Objective To evaluate the limit of detection of eight enzyme-linked immunosorbent assay (ELISA) according to hospital grade assessment and ISO15189:2012.Methods According to the new health industry standard WS/T 514-2017:"Establishment and verification of detection capability for clinical laboratory measurement procedures",the limit of detection (LoD) was established,in the sameset of detection system,using two reagent lot,each lot for 5 consecutive days 4 consecutive days to assess the value of the concentration of five specimens were detected repeatedly,calculated the corresponding hit rate,then transform into probability units,and the corresponding concentration value production regression model,the hit rate of 95 ％ corresponds to the probability unit 1.645 substituted into the equation,the resulting concentration value was LoD estimates.The detection limit values were tested for 3 consecutive days of detection of two LoD concentrations near the declared concentration of the sample (diluted by the standard material) was detected 4 times repeatedly to calculate the positive result was greater than or equal to the percentage of LoD statement,greater than or equal to the critical value of 87％,then verified success.Results HBsAg:0.100 IU/ml,HBsAb:9.642 mIU/ml,HBeAg:0.666 NCU/ml,HBeAb:3.700 NCU/ml,HBcAb:0.786 IU /ml,HCV:0.506 NCU/ml,TP:2.236 mIU/ml and HIV:0.135 NCU/ml.The detection limit estimates were passed.Conclusion The verification limit of the verification project in the testing method and detection system of the laboratory meet the requirements Objective.

Aim To identify the potential biomarkers associated with carbon tetrachloride(CCl 4 )-induced a-cute hepatic injury in rats and explore the therapeutic effect of Hugan Tablets(HGT). Methods The model was established by intraperitoneal injection of CCl4 in oil(1 : 1,V/ V)with a dosage of 1 mL·kg - 1 body weight to rats once. The levels of aspartate aminotrans-ferase(AST),alanine aminotransferase(ALT),alka-line phosphatase (ALP ) and lactate dehydrogenase (LDH)in serum of rats were determined. Moreover,a proton nuclear magnetic resonance (1 H-NMR)based metabonomic approach in combination with multivariate data analysis was applied to demonstrate CCl4-induced acute hepatic injury metabolic perturbations in rat urine and feces and identify the corresponding metabolic bio-markers. The intervention effect of HGT was evaluated based on the changes of metabolic phenotype and po-tential biomarkers related to acute hepatic injury. Re-sults The levels of AST,ALT,ALP and LDH in ser-um of rats with acute hepatic injury were significantly reduced by administration of HGT,respectively. The disturbed metabolic state associated with CCl4-induced acute hepatic injury in rat urine and feces could be re-stored by HGT. Meanwhile,five potential biomarkers (2-oxoglutarate,citrate,creatinine,trimethylamine N-oxide,hippurate)in rat urine and three potential bio-markers(butyrate,glucose,uracil)in rat feces related to acute hepatic injury were reversed by administration of HGT,respectively. Conclusion HGT exerts pro-tective effects against CCl4-induced acute hepatic inju-ry in rats,which is probably mediated by regulation of tricarboxylic acid cycle and gut microbiota metabolism.

Objective To investigate the value of diffusion weighted imaging(DWI)in predicting delayed encephalopathy of the rabbits brain after carbon monoxide(CO)poisoning.Methods Sixty healthy rabbits were put into self-made poisoning cabinet and were poisoned by inhalation of CO.Aeration of CO was stopped when the rabbits became comatous,and the cabinet was kept airpoof for 6 h.The rabbits underwent MRI before poisoning,at 1 h,3 d,5 d,7 d,15 d,30 d,45 d,and 60 d after poisoning respectively. Axial and sagittal T_2WI,axial T_1WI and DWI were performed.In the rabbits that did not show symptoms of delayed encephalopathy,the observation was discontinued on the 60~(th)day.In the rabbit that showed the symptoms,the observation was discontinued on the 30~(th)——45~(th)day.The changing pattern of cortical ADC values before and after CO poisoning was observed and its relationship with delayed encephalopathy was investigated.Results In the group without delayed encephalopathy(15 rabbits),the ADC value at 1 h after poisoning[(7.58?0.36)?10~(-4)mm~2/s]decreased significantly compared with the pre-poisoning value[(8.02?0.35)?10~(-4)mm~2/s](q=0.4441,P0.05).In the group with delayed encephalopathy(15 rabbits),the ADC value at 1 h after poisoning [(7.40?0.32)?10~(-4)mm~2/s]decreased significantly compared with the pre-poisoning value[(8.08? 0.32)?10~(-4)mm~2/s](q=0.6728,P

AIMTo investigate the effect of interleukin-2(IL-2) on the cell contractility and calcium handling in cardiomyocytes during normoxia or anoxia/reoxygenation.

METHODSChemical anoxia introduced by Krebs-Henseleit(K-H) solution containing 10(-3) mol/L sodium dithionite was used in the enzymatically isolated rat ventricular myocytes. The video-tracking system and spectrofluorometric method were employed to verify the cell contraction and calcium handling of the single myocyte.

