An improved assay for quantitation of murine macrophage-mediated antibody-dependent cellular cytotoxicity (MMADCC) by hemoglobin-enzyme release assay (Hb-ERA) has been developed The method is based on the coloration measurement by spectrophotometer,because hemoglobin has peroxidase activity capable of catalyzing the oxidation of o-phenylenediamine and producing color reaction. This method was applied to demonstrate variation of MMADCC activity with varying effector:target ratio,incubation time,antibody concentration and macrophage in different stage of activation. The method provides advantages of(l)elimination of the need for expensive and hazardous radioactive materials,(2)relative ease and rapidity, (3)sensitiyity and reproducibility.

OBJECTIVETo establish a RP-HPLC-PDA method for determination of five triterpenic acids (pomolic acid, hawthorn acid, corosolic acid, oleanolic acid and ursolic acid) in different parts of Salvia chinensis.

METHODThe isocratic elution and separation was achieved on a Kromasil C18 column (4.6 mm x 250 mm, 5 microm), using acetonitrile-water (90:10) as the mobile phase at a flow rate of 0.8 mL x min(-1). The detection wavelength and column temperature were set at 205 nm and 28 degrees C, respectively.

RESULTThe calibration curves of pomolic acid, hawthorn acid, corosolic acid, oleanolic acid and ursolic acid were linear over the ranges of 0.096 0-2.400, 0.1230-3.075, 0.2420-6.050, 0.2830-7.075 and 0.2730-6.825 microg (r = 0.9998, 0.9997, 0.9999, 0.9995, 0.9999), respectively. The average recoveries were 98.43%, 98.13%, 100.6%, 98.19% and 99.15%%, respectively, with RSD (n=6) being 1.3%, 0.67%, 1.2%, 0.87% and 0.43%.

CONCLUSIONThe proposed method is so simple and highly reproducible that it promises to be applicable for determination of major triterpenic acids in S. chinensis.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Plants, Medicinal ; chemistry ; Salvia ; chemistry ; Triterpenes ; chemistry

Objective To investigate the effect of valsartan,an angiotensinⅡtype 1 receptor (AT1 R)blocker,on radiosensitivity,invasive potential and proliferation activity of nasopharyngeal carcinoma cells(CNE-2)in vitro. Methods Radiosensitization of valsartan on CNE-2 cells in vitro was investigated by colony forming assay.Effect of ATl R blocker combined with radiation on invasive potential of CNE-2 cells was evaluated using 24-well Matrigel invasion chambers(Transwell).Apoptosis-inducing effect of valsartan combined with radiation on apoptosis of CNE-2 was identified by flowcytometry(FCM). Resuits When valsartan was given at 10-9.10-8 and 10-7 mol/L combined with radiation,sensitivity enhancement ratios (SER)were 1.10,1.20 and 1.36.and the invasive inhibition rates were 8.11%,16.49%and 16.77%,respectively.The SER of valsartan on CNE-2 distinctly increased when the exposure time was increased.After 24 h exposure to 10-8 mol/L valsartan combined witIl radiation.the apoptosis rate was 1.89%±0.09%,which was higher than 1.62%±0.06%in radiation alone group(t=4.79.P＜0.05). Conclusions AT1 R blocker valsartan combined with radiation can significantly inhibit the proliferation activity of nasophar,cngeal carcinoma cells in vitro in a dose- and time-dependent manner.Valsartan combined with radiation can potently inhibit the invasive potential of CNE-2.which may be involved in the mechanism of valsartan treatment in vivo.

Objective To setup a platform for genetic diagnosis of Cryopyrin associated periodic syndrome (CAPS) and to use it for clinical diagnosis.Methods The peripheral blood cells of the patient and the controls were collected for DNA extraction.Nine exons of CAPS associated gene NLRP3 were amplified using polymerase chain reaction (PCR) and subjected to sequencing.Blast was used to compare the sequencing results with the reference gene and to locate mutation.The clinical information of the patient was collected and the relevant literature was reviewed.Results We set up a platform for exon sequencing of NLRP3 gene.Using this platform,we identified a nonsynonymous mutation in a female patient (D303N,aspartic acid at locus 303 mutated to asparagine).Considering the clinical manifestations of the patient,chronic infantile neurologic cutaneous and articular syndrome (CINCA) was diagnosed.Conclusion The set up of the platform for NLRP3 genetic analysis will facilitate the clinical awareness and research on CAPS.

Objective:To observe clinical therapeutic effects of method for supplementing Qi,activating blood circulation and resolving phlegm and effects on vascular endothelial function and blood coagulation system in senile patient of hyperlipemia.Methods:96 senile patients of hyperlipemia were randomly divided into a treatment group(n=56)and a control group(n=40).The patients of the treatment group were treated by oral administration of Danshen Jueming Granules and the control group by oral administration of Xuezhikang Capsules.Blood lipids,vascular endothelial function,blood coagulation system and safety were observed.Results:After treatment,the treatment group in decrease of plasma total cholesterol(TC)and low density lipoprotein-cholesterol(LDL-C)was superior to the control group(P

Objective To provide the guidance for the control and treatment of blood stream infec-tion caused by Serratia marcescens strains through analyzing the homology and drug resistant genes of the iso-lates collected from the Intensive Care Unit ( ICU) of Shaoxing County Central Hospital.Methods Serratia marcescens strains were isolated from ICU patients with blood stream infection and also from the hands of health care providers in the ICU from June 1st to September 30th, 2013.The antibiotic susceptibilities of the Serratia marcescens isolates were tested.PCR was performed to amplify the common drug resistant genes. Pulse-field gel electrophoresis ( PFGE) was carried out for analyzing the homology of all isolates.The com-plete clinical data of the patients were collected and statistically analyzed with Spearman′s rank correlation coefficient.Results Seventeen strains were isolated in this study.All of the 17 strains were resistant to the first and second generation Cephalosporin, Gentamicin and Ciprofloxacin, and sensitive to Amikacin and Ceftazidime.The drug resistant rates to Carbapenems ranged from 11.76%to 35.29%.One of the isolates (5.88%) carried the TEM gene.The results of PFGE showed that the phenotypes of all isolates were identi-cal.Conclusion Serratia marcescens strains were critical hospital infectious pathogens.They were able to spread in the hospital and were resistant to multiple antibiotics.Clinical physicians should properly use anti-biotics for the patients based on the result of drug susceptibility test.A control regulation for Serratia marces-cens infection within hospital should be enforced to avoid the cross infection and the outbreak of resistant strains.

