Aged ; Aged, 80 and over ; Atrial Fibrillation ; blood ; Biomarkers ; blood ; Female ; Humans ; Male ; Middle Aged ; Natriuretic Peptide, Brain ; blood

Objective To prepare,purify and validate specific monoclonal antibodies(McAb) against fragment in C-terminal telopeptides of collagen type Ⅱ(EKGPDP)for clinical diagnostics and related research of osteoarthritis(OA).Methods 8 BALB/c mice were immunized with EKGPDP-KLH antigen complex.McAb against EKGPDP fragements were prepared by hybridoma technique.Immunoglobulin classes and subclasses were determined using an Immuno-Type~(TM)mouse McAb isotyping kit.Ascites were producted and McAb were purified by saturation ammonium sulfate(SAS)precipitation and protein G chromatography.Specificity and immunoactivity of antibodies were detected by indirect enzyme-linked imrnunosorbant assay(ELISA).Cartilage specimens were analyzed by immunohistochemical method.Results The hybridoma cells were obtained.10 IgG and 3 IgM single colonies were picked out by limiting dilution and ELISA kit.The titers of McAb in ascites were from 2.8?10~4 to 5.1?10~5.The McAb showed the characteristics of no cross reactions with KLH,BSA,cell culture supernatant,type Ⅰ,Ⅲ collagens and whole type Ⅱ collagen.The method of SAS could get better recovery,immunoactivity of the McAb than protein G chromatograply(t=25.26;P

70 bpm during scan),the proportion of segments that could not be assessed because of motion artifact were 0.1%(1/759),1.1%(7/649),2.5% (10/407),42.6%(103/242),and 75.5%(108/143),respectively.With conventional selective coronary angiography as the golden standard,the sensitivity,specificity,positive and negative prediction values to detect≥50% stenotic lesions in the assessable segments were 79.2%,96.0%,83.8%,and 94.6%,respectively.There was a significant correlation between the number of segments per patient not assessable because of motion artifact and heart rate during the scan(r=0.655,P=0.000).Conclusion MSCT is capable of achieving high accuracy for detection of coronary artery stenosis,and is a reliable technique to diagnose coronary artery disease.

ObjectiveTo investigate the differences of whole blood chemical elements and urinary fluorine between patients with endemic fluomsis and patients without endemic fluorosis,and to find out the elements associated with endemic fluorosis and further lay a theoretical basis for clarify the pathogenesis of the disease.MethodsUsing case-control study,100 children aged 8 - 12 with dental fluorosis in Wushan and Fengjie counties of Chongqing from December 2010 to February 2011,and 30 adults with skeletal fluorosis were enrolled as case group; 100 children aged 8 - 12 without dental fluorosis and 30 adults without skeletal fluorosis were enrolled as internal control group; and 50 children without dental fluorosis and 30 healthy adults were selected as external control group in non-epidemic areas in Yubei district.Whole blood copper,zinc,calcium,magnesium,iron and urinary fluorine of all subjects were determined,and differences of these indexes were compared between groups.ResultsThe levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of children in the case group were (30.08 ± 2.83),(74.04 ± 9.75)μmol/L,(1.65 ± 0.29),(1.37 ± 0.17),(6.79 ± 1.27)mmol/L,and (0.73 ±0.37)mg/L,respectively; the levels of these elements of children in internal control group were (28.65 ± 3.96),(72.83 ± 11.35)μmol/L,(1.62 ± 0.27),(1.36 ± 0.18),(6.73 ± 1.22)mmol/L,and (0.48 ± 0.21)mg/L,respectively; in external control group were (32.03 ± 2.99),(77.78 ± 10.85)μmol/L,(1.41 ± 0.11),(1.43 ± 0.13),(7.66 ±0.55)mmol/L,and (0.49 ± 0.26)mg/L,respectively(all P＜ 0.05),the comparison between any two groups indicated the levels of copper,zinc,magnesium,iron of the case group were lower than that of external control group,urinary fluorine was higher than that of internal and external control groups(all P ＜ 0.05).The levels of copper,zinc,calcium,magnesium,iron and urinary fluorine of adult case were (26.93 ± 4.37),(95.89 ± 12.45)μmol/L,(1.50 ± 1.76),(1.56 ± 1.96),(8.15 ± 1.00)mmol/L,and (2.17 ± 0.99)mg/L; internal control group were (26.26 ±4.96),(94.86 ± 12.18)μmol/L,(1.57 ± 0.12),(1.46 ± 0.16),(7.64 ± 1.00)mmol/L,and (1.44 ± 1.22)mg/L;external control group were (26.20 ± 2.96),(96.52 ± 11.11)μmol/L,(1.48 ± 0.14),(1.45 ± 0.16),(7.81 ±0.91 )mmol/L,and (0.55 ± 0.21 )mg/L,respectively.The levels of magnesium,iron and urinary fluorine of case group were higher than that of internal control group,magnesium and urinary fluorine were higher than that of external control group(all P ＜ 0.05).ConclusionsIn vivo anti-fluorine elements are deficient in the areas with endemic fluorosis.Other chemical elements,the environment and genetic factors may be related to the pathogenesisof the disease,which needs a further comprehensive analysis.

