PURPOSE: The aim of this study was to establish an in vitro method to purify human multipotent adult progenitor cells (hMAPCs) and assess their possible differentiation into hepatocytes by coculture with human hepatocyte line L02. METHODS: hMAPCs were isolated by magnetic activated cell sorting (MACS) depletion selection using CD45 and GlyA microbeads. After indirect or direct coculture of hMAPCs and human hepatocyte line L02, the expression of albumin (ALB), alpha-fetoprotein (AFP), cytokeratin (CK) 18, and CK19 by hMAPCs was detected by immunocytochemistry. RESULTS: With the MACS method, (5-10) x 10(4)/mL hMAPCs could be separated from 1 x 10(6)/mL bone marrow mononuclear cells. The purity of CD45-/GlyA- cells separated from bone marrow adherent cells was more than 98%, as determined by flow cytometry. In the coculture without cell-to-cell contact, hMAPCs expressed high AFP on day 1, and then tapered daily to low expression on day 7; ALB expression reached its peak on day 5, and remained high on day 7; CK18 was initially expressed on day 5 and was higher on day 7; CK19 was negative in all assays. In the coculture with cell-to-cell contact, ALB and CK18 were expressed by most cells while AFP appeared in only a few on day 5. CONCLUSION: hMAPCs were induced to differentiate into mature hepatocyte-like cells by coculture with a hepatocyte cell line, either with or without cell-to-cell contact, but the former seemed more effective.
Adult* ; alpha-Fetoproteins ; Bone Marrow ; Cell Differentiation ; Cell Line ; Coculture Techniques* ; Flow Cytometry ; Hepatocytes* ; Humans ; Immunohistochemistry ; Keratins ; Microspheres ; Stem Cells*

OBJECTIVETo investigate whether there is a association between estrogen receptor beta (ERb) gene polymorphism and primary hepatocellular carcinoma (HCC).

METHODS100 primary HCC patients and 100 controls from southwestern China were recruited in this study. The polymorphism of RsaI and AluI in ERb gene was analyzed by PCR- restriction fragment length polymorphism (RFLP).

RESULTSR allelic frequency was 35.0% and 51.0% in HCC patients and in control group, respectively, odds ratio (OR) was 0.517 [95% confidence intervals (CI) = 0.346-0.773], P less than 0.01. A allelic frequency was 20.5% and 11.0% in HCC patients and in control group, respectively, OR was 2.086 (95% CI = 1.191-3.654), P less than 0.01. Gene frequency of RsaI and AluI in the two groups was distributed with polymorphism.

CONCLUSIONERb gene polymorphism is associated with primary liver cancer. R allele may be the guard factor, and A allele may be its risk factor.

Carcinoma, Hepatocellular ; genetics ; Estrogen Receptor beta ; Gene Frequency ; Genetic Predisposition to Disease ; Humans ; Polymorphism, Genetic

OBJECTIVETo investigate the synergistical killing effect of docetaxel combined with ABT-737 on human prostate cancer cell line PC-3 by inducing apoptosis and further to determine the mechanism underlying such effect.

METHODSPC-3 cells were treated with various concentrations of docetaxel or (and) ABT-737. Cell viability was determined using MTT assay. Apoptosis was assessed by fluorescence microscopy analysis of cells with condensed and segmented nuclei following staining with 4',6-diamidino-2-phenylindole (DAPI). Cellular DNA was stained with propidium iodide and flow cytometric analysis was performed to analyze the cell cycle distribution. Bcl-2, Bax, Bcl-xL and Mcl-1 protein changes were detected by Western blot. The activity of caspase-3 was measured using a colorimetric assay.

RESULTSDocetaxel (20 nmol/L) combination with ABT-737 (400 nmol/L) for 48 hours, the cell viability was decreased to 19.7% ± 3.2% to compare with 44.2% ± 4.4% (t = 4.45) of docetaxel and 93.2% ± 1.8% of ABT-737 separately and there was a synergistic effect between the two drugs (CI = 0.8). Apoptosis rate of the combination group was higher than other two drugs. Docetaxel increased the cell number arrested in G(2)/M phase compared with control group (P < 0.05), but the combination treatment resulted in a significant arrest in the G(0)/G(1) phase. The combination treatment could significantly reduced the Bcl-2, Bcl-xL and Mcl-1 expression (F = 369.53, 57.89 and 32.77, all P < 0.05) and enhanced the activity of caspase-3 (419.7% ± 15.6%) (F = 207.33, P < 0.05).

CONCLUSIONSThe combination of ABT-737 with docetaxel can synergistically inhibit the proliferation of PC-3 cells through inducing apoptosis, which may be associated with cell cycle arrest, down-regulation of Bcl-2, Bcl-xL and Mcl-1 expression and activation of caspase-3.

