Denys-Drash Syndrome ; diagnosis ; Humans ; Infant ; Male

Vibrio parahaemolyticus has been considered as one of the most important foodborne bacterial pathogens.The loop-mediated isothermal amplification(LAMP) that amplifies DNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of this pathogen for the first time.A set of four primers,two outer and two inner primers,was designed specifically to recognize the thermolabile hemolysin gene(tlh) of V.parahaemolyticus.The LAMP reaction mix was optimized.The most optimal reaction temperature and time of the LAMP assay for the tlh gene were 60℃ and 60min,respectively.Genomic DNAs from 28 bacterial strains including 14 V.parahaemolyticus strains were amplified using LAMP,and no amplicon was observed in other bacterial strains.The detection limit of this LAMP assay was around 90 fg of V.parahaemolyticus genomic DNA and 24 colonies forming units for pure cultures.In addition,this method was applied to detect artificially contaminated food samples,and the detection limit was 89 cfu/g for non-cultured artificially contaminated food samples.These results suggested that detection of V.parahaemolyticus by LAMP is an effective and low-cost procedure with high specificity and sensitivity that requires no specialized equipment.This assay is expected to become a valuable tool for rapid detection and identification of V.parahaemolyticus.

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.

Objective To observe the effects of eye acupuncture and body acupuncture on tumor necrosis factor-α(TNF-α) expression in the cerebral cortex of rats after ischemia-reperfusion injury,in order to investigate the differences in therapeutic function between eye acupuncture and body acupuncture against acute cerebral ischemia-reperfusion injury. Method 48 Rats established by suture method were randomly divided into control group,sham operation group,model group,eye-acupuncture in point area group,eye-acupuncture outside point area group and body acupuncture group. After reperfusion 24 h,the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks;the levels of plasma TNF-αwere determined by ELISA method;the expression of ischemic cerebral cortex TNF-αmRNA was measured by RT-PCR method;the expression of ischemic cerebral cortex TNF-αprotein was detected by western blot. Results After reperfusion 24 h,compared with control group,neurologic impairment marks of eye-acupuncture therapy in point area group and body acupuncture group both decreased obviously (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group;the levels of plasma TNF-αin rat cerebral cortex after the eye acupuncture therapy and body acupuncture therapy were obviously decreased (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group; the expressions of TNF-α mRNA and protein in rat cerebral cortex after the eye acupuncture therapy and body acupuncture therapy were also obviously down-regulated (P<0.01),however there were no significant differences between the eye-acupuncture in point area group and body acupuncture group.Conclusion The eye and body acupuncture therapy show the same effects on treating cerebral ischemia reperfusion. The mechanisms of these two therapies may be related to up regulating TNF-αexpression in rat cerebral cortex with ischemia-reperfusion injury.

This study is designed to obtain recombinant human acetylcholinesterase (rhAChE) and apply it in screening acetylcholinesterase inhibitors. The rhAChE was overexpressed in HEK293 cells transfected by plasmid of pCMV-AChE with the cationic liposome and rhAChE was found to be secreted into cell culture medium. AChE activity was assayed according to modified Ellman method to obtain kinetic parameters. IC so50 values for donepezil compounds of rhAChE were calculated to determine their activities of inhibition. The results showed that Km value was 151.9 micromol.L-1 donepezil inhibited rhAChE in a mixed competitive-noncompetitive way (Ki= 16.03 nmol.L-1, Ki = 18.36 nmol.L-1) and that most new compounds tested exhibited high activities of inhibition on rhAChE. The study suggests that rhAChE is available to be applied in screening AChE inhibitors in vitro.
Acetylcholinesterase ; genetics ; metabolism ; Cholinesterase Inhibitors ; analysis ; pharmacology ; HEK293 Cells ; Humans ; Indans ; analysis ; pharmacology ; Inhibitory Concentration 50 ; Kinetics ; Piperidines ; analysis ; pharmacology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism ; Transfection

