1.Simultaneous detection of nine antibiotic resistance-related genes in Streptococcus agalactiae using multiplex PCR and reverse line blot hybridization assay
Xian-Yu ZENG ; Hui WANG ; Wei-Zhen WANG ; Yi-Qun DUAN ; Fan-Rong KONG ; L GWENDOLYN ;
Chinese Journal of Laboratory Medicine 2001;0(04):-
Objective To develop a multiplex PCR-based reverse line blot(mPCR/RLB) hybridization assay to detect,simultaneously,seven genes encoding AR-erm(A/TR),erm(B),mef(A/ E),tet(M),tet(O),aphA-3,aad-6 and two AR-related genes,int-Tn and mreA in group B streptococcus.Methods Nine pairs of specific primers and Oligonucleotide probes targeting erm(A/TR), erm(B),mef(A/E),tet(M),tet(O),aphA-3,aad-6 int-Tn and mreA respectively were modified according to former studies or designed in this study.The primers and probes were labeled with biotin and amino,respectively.The nine genes were amplified simultaneously in the same tube.PCR product hybridized with the probes labeled in the BiodyneC nylon membrane to detect the nine genes.To detect the sensitivity and specificity of the method developed,PCR with single pair of primer targeting each gene were tested in 318 isolates tested and the results were compared with the one abtained by RLB.Results The nine resistance-related genes could be successfully detected by mPCR/RLB assay developed in this study.Based on sequencing,21 of 22 isolates with mef had mef(E)and eight of 353 with int-Tn had an atypical sequence.Except for the above 29 genes,all the others corresponded well with the results obtained by single pair primer PCR.Conclusion The mPCR/RLB assay developed in this study is simple,rapid and suitable for surveillance of antibiotic resistance in GBS.
2.Developmental and reproductive toxicity of soybean isoflavones to immature SD rats.
Lei GUAN ; Yu HUANG ; Zhen-Yu CHEN
Biomedical and Environmental Sciences 2008;21(3):197-204
OBJECTIVETo investigate the dose-dependent toxic effect of soybean isoflavone extracts (SIE) on reproductive development in immature rats.
METHODSGrowing male and female rats (n = 50 each, 4 weeks) were divided into five groups fed with a standard cereal-based diet and gastrogavaged daily with 0, 30, 150, 300, and 600 mg SIE/kg body weight, respectively, for 12 weeks. Body weight, organ weights, and serum level of estrogen and testosterone were measured.
RESULTSOral administration of SIE had no effect on food intake but decreased food efficiency ratio (P < 0.01). Suppression on body weight gain by SIE was dose-dependent and the effect was greater on male than on female rats (P < 0.01). SIE at high doses exhibited hepatotoxicity by increasing a relative liver weight, and also caused a smaller uterus but a greater relative ovary in female rats, while leading to larger relative testis and epididymis in male rats. SIE could decrease progesterone concentrations in female rats, whereas in male rats it reduced not only total testosterone level but also sperm count compared with the control group (P < 0.05).
CONCLUSIONSIE at a range of 50-1000 times of human intake level affects not only growth but also development of reproductive system in growing rats.
Animals ; Body Weight ; drug effects ; Epididymis ; drug effects ; growth & development ; Estradiol ; blood ; Female ; Isoflavones ; toxicity ; Liver ; drug effects ; growth & development ; Male ; Organ Size ; drug effects ; Ovary ; drug effects ; growth & development ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Soybeans ; chemistry ; Sperm Count ; Testis ; drug effects ; growth & development ; Testosterone ; blood ; Uterus ; drug effects ; growth & development
3.DHA depletion in rat brain is associated with impairment on spatial learning and memory.
Ying XIAO ; Ling WANG ; Ruo-Jun XU ; Zhen-Yu CHEN
Biomedical and Environmental Sciences 2006;19(6):474-480
OBJECTIVETo examine the effect of docosahexaenoic acid (DHA) deficiency in brain on spatial learning and memory in rats.
METHODSSprague Dawley rats were fed with an n-3 fatty acid deficient diet for two generations to induce DHA depletion in brain. DHA in seven brain regions was analyzed using the gas-liquid chromatography. Morris water maze (MWM) was employed as an assessing index of spatial learning and memory in the n-3 fatty acid deficient adult rats of second generation.
RESULTSFeeding an n-3 deficient diet for two generations depleted DHA differently by 39%-63% in the seven brain regions including cerebellum, medulla, hypothalamus, striatum, hippocampus, cortex and midbrain. The MWM test showed that the n-3 deficient rats took a longer time and swam a longer distance to find the escape platform than the n-3 Adq group.
CONCLUSIONThe spatial learning and memory in adult rats are partially impaired by brain DHA depletion.
Animals ; Brain ; metabolism ; Docosahexaenoic Acids ; metabolism ; Maze Learning ; physiology ; Memory ; physiology ; Rats ; Rats, Sprague-Dawley
4.Advancement of colloidal gold chromatographic technique in screening of ochratoxin A.
