Objective To establish the amplified luminescent proximity homogeneous assay(AlphaLISA) for the detection of Sendai virus.Methods The antigen concentration,serum concentration and the donor beads/acceptor beads ratio used in the AlphaLISA method were optimumized by the phalanx experiments, then the antibodies of Sendai Virus in 40 rat sera were detected by the established AlphaLISA method and ELISA detection method.The results were compared and the differentia between the two methods was confirmed by IFA.Results The optimum concentration of SV bio-peptide in AlphaLISA assay was 250 nmol/L, the best proportion of donor beads/acceptor beads ratio was 1 ∶1, using the concentration of 20 μg/mL and the serum dilution was 1∶10000.7 of the 40 rat sera were detected SV positive by ELISA, the positive rate was 17.5%, 8 of the 40 rat sera were determined SV positive by AlphaLISA, and the positive rate was 20.0%, the AlphaLISA positive serum was confirmed by IFA.Conclusions We preliminary established the Amplified Luminescent Proximity Homogeneous Assay(AlphaLISA) for the detection of Sendai virus.The sensitivity of the method is comparable to classical ELISA method, but this method use less serum samples and without washing steps.The method has some advantages in degeneracy and accuracy.

ObjectiveTo evaluate the application of 18fluoro-deoxyglucose positron emission tomography (FDG-PET) to the staging and predicting outcome in patients with lymphoma.Methods 41 patients with newly diagnosed lymphoma(median age 57 years)were explored with FDG-PET prior to and after 4 cycles of chemotherapy.With a median follow-up of 30 months (range 10-68 months),the value of FDG-PET to staging and predicting clinical outcome was assessed. Results The maximum standardized uptake value (SUVmax) of nodal and extranodal lesions was 9.7±6.9 and 8.4±6.8 respectively prior to treatment.There were significant difference (P＜0.05) in aggressive non-Hodgkin's lymphoma and indolent non-Hodgkin's lymphoma,no significant difference(P＞0.05)in non-Hodgkin's lymphoma and Hodgkin's lymphoma(HL), B-cell neoplasms and T-cell neoplasms,germinal center B-cell-like DLBCL and activated B-cell-like DLBCL. In 41 patients, 22 patients (54 ％)were detected extranodal focus by FDG-PET before chemotherapy. FDG-PET imaging upstaged in 6(15％)of initial lymphoma patients.There were 15 patients (37 ％) in stage Ⅰ and Ⅱ and 26 patients(63 ％)in stage Ⅲ and Ⅳ by FDC-PET scan.1 patient (7 ％) in stage Ⅰ and Ⅱ,6 patient (23 ％) in stage Ⅲ and Ⅳ died of disease progression during follow-up.After 4 cycles of chemotherapy,the FDG-PET was negative in 41％(17/41),positive in 59 ％(24/41) respectively.1 patient(6 ％)died of disease relapse among 17 patients who were FDG-PET negative, 6 patient (25 ％)died of disease progression among 24 patients who were FDG-PET positive during follow-up. Conclusion FDG-PET scanning plays an important role in the pretreatment staging and prediction of the prognosis after 4 cycles of chemotherapy in patients with lymphoma.Thus it may offer the potential for change in treatment paradigms.

AIM: To investigate the change of corrected vision and stereo vision in anisometropic patients after laser in situ keratomileusis (LASIK).METHODS: The clinical data of 84 cases of anisometropic children(84 eyes) were retrospective analyzed.The changes of corrected visual acuity and stereopsis of different gender,age,type of amblyopia in children anisometropic before treatment,after treatment for 3,6mo and 1a were analyzed.The stereoscopic vision correction correlation were analyzed.RESULTS: Corrected Log MAR visual acuity and stereopsis of 84 patients after LASIK surgery treatment for 3mo,and 1a later significantly decreased than before treatment (P<0.05).The corrected Log MAR visual acuity and stereopsis of male and female after treatment for 3,6mo and 1a was not statistically significant (P>0.05).Corrected visual acuity of age <30 group for 3,6mo and 1a was significantly lower than the age ≥30 group (P<0.05).After 3,6mo and 1a stereo vision with hyperopic anisometropia amblyopia decreased more than that with myopic anisometropia amblyopia(P<0.05),but corrected Log MAR visual acuity after treatment was no statistically significant difference (P>0.05).Improved correctted vision of anisometropic patients after LASIK surgery had no connection with the decrease of stereo vision(P>0.05).CONCLUSION: LASIK surgery can improve visual acuity in patients with anisometropia amblyopia,but the course of treatment of visual acuity and stereopsis affected by age and type of amblyopia.

