Adult ; Antihypertensive Agents ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Hypertension, Portal ; drug therapy ; Liver Cirrhosis ; drug therapy ; Male ; Middle Aged ; Phytotherapy ; Propranolol ; therapeutic use ; Salvia miltiorrhiza ; chemistry

Objective To investigate the existence and variance of quinolone-resistance genes in a group of pan-drug resistant of Acinetobacter baumannii ( A.baumannii ).Methods Twenty strains of pandrug resistant A.baumannii were isolated from patients registered in Affiliated Hospital of Nantong University from January 2011 to April 2011.Drug target genes to quinolone (gyrA,parC) and quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac(6')-Ⅰ b-cr,qepA] were analyzed by PCR and verified by DNA sequencing.Results In all 20 strains of A.baumannii,the sense mutation was found in the quinolone resistance-determining region of the gyrA gene in the form of TCA to TTA at codon 83 (Ser-83-Leu).Moreover,in the quinolone resistance-determining region of the parC gene sense mutation was found in the form of TCG to TTG at codon 80 (Ser-80-Leu) and 3 synonymous mutations were CCC to CCT at codon 40,GTA to GT]T at codon 41 and CGT to CGC at codon 44.And parC gene was a new mutation.However,mutations were not found in quinolone-resistance genes mediated by mobile genetic elements [ qnrA,qnrB,qnrS,aac( 6 ' )-Ⅰ b-cr,qepA ].Conclusion Quinolone-resistance-determining region play a key role in resistance to quinolones in this group of A.baumannii.To our knowledge,this is first report about the emergence of the new mutation of parC gene in China.

Ear Canal ; Ear Neoplasms ; diagnosis ; Humans ; Myoepithelioma ; diagnosis

Filamentous fungi are widely used in industrial fermentation. Particular fungal morphology acts as a critical index for a successful fermentation. To break the bottleneck of morphological analysis, we have developed a reliable method for fungal morphological analysis. By this method, we can prepare hundreds of pellet samples simultaneously and obtain quantitative morphological information at large scale quickly. This method can largely increase the accuracy and reliability of morphological analysis result. Based on that, the studies of Aspergillus niger morphology under different oxygen supply conditions and shear rate conditions were carried out. As a result, the morphological responding patterns of A. niger morphology to these conditions were quantitatively demonstrated, which laid a solid foundation for the further scale-up.
Aspergillus niger ; cytology ; Fermentation ; Industrial Microbiology ; Reproducibility of Results

BACKGROUND:Monofocal and multifocal Toric intraocular lens that have been widely used in clinic exhibit xcelent biological and optical characteristics and have good safety and stability after implantation. OBJECTIVE:To compare the outcomes and rotation stability in patients with cataract and astigmatism after implantation of monofocal and multifocal intraocular lens. METHODS:A total of 210 patients with cataract and astigmatism who received phacoemulsification and intraocular lens implantation were included in this study. Of them, 105 patients were assigned to monofocal intraocular lens implantation and the other 105 patients to multifocal intraocular lens implantation. Uncorrected visual acuity, best corrected visual acuity, residual astigmatism were reexamined at 1, 3 weeks and 1 month after surgery. The rotation of Toric intraocular lens was determined. The incidence of complications and spectacles- independent rate were recorded. RESULTS AND CONCLUSION:Visual acuity and residual astigmatism in each group were significantly improved after 1 week of intraocular lens implantation (P < 0.05); furthermore, these two indicators became better over time. Improvement of visual acuity and residual astigmatism in multifocal intraocular lens group was more obvious than that in monofocal intraocular lens group. Postoperative intraocular lens rotation at < 5° occurred in both groups. The intraocular lens rotation degree in multifocal intraocular lens group was higher than that in monofocal intraocular lens group at different time points (P < 0.05). There were no significant differences in incidence of complications and spectacles-independent rates between two groups at 1 month after surgery. These results demonstrate that multifocal Toric intraocular lens provides better visual acuity and residual astigmatism improvement, while monofocal Toric intraocular lens provides better rotation stability.

Adult ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Peripheral Vascular Diseases ; drug therapy ; Phytotherapy ; Takayasu Arteritis ; drug therapy

BACKGROUND: Studies have found that chondroitin sulfate proteoglycans degradation with chondroitinase canpromote the migration of Müller cells on the retina, but whether it could promote the migration of adipose-derivedmesenchymal stem cells in retinal degeneration rats is unclear.OBJECTIVE: To investigate the effect of chondroitin sulfate proteoglycans degradation with chondroitinase on adipose-derived mesenchymal stem cell treatment for retinal degeneration in rats.METHODS: Human adipose-derived mesenchymal stem cells were isolated and cultured. A retinal degeneration ratmodel was established followed by administration of adipose-derived mesenchymal stem cells+chondroitinase into thesubretinal space. The migration of adipose-derived mesenchymal stem cells and retinal cell apoptosis in rats aftertransplantation were observed.RESULTS AND CONCLUSION: The human adipose-derived mesenchymal stem cells could be successfully cultured.The labeling rate of BrdU to the human adipose-derived mesenchymal stem cells was more than 90.0%. At 7 days aftermodeling, the outer nuclear layer of the retina was collapsed, and a large amount of photoreceptor cells were dissected.The outer nuclear layer was attached to the Bruch''s membrane, and the retina was arched. The central retina andperipheral retina were damaged. In normal rats, the retinal layers were clear, and the photoreceptor cells arrangedregularly; and the retinal pigment epithelium was complete. The migration rate of adipose-derived mesenchymal stemcells in adipose-derived mesenchymal stem cells+chondroitinase group was higher than that in adipose-derivedmesenchymal stem cells group (P < 0.05). There was no significant difference in the apoptotic rate of retinal cellsbetween the two groups (P > 0.05). These experimental findings show that chondroitin sulfate proteoglycans degradationwith chondroitinase can enhance the migration ability of human adipose-derived mesenchymal stem cells on the retina.

