans
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administration & dosage
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chemistry
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Rats
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Rheology
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Water
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alpha-Tocopherol