ObjectiveTo investigate the effect of mescenchymal stem cell (MSC) transplantation on Tourette syndrome(TS)model rats.MethodsStereotypies can be successfully induced in rats by intrastriatal microinfusion of TS sera.MSC suspension was bilaterally injected into the striatum.Survival and differentiation of transplanted MSC were tested through immunohistochemical analyses.ResultsFlow cytometry results demonstrated that the cells strongly expressed CD29(95.2％ ),CD105(97.2％ ),CD44(96.3％ ) and CD106 (94.1％).TS rats with MSC grafts exhibited significantly decreased stereotypic behaviors at 10 and 14 days(95.5 ±6.6,73.1 ± 6.5 vs.114.1 ± 6.0,108.0 ± 6.4).Immunohistochemistry analyses revealed survival of transplanted MSC and differentiation into neurons and astrocytes in the rat brain.ConclusionIntrastriatal transplantation of human MSC can provide therapeutic potential for TS.

Chronic pancreatitis is a chronic inflammatory disease of the pancreas,whichcan lead to irreversible damage to the morphology and function of the pancreas ultimately.So the early diagnosis and treatment for chronic pancreatitis are very important.And the differentiation between mass-forming chronic pancreatitis and pancreatic cancer is also a difficult problem in clinic.MRI as a noninvasive and nonradiative examination with excellent soft tissue resolution,is very valuable for the diagnosis of chronic pancreatitis.And the combination of functional MRI and conventional MRI is very helpful for the differentiation between pancreatic carcinoma and mass-forming chronic pancreatitis,which is of great significance to the clinical management.The application and progresses of function MRI in the diagnosis and identification of chronic pancrea titis were reviewed in this article.

Objective: To investigate whether the plasmid ?1neo-hgp100 could be expressed and presented in vitro and could protect the immunized mice from B16F10 challenge in vivo. Methods: ?1neo-hgp100 plasmid was constructed in which the DNA sequence encoding hgp100 CTL epitope inserted into CDR3 of ?1-neo vector. The expression of anti-bodized antigen and IFN-? in supernatant was measured by ELISA respectively after transfection J558L with ?1neo-hgp100 and further co-culture of J588L transfacted with ?1 neo-hgp100 and pmel TCR transgenic T cell. After introspleenic inoculation of ?1neo-hgp100, the protective efficacy of the gene vaccine was observed by means of measuring the tumor area every two days. Results: ?1neo-hgp100 could be expressed and presented in vitro, the immunogenecity of CTL epitope of hgp100 was strong enough and could activate gp100 specific T cell, the mice immunized with the gene vaccine could resist the tumor challenging in vivo. The mean survival time was prolonged to 36 days, compared to control group (P

Objective:Dipeptidyl peptidase I(DPPI),a lysosomal cysteine protease for serine proteases activation,highly expressed in granule immune cells.This study used collagen induced arthritis(CIA) rat model to investigate the effects of Triperygium Wilfordii Polyglucoside(TWP) on DPPI activity and the pharmacological mechanism in RA treatment.Methods:Rats were divided into four groups randomly,the blank control group,the CIA model group,the high dose (5.0 mg/100 g body-weight) and low dose(2.5 mg/100 g body-weight)treatment group.Bovine collagen-Ⅱ plus complete Freund′s adjuvant injected twice in rats.Physical assessments were carried out.12 days post-injections,the rats of treatment group were intragastric administered with TWP every day.The rats were killed after two week administrations.Serum and synovial membrane homogenates were collected and DPPI activity was detected by fluorescence substrate.Joint HE staining and cell counting were carried out,Zymography was used to detect the MMP-2/9 activity in synovial fluids.Total protein in synovial membrane homogenates were measured by BCA method.Results:TWP could reduce the number of CIA synovial tissue mast cells,inhibited DPPI activity in the synovial fluids and in serum.The expression levels of MMP-2/9 activity and synovium total protein content were also reduced by TWP.Conclusion:Triperygium Wilfordii Polyglucoside has inhibitory effects on DPPI activity on CIA rats,which might be the one of the pharmacological mechanisms in RA treatment.