Nutritional supplies & biodiversities of deep-sea microorganisms and correlative methodologies were introduced here in an ecological point of view, and development of their natural products was prospective in an applied point of view.

A rapid, sensitive and simple liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed and validated for the simultaneous determination of clevidipine butyrate and its primary metabolite clevidipine acid in dog blood. After one-step protein precipitation with methanol, the chromatographic separation was carried out on an Ecosil C18 column (150 mm x 4.6 mm, 5 µm) with a gradient mobile phase consisting of methanol and 5 mmol · L(-1) ammonium formate. A chromatographic total run time of 13.0 min was achieved. The quantitation analysis was performed using multiple reaction monitoring (MRM) at the specific ion transitions of m/z 454.1 [M-H]- --> m/z 234.1 for clevidipine butyrate, m/z 354.0 [M-H]- --> m/z 208.0 for clevidipine acid and m/z 256.1 [M-H]- --> m/z 227.1 for elofesalamide (internal standard, IS) in the negative ion mode with electrospray ionization (ESI) source. The linear calibration curves for clevidipine butyrate and clevidipine acid were obtained in the concentration ranges of 0.5-100 ng · mL and 1-200 ng · mL(-1), separately. The lower limit of quantification of clevidipine butyrate and clevidipine acid were 0.5 ng · mL(-1) and 1 ng · mL(-1). The intra and inter-assay precisions were all below 12.9%, the accuracies were all in standard ranges. Stability testing indicated that clevidipine butyrate and clevidipine acid in dog blood with the addition of denaturant methanol was stable under various processing and/or handling conditions. The validated method has been successfully applied to a pharmacokinetic study of clevidipine butyrate injection to 8 healthy Beagle dogs following intravenous infusion at a flow rate of 5 mg · h(-1) for 0.5 h.
Animals ; Butyrates ; blood ; pharmacokinetics ; Calibration ; Chromatography, Liquid ; Dogs ; Infusions, Intravenous ; Pyridines ; blood ; pharmacokinetics ; Tandem Mass Spectrometry

With redox-sensitive fluorescene probes DCFH-DA and DHR123, the formation of cytosolic and intramitochondrial reactive oxygen species (ROS) inside immature rat cerebellar granule cells during the apoptosis induced by nitric oxide donor S-nitroso-N-acetyl-pennicillamine (SNAP) was monitored by laser confocal scanning microscopy. The cytosolic and intramitochondrial ROS increase significantly after 0.5 mmol/L SNAP treatment for 1 h. Pre-treatment with the nitric oxide scavenger hemoglobin can effectively inhibit the formation of cytosolic and intrarnitochondrial ROS and protect neurons from apoptosis. Adding glutathione can also protect neurons from apoptosis, and the cytotoxity of nitric oxide increases significantly while the synthesis of glutathione is inhibited. The results indicated that ROS might be involved in NO-induced apoptosis in neural cells and glutathione might be the endogenesis antioxidant to protect neurons from oxidative injury.

Objective To analyze the complete genome sequence of Guangxi HEV isolate swGX32 and to compare it with other HEV isolates. Methods The overlapping fragments of HEV isolate swGX32 were amplified with reverse-transcription nested polymerase chain reaction (RT-nPCR),and the 5′ and 3′ ends of viral genome were amplified with rapid amplification of cDNA ends (RACE). The PCR products were cloned and sequenced. The sequence and phylogenetic analysis of swGX32 was performed. Results The genome of swGX32 consisted of 7240 nt excluding the polyA tail, with 4 nt overlapping between ORF1 and ORF2. ORF3 is contained in the sequence of ORF2. The complete genome sequence of swGX32 shared identity of 73%-74%, 73%, 74%-75%,83%-94% with HEV genotype 1,2,3 and 4, respectively. Among all these HEV reference sequences, swGX32 showed the highest identity with the human isolate JKO-ChiSai98C (94%). Phylogenetic tree showed that swGX32 belonged to genotype 4 and clustered with JKO-ChiSai98C in the branch of HEV subtype 4a. Conclusion The swine HEV isolate swGX32 is closely related to human strain JKO-ChiSai98C genetically and phylogenetically, which further provides molecular biology evidence of hepatitis E as a zoonosis.

