Objective To conduct the study and comparative analysis on the performance of foreign hepatitis D virus (HDV) an-tibody test kits .Methods The collected 1 000 serum samples were tested by 3 kinds of different reagent :test reagent ,reference rea-gent and third party reagent .The detection results were processed by the statistical method and the judgement was performed base on the S/Co value .Results The Kappa analysis showed that the Kappa value was 0 .950(P<0 .01) .The specificity was 100% ,the sensitivity was 97 .73% and the area under ROC was 0 .986 .Conclusion The test kit has a relatively high accuracy and high consis-tency compared with the diagnostic reagent kits now widely used in domestic .Increasing the clinical detection rate of HDV can play a helping role on prevention ,early diagnosis and treatment of the disease .

Objective To study the result of Alexandrite laser hair removal after subcutaneous in-jection of?-MSH in anagen-induced C57BL6mice.Methods Hair shafts were depilated by wax/resin mix-ture to induce hair follicles from telogen to anagen in60C57BL6mice.The mice were randomly divided in-to groups A,B,C and D.Groups A and B were injected with0.5mg/kg and0.25mg/kg of?-MSH,respec-tively,on the back skin subcutaneously once a day.Group C was injected with the same dose of normal saline.Group D was treated as blank control.Groups A,B and C were exposed to Alexandrite laser on ana-gen(substageⅣ).Biopsies were taken before treatment and0.5h,2and28days after treatment.Speci-mens were stained with Masson-Fontana method before treatment,and with haematoxylin and eosin after treatment.The cutaneous response was observed after laser hair removal.Hair regrowth was assessed28days after treatment.Results The mean gradation value of folliclar melanin was increased in the test groups than that in control group before laser hair removal.Extent of folliclar damage and cutaneous adverse reaction af-ter laser treatment was more severe in test groups than those in control group.Hair regrowth was less obvious in test groups than that in control group,while local hyperpigmentation was increased in test groups than that in control group28days after treatment.No scarring was observed in3groups.Conclusion Subentaneous injection of?-MSH could increase melanin of the hair,decrease hair regrowth,and enhance local pigmenta-tion after laser hair removal in anagen-induced C57BL6mice.

Nitrous oxide/oxygen is one of the inhaled anesthetics that receives attention by scholars at home and abroad for its unique characteristics of excellence comparing with intravenous sedation analgesia which is more commonly used. In addition, the nurses who are the main body personnels of the pain management have certain differences in the implementation of the nitrous oxide/oxygen sedation analgesia both at home and abroad. In this paper, the nitrous oxide/oxygen analgesia and sedation and its nursing implications were reviewed from the physical and chemical properties, pharmacokinetic characteristics, excellent properties, mechanism of action and clinical application, then development directions of management in the future are put forwarded so as to provide theoretical evidences and practical guidelines for domestic medical staffs cooperating to implement the technology.

BACKGROUND:An amyloid-beta (Aβ) aggregation in the brain can induce nerve cel apoptosis, loss of synapses and functional damage. However, there is stil no effective intervention. Improving the synaptic plasticity provides an important direction for the treatment of early Alzheimer’s disease. OBJECTIVE: To screen the best model of Alzheimer’s disease and to explore the expression of synapse-associated proteins in Aβ25-35-injured PC12 neurons. METHODS:PC12 cels were induced by 50 μg/L nerve growth factor to differentiate into neuronal-like cels. Then, these cels were treated with Aβ25-35 at different concentrations. Consequently, cel survival rate was detected using cel counting kit-8; neurogranin and neuregulin immunofluorescence stainings were used to observe morphological changes of model cels; western blot used to detect the expression level of neurogranin, calmodulin kinase II, postsynaptic density-95 proteins. RESULTS AND CONCLUSION:Over time, the survival rate of PC12 neurons induced by Aβ25-35 was decreased in a dose-dependent manner. Shortened synaptic length, neuronal atrophy and sparsely interconnected neurons were visible. Expression levels of neurogranin, calmodulin kinase II and postsynaptic density-95 proteins were al down-regulated. These findings indicate that to screen the cel model of Alzheimer’s disease, the optimal concentration and interventional time of Aβ25-35are 10 μmol/L and 48 hours, respectively.

AIM: To establish a PCR method for investigating the expression of clock genes in cultured rattus cardiac myocytes. METHODS: PCR was carried out using 3 primer pairs based on the published sequences of dbp, bmal1 and per2 genes of rattus. The conditions of PCR were optimized and the specificity of amplication was tested. RESULTS: In a volume of 20 ?l, the optimal PCR mixture of bmal1 gene contains 0.5 U Taq polymerase, 0.006 ?mol dNTP and 0.035 ?mol Mg 2+; the annealing temperature being 57 ℃; and circle times being 30. In a same volume, the optimal PCR mixture of dbp gene contains 0.5 U Taq polymerase, 0.006 ?mol dNTP and 0.03 ?mol Mg 2+; the annealing temperature being 58 ℃; and circle times being 32. The optimal PCR mixture of per2 gene contains 0.5 U Taq polymerase, 0.006 ?mol dNTP and 0.05 ?mol Mg 2+; the annealing temperature being 57 ℃; circle times being 30. The specificity of amplication was very high. CONCLUSION: The PCR method can successfully detect mRNA expression of clock genes in cultured rattus cardiac myocytes.