BACKGROUND:Significant increasing of intercel ular adhesion molecule 1 expression can promote the occurrence of inflammatory response and increase brain tissue injury.
OBJECTIVE:To investigate the effect of eye acupuncture point and non-point therapy on intercel ular adhesion molecule 1 expression in rat hippocampus of acute cerebral ischemia-reperfusion injury model.
METHODS:Forty Sprague Dawley rats were divided into normal group, sham-operation group, model group, eye acupuncture point group and eye acupuncture non-point group. Rats in the model group, eye acupuncture point group and eye acupuncture non-point group were used to establish the acute cerebral ischemia-reperfusion injury model with suture method, the suture was deep for 1.8-2.2 cm. In the eye acupuncture point group, the acupuncture points of hepatic region, upper energizer area, lower energizer area and kidney area were selected when cerebral ischemia-reperfusion occurred immediately and 30 minutes before drawing materials, then horizontal needling the acupuncture points around the orbit for 20 minutes. In the eye acupuncture non-point group, the puncture site was selected at 3 mm away from eye acupuncture non-point area, and then needled the puncture site with the same method in the eye acupuncture point group.
RESULTS AND CONCLUSION:After eye acupuncture point therapy, the neurological deficit scores were decreased, and the intercel ular adhesion molecule 1 expression in rat hippocampus was significantly decreased (P<0.01). After eye acupuncture non-point therapy, there were no significant changes in neurological deficit scores and the expression of intercel ular adhesion molecule 1 protein and mRNA in rat hippocampus. The results indicate that eye acupuncture can significantly improve the rat cerebral ischemia-reperfusion injury, and the mechanism may relate with reducing the intercel ular adhesion molecule 1 expression in rat hippocampus.

Objective To explore the distribution of methylenetetrahydrofolate reductase (MTHFR) C677T gene and MTHFR A1298C gene polymorphism among 1644 physical examinees in Taizhou region. Methods Fluorescence quantitative PCR method was used to detect MTHFR C677T and MTHFR A1298C genotype in 1644 physical examinees from Taizhou Central Hospital from September 2016 to September 2017. According to the characteristics of gender, the distribution features of MTHFR C677T and MTHFR A1298C genotype were described, and then were compared with data about the physical examinees in other cities in China. Results Among the 1644 physical examinees, MTHFR 677CC, 677CT and 677TT genotype frequency were 40.09％, 44.53％and 15.39％ respectively and MTHFR 677 allele frequency was 37.65％. MTHFR 1298AA, 1298AC and 1298CC genotype frequency were 65.69％, 30.47％ and 3.83％ respectively and MTHFR 1298 allele frequency was 19.07％. Statistical significance was found in genotype distribution of MTHFR C677T between males and females (P=0.036), and no statistical significance was found in genotype distribution of MTHFR A1298C between males and females (P=0.278) . As compared with the physical examinees in Henan, Wulumuqi and Beijing regions, there were statistically significance differences in the distribution of MTHFR C677T genotype and allele frequencies in Taizhou region (P <0.05) . Conclusion The distribution of MTHFR C677T gene polymorphism among physical examinees in Taizhou region is affected by gender, and the results shows certain regional specificity.

BACKGROUNDBrain-derived neurotrophic factor (BDNF) is a stress-responsive intercellular messenger modifying hypothalamic-pituitary-adrenal (HPA) axis activity. The interaction between stress and age in BDNF expression is currently not fully understood. This study was conducted to observe unpredictable stress effect on behavior and BDNF expression in CA3 subfield (CA3) and dentate gyrus of hippocampus in different aged rats.

METHODSForty-eight Wistar rats of two different ages (2 months and 15 months) were randomly assigned to six groups: two control groups and four stress groups. The rats in the stress group received three weeks of unpredictable mild stress. The depression state and the stress level of the animals were determined by sucrose preference test and observation of exploratory behavior in an open field (OF) test. The expressions of BDNF in CA3 and dentate gyrus of the hippocampus were measured using immunohistochemistry.

RESULTSAge and stress had different effects on the behavior of different aged animals (age: F = 6.173, P < 0.05, stress: F = 6.056, P < 0.05). Stress was the main factor affecting sucrose preference (F = 123.608, P < 0.05). Decreased sucrose preference and suppressed behavior emerged directly following stress, lasting to at least the eighth day after stress in young animals (P < 0.05). The older stress rats showed a lower sucrose preference than young stress rats (P < 0.05). Older control rats behaved differently from the younger control animals in the OF test, spending more time in the central square (P < 0.05), exhibiting fewer vertical movements (P < 0.05) and less grooming (P < 0.05). Following exposure to stress, older-aged rats showed no obvious changes in vertical movement and grooming. This indicates that aged rats were in an unexcited state before the stress period, and responded less to stressful stimuli than younger rats. There was significantly lower BDNF expression in the CA3 and dentate gyrus regions of the hippocampus following stress in both age groups (P < 0.05), a reduction that was still present at the eighth day after stress (P < 0.05). Stress and age were the main factors affecting the expression of BDNF (F = 9.408, P < 0.05; F = 106.303, P < 0.05). The aged stress group showed lower BDNF expression compared to the young stressed group at every testing time point.