Aim To explore the role of PI3 K/Akt/Sirt1 pathway in cardioprotection of hydrogen sulfide ( H2 S ) postconditioning against ischemia/reperfusion ( I/R) injury. Methods Langendorff perfusion appa-ratus was used to build an isolated rat myocardial I/R model. Isolated rat hearts were subjected to 30 min global ischemia followed by 60 min reperfusion after 20 min of equilibrium. 60 male SD rats were randomly di-vided into 5 groups(n=12):control group(Control), ischemia/reperfusion group( I/R) , H2 S postcondition-ing group( H2 S) , inhibitor LY294002 group( LY) and H2 S with inhibitor group( H2 S+LY) . The left ventric-ular diastolic pressure ( LVEDP ) , the left ventricular developed pressure(LVDP), the maximum rate of in-crease or decrease of left ventricular pressure ( ± dp/dtmax ) were registered at the end of 20 min equilibri-um, 30 and 60 min of reperfusion separately. Triphe-nyl tetrazolium chloride( TTC) staining was used to de-termine the myocardial infarct size. The levels of Sirt1 and PGC-1 mRNA were tested using real-time PCR. The expressions of Sirt1 and PGC-1αwere detected with Western blot analysis. Immunohistochemical method was used to determine the location of Sirt1 . Results There were no differences in equilibrium hemodynamics observed between the experimental groups(P>0. 05). At the end of reperfusion, compared with I/R group, H2 S group had obviously ameliorated functional recov-ery and significantly decreased the myocardial infarct size(26. 9 ± 4. 9)% vs(48. 9 ± 5. 6)%(P <0. 05). Meanwhile, the expression of Sirt1 and PGC-1α in-creased significantly. However,LY294002 reversed the cardioprotective effects provided by hydrogen sulfide postconditioning and reduced the level of Sirt1 and PGC-1α, the percentage of Sirt1-positive nuclei. Con-clusion PI3 K/Akt/Sirt1 signaling pathway mediates the hydrogen sulfide postconditioning-induced protec-tion against I/R injury.

AIM:To study the effects of methylene blue (MB) on myocardial ischemia/reperfusion (I/R)-induced mitochondrial injury in isolated rat hearts.METHODS:Spragure-Dawley (SD) rats were divided into 3 groups randomly (n=6): control group, I/R model group and MB treatment group (IR+MB group).The isolated rat hearts were prepared and set up to Langendorff perfusion.The rats in I/R+MB group received MB (2 mg/kg) by intraperitoneal injection 2 h before operation.The hearts in control group were perfused with K-H solution for 110 min consecutively.The hearts in I/R group and I/R+MB group were in equilibrium for 20 min, following by 45 min of global ischemia, and then reperfused for 60 min.The heart rate (HR), left ventricular developed pressure (LVDP), left ventricular pressure maximum change rate (±dp/dtmax) and left ventricular end-diastolic pressure (LVEDP) were recorded.The perfusate was collected to determine the activity of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH).The contents of reactive oxygen species (ROS), malondialdehyde (MDA) and adenosine triphosphate (ATP), and the activity of superoxide dismutase (SOD) in the myocardial tissues were all determined.Histopathological examination of left ventricle was performed.The mitochondria from the heart tissues was isolated and the mitochondrial swelling and mitochondrial membrane potential (MMP) were analyzed.RESULTS:Compared with control group, the hearts in I/R group showed poorer function, higher CK-MB and LDH levels in the perfusate, increased ROS and MDA contents, higher SOD activity and less ATP content in the heart tissues (P<0.05).Furthermore, the mitochondrial swelling level increased and MMP reduced in I/R group (P<0.05).Compared to I/R group, MB improved heart function and reduced the release of CK-MB and LDH (P<0.05).MB also decreased ROS and MDA contents, and increased the activity of SOD and the content of ATP (P<0.05).In addition, MB alleviated mitochondrial swelling and restored the reduced MMP (P<0.05).CONCLUSION: MB protects the isolated rat hearts from I/R-induced injury by attenuating the damage of mitochondria.

Ocular decompression retinopathy(ODR)is defined as multiple hemorrhagic retinopathy following abrupt lowering of intraocular pressure(IOP)that is not explained by another process. It is frequently associated with glaucoma surgeries, but also occurs in conjunction with other ocular surgical or medical procedures. Most patients are asymptomatic, so it may be difficult to detect clinically. We review the recent articles to better understand the risk factors, pathogenesis, clinical characteristics, imaging, differential diagnoses, management and prognosis of ODR.

Objective To explore the relationship between insulin resistance and other metabolic disorders in patients with essential hypertension (EH).Methods Glucose metabolism rate (GMR) was measured by euglycemic insulin clamp technique,and salt sensitivity was tested by increase in blood pressure after salt load and its decreases alter depletion of sodium in 26 healthy subjects and 84 patients with EH.Results GMR lowered significantly in patients with EH than that in healthy subjects,P