Objective The aim of this study is to evaluate clinical outcomes of patients with acute type A intranural hematoma of the aorta(IMH) received surgical treatment.Methods We analyzed 40 consecutive patients with acute type A aortic IMH in Fuwai hospital.The patients are from 2012.1.1 to 2015.12.31.The average age of patients is(56 ± 11) years.Clinical outcomes and morphological evolution by CT were analyzed for 2 years.Results Most of the patients were treated medically during their initial hospitalization.There were 2 patients died in in-hospital and no 2-year mortality.16 patients (40％) were received acute surgery,24 patients(60％)were received normal surgery.Conclusion Surgical treatment would be a favorable treatment option in type A acute IMH.

ObjectiveTo investigate the effect of mesenchymal stem cell transplantation on stereotypic behavior and level of dopamine of Tourette syndrome(TS) rats.MethodsWistar rats were randomly divided into three groups namely sham group (microinfused with normal sera),TS group (microinfused with TS sera)group and TS plus MSC transplantation group.Auto-immune TS rat model were established and MSC suspension labeled with BrdU was injected into the striatum of TS rats.DA were measured by high performance liquid chromatography and electrochemical detection.ResultsTS rats with MSC grafts exhibited significantly decreased stereotypic behaviors (7d:117.3 ±8.4,14d:94.8 ±7.9,28 d:78.8 ±7.4)and level of DA in striatum(7 d:(898.2 ± 143.2) ng/g,14d:(812.4 ±92.2)ng/g,28 d:(757.9 ± 82.4)ng/g) at 7,14 and 28 days after transplantation compared to model group( ( 129.3 ± 9.8,116.5 ± 8.5,106.8 ± 6.4 ; ( 1209.3 ± 182.3 ) ng/g,( 1033.5 ± 114.7 ) ng/g,(994.8 ±112.6) ng/g,P＜0.01).DA in plasma of TS rats with MSC grafts also decreased at 7 and 14 days(7 d:(530.2 ±86.2)ng/g vs (690.3±72.7) ng/g;14d:(562.4 ±64.2) ng/g vs (689.4±83.9) ng/g,P＜0.01).ConclusionsTransplantation of MSC could reduce stereotypic behaviors in TS rats and regulate the metabolism of DA.

Objective To construct the expressing vector of shRNA/mrp1 and study its expression in vitro. Methods 64bp oligonucleotides of pSUPER and the targeted senquence of siRNA/mrp1 were synthesized and annealed to form duplex strand, then were cloned into pSUPER to construct pSUPER-shRNA/mrp1 vector. Competenced Ecoli was transfected by vector of pSUPER-shRNA/ mrp1 to screen the positive clones for sequencing and extracting plasmid. The plasmids extracted were used to transfected HepG2/mrp1 cells with a control groups by negative vectors. The expression of mrp1 mRNA and MRP1 was measured by real-time PCR and resistance of HepG2/mrp1 by flowcytometry. Results pSUPER-shRNA/mrp1 was established successfully and was sequenced to test its accuracy. Expression of mrp1 mRNA in HepG2/mrp1-si was lower than that in HepG2/mrp1 (1-fold vs 179.76-fold, P<0.001). Compared to HepG2/mrp1, the expression of MRP1 in HepG2/mrp1-si was lower (11.2% vs 97.6%, P<0. 05). The sensitivity of HepG2/mrp1-si to adiramycin was higher than that of HepG2/mrp1(45.0-fold vs 1.2-fold, P<0.01). Meanwhile, the accumulation of DNR in HepG2/mrp1-si increased significantly as compared with the control (78.58 % vs 38.44%,P<0.05).Conclusion Vector of pSUPER-shRNA/mrp1 can be constructed by the technique of enzymatic incision. The multidrug resistance of HepG2/mrp1 can be reversed by RNA interference.

