Background and purpose:Metastatic colorectal cancer (mCRC) patients with K-ras mutation won’t benefit in the anti-epidermal growth factor receptor (EGFR) treatments. Thus K-ras mutation analysis is mandatory before this treatment. There is controversy that K-ras mutation analysis should be performed on primaries or related metastases. The aim of our study was to evaluate the concordance of K-ras status between primary and related metastases tumors, thus investigate the validity and rigorousness of clinical K-ras testing. Methods:Seventy-six patients with confirmed mCRC treated in Fudan University Shanghai Cancer Center were enrolled. After DNA extraction and PCR amplification, tumor specimens with paired primary tumors and related metastatic sites were put into sequencing analysis. And the K-ras mutation status in exon 2 was assessed. Results: K-ras mutation was detected in 31 out of 76 primary tumours (40.8%) and also 40.8%of the metastatic sites. But discordance was found between primary tumor and metastasis in 15 cases (19.7%):8 primary tumors had a K-ras mutation with a wild-type metastasis, meanwhile 7 primary tumors were wild type with a K-ras-mutated metastasis. Conclusion:Our study indicated that quite a few mCRC cases have different K-ras status between primary tumors and related metastatic sites, and it’s not very rigorous to choose the anti-EGFR treatments merely according to the primary tumor-K-ras mutation.Further study and consultation are needed on this problem.

Objective - （ ） To analyze the current situation of work related musculoskeletal disorders WMSDs in neck and low ， Methods back of acupuncturists and to explore its influencing factors. A total of 272 acupuncturists from 21 hospitals above grade B level in Guangdong Province were selected as study subjects using convenient sampling method. The revised Chinese ， version of Musculoskeletal Disorders Questionnaire was used to investigate the prevalence of WMSDs in the past one year and Results the influencing factors of WMSDs in the high incidence areas such as neck and low back were analyzed. The annual （ ）， （ ） prevalence of WMSDs among acupuncturists was 94.9%. The prevalence of WMSDs in the neck 81.6% low back 81.6% （ ） ， and shoulder 63.2% was the highest and the prevalence of WMSDs in both the neck and low back was 73.5%. The prevalence - - （ vs ，P ） of multi site WMSDs was higher than that of single site WMSDs 86.0% 8.8% <0.01 . Multivariate logistic regression ， - ， ， analysis showed that acupuncturists who were female long time sitting work repeated operations within one minute and work （ P ） changing every day were common risk factors for neck or low back WMSDs or both neck and low back all <0.05 . Keeping the ， ， same posture for a long time driving to work and personnel shortage were risk factors for low back WMSDs in acupuncturists （ P ） （P ）Conclusion all <0.05 . Uncomfortable working posture was a risk factor for WMSDs in both neck and low back <0.05 . - ， - Acupuncturists are the high risk population of WMSDs and the neck and low back are the high risk sites of WMSDs. The influencing factors of WMSDs in acupuncturists include individual factors and occupational factors such as poor ergonomics and work organization.

Objective To study the influence of mesenchymal stem cells(MSCs) implantation on ventricular remodeling and heart function after myocardial infarcion. Methods Bone marrow was aspirated from Gui-zhou Xiang swines.After being isolated,cultured and co-cultured with 5-azacytidine,either autologous MSCs(experiment group) or a comparable volume of physiologic saline(control group) were injected into the infarcted myocardium.Three and six weeks later,echocardiographic measurement was performed to assess the myocardial structure and heart function,and single photon emission computed tomography(SPECT) was employed for myocardial imaging.Implanted stem cells were detected by the anti-Brdv antibody DAB with HE staining. Results Left ventricular ejection fraction(EF),fractional shortening and wall thickening were higher in experiment group than control group.The thickness of the ventricular wall and septum was found increased while the left ventricular chamber size was smaller in experiment group.It was indicated by SPECT that three and six weeks after implantation,there was obvious image defect in control group while in experiment group there were some imaging areas in the infarcted area.Brdv-labelled cells were observed in the central part of and around the infarcted area.Conclusion Implantation of MSCs into the infarcted myocardium is believed to attenuate the remodeling process,inhibit the extent of wall thinning and dilatation of the ventricular chamber.MSCs implantation may also improve the contractile ability of the myocardium and heart function.

