BACKGROUND: Non-adherent mesenchymal stem cells (NA-MSCs) can form colony forming unit of fibroblasts and induce the differentiation into adipocytes, osteoblasts, and chondrocytes. OBJECTIVE: To determine the effect of epithelial growth factor (EGF) on colony formation of fibroblasts and differentiation into neuron-like cells from NA-MSCs.METHODS: Bilateral femur and tibia as well as total MSCs were separated, and repeated-transfer was employed to purify NA-MSCs. The fifth-passage total MSCs and NA-BMCs were induced in a medium containing EGF and basic fibroblast growth factor (b-FGF) for 2 weeks. Colony unit formation of fibroblasts, effect of EGF on colony-forming unit of fibroblasts, and relative protein expression detected by toluidine blue and immunocytochemical staining were observed. RESULTS AND CONCLUSION: Both total MSCs and NA-BMC could generate colony-forming unit of fibroblasts. After treatment of EGF, colony-forming unit of fibroblasts from NA-BMC was increased significantly. Immunocytochemical staining demonstrated that two weeks later both neuro-specific NeuN and NF-200 were observed in total MSCs and NA-BMC; while, toluidine blue staining indicated that neuron-specific Nissl body was observed in some cells. EGF can effectively promote colony-forming unit of fibroblast from NA-BMC, and repeated-transfer NA-BMC can induce differentiation into neuron-like cells.

BACKGROUND: Non-adherent mesenchymal stem cells (NA-MSCs) can form colony forming unit of fibroblasts and induce the differentiation into adipocytes, osteoblasts, and chondrocytes.OBJECTIVE: To determine whether non-adherent mesenchymal stem cells (NA-MSCs) in adult mouse bone marrow could differentiate into neuron-like cells in cerebral ischemic region.METHODS: Bilateral femur and tibia of β-Gal transgenic mice was separated, and repeated-transfer was used to collect the fifth-passage purified NA-MSCs which were adjusted at concentration of 1×10~(12)/L Middle cerebral artery occlusion was established in the two groups. After 7 days, 3 μL fifth-passaged NA-MSCs suspension was injected into cerebral ischemic region in the transplantation group, while an equal amount of saline was injected into model group. Survival, distribution, and differentiation of donor cells in cerebral ischemic region were observed at 8 weeks after transplantation.RESULTS AND CONCLUSION: LacZ staining showed that donor cells could express β-Gal protein after 8 weeks and survived in the ischemic region. Simple and double immunohistochemical staining indicated that β-Gal-positive donor cells were detected in necrotic region and at necrotic edge of ischemic model. Additionally, partial cells could express neuro-specific NeuN protein and glial cell-specific GFAP. NA-MSCs are able to survive and migrate in cerebral ischemic region; moreover, partial NA-MSCs can differentiate into mature neuron-like cells or glial cells which participate in repairing brain injury.

Objective To identify the correlativity between SCN4A V781I mutation and hypokalemic periodic paralysis(HOKPP).Methods The SCN4A V781I mutation was screened on the members of a hypokalemic periodic paralysis family(including 3 patients and 14 healthy relatives), 71 sporadical hypokalemic periodic paralysis patients and 100 healthy adults with the PCR sequencing and the incision enzyme techniques.Results The mutation existed not only in all the patients but also in 4 healthy relatives, including 1 male and 3 females, in the hypokalemic periodic paralysis family.Seven of 71 sporadieal hypokalemic periodic paralysis patients and 7 of 100 health adults indicated the SCN4A V781I mutation.There was no significant difference in the SCN4A V781I mutation rate between the hypokalemic periodic paralysis patients and the healthy adults(X~2=0.452, P=0.502).Conclusions SCN4A V781I mutation maybe is an innocent polymorphism.There is little correlativity between SCN4A V781I and hypokalemic periodic paralysis.

