Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma were widely used in strengthening spleen under different disease conditions, and were easily and often misused each other. Therefore, DNA barcode was used to distinguish Atractylodis Macrocephalae Rhizoma and Atractylodis Rhizoma from their adulterants to ensure the safe use. The sequence lengths of ITS2 of Atractylodes macrocephala, Atractylodis Rhizoma (A. lancea, A. japonica and A. coreana) were both 229 bp. Among the ITS2 sequences of A. macrocephala, only one G/C transversion was detected at site 98, and the average GC content was 69.42%. No variable site was detected in the ITS2 sequences of A. lancea. The maximum K2P intraspecific genetic distances of both A. japonica and A. coreana were 0.013. The maximum K2P intraspecific genetic distances of A. macrocephala, A. lancea, A. japonica and A. coreana were less than the minimum interspecific genetic distance of adulterants. The ITS2 sequences in each of these polytypic species were separated into pairs of divergent clusters in the NJ tree. DNA barcoding could be used as a fast and accurate identification method to distinguish Atractylodis Macrocephalae Rhizoma, Atractylodis Rhizoma, from their adulterants to ensure its safe use.
Atractylodes ; classification ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Rhizome ; classification ; genetics

Aim To monitor the whole blood trough concentration of cyclosporine A (CsA) in renaltransplant recipients reciving triple therapy with mycophenolate mofetil, cyclosporine andprednisone and to establish an optimal therapeutic window of CsA. Methods Sampleswere measured by specific fluorescence polarization immunoassay. According to the timeafter operation and different therapy plan, the whole blood trough concentration of CsA ineach group was compared with that in control group.Results The optimal therapeuticwindow of CsA with MMF plan was 150~300 ?g? L-1 (less than one month after op-eration), 120~260?g?L-1 (1~

AIM:To be one of the primary cause injury to multiple sites of ocular of glaucoma which affects over 70 million people worldwide. We applied data mining techniques, linear and the matrix operations, efficiently calculated the network and estimated the possible function of the“node” genes of the retina and optic of glaucoma, in order to provide new thought and method on the pathogenesis of glaucoma.
METHODS: The data in this study is from Gene Expression Omnibus ( GEO ) which belong to Nation Center for Biotechnology Information ( NCBI) , the quality of the raw data CEL files was processed and analyzed by the Expression software which belong to Affymetrix Inc. , Santa Clare, CA, USA. Significant analysis method ( SAM) which base on the T test was used to identified the significant genes. Based on GRNInfer and Gvedit soft we set up gene networks of optic and retina of mice and further more enriched analysis which based on DAVID and MAS3. 0 online software were processed.
RESULTS:The analysis between the group of the optic nerve heads and retinas in different stage of glaucoma showed that the amount of significant different expressed genes in the optic never head group increased significantly comparing with the group of retina in the early stage of glaucoma, the analysis of the genes network construction show that:the node genes of optic nerve heads included Unc13c、Kif5a、TRPM1、PANX; and the node genes of retina include POU4F1, NEFL, BC03870, CALB2. Metabolic pathways enrichment analysis which based on MAS3. 0 online platform show that there was mainly the amyotrophic lateral sclerosis, tyrosine metabolism, melanogenesis, Nitrogen metabolism, Gap junction, Leukocyte transendothelial migration metabolism pathway enriched out in optic nerve head; and there was mainly amyotrophic lateral sclerosis, neurodegenerative disorders, prostate cancer, leukocyte transendothelial migration metabolism pathway enriched out in retina.
CONCLUSION:By understanding bioinformatics result, it seems optic were more sensitive than the retina to high intraocular pressure, and weather high expression of TYrp1 gene can be as a sensitive diagnostic item require more evidence back up. Functional enrich analysis of node gene showed that cytoskeleton reconstructed, molecular motor and nutrients transport function improve in optic; and in retina, the most prominent finding in retina was enrichment function modules were focus on regeneration, repairing and differentiation of cells, which remind that we should reinforce research on reparation of retina of primary glaucoma. Metabolic pathways enrichment analysis show that inflammatory response plays prominent place in optic and retina of primary glaucoma, because of the optic narrow and crowed anatomic shape, nutrient metabolism and substances transfer enrichment modules play an important role in optics of primary glaucoma.

