Objective To investigate the immune modulation effect of lymphocytes co-cultured with human cord blood derived-multipotent stem cells(CB-SCs) and to explore their therapeutic potential for Alzheimer's Disease (AD) model mice.Methods CB-SCs were isolated from human cord blood.Lymphocytes were isolated from spleens of AD model mice.The lymphocytes were co-cultured with CB-SCs or cultured alone for 72 h.AD model mice were divided into experimental group and control group randomly,and then the experimental group mice were administrated with lymphocytes co-cultured with CB-SCs and control group were administrated with lymphocytes cultured alone by caudal vein injection.Then,the behavior experiment was carried out.The CD4+CD25+Foxp3+Tregswere detected by flow cytometry.The protein expression of TNF-αand IL-10 in peripheral blood was detected by ELISA,The mRNA expression of TNF-α and IL-10 in brain tissue was detected by PCR.The amyloid-β(Aβ)plaques were detected by immunohistochemistry.Results 1)There was spatial learning and memory improvement on experimental group.2)The Aβ plaques of experimental group decreased.3)The percentage of CD4+CD25+Foxp3+Tregs in peripheral blood and IL-10 level in plasma were higher in experimental group(P<0.05).The pro-inflammatory cytokines,TNF-α level in plasma of experimental group was lower than that in the control group(P<0.001).4)The mRNA expression of IL-10 in brain was higher in experimental group as compared to those in the control group(P<0.05),and the mRNA expression of TNF-α of experimental group was lower than that in the control group(P<0.05).Conclusions The lymphocytes co-cultured with CB-SCs have immunotherapeutic effect in AD model mice,which is mainly displayed with increased proportion of Tregs and enhanced anti-inflammatory function of Tregs.

OBJECTIVETo provide inspiration and ideas for clinical treatment of coronary heart disease (CHD) by data mining technology based frequency analysis and cluster analysis of medical records, prescriptions and herbs in treating CHD by distinguished veteran doctors of traditional Chinese Dedicine (TCM).

METHODSTotally 386 medical cases were retrieved from Wanfang Data, Chinese Scientific Journals Database (VIP medical information resources system, China National Knowledge Infrastructure (CNKI), and Typical Collections of Medical Cases by Contemporary Distinguished Veteran Doctors of Traditional Chinese Medicine. They input into database trimmed after unified standard. Medication laws of CHD by distinguished veteran doctors of TCM were analyzed using frequency analysis and cluster analysis, and so on.

RESULTSDistinguished veteran doctors of TCM frequently used top ten herbs in treatment of C D as Salvia miltiorrhiz , Ligusticum wallichii, Trichosanthes kirilowi, Pinellia ternat, Angelica sinensis, Poria coco stragalu , Panax ginseng, Allium macrostemon, and Radix Ophiopogonis. Cluster analysis summarized that there were 16 herb pairs commonly used, 7drug assemblies consisting of 3 herbs and 5 drug assemblies consisting of multiple herbs.

CONCLUSIONSDistinguished veteran doctors of TCM mainly used herbs assemblies capable for invigorating Pi to resolve phlegm, and promoting qi and activating blood circulation in treating CHD. Meanwhile, they concurrently used herbs combination of nourishing Xin and tranquilization, and regulating yin and yang.

China ; Cluster Analysis ; Coronary Artery Disease ; therapy ; Data Mining ; Databases, Factual ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional

OBJECTIVEThe local field potential (LFP) is a summation of dendritic potentials. The main objective of the present work is to view the features of LFP in M1 during the experimental rat pushed a paddle with the right forelimb.

METHODSFour rats were trained to press a paddle with the right forelimb for water. Then Two bundle micro-electrode with four channels were implanted into the rat's left and right primary motor cortex (M1)(AP + 3.0 mm, ML +/- 1.6 mm, H-1.6 mm) with stereotaxic apparatus. After three days recovery, 8-channel Deep-EEG and the pulse signal of paddle pressed were recorded during the rats were in operant chamber, and at the same time, the behavior were also recorded with video recorder.

RESULTSThe LFP in left M1 were defined as the substance between two channel deep-EEG. It is found that low frequency, high amplitude signal appear aligned with the paddle pressed pulse signals. With threshold detect method, about 80% press-paddle behavior could be detected.

CONCLUSIONThe result indicates that LFPs in this position in M1 are relative to forelimb's movement, and a powerful brain-computer interface system maybe developed with the LFPs.

