A strain of Mucor named M2,which could produce protease,was isolated from a traditional fermented soybean product.Culture conditions of proteases produced by M2 were studied therefore.The results showed that nitrogen source and carbon source preferred for protease production were soybean protein isolate and glucose,while inorganic salts preferred were KH2PO4,CaCl2 and MgCl2.The suitable culture conditions for protease production were as follows:culture temperature was 28℃,inoculation volume was 2%,liquid level was 100 mL in 300 mL triangle bottle at pH 5,rotating speed was 150 r/min and culture time was 48 h.The obtained protease activity in culture was about 4.35 U/mL.The protease produced by Mucor was analyzed with SDS-PAGE.The protease had a molecular weight of 36.4 kD.

AIM: To establish a method of simultaneously determining effective components in Ciwujia Naoling Oral Liquid by RP-HPLC. METHODS: Stationary phase: Alltech C_(18) column,the mobile phase was methanol(A)-0.4% HAC-water(B) with linear gradient elution.Detection was performed at wavelength of 344 nm,254 nm respectively for isofraxidin and schisandrin. RESULTS: The two effective components could be detected by this method,and their contents could be determined.A good linearity of isofraxidin and schisandrin was in the range of 0.88-17.6 ?g/mL,0.76-15.2 ?g/mL respectively.The average recoveries were 100.64%,100.33% respectively,RSD were 1.07%,0.65% respectively. CONCLUSION: The method is simple,feasible and reproducible and can be used for the quality control of Ciwujia Naoling Oral Liquid.

Objecfive To study the effects of some cytokines such as TNF-α,IL-6 and IFN-γ as well as lipopolysaccharide on CD68 expression in HaCaT cells.Methods Human HaCaT keratinocytes were randomly divided into natural proliferation group (without stimulation),IFN-γ-stimulated group,TNF-α-stimulated group,LPS-stimulated group and IL-6 stimulated group.The work concentration of TNF-α,IL-6,IFN-γ and LPS was 50 mg/L.HaCaT cells were collected after 24-hour treatment with the cytokines followed by the examination of CD68 expression with flow cytometry,immunohistochemistry and reverse transcription(RT)-PCR,respectively.Results Compared with untreated HaCaT cells,the count of CD68-positive cells was elevated in cells stimulated by TNF-α(t=3.60,P<0.01),IL-6(t=3.93,P<0.01),IFN-γ(t=2.38,P<0.05)and LPS(t=2.52,P<0.05),and the effect of TNF-α and IL-6 was stronger than that of IFN-γ and LPS.Among the four cytokines,only IL-6 enhanced the mean fluorescence intensity of CD68-positive cells (t=8.34,P<0.01).After 24-hour treatment with TNF-α,IFN-γ and IL-6,CD68 expression was observed in the cytoplasm and on the membrane of HaCaT cells and was stronger in cells treated with TNF-α and IL-6 than in those with the other cytokines.A significant increase was observed in the CD68 mRNA expression after 24-hour treatment with TNF-α (t=4.34,P<0.01),IL-6 (t=7.52,P<0.01)and IFN-γ (t=2.81,P<0.05);TNF-α and IL-6showed a stronger promotive effect than IFN-γ.Conclusion IL-6,TNF-α,IFN-γ and LPS can upregulate the CD68 expression in HaCaT cells.

The effect of phenol concentration on the structure and function of microbial communities,which were cultured in different conditions using coking wastewater biofilm as seeding,was investigated by Biolog and denaturing gradient gel electrophoresis(DGGE)methods.The less number of bands of cultivated sam-ples on the denaturing gradient gel electrophoresis fingerprint of 16S rRNA gene indicated reduction of di-versity after enrichment and cultivation.Some bands on the DGGE gel were significantly influenced by the phenol concentration in medium.The results of Biolog showed that the original biofilm sample had the highest substrate utility capacity as measured by average well color development(AWCD).But low concen-tration of phenol enriched sample S7 showed more diverse activity on the utility of Carboxylic acids.The principal component analysis(PCA)of Biolog data revealed that the metabolic patterns were similar when using the same phenol concentration,although the sample S7 much less similar to other cultivated samples.These results suggested that the enrichment and cultivation with phenol supplemented decreased the diver-sity and also changed the metabolic function of the microbial community.Lower phenol concentration in-creased the microbial community metabolic activity.The phenol degrading capacity of isolates from each samples indicated that the enrichment and cultivation condition had changed the type and property of cul-truable bacteria.Based on these results,we concluded that the different microorganisms will be isolated un-der different cultivation condition.