Objective To summarize the clinical characteristics and make genetic diagnosis in the patients with hereditary spinocerebellar ataxia type 7 (SCA7).Methods Pedigree analysis and clinical examination were performed in one family with SCA7 by clinical findings,of which retinal morphology and visual electrophysiology were available on part numbers.The polymorphic cytosine adenine guanine (CAG) repeats in the encode region of SCA7 gene were detected by combining polymerase chain reaction with deoxyribonucleic acide (DNA) sequencing on 19 familial numbers and 12 controls.Results 6 patients were identified,who manifesting cerebellar ataxia,decreased visual acuity and colour vision defect,as was pigmentary retinopathy on fundoscopy;The 6 patients had not only extinction of the electroretinogram (ERG) but also remarkably reduced amplitudes of oscillatory potentials and flash-visual evoked potentials. On normal alleles CAG repeat size ranges from 8 to 25 repeats,wherease on mutated alleles of the 6 numbers it ranges from 50 to 97 repeats.The 6 numbers were diagnosised as SCA7 patients.One asymptomatic individual of this family,who displayed a normal allele with 18 CAG repeats and another containing abnormal expantion of 56 repeats,was diagnosised as a asymptomatic carrier whose age maybe still below the age of onset.Conclusion The clinical manifestations of SCA7 are heterogeneous,and the detection of CAG repeats can provide an effective way for the gene diagnosis and the prediction of asymptomatic patients.

OBJECTIVETo study the clinical features and proper treatment of 38 elderly patients with early double primary cancers.

METHODSThirty-eight elderly patients with early double primary cancers treated from January 1980 to March 2003 were retrospectively reviewed for involved organs, treatment and prognosis.

RESULTSDigestive tract was the most frequently involved, followed by urogenital system and lung. Long-term results of endoscopic mucosal resection (EMR), operation and radiotherapy were superior to other methods. The prognosis of gastrointestinal carcinoma was better than that of prostate carcinoma and hematopoietic system. The operation rate decreased with increasing age. The 5-year survival rates of EMR, operation and radiotherapy were 85.7%, 71.1% and 75.0%, respectively. The medium survival time was 120 months in first cancer and 39 months in the second primary cancer. The 5-year survival rates of the first cancer and second primary cancer were 88.6% and 53.8%.

CONCLUSIONYearly follow-up for elderly patients with endoscopy, beta ultrasonic scan and X-ray contribute to finding of early double primary cancers. Operation is the best treatment of early double primary cancers. Endoscopic mucosal resection is especially suitable for old patients with digestive tract and bladder cancer.

Aged ; Aged, 80 and over ; Colorectal Neoplasms ; diagnosis ; radiotherapy ; surgery ; Endoscopy, Digestive System ; Female ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; surgery ; Male ; Middle Aged ; Neoplasms, Multiple Primary ; Prostatic Neoplasms ; radiotherapy ; surgery ; Retrospective Studies ; Stomach Neoplasms ; radiotherapy ; surgery ; Survival Rate

AIMTo identify up-regulated genes specific to human normal gastrointestinal tissues.

METHODSStudy was made on human normal tissue gene expression database open to the public. Tissue-specific genes were identified using one-tailed student T test. Online software including Ingenuity and KEGG were applied for physiological function analyses. Unsupervised two-way hierarchical clustering method was used to analyze the expression profile of stomach-specific genes in gastric cancer gene expression datasets.

RESULTSThe analyses identified 196 stomach-specific genes, 203 ileum-specific genes and 224 colon-specific genes, respectively. The gene expression profiles reflect major organ-specific physiological functions on the molecular level. Some putative oncogenes and tumor suppressor genes were found in the tissue-specific gene list. Hierarchical clustering analysis revealed that the stomach-specific genes were up-regulated in normal stomach tissues but down-regulated in stomach cancer tissues. The normal tissues clustered together, so did the cancer tissues. At the meantime, clustering could also distinguish the moderate and severe differentiated stomach cancer.

CONCLUSIONHuman normal stomach, ileum and colon possess tissue-specific up-regulated genes, which are closely associated with physiological functions.

Cluster Analysis ; Colon ; metabolism ; Computational Biology ; Databases, Genetic ; Gastrointestinal Tract ; metabolism ; Gene Expression Profiling ; Gene Expression Regulation ; Humans ; Ileum ; metabolism ; Stomach ; metabolism ; Stomach Neoplasms ; genetics ; Transcriptome

OBJECTIVESome members of the S100 gene family have been suggested to be associated with cancer development and metastasis. Our previous cDNA micro-array studies have showed S100A6 expression is elevated in gastric cancer compared with that in paired normal mucosa. To validate our previous results and further investigate the possible role of S100A6 gene in gastric cancer, we carried out this detailed S100A6 expression analysis in more matched gastric cancer samples.