Through scale-up cultivation of Eriobotrya japonica suspension cells using WAVE bioreactor, the cell growth and ursolic acid (UA) accumulation were studied. The comparison test was carried out in the flask and the reactor with cell dry weight (DW) and UA content as evaluation indexes. The culture medium, DW and UA content were compared in 1 L and 5 L working volumes of bioreactor. The orthogonal test with main actors of inoculation amount, speed and angle of rotation was developed to find the optimal combination, in 1 L working volume of bioreactor. DW of the cell growth and the UA content in bioreactor were higher than those of the shaker by 105.5% and 27.65% respectively. In bioreactor, the dynamic changes of elements in the fluid culture, the dry weight of the cell growth and the UA content in 1 L and 5 L working volumes were similar. Inoculation of 80 g, rotational speed of 26 r · min(-1), and angle of 6 ° was the optimal combination, and the cell biomass of 19.01 g · L(-1) and the UA content of 27.750 mg · g(-1) were achieved after 100 h cultivation in 1 L working volume of bioreactor. WAVE Bioreactor is more suitable than flasks for the E. japonica cell suspension culture, and culture parameters can be achieved from 1 L to 5 L amplification.
Biomass ; Bioreactors ; Cell Culture Techniques ; instrumentation ; methods ; Culture Media ; chemistry ; metabolism ; Eriobotrya ; chemistry ; growth & development ; metabolism ; Triterpenes ; analysis ; metabolism

Sodium houttuyfonate (SH) is a derivative of effective component of a Chinese material medica, Houttuynia cordata, which is applied in anti-infection of microorganism. But, the antimicrobial mechanisms of SH still remain unclear. Here, we firstly discovered that SH effectively inhibits the three types of virulence related motility of.Pseudomonas aeruginosa, i.e., swimming, twitching and swarming. The plate assay results showed that the inhibitory action of SH against swimming and twitching in 24 h and swarming in 48 h is dose-dependent; and bacteria nearly lost all of the motile activities under the concentration of 1 x minimum inhibitory concentration (MIC) (512 mg x L(-1) same as azithromycin positive group (1 x MIC, 16 mg x L(-1)). Furthermore, we found that the expression of structural gene flgB and pilG is down-regulated by SH, which implies that inhibitory mechanism of SH against motility of P. aeruginosa may be due to the inhibition of flagella and pili bioformation of P. aeruginosa by SR Therefore, our presented results firstly demonstrate that SH effectively inhibits the motility activities of P. aeruginosa, and suggest that SH could be a promising antipseudomonas agents in clinic.
Alkanes ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Biofilms ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Fimbriae, Bacterial ; drug effects ; genetics ; metabolism ; Houttuynia ; chemistry ; Pseudomonas aeruginosa ; cytology ; drug effects ; genetics ; pathogenicity ; Sulfites ; pharmacology ; Virulence ; drug effects

To study the effect of Fuzheng Huayu recipe (FZHY) on five types of isozymes of cytochrome P450 (CYP450) of normal and liver fibrosis rats by using the cocktail probe method. Dimethylnitrosamine ( DMN) was injected to induce the liver fibrosis model. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the plasma concentrations of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK solutions 2. After the oral administration with FZHY, normal rats given phenacetin, omeprazole, tolbutamide and dextromethorphan showed increase in AUC(0-t) and decrease in CL to varying degrees, indicating that FZHY obviously inhibited the activities of CYP1A2, CYP2C9, CYP2C19 and CYP2D6 in normal rats, but with no obvious effect on the activity of CYP3A4. After the oral administration with FZHY, liver fibrosis rats treated with CYP2C9 showed the significant increase in AUC(0-t) and significant decrease in Vd, hut with no obvious changes in the pharmacokinetic parameters of other four types of prove substances, suggesting that FZHY could significantly inhibit the activity of CYP2C9 in rats but had no effect on the activities of CYP1A2, CYP2C19, CYP2D6 and CYP3A4. The changes in the activity of CYP450 isozymes in liver fibrosis rats may be the reason for FZHY's different effects on CYP450 isozymes in normal and liver fibrosis rats.
Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Humans ; Isoenzymes ; genetics ; metabolism ; Liver Cirrhosis ; drug therapy ; enzymology ; genetics ; Male ; Mass Spectrometry ; Rats ; Rats, Wistar