Objective To investigate the effects of andrographolide on the expressions of Th1 cytokine [interferon (IFN)-γ] mRNA and Th2 cytokines [interleukin (ID-4, IL-10] mRNA of peripheral blood mononuclear cells (PBMCs) from patients with chronic hepatitis B (CHB) and its anti-hepatitis B virus (HBV) activity. Methods PBMCs from CHB patients were cultured with 10 mg/L andrographolide (experimental group) or 0.1% DMSO (control group). HepG2. 2. 15 cells were stimulated with andrographolide of different concentrations (experimental group) or adefovir (control group). The expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA of PBMCs and the replication of HBV DNA in HepG2. 2. 15 cell line were detected by fluorescence quantitative PCR. Results The expression of IFN-γ mRNA in PBMCs cultured with 10 mg/L andrographolide for 16 h was higher than that in control group (Z=-2. 78, P=0. 05), and the expressions of IL-4 and IL-10mRNA in experimental group were lower than control group (Z= -3. 82, P<0. 01), while the ratio of IFN-γ/IL-4 mRNA was higher than control group (Z= - 3. 82, P<0. 01). Andrographolide with different concentrations had no effect on the replication of HBV DNA in HepG2. 2.15 cells ((=11. 88, P>0.05). However, adefovir had inhibitory effect on the replication of HBV DNA (t =15. 95,P< 0. 05). Conclusion Andrographolide can regulate the expressions of IFN-γ mRNA, IL-4 mRNA and IL-10 mRNA in PBMCs from CHB patients and improve Thl/Th2 balance, while it has no effect on the replication of HBV DNA.

AIM:To investigate whether early endothelial progenitor cells (early-EPCs) expressβ2-adrenergic receptor (β2 AR) in the chronic obstructive pulmonary disease ( COPD) patients and the effect of β2 AR expression on the migration of early-EPCs.METHODS:Venous blood samples (20 mL) were obtained from antecubital vein of COPD pa-tients or healthy controls .Peripheral blood mononuclear cells were isolated by standard Ficoll gradient centrifugation , and purified by CD34 positive selection cocktail .The mRNA expression of β2 AR in the early-EPCs was detected by RT-PCR. The protein levels of β2 AR were assessed by Western blotting and flow cytometry .Chemotaxis was studied by Transwell as-say.Cultured early-EPCs were treated with ICI118551, norpinephrine (NE) or monoclonal antibody of β2AR (mAb-β2 AR) for 24 h.The number of migratory cells was counted under a light microscope .RESULTS:The level of β2 AR ex-pression in the COPD patients was higher than that in the controls .The number of migratory early-EPCs to stromal cell-de-rived factor 1αwas significantly improved by ICI 118551 compared with other COPD groups .When early-EPCs from the COPD patients or the controls were treated with different concentrations of mAb-β2 AR for 24 h, the number of migratory early-EPCs from the COPD patients and the controls treated with NE at concentration of 100 nmol/L was significantly re-duced.However, a marked decrease in the number of migratory early-EPCs from the COPD patients treated with NE was observed compared with control group .Before treated with ICI118551 or NE for 24 h, the early-EPCs were co-incubated with mAb-β2 AR for 40 min, and the number of migratory early-EPCs was not significantly different between COPD group and control group .Genetic down-regulation of β2 AR promoted the migration of early-EPCs in COPD group .CONCLU-SION:The level of β2 AR expression in the COPD patients is increased compared with the controls .The down-regulation ofβ2 AR improves the migration of early-EPCs.