Objective To study the content of monoamine neurotransmitters and neurotrophic factor in the hippocampus, amygdala and prefrontal cortex in anxious depression rats, and explore the possible pathogenesis.Methods 60 SD rats were randomly divided into normal group, vehicle group, anxiety group, depression group, and anxious depression group, 12 rats in each group.Chronic restraint stress combined with corticosterone injection was used to establish anxiety and depression model, the modeling time was 21 d.After modeling, elevated plus maze test, open field test, and forced swimming test were used to evaluate the anxiety and depression-like behavior, HPLC-ECD was used to detect the content of 5-HT, NE, and DA in the hippocampus, amygdala, and prefrontal cortex of rats.Western-blotting was used to detect the expression of BDNF and NT-3 in rats.Results Rats in anxious depression model group were comparable to the anxiety group in time and frequency entering open arm time, and number of locomotor activity in open field, and it had a significant difference when compared with the control and depression groups (P<0.01 or P<0.05).Immobile time in anxious depression model rats was increased significantly when compared with the control and anxiety groups (P<0.01).Meanwhile, compared with the control group, 5-HT in hippocampus and 5-HT, NE in amygdala or prefrontal cortex were significantly decreased in the depressive rats with anxiety (P<0.01 or P<0.05).Moreover, the content of BDNF and NT-3 was significantly decreased in each brain regions compared with the control group (P<0.01 or P<0.05), and BDNF levels were obviously decreased compared with the anxiety group (P<0.05).Conclusions Rats of anxious depression have significant anxiety and depression-like behaviors.Its mechanism may be associated with the down-regulation of monoamine neurotransmitters and neurotrophic factors BDNF and NT-3 in hippocampus, amygdala, and prefrontal cortex region.

Ethyl glucuronide is a specific metabolite of ethanol. There have been plenty of articles referring its pharmacokinetics, detection and application as a specific bio-marker of alcohol intake. This article reviews various analytical methods of EtG, relationship between EtG quantification and ethanol intake, and criteria for determining chronic alcohol abuse, and origin of ethanol found in the cadavers by EtG analysis. EtG has its potential application in forensic toxicology.
Alcoholism/metabolism* ; Forensic Toxicology/methods* ; Glucuronates/urine* ; Hair/chemistry* ; Humans

OBJECTIVE: To develop a method for determining ethyl glucuronide (EtG) in blood and urine by liquid chromatograph coupled with tandem mass spectrometer (LC-MS/MS). METHODS: After blood and urine de-proteined by acetonitrile, the supernate obtained from a centrifuge was analyzed by LC-MS/MS. RESULTS: Determination limit of EtG in both blood and urine was 0.05 pg/mL, with a linear range of 0.10-5.00 microg/mL (r > 0.999). Accuracy in both matrixes was 95%-109%. Inter- and intra-day RSD were less than 12%. The method showed an excellent performance when it was used to analyze authentic blood and urine samples for EtG. CONCLUSION The method is capable for blood and urine EtG analysis.
Alcoholism/urine* ; Biomarkers/urine* ; Chromatography, Liquid/methods* ; Ethanol/metabolism* ; Forensic Toxicology/methods* ; Glucuronates/urine* ; Humans ; Reproducibility of Results ; Sensitivity and Specificity ; Substance Abuse Detection/methods* ; Tandem Mass Spectrometry/methods*

This article presented the inhibitory activity of methyl 3, 4-dihydroxyphenylacetate on the enterovirus 71 (EV71) infection. The EV71 VP1 capsid protein expression levels were analyzed with Western blotting. Results revealed that the compound is able to inhibit EV71 replication in rhabdomyosarcoma (RD) cells. After being incubated with the compound at a concentration of 0.01 microg x microL(-1) for 48 h, the level of EV71 vp1 mRNA in RD cells decreased by (76.83 +/- 2.47)%. The cytotoxic activity of the compound was evaluated against RD cells by a MTT assay. The results showed that the compound had low toxicity with a CC50 of 0.072 6 microg x microL(-1). These findings suggest that methyl 3, 4-dihydroxyphenylacetate is a novel compound for antiviral therapies against EV71, which merited further investigation.
Antiviral Agents ; administration & dosage ; metabolism ; pharmacology ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Enterovirus A, Human ; physiology ; Humans ; Phenylacetates ; administration & dosage ; metabolism ; pharmacology ; RNA, Messenger ; metabolism ; Rhabdomyosarcoma ; metabolism ; pathology ; virology ; Virus Replication ; drug effects

OBJECTIVETo evaluate the skin irritation and sensitization potential of the swine acellular dermal matrix treated with hyaluronic acid (SADM-HA).

