Objective To explore the differential diagnostic significance of clone analysis for multicentric occurrence (MO) and intrahepatic metastasis (IM) in hepatocellular carcinomas (HCCs).Methods Loss of heterozygosity (LOH) and microsatellite instability (MSI) were analyzed by microsatellite polymorphism test and the integration sites of HBV were assessed by Southern blot in each nodule of the HCCs. The clonalities were then compared between each nodule of the same patient and the diagnosis of MO or IM was concluded. Finally, the results based on clonality analysis were compared with those according to clinicopathological and imaging data. Results According to the results of LOH and MSI in 79 nodules and nontumorous tissue from 35 cases of mutiple HCCs, 5 (14.3%)and 29 cases (82.9 %) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The integration sites of HBV could be analyzed in 77 nodules from 34 multiple HCCs. Among them, 6 (17. 6%) and 27 cases (79.4%) were divided into MO and IM, respectively. Both MO and IM presented simultaneously in 1 patient (2.9%). The classification results of microsatellite polymorphism test and HBV integration sites analysis demonstrated a significant positive correlation (rs = 0.909, P＜0.001). Nevertheless, neither the classification of microsatellite polymorphism test nor that of HBV integrate sites analysis was correlated with the discrimination according to clinicopathologic and imaging data (rs=0. 133, P=0. 468, rs =0. 262, P=0. 155, respectively). Recurrence in patients in the MO group occurred significantly later than that in IM cases who were diagnosed by clonality analyses (P=0. 001). Conclusion The clonality analysis based on the results of LOH and MSI or assessments of HBV integrate sites is useful for the differential diagnosis of MO and IM and their treatment and prognosis.

BACKGROUND:Under co-culture conditions, mesenchymal stem cel s could regulate osteogenic differentiation and osteogenesis of osteoblasts. OBJECTIVE:To observe the osteogenic efficiency of osteoblastic precursor cel s co-cultured with undifferentiated bone marrow-derived mesenchymal stem cel s, umbilical cord-derived mesenchymal stem cel s, or placenta-derived mesenchymal stem cel s in mineralization medium. METHODS:Adipose-derived stem cel s were induced in osteogenic differentiation medium for 7 days before being indirectly co-cultured with undifferentiated mesenchymal stem cel s isolated from different tissues (bone marrow group, umbilical cord group and placenta group) in Transwel plates. Induced adipose-derived stem cel s cultured alone served as control group. At different experimental intervals, quantitative analysis of alkaline phosphatase activity and calcified matrix was preformed to observe the effects of mesenchymal stem cel s from different sources on the osteogenic efficiency of induced adipose-derived stem cel s. RESULTS AND CONCLUSION:Expression of alkaline phosphatase was significantly higher in different experimental groups than the control group (P<0.05), and it was also higher in the bone marrow group than the umbilical cord and placenta groups (P<0.05). Quantitative analysis of calcified matrix revealed that the experimental groups were significantly higher than the control group (P<0.05);and in experimental groups, the umbilical cord group was higher than bone marrow group and placenta group (P<0.05). These findings indicate that the osteogenic efficiency of induced adipose-derived stem cel s is improved dramatical y under co-culture conditions.

As the human population ages and the life expectancy increases,tendon injuries will become more prevalent,especially among young individuals.Though the traditional operative therapy for tendon lesion can relieve the suffering of patients,the functional reconstruction is usually not optimistic.Tissue engineering is an advancing field,as the technology of construction in vitro and application in vivo matures,that can provide a more promising approach for tendon repair without tendon autograft.Challenges and future directions in the field of tendon tissue engineering focusing on four key parameters:seed cells,novel scaffolds,and mechanical stimulation.Recently,the discovery of TDSCs (tendon-derived stem cells) provides new ideas for the selection of seed cells and effect of mechanical stimulation on the tendon tissue engineering has become a hot spot.This article provides a review of recent progress in research about seed cells,scaffolds and mechanical stimulation for tendon engineering,and also speculates on the development in the future.

OBJECTIVE:To prepare Frovatriptan succinate film-coated tablet,and investigate its in vitro dissolution behavior. METHODS:Using lactose monohydrate,microcrystalline cellulose,dioxide,silica,sodium carboxymethyl starch and magnesium stearate as accessories,Frovatriptan succinate tablet was prepared. Using opadry premix spray-coating liquid,Frovatriptan succinate film-coated tablet was prepared. Single factor test was used,using moisture,angle of repose,rigidity,friability,disintegration time and dissolution rate as indexes,to screen the formulation;using dissolution degree as index,coating material dosage was screened. The dissolution curves in vitro of self-made tablets and imported tablets in water,0.1 mol/L HCL,pH of 5.5,6.8 phos-phate buffer solutions were compared. RESULTS:The optimal formulation of Frovatriptan succinate uncoated tablet was as follow as frovatriptan succinate 3.91 mg,lactose monohydrate 99.18 mg,microcrystalline cellulose 33.06 mg,magnesium stearate 1.40 mg,sodium carboxymethyl starch 1.05 mg,silica 1.40 mg;optimal coating weighed quality was 2.0%-4.0%. In the 4 mediums, the dissolution behavior of self-made tablets and imported tablets were similar. CONCLUSIONS:Frovatriptan succinate film-coated tablet is prepared successfully,and its in vitro dissolution behavior is similar to the imported preparations.