Objective: To study the relationship between UPLC and anti-inflammatory effects of different extracts of Salvia miltiorrhiza, and to provide a basis for clarifying the material basis of anti-inflammatory effects of S. miltiorrhiza. Methods: UPLC was used to analyze the different extraction sites of S. miltiorrhiza. The anti-inflammatory effect of anti-bronchial epithelial cell inflammation model was studied by different extraction sites. The spectral relationship was established by gravitational correlation analysis (GRA). Results: 50% grams of ethanol extract (S3), 80% ethanol eluent (S4), filtered precipitate (S5), and extracted aqueous layer extract (S9) had strong anti-inflammatory ability to inflammatory cells. The 50% ethanol eluate (S3) of the ethanol extract of Danshen was the strongest. The effect of chemical composition on the anti-inflammatory efficacy of each characteristic peak was as the following order: A9>A1>A2>A6>A15>A11>A12>A7>A10>A5>A14>A3>A18>A4>A13>A16>A8, and the top four peaks with strong contribution to anti-inflammatory effects were peaks 9, 1, 2, and 6, respectively. The results were as follows: tanshinone IIA, danshensu, salvianolic acid B, and 3’-methyl salvianolic acid B. Conclusion: The anti-inflammatory efficacy of S. miltiorrhiza is the result of combination of various components. It is clear that the four components are tanshinone IIA, danshensu, salvianolic acid B, and 3’-methyl salvianolic acid B. The efficacy of the largest contribution to the future to explore the efficacy of anti-inflammatory effects of S. miltiorrhiza provides new ideas.

Objective To investigate the curative effect of brucellar spondylitis,so as to provide scientific proof for improving the curative level of the disease.Methods Epidemiological information was collected from 113 patients diagnosed as brucellar spondylitis,who were divided into 5 groups according to different drugs and drug combinations of doxycycline,gentamicin,sulfamethoxazole and rifampicin.Then the curative effect was investigated.Twenty-one patients who had greater paoas muscle abscess or Para vertebral abscess,intraspinal abscess of spinal canal,necrotic intervertebral disk and major osteolasia received the minimal invasive surgery and the focus removal surgery.Results The occurrence of the disease in female was much higher than that in male.Grazing and breeding beasts was the principal route of infection.Lumbars was mostly involved.they usually was infected in the adjacent 2 piece.L4 was the most common and seriuous one.The curative effect of doxycycline group was better than that without doxycycline(72.60% vs 35.00%,X2=15.14,P＜0.05).Doxycyeline+gentamicin+sulfamethoxazole was reeommended as the first choice.However,the curative effect did not increase despi~course of the treatment prolonged.The heMing rate and effective rate after 1 course was 52.21%(59/113)and 92.04%(104/113).that after 2 courses 58.41%(66/113)and 95.58%(108/113),that after 3 courses 59.29%(67/113)and 95.58%(108/113).The healing rate in different course did not present differences(P＞0.05).21 patients undergoing surgery were followed-up,12 patients were after 2 years and 9 patients were between 1-2 years.The healing rate was 95.24%(20/21),1 case was healed basically,the effective rate was 100%.None reoccurred.Conclusions There are characterized features in clinical epidemiology of the brucell spondylitis.Long term,adequate in dosage,combination and multi-approach use of antibiotics is the most reliable way to treat and prevent it from recurring.But fof the patients soitable for surgery.the minimal invasive surgery or the focus removal could shorten the course of therapy,decrease the complications and increase the cure rate.

Objective To explore the residual undifferentiated mouse embryonic stem cells (ESCs) in embryoid bodies. Methods Mouse R1 and Oct-4-GFP transgenic ESCs were firstly cultured in suspension to form embryoid bodies (EBs). Twenty days later, EBs were digested into single cells and then re-plated in standard ESC culture condition. The morphology of residual undifferentiated cells in EBs was observed, and surface makers and in vitro redifferentiation potency of residual cells were examined by flow cytometry and immunofluoreseent staining. The residual cells were expanded and subcutaneously injected into nude mice, and the specimens were harvested from the injection site for histological analysis 6 weeks after injection. Results There were residual undifferentiated ESCs in EBs differentiated for 20 days, which displayed clonal morphology and expressed undifferentiated cell markers of ESCs, including SSEA1, CD31, CD9 and Oct-4. The cells could be differentiated to form EBs again, and could be re-expanded from secondary EBs. The residual cells were able to form teratoma at the injection site, and mature endoderm, mesoderm and ectoderm tissues could be found in teratoma tissues. Conclusion There are residual undifferentiated ESCs after differentiation of ESCs into EBs. The residual ESCs can differentiate again in vitro and in vivo, and can residue again in the in vitro differentiation.