OBJECTIVETo explore the causes, clinic diagnosis, treatment and prognosis of inflammatory myofibroblastic tumor (IMT) of the chest.

METHODSThe clinical data of 12 patients with IMT were retrospectively analyzed including 9 male and 3 female patients aged 36 to 81 years (mean 60.08 years). Five patients underwent pulmonary lobectomy and mediastinal lymph node dissection, 1 had partial lobectomy, 1 had mediastinal tumor resection and costectomy, and 2 received conservative treatment.

RESULTSAll the surgical patients recovered after surgery without lymph node metastasis detected by pathological examinations. In the follow-up lasting for 2 months to 5 years, no recurrence or metastasis occurred in these cases. In the 2 cases having conservative treatment, the tumor remained unchanged in one case at 3 month of follow-up and disappeared in the other case.

CONCLUSIONIMT in the chest is a rare benign tumor with a low incidence rate and malignancy. Pathological examination is reliable for definite diagnosis. Surgical operation is the primary therapeutic approach. All IMT patients should be closely followed up after the treatment.

Aged ; Aged, 80 and over ; Female ; Humans ; Lung Neoplasms ; diagnosis ; surgery ; Male ; Mediastinal Neoplasms ; diagnosis ; surgery ; Middle Aged ; Neoplasms, Muscle Tissue ; diagnosis ; surgery

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

Objective:To screen the common low-frequency mutation sites in primary biliary cholangitis (PBC) by whole exome sequencing (WES), in order to find PBC-related new susceptibility genes.Methods:From January 2000 to December 2017, the clinical data of seven patients with PBC of three PBC families diagnosed at General Hospital of Tianjin Medical University and two healthy controls were collected. The DNA blood samples were extracted and analyzed by WES. SAMtools 1.3 software was used to detect gene single nucleotide polymorphism (SNP) and indel sites, and gene mutation sites were screened from known databases of 1000 Genome, ExAC, ESP6500 and Novo-Zhonghua gene database. Pymol V2.3.2 software was performed to simulate the three-dimensional structure of major histocompatibility complex-Ⅱ (MHC-Ⅱ), and the amino acid position corresponding to the common mutation sites among families were observed.Results:The age of first diagnosis of seven PBC patients was (61.2±10.2) years. The results of serum test of seven patients indicated that alkaline phosphatase (ALP) level was (306.9±242.5) U/L, γ-glutamyltranspeptidase (GGT) level was (121.7±85.9) U/L, alanine aminotransferase (ALT) level was (47.6±33.1) U/L, aspartate aminotransferase (AST) level was (55.7±34.1) U/L and immunoglobulin G level was (14.9±3.1) g/L. The antinuclear antibody were all cytoplasmic granule types and anti-mitochondrial antibody were all positive. Five PBC patients developed intra-abdominal lymphadenopathy; two patients had extrahepatic autoimmune diseases and the pathological results of liver biopsy of two patients both showed interface hepatitis and small bile duct lesions. Eighteen SNPs were common in three PBC families, which were located in the gene of OTOA, OBSCN and human leucocyte antigen- DRB1( HLA- DRB1). rs200988634 located in OTOA gene was a common polymorphic locus among the three families. rs746424683, rs545316651, rs553144914, rs533059830 and rs56087721 located in OBSCN caused the changes of nine amino acids of different location. There were 12 SNP variations located in HLA- DRB1 gene, which leaded to the changes of 12 amino acids of different location, among them rs16822698, rs112796209 and rs11554463 mutation induced G154A, Y152C and Y107X amino acid variation of MHC-Ⅱ beta chain, and Y107X amino acid was located in the groove region of MHC-Ⅱ binding with peptide. Conclusions:WES in PBC families is a good strategy to elucidate the candidate deleterious mutation genes OBSCN and OTOA. HLA- DRB1 which is a susceptible gene of PBC may affect MHC-Ⅱ mediated antigen presentation process by the changing amino acid sequence.