RESULTSDuring anoxia, the cell contraction, amplitude of calcium transient induced by electrical stimulation and Ca2+ release induced by caffeine were depressed while resting calcium level was elevated, but the activity of the L-type calcium channels were not changed. All the parameters could not return to the pre-anoxia level during reoxygenation. IL-2 at 2 x 10(5) U/L administrated during anoxia aggravated the effect of reoxygenation on cell contraction and the calcium handling.

CONCLUSIONCoexistence of IL-2 during anoxia aggravated the effect of reoxygenation on the cell contraction and calcium handling in the isolated rat ventricular myocytes, in which the reduced calcium release from sarcoplasmic reticulum was involved.

Animals ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Heart Ventricles ; cytology ; Interleukin-2 ; pharmacology ; Male ; Myocardial Contraction ; drug effects ; Myocytes, Cardiac ; drug effects ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sarcoplasmic Reticulum ; metabolism

Objective To evaluate the clinical significance of bone scintigraphy in the preoperative diagnosis of sacral tumor. Methods Preoperative 99Tcm-methylene diphosphonate (MDP) whole body bone scintigraphy was performed in total of 103 patients with sacral tumor for whole body survey and radionuclide uptake in the sacral tumor. Of these 103 patients,39 had SPECT. According to the osteoblastic reaction in bone SPECT studies,patterns of tumor with a "hot" lesion was defined as type Ⅰ,a "cold" lesion accompanied with partial uptake was defined as type Ⅱ,a purely "cold" lesion was defined as type Ⅲ,and a "cold" lesion with marginal uptake which produced "doughnut sign" was defined as type Ⅳ. Imaging interpretation was correlated with the final pathologic diagnosis. Results Of the 103 patients,18 ( 17.5% ) had polyostotic involvement. About 46.6% (48/103 ) in planar and 84.6% ( 33/39 ) in SPECT showed decreased uptake at sacrum. Of the bone metastatic patients (n =21 ) ,12 (51.7%) had sole metastasis to sacrum. Tumor with type Ⅰ (6/6) or type Ⅱ (16/19) uptake was likely to be a malignancy,whereas type Ⅲ uptake tended to occur in the benign disease in those patients without polyostotic involvement( 5/7 ),and type Ⅳ was all appeared in giant cell tumors( n = 5 ). Conclusions Preoperative bone scintigraphy is useful in examination of polyostotic involvement for the patients with sacral tumor,but it is limited for diagnosing isolated sacral metastatic disease. Tumor uptake on bone scintigraphy can be helpful in differential diagnosis of sacral tumor.

Objective:To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g (SAMPg) mice models with Alzheimer disease (AD),and to explore the possible protective mechanism of acupuncture on mitochondria.Methods:Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group,an acupuncture at non-acupoint group and a model group,20 mice in each group.The 20 male senescence-accelerated mouse/resistance 1 (SAMR1) mice of the same age were used as a normal control group.Shenshu (BL 23),Baihui (GV 20),Xuehai (SP 10) and Geshu (BL 17) were selected for acupuncture intervention in acupuncture at acupoint group.After an 8-week intervention,mitochondrial tissues were extracted from the hippocampus.Differentially expressed proteins were identified by subcellular organelle proteomics.Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.Results:Compared with the model group,there were 13 differentially expressed protein spots in the acupuncture at acupoint group,of which,9 were up-regulated,including neurofilament light polypeptide (NFL),actin (cytoplasmic 1,database ID:ACTB),tubulin beta-2A chain (TBB2A),tropomodulin-2 (TMOD2),pyruvate dehydrogenase E1 component subunit beta (PDHE1-β),NADH-ubiquinone oxidoreductase 75 kDa subunit (database ID:NDUS1),heat shock cognate 71 kDa protein (HSC71),pyruvate dehydrogenase E1 component subunit alpha (PDHE1-α) and ATP synthase beta subunit (ATP-β);4 were down-regulated,including glial fibrillary acidic protein (GFAP),pyruvate dehydrogenase phosphatase 1 (PDP1),mitochondrial-processing peptidase subunit alpha (MMP-α) and adenosine kinase (ADK).According to the information provided in the protein database,most of the differentially expressed proteins involve the regulation of mitochondrial function and structure.The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group (P＜0.05).ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group (P＜0.05);there was no significant difference between the acupuncture at non-acupoint group and the model group (P＞0.05).Conclusion:Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.

OBJECTIVETo observe the effect of synchronous perfusion of specific respiratory chain complex IV inhibitor sodium azide (NaN3) in brain on rat ventromedial prefrontal cortex (mPFC) and acetylcholine (ACh) and choline (Ch) contents in hippocampal extra-cellular fluid, and establish the AD rat model induced by mitochondrial acute injury.