Objective To explore the possible mechanisms of Arsenic Trioxide in treating rheumatoid arthritis(RA)by observing the changes of HE staining and NF-KB expression as well as the apoptosis of syn- oviocytes in adjuvant-induced arthritis rats.Methods After the animal model was set up on Wistar rats sue- cessfully,they were randomly divided into AA model group and arsenic trioxide treatment group.The treat- ment group were injected with 4 mg'kg~(-1)9?d~(-1)arsenic trioxid fluid for 7 days.All of the rats were killed 3 days after the complete of injections.The joint specimens were exposed,fixed,decalcified,wrapped and cut into slices.All slices were examined by HE stain and immunohistological evaluation.Results HE staining showed that when compared with the normal control group,the layers of synoviocytes of the AA group were increased to 6-8,and the arrangement of synoviocytes was disordered and heavy inflammatory cell infiltration were found in the AA group.In the arsenic trioxide treatment group,the layers of synoviocytes increased to 3～4,and medi- um amount of inflammatory cell infiltration were found.The intensity of synovial NF-kB(p65)positive stain in AA model group was significantly higher than that in the normal control group.The synovial expression and ac- tivation of NF-kB in the treatment group were decreased markedly,and did not return to normal level.The average gray scale calculation showed that there were significant differences between the three groups(P

Five compounds (tenuifoliside C, tenuifoliside D, telephiose A, telephiose C and polygalaxanthone III) from polygala tenuifolia wild were incubated together with CYP probe substrate in human liver microsomes to investigate the inhibitory effect towards CYP450 enzyme. Phenacetin (CYP1A2), coumarin (CYP2A6), paclitaxel (CYP2C8), diclofenac (CYP2C9), S-mepheriytoin (CYP2C19), dextromethorphan (CYP2D6), chlorzoxazone (CYP2E1), midazolam (CYP3A) were selected as the isoforfn specific substrate. And the formation of paracetamol, 7-hydroxycoumarin, 6alpha-hydroxy paclitaxel, 4'-hydroxydiclofenac, dextrorphan, 6-hydroxychlorzoxazone, 1'-hydroxymidazolam, 4'-hydroxymephenytoin were detected respectively to measure the effect towards CYP450 by high-pressure liquid chromatography (HPLC). The result shows that five compounds from polygala tenuifolia willd significantly inhibit chlorzoxazone 6-hydroxylation catalyzed by CYP2E1, while showed no effect towards CYP1A2, CYP2A6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A. And IC50 value was 38.73, 54.14, 61.77, 62.22, 50.56 micromol x L(-1), respectively.
Cytochrome P-450 Enzyme System ; metabolism ; Esters ; pharmacology ; Glycosides ; pharmacology ; Humans ; Microsomes, Liver ; drug effects ; enzymology ; Oligosaccharides ; pharmacology ; Polygala ; chemistry ; Xanthones ; pharmacology

OBJECTIVETo evaluate the role of magnetic resonance imaging (MRI) in the diagnosis of lesions in the sellar region.

METHODSThe MRI data of 142 patients with surgically and pathologically proved lesions in the sellar region were retrospectively analyzed. Based on the MRI findings, the lesions were divided into pituitary adenoma and sellar region diseases except pituitary adenoma. According to the pathologic features, the sellar region diseases except pituitary adenoma were further divided into cystic lesions, parenchymatous lesions, and cystic and parenchymatous lesions.

RESULTSOf these 142 patients, pituitary adenoma was found in 30 cases, cystic lesions of sella region diseases except pituitary adenoma in 24. Further classification resulted in 66 cases of parenchymatous lesions and 22 cases of cystic and parenchymatous lesions.

CONCLUSIONMRI-based classification of lesions in the sellar region is useful for the diagnosis and differential diagnosis.