METHODSS100A6 expression was detected in 20 paired fresh surgical samples of gastric tumor tissue and matched non-cancerous mucosa by QRT-PCR. A gastric cancer tissue microarray (TMA) containing 1020 duplicate matched normal mucosa, gastric cancer tissue and metastatic lymph node tissue cores from 208 gastric cancer patients was constructed. S100A6 expression was detected by immunohistochemistry and the correlation between S100A6 expression with clinicopathological factors and survival was analyzed.

RESULTSAs quantitated by QRT-PCR, S100A6 transcript level was elevated in 73.7% of the primary cancer lesions with an average 2.25-fold up-regulation than that in matched non-neoplastic mucosa. As displayed by immunohistochemistry, the positive rate of S100A6 in non-neoplastic mucosa, tumor lesions and metastatic lymph nodes was 34.3%, 84.1% and 90.9%, respectively. S100A6 expression level in cancer and metastatic lymph node was significantly higher than their matched non-neoplastic mucosa (P < 0.05). 65.5% of patients showed an increased S100A6 expression in cancer tissue compared with that in matched normal mucosa. S100A6 overexpression was associated with larger tumor size and deeper invasion (P = 0.022 and P = 0.009). No evidence was found for an association between S100A6 expression level and other variables, including tumor grade, nodal metastases, and TNM stage. There was no association between S100A6 expression level and survival. But compared with paired non-neoplastic mucosa, an increased S100A6 expression in tumor lesion predicated a decreasing suvival if compared with a decreased S100A6 expression, though the difference was statistically not significant.

CONCLUSIONElevated expression of S100A6 gene may be an early event in the development and progression of gastric cancer. Further study of this gene may be helpful for understanding the nature of gastric carcinoma.

Cell Cycle Proteins ; metabolism ; Follow-Up Studies ; Gastric Mucosa ; metabolism ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Staging ; RNA, Messenger ; metabolism ; S100 Calcium Binding Protein A6 ; S100 Proteins ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; surgery ; Survival Rate ; Tumor Burden ; Up-Regulation

OBJECTIVETo investigate whether oxymatrine (OM) could promote mesenchymal stem cell (MSC) therapy in CCl4-induced hepatic fibrosis (HF) in rats and to initially explore its mechanisms.

METHODSTotally 50 male SD rats were randomly divided into five groups,i.e., the normal control group, the model group, the MSC therapy group, the OM therapy group, and the MSC combined OM therapy group, 10 in each group. Except the normal control group, the HF model was duplicated by CCl4 induction. After successful modeling, rats in the MSC therapy group received 5 x10(6) MSCs by intravenous injection via caudal vein, once a week. Rats in the OM therapy group received 50 mg/kg OM by intramuscular injection, three times a week. Rats in MSC combined OM therapy group received 5 x 10(6) MSCs by intravenous injection via caudal vein, once a week and 50 mg/kg OM by intramuscular injection three times a week. Equal volume of normal saline was given to those in the normal control group and the model group. All medication lasted for 8 weeks. Serum levels of ALT and AST were detected 8 weeks later. The hepatic histopathological injury and extracellular matrix deposit were assessed using HE and Masson staining. Expressions of serum interleukin-4 (IL-4) and interleukin-10 (IL-10) were detected using enzyme linked immunosorbent assay (ELISA).