Objective To explore the efficacy of postoperative transarterial chemoembolization (TACE) and portal vein chemotherapy (PVC) for hepatocellular carcinoma (HCC) patients complicated with portal vein tumor thrombus (PVTT). Methods One hundred and eleven HCC patients with PVTT were randomly divided into three groups receiving respectively tumor resection only ( group A) , resection plus TACE ( group B) , and resection plus TACE and PVC ( group C). Results (1) Group B had significantly lower recurrence rates at 0. 5- and 1-year, and higher survival rates at 0. 5-year compared with group A (P

BACKGROUND:Present studies mainly focused on in vitro culture of bone marrow mesenchymal stem cells(BMSCs)and cell transplantation for treating intracalvarium diseases.However,the understanding of survival,differentiation,migration and structure of transplanted cells in the damaged spinal cord is limited.OBJECTIVE:To explore effects of local BMSC transplantation in repair of spinal cord damage and feasibility of replacement therapy of BMSCs.METHODS:Adult healthy female Sprague-Dawley rats were randomly assigned to cell transplantation and control groups.Rat models of spinal cord transection damage were established.Rat BMSC suspension or calcium and magnesium phosphate buffer were transplanted immediately after injury to the damage zone.At 1 day,1,2,3,4 and 8 weeks before and after transplantation,BBB score motor function was observed in rats,and at 1 week after transplantation,immunohistochemical staining was utilized to observe BrdU-labeled BMSC survival in the spinal cord damaged site.At 4 weeks after transplantation,the general observation and histological detection were observed.RESULTS AND CONCLUSION:At 1-8 weeks after transplantation,BBB scores were higher in the cell transplantation group than in the control group.At 1 week following surgery,immunohistochemical staining showed that BrdU-positive cells were detected in the distal end of rat spinal cord in the cell transplantation group.At 4 weeks following surgery,nerve fibers were found in the damaged spinal cord.These verified that BMSCs were transplanted into rat damaged spinal cord immediately following damage,and the transplanted cells could survive.Living BMSCs can differentiate into neurons,and formed neuron pathway in the local region of damage,which will promote the recovery of conduction function of spinal nerve fibers,and contribute to the recovery of rat hindlimb motor function following high-level spinal cord injury.

BACKGROUND: Acidic fibrobiast growth factor can regulate cell proliferation, migration, differentiation and survival, also can down-regulate the known inhibitor of axon regeneration, such as proteoglycan, help axons overcome these inhibitory factors, and have significant role on the regeneration of nerve fibers.OBJECTIVE: To study the feasibility and effect of the acidic fibroblast growth factor combined with peripheral nerve transplantation in the treatment of high-level spinal cord injury in rats.METHODS: A total of adult 108 female SD rats were randomly divided into autologous nerve group, autologous nerve combined .with acidic fibroblast growth factor group, and high-level spinal cord injury group. The rat T_(8-10) spinous process and lamina were bite, revealing dural sac, high-level spinal cord was resected at a horizon level, cutting 3 mm, no nerve fibers were confirmed to be attached under the microscope. In the autogenous nerve group and autologous nerve combined with acidic fibroblast growth factor group, bilateral the 8~(th) to 10~(th) pairs of intercostal nerves were harvested 2 cm, then cross-transplanted into high-level spinal cord defect (proximal white matter and distal gray matter, distal white matter and proximal gray matter), fibrin gel and fibrin gel containing acidic fibroblast growth factor were used respectively to fix the implanted intercostal nerve, followed by dural suture.High-level spinal cord transection group was subjected to exclusion between stumps. At 90 days postoperation, somatosensory evoked potential and motor evoked potential were used to test nerve electrophysiological recovery. At 76 days postoperation,biotinylated dextran amine anterograde tracing was applied to observe the motor conduction bundle recovery. At 60 days postoperation, hindlimb motor function recovery was assessed by BBB score.RESULTS AND CONCLUSION: The somatosensory and motor evoked potential waveforms were not elicited in rats of high-level spinal cord transaction group, but did elicit in autogenous nerve group and autologous nerve combined acidic fibroblast growth factor group. The average latency and amplitude of somatosensory and motor evoked potentials, as well as BBB scores in autologous nerve combined acidic fibrobiast growth factor group were significantly superior to autologous nerve group (P ＜ 0.01).In the autogenous nerve group and autologous nerve combined acidic fibroblast growth factor group, many more biotinylated daxtran amine-positive nerve fibers passed in the damage zone, compared with high-level spinal cord transection group (P ＜0,01), the autologous nerve combined acidic fibrobiast growth factor group was more than autogenous nerve group (P ＜ 0.01). It is indicated that autologous peripheral nerve graft acidic flbroblast growth factor can better restore the limb motor functions of rats after high-level spinal cord injury.

