Aged ; Cell Dedifferentiation ; Chondrosarcoma ; pathology ; Humans ; Ribs ; pathology

OBJECTIVETo obtain more information on DNA fingerprintings and to confirm the genetic relationships of ten wide populations of Pinellia tornata.

METHODCollect wide-type or cultivated P. tornata from ten major production areas nation-wide as material. Twelve combinations of EcoR I and Mse I primers were used to asses divergence among ten populations. The data were analyzed using unweighted pair-group method, based on arithmetic averages (UPGMA) bootstrap analysis. Cluster analyses were performed by using NTSYSpc software version 2.1. The alkaloid was extracted from P. ternate with chlorolform and determined in UV absorption spectroscop.

RESULT1,673 bands were applied with 12 combinations. These results of cluster were congruent with differences in total alkloids.

CONCLUSIONThe results of the study showed a potential application of AFLP fingerprinting for identification of P. ternata.

Alkaloids ; analysis ; China ; Cluster Analysis ; DNA Fingerprinting ; DNA, Plant ; genetics ; Phylogeny ; Pinellia ; chemistry ; genetics ; Plants, Medicinal ; chemistry ; genetics ; Quality Control

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

Abstract?AIM: To investigate the influence of propylene glycol mannite sulfate ( PGMS ) on the expression of tumor necrosis factor -α( TNF-α) and interleukin-1β( IL-1β) , in diabetic retinopathy by a rat model, to study the mechanism of PGMS against diabetic retinopathy, and provide a reliable theoretical and experimental evidence for the PGMS to be applied to clinical prevention and treatment of diabetic retinopathy.?METHODS: Male Wistar rats were randomized into 4 groups, normal control group, diabetic control group and PGMS in group, the PGMS in groups included the doses of 50mg/kg and 100mg/kg. 1% streptozotocin ( STZ) of 60 mg/kg was intraperitoneally injected in rats to establish the diabetic models. The PGMS with the doses of 50mg/kg and 100mg/kg were used to gavage in different groups of models for 12wk.Twelve weeks later, the animals were sacrificed and retinas were isolated. The aqueous humors and serums were taken, expressions of TNF-αand IL-1βprotein in retinas, aqueous humors and serums were detected by enzyme-linked immunosorbent assay ( ELISA) , respectively.The location and the expression of TNF-αand IL-1βprotein in retina tissue was detected by immunohistochemistry.?RESULTS: Twelve weeks after the use of PGMS, the level of blood glucose was not changed.ELISA showed that the expression of TNF-αand IL-1βprotein in serum and retina was significantly increased in diabetic control group than in normal control group(P<0.05), but in the groups which PGMS was given reduced, lower than those in diabetes mellitus( DM) group, especially as the concentration of PGMS increased ( P<0.05 ).But the levels of aqueous humor's TNF-αand IL-1βproteins in PGMS group were not reduced.Immunohistochemistry showed that the TNF -α protein was almost not expressed in normal control group. But the TNF-αprotein was highly expressed in diabetic control group. The expression mainly located in the ganglion cell layer, the inner plexiform layer, outer plexiform layer and pigment epithelium. The TNF-αprotein was weakly expressed at the group of 50mg/kg PGMS, the TNF-αprotein was almost not expressed at the group of 100mg/kg PGMS.When the normal control group was detected, the IL-1βprotein was weakly expressed in the outer plexiform layer.But the IL-1βprotein was also highly expressed in diabetic control group.The expression mainly located in the inner plexiform layer, outer plexiform layer and pigment epithelium. The IL -1βprotein was weakly expressed at the group of 50mg/kg and 100mg/kg PGMS.?CONCLUSION:PGMS can treat the diabetic retinopathy by downregulating the expressions of TNF-αand IL-1βin early diabetic retinopathy.PGMS maybe have a good control effect on early diabetic retinopathy.

Cathepsin B from Helicoverpa armigera (HCB) belongs to the group of cysteine proteinases. HCB is proved being involved in the degradation of yolk proteins during embryonic development,which is an acidic preferring enzyme and is resistant to SDS. The expression of the proenzyme may offer a model for investigating the activation of the enzyme. The HCB gene was constructed into pPIC9K and expressed in Pichia pastoris KM71 strain . After induction by methanol, HCB was expressed and secreted into the medium. The molecular weight of the recombinant procathepsin B was determined as about 38 kDa. The expressed product was confirmed to be HCB by immunoblotting assay using specific rabbit anti-HCB polyclonal antibody. The activity of the product was assayed by in situ hydrolysis (gelatin-SDS-PAGE). These results showed that HCB with proteolytic activity was expressed in P. pastoris KM71. This proenzyme can be used for further research on the activation of the proenzyme or industrial production.