RESULTS(1) Compared with the normal control group, serum levels of ALT and AST significantly increased in the model group (P < 0.05). Compared with the model group, serum levels of ALT and AST significantly decreased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group at the end of 8 weeks of treatment (P < 0.05). But serum levels of ALT and AST were significantly lower in the MSC combined OM therapy group than in the OM therapy group and the MSC therapy group (P < 0.05). (2) Compared with the model group, the hepatic injury was significantly lessened and the area of extracellular matrix deposit was significantly reduced in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05). Besides, they wer more significant in the MSC combined OM therapy group (P < 0.05). (3) Compared with the model group, the serum IL-4 level was significantly higher in the MSC therapy group and the MSC combined MO group (P < 0.05). It was higher in the MSC combined MO group (P < 0.05). Although the serum IL-4 level also increased in the OM therapy group, but with no statistical difference (P > 0.05). (4) The serum IL-10 level significantly increased in the OM therapy group, the MSC therapy group, and the MSC combined OM therapy group (P < 0.05), and it was the highest in the MSC combined OM therapy group among the three groups (P < 0.05). (5) Two-photon fluorescence imaging showed no signals of MSCs in liver with or without OM injection.

CONCLUSIONOM could promote mesenchymal stem cell therapy in hepatic fibrosis rats, which might be involved in increasing serum levels of IL-4 and IL-10.

Alkaloids ; therapeutic use ; Animals ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Liver Cirrhosis, Experimental ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; Quinolizines ; therapeutic use ; Rats ; Rats, Sprague-Dawley

OBJECTIVEOptimization of combining electroencephalography (EEG), short latency somatosensory evoked potentials (SLSEP) and transcranial Doppler (TCD) techniques to diagnose brain death.

METHODSOne hundred and eleven patients (69 males, 42 females) from the major hospitals of Zhejiang Province were examined with portable EEG, SLSEP and TCD devices. Re-examinations occurred < or =12 h later.

RESULTSThe first examination revealed that the combination of SLSEP and EEG led to more sensitive diagnoses than the combination of SLSEP and TCD. Re-examination confirmed this and also revealed that the combination of TCD and EEG was the most sensitive.

CONCLUSIONThe results show that using multiple techniques to diagnose brain death is superior to using single method, and that the combination of SLSEP and EEG is better than other combinations.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Brain Death ; diagnosis ; diagnostic imaging ; Child ; Electroencephalography ; methods ; Evoked Potentials, Somatosensory ; Female ; Humans ; Male ; Middle Aged ; Ultrasonography, Doppler, Transcranial ; methods ; Young Adult

OBJECTIVETo study the relation between dendritic cell (DC) infiltration and clinicopathologic parameters, biologic characteristics and prognosis of progressing gastric cancer.

METHODSThe development of apoptotic cell death (apoptotic index, AI) in 61 progressing gastric carcinoma tissues was analyzed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick end labeling (TUNEL) method. The PCNA labeling index (PCNA-LI), density of dendritic cells in the tumor were detected by immunohistochemical method by the LSAB kit using antibody against S-100 protein and PC-10.

RESULTSDC infiltration was negatively correlated with lymph node metastasis, clinical stage and PCNA-LI, but positively with AI. The DCs in gastric cancer groups with and without lymph node metastasis were (5.63 +/- 4.37)/HPF and (8.51 +/- 5.57)/HPF with difference significant (P < 0.05). The DC infiltration in I, II, III stage lesions were (11.23 +/- 6.05)/HPF, (6.28 +/- 4.37)/HPF and (5.53 +/- 5.19)/HPF also with differences significant (P < 0.01). The PCNA-LI was significantly higher in the low DC group (57.10% +/- 14.18%) than that of high DC group (48.15% +/- 10.59%, P < 0.01). AI findings were 3.77% +/- 1.26% and 2.95% +/- 1.07% in the high and low DC groups (P < 0.01). A positive correlation was observed between DC infiltration and AI (r = 0.39, P < 0.01) whereas a negative correlation between DC infiltration and PCNA-LI (r = -0.47, P < 0.01). The prognosis of high DC infiltration patients was significantly better than those with low ones.

CONCLUSIONThe infiltrating dendritic cells in and around tumor, representing the local immune status of the host, may play an important role in immunological defense mechanism of host versus tumor. Dendritic cells may inhibit the proliferation and induce the apoptosis of the tumor cells, thus affecting the clinical features and improve the prognosis of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Apoptosis ; Cell Division ; Dendritic Cells ; physiology ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Proliferating Cell Nuclear Antigen ; analysis ; Stomach Neoplasms ; mortality ; pathology ; Survival Rate