Animals ; Brain-Computer Interfaces ; Evoked Potentials, Motor ; physiology ; Male ; Microelectrodes ; Motor Cortex ; physiology ; Movement ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVE To investigate the effect of overexpression of vascular cell adhesion molecule-1(VCAM-1)on the migration in vitro of the murine mesenchymal stem cells(MSCs)and its possible mechanism. METHODS The migration ability of normal mouse MSC (C3) ,empty vector-transfected MSC(C3+N) and VCAM-1 transfected MSC(C3+VCAM-1)was assessed by Transwell culture system in vitro after incubation for 8 and 12 h,respectively. The fetal bovine serum (FBS) was used as the chemotactic agent to induce MSC migration. The transmigrated cells were detected with methylosaniliam chloride(crystal violet)as well as DAPI staining.Furthermore,the specific chemical inhibitors of mitogen-activation protein kinase (MAPK) pathway ( SB203580,PD98059 and JNK inhibitorⅡ)were added to the Transwell system for 12 h and the alteration of the MSC migration ability was evaluated. RESULTS After incubation with FBS for 8 and 12 h,the absolute migrated cell number(7467 ± 485 and 8795 ± 255)and migration rate〔(14.9 ± 1.0)% and(17.6 ± 0.5)%〕of MSC in C3+VCAM-1 group were significantly increased compared with C3 group〔2731±562 and 4779±224, (5.5 ± 1.1)%and(9.6 ± 0.4)%〕and C3+N group〔2539 ± 321 and 5645 ± 1080,(5.1 ± 0.6)%and(11.3 ± 1.1)%〕(P<0.05,P<0.01),but there was no significant difference between C3 and C3+N groups. Moreover,the MSC migration ability of C3+VCAM-1 group was partially suppressed by addition of JNK inhibitorⅡ. The transmigrated cell number(4843 ± 167)and migration rate〔(9.7 ± 0.3)%〕were decreased compared with those of C3+VCAM-1 group without JNK inhibitorⅡ(P<0.01). SB203580 and PD98059,as specific chemical inhibitors of MAPK pathway,had no effect on MSC migration. CONCLUSION VCAM-1 can enhance mouse MSC migration in vitro and th4e mechanism may be related to JNK/MAPK pathway activation.

Nowadays the role of genetic findings in determining the diagnosis, therapy and prognosis of acute myeloid leukemia (AML) has become more valuable. To improve and validate the detection of clonal chromosomal aberrations in leukemia, we designed a combined application of karyotyping with multiplex reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH), and addressed the expression and distribution of fusion genes among the subtypes of Chinese adult patients with de novo AML. Multiplex RT-PCR assays were performed on 477 samples from newly diagnosed AML patients, and cytogenetic data were obtained from 373 of them by R or G banding techniques and those in some cases were confirmed by FISH. The PCR products in some suspected cases were tested by two-directional sequencing. The results showed that except unqualified samples, fusion genes were detected by multiplex RT-PCR in 211 of 474 patients (44.51%), including AML1-ETO, CBFβ-MYH11, PML-RARα, PLZF-RARα, NPM-RARα, MLL rearrangements, BCR-ABL, DEK-CAN, SET-CAN, TEL-PDGFR, TLS-ERG, AML1-MDS1 (EVI-1). In 373 patients, who took both multiplex RT-PCR and karyotype analysis, the detection rate of chromosomal aberrations by using multiplex RT-PCR and karyotyping was 160/373 (42.89%) and 179/373 (47.98%) respectively, and the combination could optimize the detection rate of clonal genetic abnormalities to 216/373 (57.90%). The PCR results from 11 cases "normal" in karyotyping but abnormal in RT-PCR for MLL rearrangements were confirmed by two-directional sequencing. It is concluded that karyotype studies remain the cornerstone for genetic testing; conventional cytogenetics and molecular-based methods are complementary tests for the detection of clonal genetic aberrations in AML, especially for the cryptic or submicroscopic aberrations. Once a genetic marker has been identified by combined analysis, it could be used to monitor residual disease during/after chemotherapy, by quantitative RT-PCR and/or FISH.

Objective To study the effect of autoantibody test which includes antinuclear antibodies(ANA),antinuclear antibody fluorescence patterns,anti-extractable nuclear antigen(ENA) antibodies and anti-double strands DNA(ds-DNA) antibodies in the diagnosis of pediactic autoimmune diseases.Methods Of all the inpatients which had positive results of autoantibody tests,135 cases were reviewed.The autoantibody assay,positive value(PV) analysis were performed respectively.Results PV of ANA test to autoimmune diseases was 0.36 which was proportional to the intensity of fluorescence;Of all the fluorescence patterns,speckled(fine) had a relatively high PV;Anti-ENA and anti-dsDNA antibody tests had higher PV than ANA test.Conclusion Fluorescence intensity,(anti-ENA) antibody test and anti-dsDNA antibody test may be useful in identifying autoimmune diseases in clinic.