Apoptosis ; drug effects ; Biphenyl Compounds ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Drug Synergism ; Humans ; Male ; Myeloid Cell Leukemia Sequence 1 Protein ; metabolism ; Nitrophenols ; pharmacology ; Piperazines ; pharmacology ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Sulfonamides ; pharmacology ; Taxoids ; pharmacology ; bcl-X Protein ; metabolism

Animals ; Cadherins ; metabolism ; Catenins ; metabolism ; Estradiol ; pharmacology ; Estrogen Receptor Modulators ; pharmacology ; Female ; Ovariectomy ; Rats ; Uterus ; drug effects ; metabolism

OBJECTIVETo investigate the clinical outcomes of semicircular decompression in treating old thoracolumbar fractures and intractable neuropathic pain.

METHODSFrom September 2009 to September 2013, 21 patients with old thoracolumbar fracture and intractable neuropathic pain were treated with semicircular decompression. Among initial surgery, posterior pedicle screw fixation was used in these patients, with or without laminectomy. All patients were male, range in age from 20 to 28 years old with an average of (25.00±2.38) years. Vertebral body residual bone block resulted in intra-spinal placeholder more than 50%. All patients were complete spinal cord injury (ASIA grade) or cauda equina injury. VAS scores was from 6 to 10 points with the mean of 7.14±0.91. In these patients, MRI, CT, X-rays were performed; denomination and dosage of analgesics were recorded; nerve function and pain status were respectively evaluated by ASIA grade and VAS score before and after operation.

RESULTSAll patients were followed up from 8 to 32 months with an average of (17.29±6.02) months. All bone fragments of spinal canal were removed and spinal cord decompressions were achieved. At final follow-up, VAS scores were from 0 to 8 points with an average of (2.43±2.46) points, and were obviously reduced than peroperative data (P<0.05). Eleven cases of them stopped analgesic intake and 7 cases reduced using. Three patients' symptoms and VAS scores were not improved.

CONCLUSIONOld thoracolumbar fractures and intractable neuropathic pain need receive imaging examination as soon as possible and consider semicircular decompression therapy if bone fragments were in vertebral canal and spinal canal stenosis existed. This therapy can effectively relieve pain and profit nerve functional recovery.

Adult ; Decompression, Surgical ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Neuralgia ; etiology ; surgery ; Pain, Intractable ; etiology ; surgery ; Spinal Fractures ; physiopathology ; surgery ; Thoracic Vertebrae ; injuries ; surgery ; Visual Analog Scale ; Young Adult

Objective To evaluate the clinical value of multi-slice CT(MSCT)angiography in the diagnosis of Takayasu's arteritis.Methods Fourteen patients underwent MSCT angiography with use of 90 ml contrast media at a rate of 3.5 ml/s.Images were generated for vascular assessment with various reconstructional techniques,including multi-planar reformat(MPR),maximum intensity projection(MIP), volume rendering(VR),CT virtual endoscopy(CTVE)and advanced vessel analysis(AVA).Results MSCT angiography findings was consistent with the clinical changes of Takayasu's arteritis in all cases.The integrated information including reconstructional and transverse images clearly presented various luminal abnormalities,such as stenosis,occlusion and dilatation in the affected aorta,as well as its major branches and collateral circulation.Furthermore,it depicted mural abnormities,especially concentric arterial wall thickening.Steno-occlusive type(8/14)was more common than mixed type(6/14).According to Lupi- Herrea's classification,type Ⅰ in 5 cases,type Ⅱ in 2,type Ⅲ in 6 and type Ⅳ in 1 were recorded in the present study.Conclusion MSCT angiography is reliable imaging modality for the detection of Takayasu's arteritis.It is useful for early diagnosis because it allows evaluation of wall thickness rather than merely the luminal diameter,which is especially important for prognosis.It might be used as the first choice in the diagnosis of this disease as a simple,convenient and noninvasive method.

Antiviral Agents ; therapeutic use ; Base Sequence ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Lamivudine ; therapeutic use ; Molecular Sequence Data ; Mutation

OBJECTIVETo investigate the effects of Tianzhi Keli (TZ) on acetylcholine (ACh) and catecholamine levels in striatum of rats with neuromitochondrial impairment, and try to find out the neuroprotective mechanism of TZ.

METHODThe microdialysis and high performance liquid chromatography (HPLC)-post column Immobilized enzyme reactor (IMER)-electrochemical detection (ED) were used to establish a model of mitochondrial energy metabolism impairment which induced by perfusion with sodium azide (NaN3), and measure continuously the effects of TZ on extracellular ACh, choline (Ch) and catecholamine of model rats.

RESULTAfter perfusion with NaN3, ACh, noradrenalin (NE), adrenaline (E), dopamine (DA), 3,4-Dihydroxyphenyl-aletic (DOPAC), and homovanillic acid (HVA) levels were decreased obviously (P < 0.05-0.01), while Ch level was increased distinctly (P < 0.01). Transmitters levels were recovered individually after stop the perfusion with NaN3. TZ can postpone the decrease of ACh and advance the recover of Ch. The effect of TZ coupled with duxil on increasing ACh level is more obviously than effect of TZ or duxil. TZ is also showing a tendency to postpone the decrease of catecholamine and advance its recovery. TZ coupled with duxil can advance the recovery of DOPAC and adjust the metabolic abnormity positively.

CONCLUSIONTZ has effect on protecting impairment of choline neurosystem, which induced by damage of mitochondrion and abnormity of energy metabolism; coupled with duxil have synergistic action. TZ also has tendency to protect the impairment of epinephrine and dopamine neurosystem.

3,4-Dihydroxyphenylacetic Acid ; metabolism ; Acetylcholine ; metabolism ; Animals ; Catecholamines ; metabolism ; Corpus Striatum ; metabolism ; Dopamine ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Extracellular Space ; metabolism ; Gastrodia ; chemistry ; Male ; Microdialysis ; Mitochondrial Diseases ; chemically induced ; metabolism ; Norepinephrine ; metabolism ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sodium Azide ; Uncaria ; chemistry

OBJECTIVETo investigate the effect of Tianma Gouteng recipe (TGR) on interfering left ventricular (LV) and aortic hypertrophy and tissue angiotensin II (Ang II) in rats with renovascular hypertension.

METHODThe animal model of renovascular hypertension was used in this experiment. Hypertensive rats were randomly allocated into model group, Enalapril group and TGR group, and the drugs were used for 6 weeks continuously. During this period, the blood pressure of rats was measured every two weeks. After rats were sacrificed, the wet weight, tissue Ang II level of LV and aorta, and the cardiac index were measured.

RESULTOne week after renovascular stenosis, the systolic blood pressure (SPS) of model group was increased by 37.4 mmHg, and 7 weeks after stenosis, the LV and aortic hypertrophy was obvious increased, meanwhile, tissue Ang II of LV and aorta was raised markedly (P < 0.01). Contrasting with the model group, blood pressure was reduced and the morphological index was improved in Enalapril group respectively (P < 0.05 or P < 0.01); the wet weight of LV and aorta were reduced, the morphological index was improved, the rise of Ang II in tissue was suppressed, in TGR group significantly.

CONCLUSIONTGR can attenuate myocardial and aorta hypertrophy induced by renovascular hypertension, and suppress the rise of Ang II in tissue significantly. This suggests that TGR has the effects on interfering LV and aortic hypertrophy by an independent-antihypertensive way.

Angiotensin II ; metabolism ; Animals ; Antihypertensive Agents ; pharmacology ; Aorta ; metabolism ; pathology ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Enalapril ; pharmacology ; Gastrodia ; chemistry ; Hypertension, Renovascular ; complications ; physiopathology ; Hypertrophy, Left Ventricular ; etiology ; metabolism ; pathology ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Uncaria ; chemistry

OBJECTIVETo investigate the clinical significance of resection of the pancreatic capsule and anterior layer of transverse mesocolon in radical gastrectomy.

METHODSBetween January 2007 and July 2008, a total of 213 gastric cancer patients enrolled in the study. These patients were randomly assigned into two groups: 105 in group R and 108 in group N. Only in group R were the pancreatic capsule and anterior layer of transverse mesocolon resected during radical gastrectomy. The pancreatic capsule and anterior layer of transverse mesocolon were histologically analyzed for metastasis. The data including blood loss during operations, number of dissected lymph nodes and postoperative complications were analyzed in both groups.

RESULTSThere were no significant differences between the two groups in blood loss during operation and postoperative complications, but the differences in operation time and number of dissected lymph nodes between the two groups were significant. Metastases to the pancreatic capsule and/or anterior layer of transverse mesocolon were diagnosed in nine (8.6%) patients of group R. The metastases to the pancreatic capsule and/or anterior layer of transverse mesocolon were found to be associated with tumor invasion depth, anterior or posterior gastric wall, clinical staging and perigastric lymph node metastasis extent (P<0.05), but not with age, gender, tumor location, size, Borrmann type and pathological classification (P>0.05).

CONCLUSIONSResection of pancreatic capsule and anterior layer of transverse mesocolon in group R does not increase postoperative complications in comparison with group N. The resection is beneficial to the patients with advanced gastric cancer staging relatively late because of potential metastasis to pancreatic capsule and anterior layer of transverse mesocolon.

Adult ; Aged ; Aged, 80 and over ; Colon, Transverse ; Female ; Gastrectomy ; methods ; Humans ; Male ; Mesocolon ; surgery ; Middle Aged ; Pancreas ; surgery ; Stomach Neoplasms ; surgery