METHODThe synchronous dual-probe dual-channel brain microdialysis sampling technology was applied to synchronously perfuse modified Ringer's solution containing NaN3 (50 micro mol L-1) and neostigmine (2 micro mol L-1) into mPFC and hippocampus of conscious, freely moving normal rats, and continuously collect dialysates from different encephalic areas. Dynamic contents of ACh and Ch were determined by high performance liquid chromatography-post-column immobilized enzyme reactor-electrochemical process.

RESULTACh and Ch contents in mPFC extracellular fluid of normal rats were higher than that in hippocampus. During the process of perfusion, NaN3 could significantly reduce ACh in mPFC/hippocampal extra-cellular fluid, but remarkably increase Ch, and constantly inhibit the recovery of ACh and Ch contents in mPFC/hippocampus.

CONCLUSIONThe synchronous perfusion of NaN3in rat mPFC and hippocampus can injure functions of the cholinergic nerve projection area, and cause the acute AD model with ACh and Ch metabolic disorders. This model can be used in pathogenetic and pharmacological studies.

Acetylcholine ; metabolism ; Animals ; Choline ; metabolism ; Extracellular Fluid ; drug effects ; metabolism ; Hippocampus ; cytology ; Male ; Neurotransmitter Agents ; metabolism ; Perfusion ; Prefrontal Cortex ; cytology ; Rats ; Rats, Sprague-Dawley ; Sodium Azide ; administration & dosage ; pharmacology ; Time Factors

Alanine Transaminase ; blood ; DNA, Viral ; blood ; Female ; Hepatitis B Antibodies ; blood ; Hepatitis B Surface Antigens ; blood ; Hepatitis B, Chronic ; virology ; Humans ; Male

AIMTo investigate the antiarrhythmic effect of jumi (JM) extraction.

METHODSThe conventional antiarrhythmic methods were used.

RESULTSAdministration of JM extraction reduced the occurrence of ventricular fibrillation induced by chloroform in a dose-dependent manner in mice. Quinidine significantly decreased the number of ventricular premature beats and ventricular tachycardia, shortened the duration of arrhythmia in aconitine-treated rats. But JM extraction had no effect on aconitine-induced arrhythmia. Compared with control, arrhythmia score was lower in ischemia/reperfusion rats which pretreated with 2.0 g/kg of JM extraction.

CONCLUSIONJM extraction has obvious protection effects in chloroform- and ischemia-induced arrhythmia, but has no effect in aconitine-induced arrhythmia.

Animals ; Anti-Arrhythmia Agents ; therapeutic use ; Arrhythmias, Cardiac ; chemically induced ; drug therapy ; Female ; Male ; Mice ; Plant Extracts ; therapeutic use ; Rats ; Rats, Sprague-Dawley

The aim of the present study was to explore the effect of nitric oxide (NO) on iron-induced toxicity in rat hearts. Langendorff perfused rat heart and enzymatically isolated cardiomyocytes were used. It was shown that lipophilic Fe-HQ reduced the contractile amplitude, velocity and end-diastolic cell length in the cardiomyocyte, while the left ventricular developed pressure (LVDP), +/-dp/dt(max), heart rate and coronary flow showed biphasic alterations, which increased in the first 2 min and then was followed by a decline in isolated perfused rat heart; the contents of lactate dehydrogenase (LDH) and creatine kinase (CK) in the coronary effluent and the malondialdehyde (MDA) in the myocardium were increased. L-arginine (L-Arg), an NO precursor, reduced the contractile amplitude and end-diastolic cell length in the cardiomyocyte; but reversibly increased LVDP, +/-dp/dt(max), and coronary flow in isolated perfused rat heart. Pretreatment with L-Arg aggravated the Fe-HQ-induced decrease in contractile amplitude, velocity and end-diastolic cell length in the cardiomyocyte; LVDP, +/-dp/dt(max), heart rate and coronary flow were significantly reduced in the perfused heart, and the levels of LDH and CK increased in the coronary effluent. In contrast, the NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME) blocked the Fe-HQ induced change in contractile amplitude, velocity and end-diastolic cell length in the cardio- myocyte; it inhibited the decrease in LVDP, LVEDP and +/-dp/dt(max), and reduced the LDH and CK. Removing endothelial cells in coronary vessels attenuated the increase in LVDP and +/-dp/dt(max) at the beginning of Fe-HQ perfusion. It is suggested that L-Arg aggravates the iron-induced cardiac dysfunction, NO can mediate the iron-induced toxicity in heart, and endothelial cells in coronary vessels play an important role in the early stage of the effect of iron.
Animals ; Arginine ; pharmacology ; Coronary Vessels ; cytology ; Creatine Kinase ; metabolism ; Endothelial Cells ; drug effects ; Heart ; drug effects ; Iron ; toxicity ; L-Lactate Dehydrogenase ; metabolism ; Malondialdehyde ; metabolism ; Myocardium ; metabolism ; Myocytes, Cardiac ; cytology ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; metabolism ; Rats