Objective: To establish a mouse hepatocellular carcinoma cell line that can produce mMIP-1α and to evaluate the possibility of cancer gene therapy by mMIP-1α. Methods: mMIP-1α cDNA was cloned into retrovirus vector pBabe puro and pBabe puro-mMIP-1α was constructed, then pBabe puro-mMIP-1α was used to transfect packaging cells, anti-puromycin cells was proliferated, the supernatant was used to infect hepa1-6, the anti-puromycin clone (hepa1-6 mMIP-1α) and hepa1-6 were analysed for the expression of mMIP-1α mRNA and protein by RT-PCR and immunohistochemistry respectively. The growth curve of hepa1-6 and hepa1-6 mMIP-1α was drawn. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed on agarose gel. C57B/L mouse was inoculated with the tumor cell and the tumorigenicity was studied. Results: Recombinant retrovirus vector pBabe puro-mMIP-1α with mMIP-1α cDNA was constructed. Hepa1-6 did not produce mMIP-1α mRNA and protein, while hepa1-6 mMIP-1α could produce mMIP-1α mRNA and protein. The growth curve of hepa1-6 and hepa1-6 mMIP-1α showed no difference. The chemotaxis of mMIP-1α produced by hepa1-6 mMIP-1α to mouse spleen cells was observed. The tumorigenicity was reduced. Conclusion: A mouse hepatocellular carcinoma Hepa1-6 mMIP-1α is established and mMIP-1α can affect the tumorigenecity of hepa1-6.

OBJECTIVETo investigate the specific antitumor immune response induced by idiotype protein (Id)-pulsed dendritic cells (DC) in vitro.

METHODSDC was generated from peripheral blood monocytes of the multiple myeloma (MM) patients using GM-CSF, IL-4, and TNF-alpha. The DCs were pulsed with idiotypic fragment, the F(ab')2 fragment of M protein from MM patient at the immature stage. The morphologic characteristics of the cells were observed with light and electron microscopes. The phenotypic features were analyzed with FACS, MTT assay was employed to evaluate the proliferation of autologous T cells and the inhibition rate of MM cells.

RESULTSDC precursors in peripheral blood could be induced to typical mature DC in medium containing GM-CSF, IL-4 and TNF-alpha. Mature DC with Id could increase the proliferation of the autologous T cells and activate naive T cells to become tumor specialized cytotoxic T lymphocytes (CTL). The CTL at different doses showed significant inhibition on or killing ability to autologous MM cells in vitro.

CONCLUSIONSIn a suitable cytokine environment, the DC precursors from peripheral blood of MM patients could be induced to functional DC, and vaccination of Id-pulsed DC could induce active antitumor immune response.

Adult ; Aged ; Antibodies, Anti-Idiotypic ; immunology ; Cancer Vaccines ; immunology ; Cells, Cultured ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy, Active ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo investigate the effects of C-reactive protein (CRP) on monocytes chemotaxis ability in vitro.

METHODSTranswell chemotaxis assay was used to evaluate the changes of chemotactic ability of THP-1 monocytes in each group treated with CRP in different concentration.

RESULTSCRP increased the number of attracted monocytes in response to MCP-1 (monocyte chemoattractant protein-1). When treated with CRP concentration at 2 microg x mL(-1), the number of chemotactic monocytes increased (P < 0.05). The number of attracted monocytes increased as CRP concentration was elevated (P < 0.05).

CONCLUSIONCRP can increase chemotactic ability of THP-1 monocytes in concentration dependent manner.

C-Reactive Protein ; Chemokine CCL2 ; Chemotaxis ; Humans ; In Vitro Techniques ; Monocytes

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin