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

OBJECTIVETo investigate the antitumor activity of Diosgenin in vivo and in vitro.

METHODS-180, HepA, U14 and EAC transplant mice were given Diosgenin ig or i.p. everyday for 10 days, from the next day when they were inoculated in axilla. Tumor growth inhibit rates were calculated. Four kinds of cells, MCF, L929, A375-S2 and HeLa, were incubated respectively with Diosgenin in vitro. Tumor growth inhibit rates were also calculated.

RESULTIn vivo, both ig and i.p., Diosgenin inhibited S-180, HepA, U 14 mice transplant tumor, the inhibit rates being 30%-50%, but it did not inhibit the EAC mice transplant tumor. In vitro, Diosgenin inhibited L929, HeLa, MCF cell growth, and IC50 were 1.2, 18.2, 19.8 micrograms.mL-1 respectively, but it did not significantly affect A375-S2 cells.

CONCLUSIONDiosgenin has an obvious antitumor activity on S-180, HepA, U14 transplant mice in vivo and L929, HeLa, MCF cells in vitro.

Animals ; Antineoplastic Agents, Phytogenic ; therapeutic use ; Carcinoma, Ehrlich Tumor ; drug therapy ; Dioscorea ; chemistry ; Diosgenin ; isolation & purification ; therapeutic use ; Female ; Humans ; Inhibitory Concentration 50 ; Male ; Mice ; Neoplasm Transplantation ; Phytotherapy ; Plants, Medicinal ; chemistry ; Sarcoma 180 ; drug therapy ; Tumor Cells, Cultured ; drug effects

BACKGROUNDAn apnea test is essential in the clinical determination of brain death. This study was conducted to analyse complications associated with the apnea test in the determination of the brain death.

METHODSOn 93 adult patients in coma in Zhejiang Province of China from January 2003 to December 2006, 179 apnea tests were performed as a part of the determination of brain death. Potential risk conditions and complications were analysed during apnea tests.

RESULTSDuring apnea, serious cardiac arrhythmia did not occur in all patients. Complications occurred in 37 of 179 (21%) apnea tests. Hypotension occurred in 30 patients (17%) and it was observed in 8/94 (9%) tests with baseline value of systolic arterial blood pressure not less than 120 mmHg, and 22/85 (26%) less than 120 mmHg (P < 0.05). Severe hypoxaemia occurred in 10 patients (6%) of which 3/138 (2%) tests with baseline value of arterial oxygen pressure not less than 200 mmHg, and 7/41 (17%) less than 200 mmHg (P < 0.05).

CONCLUSIONSThis study demonstrated that complications occurred mostly in patients with inadequate baseline systolic arterial blood pressure and preoxygenation. Adequate precautions during the apnea tests may reduce the risk of cardiovascular and oxygenation complication.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apnea ; physiopathology ; Brain Death ; diagnosis ; Carbon Dioxide ; blood ; Female ; Humans ; Hypotension ; etiology ; Hypoxia ; etiology ; Male ; Middle Aged

Objective To investigate the abdomen CT characteristics of patients with ischemic bowel disease (ICBD).Methods CT imaging data of ICBD patients from January 2008 to December 2013 in the Chinese PLA General Hospital were retrospectively analyzed to find CT imaging features in intestinal lesions associated with ICBD death.Results In CT imaging analysis,151 patients including acute superior mesenteric artery thromboembolism (ASMATE,n=51),acute superior mesenteric vein thrombosis (ASMVT,n=53),non-occlusive mesenteric ischemia (NOMI,n=8),chronic mesenteric ischemia (CMI,n=10) and ischemic colitis (IC,n=29) were divided into survival group (n=115) and death group (n=36).In comparison with the survival group,the death group had higher incidence of abdomenal effusion,portomesenteric gas,pneumatosis intestinalis and pneumoperitoneum (P＜0.001,P＜0.001,P＜0.001,P=0.003).Conclusion The ascites,portomesenteric vein gas,pneumatosis intestinalis and pneumoperitoneum in bowel CT may be associated with the death of ICBD patients.

Objective To observe the effect of Yang-warming and Qi-tonifying Recipe (YQP)on aquaporin 3(AQP3) and AQP8 in rats with slow transit constipation,and to explore its therapeutic mechanism. Methods Forty rats were randomly divided into modeling group(N=30)and normal control group(N = 10). After successful modeling by gastric gavage of loperamide,the modeled rats were randomly divided into model group,YQP group and Mosapride group,10 rats in each group,and were separately treated with corresponding medicine for 2 weeks. After treatment, the colonic transit function was measured by carbon propelling test. The protein levels of AQP3 and AQP8 were detected by immunohistochemistry and their mRNA expression levels were detected with real-time fluorescence polymerase chain reaction (qPCR). Results Compared with the normal control group,the propelling rate of carbon particle in the model group was decreased,and the protein and mRNA expression levels of AQP3 and AQP8 were significantly increased(P<0.05 or P<0.01). Compared with the model group,the propelling rate of carbon particle of YQP group and mosapride group was significantly increased, and the protein and mRNA expression levels of AQP3 and AQP8 were significantly decreased (P<0.05),but there was no significant difference between YQP group and mosapride group (P >0.05). Conclusion YQP had therapeutic effects on loperamide-induced constipation through decreasing the expression of AQP3 and AQP8 in the intestine,reducing the reabsorption of intestinal fluid, and increasing the fecal water content.