Since HantavaxTM, formalin inactivated Hantaan virus vaccine (10,240 ELISA units/ml), has been developed in 1990 to prevent against haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan or Seoul virus, it has been commercially available in Korea. Twenty-one healthy people were booster shot once and twice after primary basic vaccination with HantavaxTM. Seroconversion rates were measured by immunofluorescent antibody technique (IFAT), enzyme-linked immunosorbent assay (ELISA), high density composite particle agglutination (HDPA), and plaque reduction neutralization test (PRNT). Seroconversion rates of 21 vaccinees at one year after primary basic vaccination were 52.3%, 95.2%, 0.0%, 47.6%, and 28.6%, and 13 vaccinees of one month after 1st booster vaccination were 100%, 100%, 30.7%, 100% and 100% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates declined slightly by twenty months, and they were 84.6%, 92.3%, 0.0%, 84.6% and 69.2% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Seroconversion rates of 9 vaccinees at three months after 2nd booster vaccination were 100%, 100%, 0.0%, 100%, and 88.9%, and 16 vaccinees at one year after the 2nd booster vaccination were 87.5%, 93.8%, 0.0%, 87.5% and 81.3% by IFAT, ELISA (IgG, IgM), HDPA and PRNT, respectively. Based on the above result HantavaxTM has proved a vigorous anamnestic response after the 1st and the 2nd booster vaccination and has persisted higher fluorescence, agglutination and neutralizing antibody titers in vaccinees.
Agglutination ; Antibodies, Neutralizing ; Enzyme-Linked Immunosorbent Assay ; Fever* ; Fluorescence ; Formaldehyde ; Hantaan virus ; Korea ; Neutralization Tests ; Seoul virus ; Vaccination

OBJECTIVETo study the activities of the key enzymes in reteplase production by Pichia pastoris.

METHODSIn shaking flasks, a series of samples were maintained after methanol induction. The cells were sonicated to prepare cell-free suspensions, in which the activities of AOX, FAD, PDC, G-6-PD, ID, alpha-KGD and SD were measured.

RESULTSThe specific activity of AOX increased during the initial 6 h, reaching the maximum of 44.5 U/mg protein. The activity decreased quickly between 6 and 24 h, followed by increment in the following 24 h and decreased afterwards. The specific activity of FAD increased gradually in the initial 48 h and then decreased, with the peak level of 6.72 U/mg protein occurred at 48 h. The specific activity of G-6-PD increased at in 2-6 h and 24-48 h, but decreased in 6-24 h and after 48 h. The specific activity of PDC decreased during the initial 6 h and increased slowed afterwards. The specific activities of ID, alpha-KGD and SD all showed a rapid decrease in the initial 6 h and a slow decrease in 6-24 h. After 24 h, the activity of ID continued to decrease, but the other two increased in the following 24 h and then decreased, reaching the maximum at 48 h.

CONCLUSIONSAccording to the changes of these enzyme activities, the whole induction phase can be divided into 4 stages: the methanol-adaptive period in the initial 6 h, the fast growth period between 6 and 24 h, the product accumulation period in 24-48 h and the metabolism lag period in 48-72 h. In the methanol-adaptive period, complete oxidation of methanol is the dominant pathway. But in the following two stages, the metabolic pathway shifts towards glycolysis and TCA cycle.

Alcohol Oxidoreductases ; metabolism ; Fermentation ; Genetic Vectors ; Glucosephosphate Dehydrogenase ; metabolism ; Methanol ; pharmacology ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics ; metabolism ; Tissue Plasminogen Activator ; biosynthesis ; genetics ; metabolism

A rapid and accurate determination method of lipids in microalgae plays a significant role in an efficient breeding process for high-lipid production of microalgae. Using low field nuclear magnetic resonance (LF-NMR), we developed a direct quantitative method for cellular lipids in Chlorella protothecoides cells. The LF-NMR signal had a linear relationship with the lipid content in the microalgae cells for both dry cell samples and algal broth samples (R2 > 0.99). These results indicated that we could use this method for accurate determination of microalgal lipids. Although LF-NMR is a rapid and easy lipid determination method in comparison to conventional methods, low efficiency would limit its application in high throughput screening. Therefore, we developed a novel combined high throughput screening method for high-lipid content mutants of C. protothecoides. Namely, we initially applied Nile red staining method for semi-quantification of lipid in the pre-screening process, and following with LF-NMR method for accurate lipid determination in re-screening process. Finally, we adopted this novel screening method in the breeding process of high-lipid content heterotrophic cells of C. protothecoides. From 3 098 mutated strains 108 high-lipid content strains were selected through pre-screening process, and then 9 mutants with high-lipid production were obtained in the re-screening process. In a consequence, with heterotrophical cultivation of 168 h, the lipid concentration could reach 5 g/L, and the highest lipid content exceeded 20% (W/W), which was almost two-fold to that of the wild strain. All these results demonstrated that the novel breeding process was reliable and feasible for improving the screening efficiency.
Chlorophyta ; chemistry ; Heterotrophic Processes ; High-Throughput Screening Assays ; Lipids ; analysis ; Magnetic Resonance Spectroscopy ; Microalgae ; chemistry ; Staining and Labeling