METHODS(1) Skin irritation test. Twelve New Zealand rabbits were divided into SADM-HA group, allogeneic skin group, and (human) xeno-skin group according to the random number table, with 4 rabbits in each group. Four test sites were designed on the back of each rabbit. Two test sites of each rabbit in the three groups were covered with SADM-HA, allogeneic skin, and xeno-skin, respectively. Another test site was covered with gauze containing 200 g/L sodium dodecyl sulfate solution as positive control. The last test site was covered with gauze containing normal saline as negative control. The primary irritation index and cumulative irritation index of each material were calculated. (2) Skin closed-patch test. Sixty guinea pigs were used. Fifty-four guinea pigs were divided into SADM-HA group, allogeneic skin group, and (human) xeno-skin group according to the random number table, with 18 guinea pigs in each group. Twelve guinea pigs in each of the three groups were correspondingly induced and stimulated by SADM-HA, allogeneic skin, and xeno-skin, with 6 guinea pigs in each group treated with ethanol-soaked gauze to serve as negative control. The remaining 6 guinea pigs were treated with gauze containing 25% α-hexylcinnamaldehyde ethanol solution as positive control. The rating scales of Magnusson and Kligman were used to grade the condition of skin after being treated with above-mentioned materials to evaluate skin sensitivity to them at post stimulation hour 24 and 48. Data were processed with the non-parametric test of independent samples.

RESULTS(1) In the skin irritation test, the primary irritation indexes of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were respectively -0.04, 0.13, and 0.08. The cumulative irritation indexes of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were respectively 0.27, 0.10, and 0.25, which were close to those of negative control within the three groups. The skin irritation of each of the three materials was negligible. (2) In the skin closed-patch test, all scores of the three dressings in SADM-HA group, allogeneic skin group, and xeno-skin group were between 0 and 1. The scores of SADM-HA group and allogeneic skin group were close to those of negative control within the two groups (with U values respectively 188.00 and 90.00, P values both above 0.05). The differences were statistically significant between each material of the three groups and positive control (with U values respectively 19.00, 59.00, 21.50, P values all below 0.01).

CONCLUSIONSThe SADM-HA is safe and reliable without skin irritation and sensitization, and it has encouraging prospect in clinical application.

Acellular Dermis ; adverse effects ; Animals ; Guinea Pigs ; Hyaluronic Acid ; adverse effects ; Rabbits ; Skin ; Skin Irritancy Tests ; Skin Transplantation ; methods ; Swine

Objective To observe the effects of Zuogui Jiangtang Jieyu Prescription (ZGJTJYP) on pathological morphology of hippocampal neurons in rats of diabetes mellitus with depression (DD); To discuss its protective mechanism of action. Methods DD rat model was established by tail vein injection of STZ combined with chronic stress. Totally seventy-two SPF rats were randomly divided into normal group, model group, positive medicine group, ZGJTJYP low-, medium-, and high-dose groups, with 12 cases in each group, and were given corresponding medicine. After the last administration, blood glucose and behavioral changes were measured in each group. HE staining, Nissl staining, and Golgi staining were used to observe the changes of pathological morphology in rat hippocampus. The expression of Caspase-3 in hippocampus was detected by immunohistochemistry. Results Compared with the normal group, blood glucose level was significantly increased in the model group; the number of neurons in the hippocampus decreased in the model group; the apoptosis increased in the model group; the neuronal soma showed deformation and reduced dendritic spines and other pathological damages in the model group. After intervention of ZGJTJYP, hyperglycemia and depression-like status of the model rats were relieved; the number of hippocampal neurons increased significantly; neuronal morphology basically returned to normal. The expression of Caspase-3 in ZGJTJYP high-and medium-dose groups was significantly lower than that in model group. Conclusion ZGJTJYP can significantly improved blood glucose and learning and memory abilities in DD rats, which may be related to slowing hippocampal injury and apoptosis, and restoring neuronal quantity and morphology.