CONCLUSIONStress has age-dependent effects on behavioral responses and hippocampal BDNF expression in rats.

Age Factors ; Animals ; Behavior, Animal ; Brain-Derived Neurotrophic Factor ; analysis ; genetics ; Chronic Disease ; Dentate Gyrus ; chemistry ; Exploratory Behavior ; Hippocampus ; chemistry ; Immunohistochemistry ; Rats ; Rats, Wistar ; Stress, Psychological ; metabolism ; psychology ; Sucrose ; administration & dosage

OBJECTIVETo explore the role of toll-like receptor 2 (TLR2) and toll-like receptor 4 (TLR4) in myocardial ischemia/reperfusion injury (MI/RI) by observing the dynamic expression changes at mRNA and protein levels early after myocardial ischemia/reperfusion (I/ R).

METHODSThe Wistar rats were randomly divided into Sham and I/R group (n = 42), and killed according to different reperfusion time (1, 2, 4, 6, 12, 24 h and 7 d). Structural and morphous changes of myocytes were observed under optical microscope. The mRNA and protein levels of TLR2 and TLR4 were detected using real-time PCR (RT-PCR). Monocyte chemokine protein-1 (MCP-1) and interleukine-6 (IL-6) mRNA levels were measured by reverse transcriptase-polymerase chain reaction (rt-PCR).

RESULTS(1) With the extension of reperfusion time, the myocardial infarct size increased smoothly, and reached the plateau at 4 h, then stayed in the platform. After reperfusion for 7 d, the ventricular had been remodeled. (2) At the beginning of reperfusion, myocardial structure showed no significant change in Sham group, but had different degrees of injury in I/R group. In rats of the group reperfused for 7 d the left ventricular remodeling could be visible. (3) Compared to sham group,TIR2, TLR4, MCP-1, IL-6 mRNA level were increased in myocardium in I/R group. TLR2 and TLR4 both peaked at 4 h of reperfusion, IL6 peaked at 6 h, followed by a gradually decrease. TLR4 and IL-6 mRNA levels rose again at 7 d. MCP-1 level in I/R group remained fairly with sham group at the beginning of reperfusion, and markedly elevated at 7 d.

CONCLUSIONExpression of TLRs mRNA in myocardium during early after myocardial ischemia/reperfusion increased rapidly and activated TLRs might play an important role in MI/RI through promoting the generation of inflammatory factors. At the late reperfusion, TLRs levels raise again and the expression of inflammatory factors increase once again, Those may probably affect the remodeling of ventricular, and injure myocardial structure and function.

Animals ; Chemokine CCL2 ; metabolism ; Disease Models, Animal ; Interleukin-6 ; metabolism ; Male ; Myocardial Reperfusion Injury ; metabolism ; Rats ; Rats, Wistar ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism

Haizao Yuhu decoction (HYD) is a formula that has been used for approximately 500 years and famous for its efficiency in treating thyroid-related diseases in traditional Chinese medicine (TCM). HYD was first presented by Chen Shi-gong in a famous surgical monograph named Waike Zhengzong during the Ming Dynasty. We conducted the research to investigate the possible pharmacokinetic profile of different prescriptions of HYD in rats, in order to reveal the interactions of Haizao and Gancao drug pair with other herbs in HYD. Liquiritin, naringin, besperidin, peimine, peiminine liquiritigenin, glycyrrhizic acid, hergapten, nobiletin, osthole, glycyrrhetinic acid in blood samples were determined by UPLC-MS/MS. The result revealed tbat Haizao could enhance the peak concentration of glycyrrhizic acid. The other herbs in HYD may promote'the absorption of flavonoids in Gancao in normal rats, but inhibit the absorption of saponins and accelerate their metabolism. Gancao and Haizao drug pair could enhance the bioavailability of hesperidin, peimine, bergapten, nobiletin and osthole and prolong the elimination of peimine and naringin.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Magnetic Resonance Spectroscopy ; Male ; Mass Spectrometry ; Plasma ; chemistry ; Rats ; Rats, Wistar