Wei-lu ZHOU ; Yu-ting WANG ; Wei-jun KONG ; Mei-hua YANG ; Ming ZHAO ; Zhen OU-YANG
China Journal of Chinese Materia Medica 2015;40(15):2945-2951
Ochratoxin A (OTA) is a toxic secondary metabolite mainly produced by Aspergillus and Penicillium species, existing in a variety of foodstuffs and Chinese medicines. OTA is difficult to be detected in practice because of the characteristics such as trace amounts, toxicity, existing in complex matrices. In the numerous detection technologies, colloidal gold chromatographic techniques are highly sensitive, specific, cost-effective and user-friendly, and are being used increasingly for OTA screening. Recently, with the development of aptamer technology and its application in chromatographic technique, a newly colloidal gold aptamer chromatographic technique has been developed. This review elaborates the structures and principles of both traditional and newly colloidal gold chromatographic techniques, focuses on newly colloidal gold aptamer chromatographic technique, summarizes and compares their use in rapid detection of OTA. Finally, in order to provide a reference for better research of related work, the development trends of this novel technique are prospected.
Base Sequence
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Chromatography
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methods
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Gold Colloid
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chemistry
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Molecular Sequence Data
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Ochratoxins
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analysis
5.Treatment of hepatic cysts with dehydrated alcohol sclerosing agent guided by CT
Jian KONG ; Yong-Chong DOU ; Yan-Fang ZHANG ; Yu-Xuan WU ; Xin-Ying SHEN ; Zhen-Wen LIN ;
Journal of Interventional Radiology 2006;0(12):-
Objective To evaluate the clinical effects of CT guided percutaneous aspiration and sclerotherapy in treatment of hepatic cysts.Methods Sixty three patients with single(n=41)and muttiple(n= 22)hepatic cysts were undertaken CT guided pereutaneous aspiration and sclerotherapy with injection of absolute alcohol.Results Sixty three patients underwent follow-up for 3-15 months after the operation showing effective indexes as grade 0 for 4(6.39%),gradeⅠfor 8(12.69%),gradeⅡfor 23(36.51%)and gradeⅢfor 28(44.44%)cases.The total effective rate reached 93.61%.No serious complications occurred. Conclusion Sclerosing therapy with absolute alcohol is safe,economic,simple and effective for treating hepatic cysts.(J Intervent Radiol,2007,16:850-852)
6.Study on prevalence and correlation factors of bronchial asthma in Zaozhuang area, Shandong province.
Shou-zhen ZHANG ; Qiang XI ; Wei-shun KONG ; Zi-hong LI ; Xiang-tai KONG ; Ling-yu KONG ; Ai-hua QIAN ; Jia-ling LI
Chinese Journal of Epidemiology 2005;26(4):273-276
OBJECTIVETo study the prevalence of asthma and its correlated factors in Zaozhuang area in 2003, to provide a basic consideration for prevention/treatment and control policy.
METHODS6 points were selected by stratified-clusterd-random sampling with a total of 16,725 persons expected, but only 10,610 subjects investigated.
RESULTSIn this survey, 128 asthma cases were identified with a overall prevalence of 1.21%. The prevalence for children was 2.02%, and for adult was 0.90% with the former significantly higher then the latter (chi(2) = 21.39, P < 0.01). Rates for male and female were 1.08%, 1.32% with a ratio of 1:1.22. For 77.97% of children with asthma. The initiative age of asthma was before 7 years old among children while among 36.23% of the adults, it was before 15 years of age. Correlation analysis showed that upper respiratory tract infection (OR = 17.81, 95% CI: 12.25-25.89), cold air exposure (OR = 3.43, 95% CI: 2.41-4.90), stimulation through cooking and by harmful gases (OR = 2.56, 95% CI: 1.80-3.63), allergic materials (OR = 2.74, 95% CI: 1.80-4.17) were main inducing factors. 65.63% of the asthma cases having had history of allergic disease while 25.78% having had family history with the OR of allergic history and family history as 21.69 vs. 73.96.
CONCLUSIONThe epidemic status of bronchial asthma was serious, with an assumption that asthma cases might have reached the number of 43 thousand in Zaozhuang area.
Adolescent ; Adult ; Aged ; Asthma ; epidemiology ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Mass Screening ; Middle Aged ; Prevalence ; Risk Factors
7.Red yeast rice increases excretion of bile acids in hamsters.
Ka-Ying MA ; Ze-Sheng ZHANG ; Shu-Xin ZHAO ; Qi CHANG ; Yin-Mei WONG ; Sai Ying Venus YEUNG ; Yu HUANG ; Zhen-Yu CHEN
Biomedical and Environmental Sciences 2009;22(4):269-277
OBJECTIVETo investigate the hypocholesterolemic activity of red yeast rice (RYR) and its underlying mechanism.
METHODSThree groups of hamsters were fed either the control diet or one of the two experimental diets containing by weight 0.1% RYR (0.1RYR) or 0.3% RYR (0.3RYR). Blood (0.5 mL) was collected from the retro-orbital sinus into a heparinized capillary tube at the end of week 0, 3, and 6. Plasma lipoproteins were measured using enzymatic kits, while fecal neutral and acidic sterols were quantified using a gas-liquid chromatography.