Adult ; Calcaneus ; injuries ; surgery ; External Fixators ; Female ; Fractures, Bone ; surgery ; Humans ; Ilizarov Technique ; Middle Aged ; Treatment Outcome ; Young Adult

Objective To investigate whether elevated pre-procedural C-reactive protein (CRP) and Interleukin-6 (IL-6) concentrations may be relevant to early outcome in patients undergoing PCI.Method 100 consecutive patients undergoing pereutaneuous coronary intervention (PCI) were included in our study.Peripheral blood samples for CRP and IL-6 testing were withdrawn before PCI.Acute vascular complications resulted from PCI were determined by subsequently coronary angiography.The early coronary events during hospitalization were clinically followed.Results Thirty patients developed acute vessel occlusion,and another one developed subacute coronary thrombosis at 2 days after PCI.Increased levels of CRP correlated well with the occurrence of vascular complications as regards the significant difference existing amongⅠvsⅢandⅠvsⅣquartile groups,P

Objective To explore human umbilical cord blood mesenchymal stem cells(MSCs)colonization in hypoxic-ischemic encephalopathy(HIE)rats brain.Methods Models of 7-day-old newborn rats with HIE brain injury were established.Meanwhile,on the same day,MSCs were transplanted with Hoechst 33258 for 24 hours into rats models marked by flurescent nucleotide dye injected through caudal vein or with stereotactic instrument.After 15-30 days,then MSCs were detected with fluorescene microscope.Results With the improved rice methods,HIE animal model was successfully attained.Majority of MSCs were distributed in the cortex,hippocampus of the lesioned hemisphere,especially in the forehead.And abtained a good fusion with HIE rats brain tissue.Conclusion Human umbilical cord blood MSCs can be cultured,when transplanted into the HIE rats model,they can move into intracranial,and integer with rats brain tissue.

Background:The preferred treatment for uncomplicated type B dissection (thoracic endovascular aortic repair [TEVAR] or medical) is still under debate.Since 2001,our center has performed TEVAR for uncomplicated type B dissection.Based on our data,5-and 10-year survival rates among patients with uncomplicated type B dissection after TEVAR were 96.5％ and 83.0％,respectively.We,therefore,believe that TEVAR is preferable for uncomplicated type B dissections.This study analyzed the impact of a pre-operative smoking history on long-term survival after TEVAR in patients with uncomplicated type B dissections.Methods:From May 2001 to December 2013,data from 751 patients with type B dissections were collected and analyzed.Patients were divided into two groups (337 smoking patients and 414 non-smoking patients).The Kaplan-Meier method and log-rank test were used to compare survival curves of the two groups.Multivariable analyses using the Cox proportional hazards model were used to estimate the effects of smoking on survival rates.Results:The 5-and 10-year survival rates of non-smokers were 97.6％ (95％ confidence interval [CI],96.0％-99.2％) and 87.0％ (95％ CI,81.6％-92.7％),respectively,and 94.9％ (95％ CI,92.2％-97.7％) and 73.8％ (95％ CI,62.3％-87.5％) for smokers,respectively (Log-rank test,P =0.006).Multivariable analyses showed that smoking increased the risk of death during follow-up,2.1-fold when compared to non-smokers (P =0.039).Conclusion:A pre-operative smoking history increases long-term mortality rates after TEVAR in patients with uncomplicated type B dissections.

BACKGROUNDNeuropathic pain results from a lesion or disease affecting the somatosensory system at either the peripheral or central level. The transmission of nociception within the central nervous system is subject to modulation by release and reuptake of neurotransmitters, which maintain a dynamic balance through the assembly and disassembly of the SNARE complex as well as a series of neurotransmitter transporters (inhibitory GABA transporters GAT and excitatory glutamate transporters GT). Neuronal hyper-excitability or defected inhibition involved in neuropathic pain is one of the outcomes caused by imbalanced neurotransmission. SNAP-25, which is one of the SNARE complexes, can modulate the release of neurotransmitters. Glia glutamate transporter (GLT) is one of the two glutamate transporters which account for most synaptic glutamate uptake in the CNS. The role of SNAP-25 and GLT as well as GAT is not clearly understood.

METHODSWe used the rat chronic constriction injury (CCI) model for research, and degraded SNAP-25 by a single intrathecal administration of BoNT/A. The mechanical (MWT) and thermal withdrawal latency (TWL) were tested. The level of SNAP-25, GLT, and GAT-1 were assayed using RT-PCR and Western blotting.

RESULTSSNAP-25 was suppressed by a single intrathecal administration of 0.01U BoNT/A and the reduction of SNAP- 25 was correlated with the relief of nociceptive responses in CCI rats. MWT and TWL returned to normal from the 5th to 14th day (P < 0.05) after the administration. On the 14th day after surgery, compared to the sham group, the upregulation of SNAP-25 in CCI rats was reversed after BoNT/A treatment (P < 0.05). The decreased GLT was reversed after BoNT/A treatment but increased GAT-1 was not influenced by BoNT/A treatment.

CONCLUSIONSSNAP-25 and GLT play important roles in the development of neuropathic pain, and the mechanism may involve the imbalance of neurotransmission after peripheral nerve injury. Intrathecal administration of BoNT/A reversed the upregulation of SNAP-25 and downregulation of GLT after CCI, but had no significant effect on the expression of GAT-1.