Objective: To propose a method for making organ-on-a-chip based on 3D printing, and study the relationship between cell growth on the chips and various factors. Methods: Through 3D printing technology and surface microstructure transfer method, ulcer-like and ridge-like mi-crostructures of the tumor surface and the intestinal villi were fabricated on a polydimethylsiloxane (PDMS) chip. Combined with fluorescence imaging, the effects of surface modification, shapes and heights of microstructures, and culture time on the surface coverage and density of Caco-2 cells on the chip were measured. Results:The PDMS chip was more likely to induce cell adhesion and growth rather than the 3D printing resin chip. On the surface of three-dimensional structure, cell surface coverage and cell density increased after the surface was treated with rat tail collagen (P<0.05). The height of surface structure of the chip affected the cell density (P=0.000). After surface modification, there was no significant difference in cell density at the same height of different steric structure (P>0.05). Conclusion: The intestinal villi and tumor topological organ chips can be fabricated by 3D printing technology and surface microstructure transfer method. The surface modification and microstructure height affect the cell growth on the surface.

OBJECTIVETo discuss the feasibility of vascular bundle implantation combined with allogeneic bone marrow stromal cells (BMSCs) transplantation in treating rabbit femoral head osteonecrosis and bone defect, in order to explore a new method for the treatment of femoral head necrosis.

METHODSThirty-six New Zealand rabbits were randomly divided into three groups,with 12 rabbits in each group. Bilateral femoral heads of the rabbits were studied in the experiment. The models were made by liquid nitrogen frozen, and the femoral heads were drilled to cause bone defect. Group A was the control group,group B was stem cells transplantaion group of allograft marrow stromal,and group C was stem cells transplantation group of allograft marrow stromal combined with vascular bundle implantation. Three rabbits of each group were sacrificed respectively at 2, 4, 8, 12 weeks after operation. All specimens of the femoral heads were sliced for HE staining. Furthermore ,vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area were measured and analyzed statistically.

RESULTSIn group C,new bone trabecula and original micrangium formed at the 2nd week after operation; new bone trabecula was lamellar and interlaced with abundant micrangium at the 8th week;at the 12th week,the broadened,coarsened bone trabecula lined up regularly,and the mature bone trabecula and new marrow were visible. At the 2nd week after operation,there was no statistical significance in the percentage of new bone trabecula of femoral head coronary section in defect area between group B and C. While at 4, 8, 12 week after operation, vascular density and the percentage of new bone trabecula of femoral head coronary section in defect area of group C was higher than that of group B.

CONCLUSIONAllogeneic bone marrow stromal cells cultured in vivo can form new bone trabecula, and can be applied to allotransplant. Vascular bundle implanted into the bone defect area of femoral head necrosis could improve blood supply, and promote the formation of bone trabecula.

Animals ; Blood Vessels ; transplantation ; Combined Modality Therapy ; Female ; Femur Head Necrosis ; pathology ; surgery ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; Rabbits ; Transplantation, Homologous

MicroRNAs (miRNAs) are small, single-stranded and noncoding RNA molecules of about 22 nucleotides (nt) in length that regulate mRNA by binding to 3' untranslated regions (3'UTR) of target mRNA, inducing digestion, degradation and/or translational repression of the latter. Posttranscriptional regulation of gene expression by miRNA is critical for a wide range of physiologic and pathologic processes, including cell proliferation, differentiation, apoptosis, development and oncogenesis. Dendritic cells (DC) are professional antigen presenting cells that have a pivotal role in controlling immune responses. The latest studies indicated that miRNA are indispensable in regulation of development, differentiation and functions of DC. This review discusses the latest studies of miRNA controlling DC biological properties in order to deep understand the regulatory mechanism of DC, therefore, provide a new thinking for the therapeutic strategies of DC-associated immunological disorders.
Animals ; Cell Differentiation ; Cell Proliferation ; Dendritic Cells ; cytology ; metabolism ; Humans ; MicroRNAs ; metabolism

OBJECTIVETo screen the active ingredients of Chailing decoction (CLD) by using rat nephritis model induced by mono-colonal antibody 1-22-3 (mAb) injection.

METHODSThe active ingredients of CLD was screened by 5 successive times of experiment. In each time, 28 rats were randomly divided into 4 groups, 7 in each. Group 1 was treated with PBS as control, Groups 2-4 were treated separately with CLD and its various ingredients, the medication was started 5 days before and lasted to 8 days after modeling by peritoneal injection, 13 days totally. All the rats were killed 8 days after modeling to observe the effect of various drugs on proteinuria, morphological change of kidney and biochemical parameters.

RESULTSCLD, Xiaochaihu decoction, various combination of thorowax root and its extract (saikosaponin-d) could reduce urinary protein, inhibit the proliferation of mesangial cell and expansion of extracellular matrix.

CONCLUSIONCLD and its active ingredients had inhibition on mAb induced rat model of nephritis, the active is saikosaponin-d.

Animals ; Antibodies, Monoclonal ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Nephritis ; chemically induced ; pathology ; Oleanolic Acid ; analogs & derivatives ; pharmacology ; Proteinuria ; pathology ; Rats ; Rats, Wistar ; Saponins ; pharmacology