RESULTSPlasma total cholesterol was reduced by 12% in 0.1RYR group and by 18% in 0.3RYR group compared with the control value. Similarly, plasma triacylglycerol was decreased by 11% in 0.1RYR group and by 24% in 0.3RYR group. Western blotting analysis demonstrated that RYR had no effect on sterol regulatory element binding protein 2, liver X receptor, 3-hydroxy-3-methylglutary-CoA reductase, LDL receptor, and cholesterol-7alpha-hydroxylase. HPLC analysis confirmed that RYR contained 0.88% monacolin K. It was recently found that RYR supplementation increased excretion of fecal acidic sterols by 3-4 folds compared with the control value.
CONCLUSIONHypocholesterolemic activity of RYR is mediated at least partially by enhancement of acidic sterol excretion.
Animals ; Bile Acids and Salts ; secretion ; Biological Products ; pharmacology ; Blotting, Western ; Body Weight ; drug effects ; Cholesterol ; metabolism ; Cholesterol 7-alpha-Hydroxylase ; metabolism ; Cricetinae ; Dietary Supplements ; Feces ; chemistry ; Feeding Behavior ; drug effects ; Hydroxymethylglutaryl CoA Reductases ; metabolism ; Lipoproteins ; blood ; Liver ; enzymology ; Liver X Receptors ; Naphthalenes ; analysis ; Organ Size ; drug effects ; Orphan Nuclear Receptors ; metabolism ; Receptors, LDL ; metabolism ; Sterol Regulatory Element Binding Protein 2 ; metabolism ; Weight Gain ; drug effects
8.Evaluation of a self-prepared anti-WNV-IgG diagnostic ELISA kit with a panel of serum samples collected from the people from areas in which West Nile fever is endemic.
Yu-Chun WANG ; Qiang SHAO ; Li-Ping ZHANG ; Wei ZHEN ; Xue-Min WU ; Xue-Zheng MA ; Yong ZHAO ; Kong-Xin HU
Chinese Journal of Virology 2014;30(5):549-553
In view of that there is no report of west Nile virus infection cases in our country, evaluation the self-prepared anti-WNV-IgG diagnostic ELISA kit should be employed with the establishment of the serum sample panel collected from the entry personnel. All individuals of entry personnel were traveled from epidemic area of infectious west Nile disease. In our study, the serum samples were both detected by self-prepared anti-WNV-IgG diagnostic ELISA kit and the FDA certified kits ,which are FOCUS West Nile Virus IgG Dxselect and Panbio Dengue IgG Capture ELISA kits. The self-prepared kit and FDA certified kits were compared and assessed simultaneously. Furthermore, the specificity, repeatability and stability of the kits were also evaluated. The results indicated that no significant difference of detective rates (35. 6% for self-prepared kit vs. 32.5% for FOCUS kit, χ2 = 3. 05, P > 0.05) and good consistency (Kappa = 0.8372) between the self-prepared kit and FDA certified kits. Also, the positive coincidence rate, the negative coincidence rate and the total coincidence rate were calculated as 91.18%, 95.34% and 92.66%, respectively. The laboratory self-developed kit presented similar quality as the counterpart kits with FDA certificate. The development of our self-prepared anti-WNV-IgG diagnostic ELISA kit will provide technical support for the prevention and control of west Nile virus entry.
Endemic Diseases
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Enzyme-Linked Immunosorbent Assay
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Humans
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Immunoglobulin G
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blood
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Reagent Kits, Diagnostic
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West Nile Fever
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epidemiology
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West Nile virus
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immunology
10.Determination of ochratoxin A in human urine by HPLC-FLD after cleaned-up by molecularly imprinted polymer solid phase extraction column.
Li-Wei XIE ; Xiang-Sheng ZHAO ; Wei-Jun KONG ; Yu-Ting WANG ; Yi-Chen HU ; Zhen OU-YANG ; Mei-Hua YANG
Acta Pharmaceutica Sinica 2014;49(4):517-523
A method was developed for the determination of ochratoxin A (OTA) in human urine by HPLC-FLD after molecularly imprinted polymer solid phase extraction (MIP-SPE) column. After the pH being adjusted to 2.5 with 0.1 mol x L(-1) HC1, sample was cleaned up with MIP-SPE column for ochratoxin A, the analyte was analyzed by high performance liquid chromatography coupled with fluorescence detection (HPLC-FLD), and finally all the positive results were confirmed by LC-MS/MS. Recoveries from urine samples spiked with OTA at levels ranging from 2 to 20 ng x mL(-1) were 90.6%-101.9%, and RSDs were 0.1%-1.6%. Sixty-five volunteers living in Beijing took part in the study, of which 5 were found containing OTA in their urine and the highest value was 0.091 ng x mL(-1). The MIP-SPE column was firstly applied to purify and concentrate OTA in human urine, this method is simple, rapid and reliable and can be used to determine the contents of OTA in human urine.
Chromatography, High Pressure Liquid
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methods
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Female
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Humans
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Male
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Molecular Imprinting
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Ochratoxins
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urine
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Polymers
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Reproducibility of Results
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Sensitivity and Specificity
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Solid Phase Extraction