This study aimed to simultaneously determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin by quantitative analysis of multi-components by single marker (QAMS), with astilbin as the internal standard substance. On UPLC and HPLC chromatograms, different models of instruments were used to investigate relative correction factors (RCF), in order to discuss the interoperability of RCFs established in different chromatographic systems. The engeletin content was calculated based on the established RCFs and compared by the one point external standard method and the external standard working curve method, in order to verify the accuracy of QAMS. According to the result, in different chromatograms, the ratios between RCF and retention time of engeletin and astilbin had a good reproducibility, with RSD between 2.0% and 1.8%, both were less than 3%. The relative differences among results of QAMS, the external standard working curve method of dong medicine "sunl gaems" ranged between 1.6% and 3.9%, with RSD between 2.02%-0.80% in line with relevant requirements and Pearson correlation coefficient at 0.9998 (P <0.01). The findings showed that QAMS was an accurate, reliable and highly reproducible method to determine the contents of astilbin and engeletin in dong medicine "sunl gaems" of Guizhou origin and so could be used to control the inherent quality of the herb.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Flavonols ; analysis ; Glycosides ; analysis

OBJECTIVETo observe the angiogenesis promoting effects of clinical common used Chinese herbal medicines (CHM) for activating blood circulation to remove stasis on chick embryo chorio-allantoic membrane (CAM).

METHODSChicken CAM model was established and mice blood serum containing different kinds of medicines, including Radix Peaoniae rubra, Radix Angelicae sinensis, Flos Carthami, Rhizoma Chuanxiong, Radix Salviae miltiorrhizae, Astragalus membranaceus, and their complex prescriptions, Danggui Buxue Decoction, Xuefu Zhuyu Decoction, Xiongshao Capsule, was applied on it respectively to observe the condition of angiogenesis 72 hrs after incubation. Besides, the normal saline group, blank serum group, blank group and basic fibroblast growth factor (bFGF) group were set up for control.

RESULTSAll the CHM applied and bFGF had the CAM angiogenetic promoting effect, among them, Radix Salviae Miltiorrhizae and the three complex prescriptions showed better effects than the three negative control groups in capillary formation and count, with the efficacy similar to that of bFGF. The effect of complex prescriptions was superior to that of single herb except Radix Salviae miltiorrhizae.

CONCLUSIONRadix Salviae miltiorrhizae, Danggui Buxue Decoction, Xuefu Zhuyu Decoction and Xiongshao Capsule have good angiogenesis promoting effect on CAM. This study elucidated, from a certain aspect, the mechanism of action of CHM on ischemic diseases, and unfolded the scientific evidence of applying complex prescription.

Angiogenesis Inducing Agents ; pharmacology ; Animals ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; Chorion ; blood supply ; Drugs, Chinese Herbal ; pharmacology ; Mice ; Neovascularization, Physiologic ; drug effects ; Random Allocation ; Salvia miltiorrhiza

BACKGROUNDHuman piebaldism is a rare autosomal dominant condition characterized by congenital white forelock and depigmented patches of skin, typically on the forehead, anterior trunk and extremities. Mutations in the KIT gene have been proposed to be responsible for the underlying changes in this disorder. The aim of this study was to identify gene mutation in a Chinese family with piebaldism.

METHODSA Chinese family with piebaldism presenting with white forelock and large depigmented skin macules on the abdomen, arms and legs was collected. DNA was isolated from peripheral blood of the family members. The encoding exons with flanking intron regions of the KIT gene were analyzed by polymerase chain reactions (PCR) and direct DNA sequencing. Besides, DNA extracted from 100 ethnically matched population individuals was as controls.

RESULTSA heterozygous missense mutation c.2590T > C was identified in the patients of the family. This mutation converted a serine residue to proline (p.Ser864Pro). The mutation was not found in their unaffected family members or normal controls.

CONCLUSIONA novel missense mutation c.2590 T > C was found and it might play a significant role in the piebaldism phenotype in the family.

Child ; Humans ; Male ; Mutation, Missense ; Piebaldism ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; physiology

To conduct sequencing of full-length genomes of two Japanese encephalitis virus strains (JEV) newly isolated in 2009 in China and analyze the characteristics of complete nucleotide sequences. The complete genomic sequences were obtained by RT-PCR and sequencing directly. Bioinformatics was used to analyze the nucleic acid data, deduced amino acid sequence and phylogenetic trees. The result of sequence analysis showed that the genomes of YN0911 and YN0967 strains were both 10965nt in length, which coded 3432 amino acid polyprotein. The homology of genome ranged from 83.3% to 98.9% in nt and from 94.8% to 99.7% in aa, respectively, when compared with selected JEV strains in GenBank. There were 13 amino acid divergences which were not the key virulence sites in E protein when compared with vaccine strain SA14-14-2. There were 11nt deletions in the 3' UTR region. Phylogenetic analyses based on C/ PrM, E gene and full-length genome all showed that YN0911 and YN0967 strains belonged to genotype I. The result also showed that two new JEVs had close phylogenetic relationship with the strains from Viet Nam, Sichuan Province, Guizhou Province, Guangxi Province, China. This study indicated that JEV strains newly isolated in 2009 in China were the members of JEV genotype I. The key virulence sites in E protein did not change.
Amino Acid Sequence ; Base Sequence ; China ; Encephalitis Virus, Japanese ; classification ; genetics ; isolation & purification ; Encephalitis, Japanese ; virology ; Genome, Viral ; genetics ; Humans ; Molecular Sequence Data ; Phylogeny ; Sequence Analysis, DNA