Objective To evaluate the concordance of two-dimensional echocardiography (2DE) and intelligent spatiotemporal image correlation (iSTIC) in measuring fetal aortic and aortic arch diameters during the second and third trimesters.Methods Data were collected by a prospective cross-sectional study of 140 normal singleton fetuses with the gestational age from 22 to 32 weeks.A total of 6 dimensions of the fetal aortic and aortic arch,including aortic annular diameter (AO),ascending aorta diameter (AAO),aortic arch diameter [AO Arch (INA to LCCA)],aortic arch diameter [AO Arch (LCCA to LSA)],aortic isthmus diameter and descending aorta diameter (DAO),were measured by two different methods.Concordance was assessed by comparing the measurements acquired by iSTIC with those determined by 2DE and depicted by Bland-Altman plots.Inter-and intra-observer variability was evaluated by the intraclass correlation coefficient (ICC) test.Results A total of 137 iSTIC volumes in 140 cases were found to be suitable for further analysis.Good correlation was observed in the measurements that determined by 2D or iSTIC (Pearson's R2 =0.977-0.983).There was no significant difference in the mean values of all the parameters that measured by two methods.Bland-Altman plot showed that the 95％ limits of agreement (LOA) in AO,AAO,AO Arch (INA to LCCA),AO Arch (LCCA to LSA),aortic isthmus diameter and DAO were (-0.1260/+ 0.2299),(-0.1707/+ 0.2241),(-0.1547/+ 0.2190),(-0.1736/+ 0.2024),(-0.1514/+ 0.2039) and (-0.1485/+ 0.2228),respectively.The points in the outside of LOA were 5.11％ (7/137),4.38％ (6/137),5.11％ (7/137),5.84％ (8/137),4.38％ (6/137)and 4.38％ (6/137),respectively.Conclusions iSTIC has a good agreement with 2DE in measuring fetal aortic and aortic arch dimensions during the second and third trimesters.

Objective To construct Z-score models for normal fetal heart size measurements derived from fetal echocardiography.Methods Fetal echocardiography were performed in 910 normal singleton fetuses from 14th to 40th gestational weeks.Fetal transverse heart diameter (HD),heart length (HL),heart circumference (HC) and heart area (HA) were derived from a standard four-chamber view during end diastole.Using fetal somatic sizes as independent variables and heart sizes as dependent variables,the regression analyses of the mean (M) and the standard deviation (SD) for each parameter were calculated separately.A group of fetal heart diseases were assessed using these models.Results Strong correlations were found between fetal heart sizes and somatic sizes.Linear-cubic regression equations were each fitted to the models of the means of the heart sizes,whereas linear-quadratic equations were fitted to the models of the SDs.HD (r =0.984-0.986) was a dependent variable that provided the highest correlation coefficient with all of the fetal sizes,followed by HL (r =0.981-0.984),HC (r =0.981-0.982) and HA (r =0.978-0.979).All fetuses with Ebstein' s anomaly and most with homozygous α thalassemia-1 demonstrated Z scores reflective of increased heart sizes.Conclusions The fetal heart sizes Z-scores models had been constructed.The calculation of Z-scores for heart sizes as a function of fetal somatic size is feasible and simple.They might be useful for quantitative assessment of some cardiac diseases and used as new predictive indicators for homozygous α-thalassemia-1 particularly.

Objective To establish a novel and convenient method to study the phenotype of drug resistant hepatitis B virus (HBV) isolates,and to analyze the drug susceptibility by replacing the reverse transcriptase (RT) domain of wild-type HBV with that of the drug resistant HBV isolates.Methods Full length of HBV isolates was amplified and cloned from the sera of patients prior to nucleoside/nucleotide analogues (NA) treatment.Wild-type full-length HBV genomes was used to construct the recombinant expression plasmids PHY536207 (genotype B) and PHY97 (genotype C).The restriction enzyme sites were introduced in the upstream and downstream region of reverse transeription (RT) domain to construct plasmid,which were named as mPHY536207 and mPHY97,respectively.Lamivudine (LAM) resistant mutant and adefovir (ADV) resistant mutant were isolated and cloned to construct recombinant expression plasmids PHY634 and PHY6923,respectively.Subsequently,the RT domain of mPHY536207 was replaced by that of drug resistant mutant to construct the plasmids RT634 (LAM-resistant) and RT6923 (ADVresistant).The HBV constructs were transfected into Huh7 cells.The HBsAg levels in supernatant were determined by enzyme-linked immunosobent assay (ELISA),and the amount of intracellular HBV DNA was assayed by real-time polymerase chain reaction and Southern blot analysis.Results The plasmids PHY536207 and PHY97 containing genotype B and genotype C wild-type fulllength HBV genomes were constructed successfully,both of which could replicate in Huh7 cells.Intracellular HBV DNA extracted from cells in each of six-well culture plates was more than 1 × 107 copy/ mL,and the introduction of Pst Ⅰ restriction enzyme site did not affect the viral replication and HBsAg secretion.PHY634 and RT634,in which mutant RT domain was replaced into a wild type HBV expressing vector,exhibited the same HBV DNA replication under the treatment with different doses of LAM,the value of 50％ inhibitory concentration (IC50) was ＞100 μmol/L,while the IC50 of mPHY536207 was 0.18μmol/L.Moreover,wild-type isolate was sensitive to ADV (IC50 =1.2 μmol/L),while PHY6923 and RT6923 were resistant to ADV treatment (IC50 ＞100 μmol/L).Conclusion The phenotypic assay is successfully developed in this study based on replacing RT domain of wild-type HBV strains with that of clinical isolated drug resistant strain.