Adolescent ; Adult ; Aged ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pituitary Diseases ; diagnosis ; Retrospective Studies ; Sella Turcica ; Young Adult

italic>Alangium chinense is a commonly used medicinal plant of Alangiaceae Alangium, one of the characteristic Miao medicines in Guizhou. The genus is strongly differentiated and has complex morphological variation, with significant differences in active ingredients and potency among herbs. In this paper, we sequenced the chloroplast genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib using Illumina high-throughput sequencing technology. The genomes of Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplasts were sequenced, their assembly annotation and structural characterization were completed, and the genomes of the congeners Alangium chinense and Alangium alpinum were downloaded from NCBI for comparative analysis. Finally, the phylogenetic tree was constructed by selecting published species with close affinity to Alangium chinense family from NCBI. The results were as follows: Alangium chinense subsp. Pauciflorum, Alangium chinense subsp. Strigosum and Alangium kurzii Craib chloroplast genomes were 156 670, 156 672 and 156 871 bp in length, with a typical four-segment structure, all containing one long single copy region (LSC), one short single copy region (SSC) and two inverted repeat regions (IRa and IRb). All of them were annotated to 133 genes, including 88 protein-coding genes, 37 tRNA genes and 8 rRNA genes. Three types of long repeat sequences and SSR loci, forward repeats, palindrome repeats and reverse repeats, were found in all chloroplast genomes. Comparative genomic analysis showed that there was diversity in the boundary transition regions of the five species, no rearrangements or inversions were found in the chloroplast genome, and there were significant differences in the coding regions of ndhA, ycf1, rpl16, ycf2, and petD genes, which provided new loci for molecular identification of Patagonia. In the phylogenetic analysis, Alangium kurzii var. kurzii clustered as a single species, and Alangium chinense subsp. Pauciflorum and Alangium chinense subsp. Strigosum clustered as a single species, with a support rate of 93 and close affinity, indicating that the phylogenetic tree can be used for the species identification. This paper can provide a basis for the taxonomic identification of Alangium, ensure the safety of clinical use of anise herbs and regulate the market of Alangium chinense herbs.

Objective To study the relationship of clinical manifestation and pathological changes and prognosis in Henoch-Schonlein purpura nephritis(HSPN)in children.Methods Clinical and pathological characteristics of 42 children with HSPN were analysed.Among them,40 children were detected of angiotensin-convertion enzyme(ACE)gene and had been followed up.Results Among them,there were 9 cases of level Ⅰof pathological types,21 cases of level Ⅱ,12 cases of level Ⅲ,but no cases of level Ⅳ.Ⅰand Ⅱ level were found in those cases of clinical manifestation with solitary hematuria and albuminuria.Pathological grades were Ⅰ,Ⅱ and Ⅲ levels in the cases of hematuria and albuminuria.Pathological types of nephrotic syndrome(NS)were Ⅱ and Ⅲ level,which were of more gross hematuria than those of other grades.ACE gene DD had serious pathological damnification.Conclusions Change of pathology cannot only be anticipated by clinical manifestation of HSPN.But if pathological damnification gets more serious,the albuminuria gets more serious.Gross hematuria and albuminuria can serve as indicators of biopsy.NS of ACE DD type have serious pathological damnification.Children with HSPN has favourable prognosis in the future.

BACKGROUNDWith features of high tissue contrast, MRI can be used for the qualitative and quantitative evaluation of atherosclerosis plaques. In this study we investigated the development of atherosclerosis plaque with high resolution 3T MRI in a rabbit model and compared the findings with the histopathological results.

METHODTwenty male New Zealand white rabbits were randomly allocated into an experimental group (n = 16) and a control group (n = 4). Atherosclerotic lesions were induced in the abdominal aorta by balloon injury and cholesterol feeding. Multiple sequences MRI examination (ToF, T1WI, T2WI, and CE T1WI) were performed at the 2nd, 3rd, and 4th months after aortic denudation. Vessel wall thickness, total vessel area, lumen area, and vessel wall area were recorded. Plaque components were analyzed using histological results as a standard reference.

RESULTSSeventeen rabbits (14 in the experimental group and 3 in the control group) received all three MR examinations. Gradually, from 2 months to 4 months, vessel wall thickness and area in the experimental group increased significantly compared with the control group (P < 0.01). In the lumen area progressive stenosis was not found, even a slight dilation had developed in the experimental group. Lipid, fibrotic and calcified plaques can be differentiated by MR image. According to histological results, MRI had good performance in detection of lipid plaque.

CONCLUSIONMRI can be used to monitor progression of atherosclerosis and differentiate plaque components.

Animals ; Aorta, Abdominal ; pathology ; Magnetic Resonance Imaging ; methods ; Male ; Plaque, Atherosclerotic ; pathology ; Rabbits ; Random Allocation