AIM:To investigate the effects of osteogenic induction media and the medias containing different concentration of calcium on the induction of osteogenic differentiation of human renal fibroblasts in vitro.METHODS: Culturedhuman renal fibroblasts were divided into 5 groups in this experiment: osteogenic induction group (osteogenic inductionmedia), CaⅠgroup (0.5 mmol/L Ca2 + media), CaⅡgroup (1.5 mmol/L Ca2 + media), Ca Ⅲ group (2.5 mmol/LCa2 + media) and control group (PBS).The cell activity in each groups was measured by MTT assay .At 9th day, the cellcalcium Alizarin red S staining and alkaline phosphatase (ALP) Gomori calcium cobalt staining were performed respectivelyto observe the formation of calcium nidus and the expression of ALP .In addition, the expression of Runt-related transcriptionfactor 2 (Runx2) at mRNA and protein levels was determined by real -time PCR and Western blot, respectively.RE- SULTS: The remarkable positive signs which represented the formation of calcium nidus and the deposit of calcium objectsin all experiment groups were observed .The mRNA and protein expression of Runx2 in osteogenic induction group increasedin accordance with the induction time .Compared with control group, the mRNA and protein expression of Runx2 inthe CaⅠ ~Ⅲ groups increased gradually in a calcium concentration dependent manner at the 9th induction day.CON- CLUSION: Human renal interstitial fibroblasts show the potential activity in osteogenic differentiation induced by osteogen -ic induction media or high level calcium in vitro, which may be account for the cytological formation of the Randall ’splaque in the kidney.

PurposeTo investigate the feasibility and clinical value of 3.0T MRI iterative decomposition of water and fat with echo asymmetry and least-squares estimation (IDEAL) technique in imaging normal lumbosacral nerve roots.Materials and Methods Twenty-five healthy volunteers and 15 patients with lumbosacral pain were examined on MR scanner using IDEAL sequence thin-layer coronal scanning and reconstruction technique. The evaluation of bilateral lumbosacral nerve roots from L1 to S1 included the detection rate of lumbosacral nerve roots, the length of preganglionic nerve, the width and length of dorsal root ganglia (DRG) and the length of nerve root sheath.Results The anatomic structure and the contour of spinal nerve roots were well demonstrated on IDEAL sequence imaging, with 100% detection rate for preganglionic nerve and DRGs of nerves from L1 to S1 level, as well as the structures posterior to the ganglions from L3 to S1 level. The structures posterior to the ganglions at L1 to L2 level were shown as grade I in 12% and 42%, grade II in 16% and 32%, grade III in 72% and 26%. The length of preganglionic nerve, DRGs and nerve root sheath from L1 to S1 were gradually increased (P<0.05 orP<0.001). Normal lumbosacral nerve roots appeared as isointense linear structure in the nerve canals.ConclusionIDEAL sequence can depict the anatomic structure of the lumbosacral nerve roots and provide reference standards for lumbosacral nerve roots compression.

Purpose To evaluate the diagnostic significance of non-contrast enhanced magnetic resonance angiography (NCE-MRA) for lower extremity arterial stenosis on a 3.0T MR scanner, in order to provide a reliable method for clinical application. Materials and Methods Thirty patients with arterial disease in lower extremity underwent NCE-MRA before contrast enhanced magnetic resonance angiography (CE-MRA). Image quality of the two methods was compared. The diagnostic accuracy for significant stenosis ( ≥50%) of NCE-MRA was assessed using CE-MRA as a golden standard. The consistency of the two methods in diagnosis of significant stenosis ( ≥ 50%) was analyzed. Results All patients successfully underwent both NCE-MRA and CE-MRA examination. There were 532 arterial segments detected by NCE-MRA. In the calf region, venous artifacts presented more frequently on CE-MRA (Z=4.92, P<0.01), while in the abdominal and the femoral regions, venous artifacts presented more frequently on NCE-MRA (Z=4.58 and 3.56, P<0.01). The sensitivity, specificity, accuracy, positive predictive value and negative predictive value of NCE-MRA for the diagnosis of significant stenosis ( ≥ 50%) were 97.89%, 97.69%, 97.74%, 93.92% and 99.22%, respectively. There was good agreement (Kappa=0.94, P<0.05) between the two methods. Conclusion For the imaging of lower extremity arterial stenosis, NCE-MRA shows similar image quality and diagnostic accuracy with CE-MRA, thus can be used as an alternative method for lower extremity arterial stenosis in patients who have renal insufficiency or other contraindication of contrast media.