Amino Acid Transport System X-AG ; genetics ; metabolism ; Animals ; Disease Models, Animal ; GABA Plasma Membrane Transport Proteins ; Male ; Neuralgia ; genetics ; metabolism ; Neuroglia ; metabolism ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Synaptic Transmission ; genetics ; physiology ; Synaptosomal-Associated Protein 25 ; genetics ; metabolism

OBJECTIVEChemokine receptor CXCR4 and its ligand stromal-derived factor 1 alpha (SDF-1alpha) have been paid increasing attention for their involvement in megakaryocytic hematopoiesis. It has been revealed in recent years that they can induce mature and immature megakaryocytes (MKs) to migrate through bone marrow endothelial cells (BMEC) by increasing the affinity of MKs for BMEC. Thus MKs maturity and eventual release of platelet from MKs ensues. While maturity disturbance of MKs and impaired production of platelets have been regarded as the main pathogenesis of ITP, the mechanism of which still remains unclear. Therefore, a clear understanding of the levels of CXCR4 and SDF-1alpha within bone marrow in children with ITP will help us to elucidate further the mechanism of ITP as well as to provide direct theoretical evidence for predicting treatment effect and evaluating prognosis.

METHODSBone marrow were aspirated from 28 children with AITP and 12 normal children. Percoll density gradient and immunomagnetic beads method were used to purify megakaryocytes from the bone marrow. The immune cytochemistry was used to detect CXCR4 on megakaryocytes. The levels of SDF-1alpha were detected by ELISA. SPSS10.0 statistical software was used to deal with the experimental data.

RESULTSBefore the treatment in children with AITP, both the CXCR4 expression on megakaryocytes and the SDF-1alpha level in bone marrow plasma were markedly decreased compared with the normal controls (P < 0.05). As to the cases who were sensitive to the high-dose intravenous immunoglobulin (HDIVIgG), the CXCR4 and SDF-1alpha levels were much higher in children after the treatment than those before the treatment (P < 0.05). In 6 cases insensitive to HDIVIgG, before the treatment the CXCR4 level was much lower than the children sensitive to HDIVIgG (P < 0.05).

CONCLUSIONSThe low levels of CXCR4/SDF-1alpha system in bone marrow may be one of the factors which contribute to the maturity disturbance of megakaryocytes and disturbance of platelets production in AITP, while decreased CXCR4/SDF-1alpha system may be caused by the effect of autoantibody against platelet. The mechanism of HDIVIgG in the treatment of AITP may involve in the increasing expression of CXCR4/SDF-1alpha system. The level of CXCR4 on megakaryocytes may play a certain role in predicting the treatment effect of immunoglobulin.

Adolescent ; Bone Marrow ; metabolism ; Chemokine CXCL12 ; Chemokines, CXC ; blood ; Child ; Child, Preschool ; Enzyme-Linked Immunosorbent Assay ; Humans ; Infant ; Ligands ; Megakaryocytes ; metabolism ; Purpura, Thrombocytopenic, Idiopathic ; blood ; Receptors, CXCR4 ; biosynthesis

Programmed death-1 ligand-1(PD-L1) is a recently identified member of the B7 family molecules and is shown to mediate the inhibition of immune responses. This study was purposed to enhance the weak immunological function of dendritic cells (DCs) derived from the patients with chronic myelocytic leukemia (CML) by blockade of the expression of PD-L1. Bone marrow mononuclear cells (BMMNCs) of CML patients were induced into DCs in the presence of cytokine cocktail of rhGM-CSF, rhIL-4 and TNF-alpha. The phenotypes of DCs were detected by flow cytometry, mixed lymphocyte reaction was analyzed by MTT assay and IFN-gamma, IL-2 and IL-10 in the cell culture supernatant were detected by ELISA. The results showed that the expression of PD-L1 on CML-DCs was upregulated with the maturation of CML-DCs. PD-L1-blockaded DCs could enhance T lymphocyte proliferation, increase the secretion of IL-2 and IFN-gamma, and inhibit the production of IL-10. Taken together, PD-L1-blockaded DCs originated from CML cells had more potent immunostimulatory capability. It is concluded that PD-L1 blockaded can enhance the function of CML-DCs. This approach presents new possibilities for achieving anti-tumor immunity by DC-based vaccination.
Antigens, CD ; metabolism ; B7-H1 Antigen ; Dendritic Cells ; cytology ; immunology ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interferon-gamma ; metabolism ; Interleukin-10 ; metabolism ; Interleukin-2 ; metabolism ; Interleukin-4 ; pharmacology ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; immunology ; metabolism ; Recombinant Proteins ; Tumor Necrosis Factor-alpha ; pharmacology