BACKGROUND: Our previous studies have demonstrated that cryopreserved bone marrow stromal cells (BMSCs) still maintain high survival rate, cell proliferation and osteogenic differentiation potentials after thawing. However, this result needs confirmed in vivo environment. OBJECTIVE: To explore the effects of cryopreserved BMSCs and collagenic membrane BME-10X complex on type Ⅰ collagen synthesis in vivo. METHODS: Beagle dog BMSCs were cultured in vitro and cryopreserved for 12 months, which were thawed and prepared complexes with collagenic membrane. The complexes were cultured with mineralization induction medium or normal medium for 5 days, followed by implanting into nude mice. The specimens were harvested and analyzed by gross observation, histopathological and immunohistochemistry at 4 weeks after implantation. The collagenic membrane cultured with mineralization induction medium served as controls. RESULTS AND CONCLUSION: In the control group, the boundary of collagenic membrane was distinctly, without cell growth around boundary or intra collagenic membrane, additionally, there was little type Ⅰ collagen. In the non-induction group, cells grew into collagenic membrane, trabes-like collagen formed, and type Ⅰ collagen distribution increased at 4 weeks. In the induction group, scaffold degraded, more cells grew, and plenty of collagen formed osteoid-like tissues. The distribution of typeⅠcollagen was obviously increased than that of other groups. The findings demonstrated that cryopreserved BMSCs possess strong osteogenic differentiation potentials after proliferation and induction combined with collagenic membranes in vitro.

BACKGROUND: The biological functions of platelet-rich plasma (PRP) are affected by multiple factors, such as individual difference, PRP concentration, PRP carder, PRP-activated methods and so on. OBJECTIVE: To evaluate the effect of PRP, activated by different concentrations of thrombin, on the repair of cranial defects. METHODS: Whole blood of the central artery of rabbit ears was extracted to prepare PRP, which was then diluted so that the final platetet count was about 5 times of the whole blood. Four whole-thickness layer of cranial defects at an 8-mm diameter were created in 16 New Zealand rabbits and randomly grafted with β-tdcalcium phosphate (β-TCP) and PRP, activated by 60 U/mL. thrombin; β-TCP and PRP, activated by 1 000 U/mL thrombin; β-TCP and PRP; β-TCP alone. At 1 and 3 months following implantation, X-ray analysis and microscopic observation were performed to onserve cranial repair, the area percent of new bone formation was calculated. RESULTS AND CONCLUSION: At one month post-surgery, the edge of defects was clear in each group, with varying degrees of new bone formation surrounding the defects, β-TCP particles partially degraded and the degradation lesion was replaced by new bone, only a small amount of bone lacunae was seen, fiber wrapped around the defect center β-TCP, only a small number of specimens showed new bone formation; X-ray showed a clear boundary and uniform defect density; the percentage of new bone formation in the PRP groups were higher than β-TCP groups (P < 0.05). However, there was no significant difference between PRP group groups (P > 0.05). At 3 months post-surgery, the defect boundary was unclear in each group, the new bone formation increased, the β-TCP particles surrounding defects partially or all degraded and were replaced by new bones, some regions appeared trabecular bone, bone lacuna in new bone was increased, the central defect of the majority of specimens exhibited new bone formation; X-ray showed defect boundary was unclear in each group, defect surrounding density was higher than the center defect, and bone mineral density was equivalent to other normal parts; the percentage of new bone formation in the PRP groups was significantly higher than that in the β-TCP groups (P < 0.05), PRP +β-TCP group was higher than the other 3 groups (P <0.05), there was no significant difference between two thrombin groups (P > 0.05). It is indicated that although PRP improves the repair of cranial defects, 60 and 1 000 U/mL of thrombin has no effects on PRP rapairing cranial defects in New Zealand white rabbits, compared with PRP+β-TCP group, possible the absence of the optimal concentration of thrombin.

A method for the determination of bisphenol A, octyphenol, nonylphenol in water samples was developed using stir bar sorptive extraction (SBSE) based on poly (vinylimidazole-divinylbenzene) monolithic material (SBSEM) combined with high performance liquid chromatography with diode array detection.To achieve the optimum extraction performance, several main extraction parameters, including extraction and desorption time, pH value and contents of inorganic salt in the sample matrix, were investigated.Under the optimized experimental conditions, the method showed good linearity and repeatability, low detection limits (S/N = 3) and quantification limits (S/N = 10) of the proposed method for the target compounds were achieved within the range of 0.13-0.66 and 0.44-2.19 μg/L, respectively.The extraction performance of SBSEM to the target compounds was also compared with commercial SBSE which used polydimethylsiloxane as coating.The proposed method was successfully applied to the determination of the target compounds in water samples.The recoveries of spiked target compounds in real samples ranged from 37.8%-101.1%.The results indicated that the developed method possessed advantages such as sensitivity, simplicity, low cost and high feasibility.

Objective To investigate the imaging features of focal nodular hyperplasia and hepatocellular carcinoma on DWI. Methods The data of patients with histopathologically confirmed FNHs and HCCs between August 2008 and November 2010 were collected. A total of 24 patients with 26 FNH lesions and 36 patients with 39 HCC lesions were included in our study. All patients underwent breath-hold DWI with b = 500 s/mm2 and dynamic contrasted-enhanced (DCE) MRI. The imaging findings of FNHs and HCCs were retrospectively analyzed and compared. The signal intensity (SI) of the lesions on DWI were classified as iso-, slightly high, high SI and the distribution of SI between FNHs and HCCs was compared with Fisher exact test. ADC value and lesion-to-liver ADC ratio of FNHs and HCCs were measured and compared by using independent sample t test. ROC was performed to assess the diagnostic value of ADC value and lesion-liver ADC ratio in the characterization FNHs versus HCCs. Results Of 26 FNHs,23 manifested as isointensity or slightly high SI on DWI, but most 25 out of 39 HCCs showed high SI. The distribution of SI between FNHs and HCCs had significant difference ( P = 0. 000). The mean ADC value and lesion-liver ADC ratio for FNHs [ (1.76 ± 0. 62 ) × 10-3 mm2/s and 1.06 ± 0. 18, respectively ] were significantly higher ( P = 0. 001, P = 0. 000, respectively ) than those for HCCs [ ( 1.26 ± 0. 46 ) × 10-3mm2/s and 0. 79 ±0. 12, respectively]. The area (Az) under the ROC for the ADC value and lesionliver ADC ratio for the differentiation of FNHs versus HCCs were 0. 79 ± 0. 05 and 0. 85 ± 0. 05,respectively, with no significant difference (P =0. 270). The specificity of the two measures was 69. 23% and 97.44%, respectively, with significant difference (P = 0. 001 ). Conclusion FNH shows isointensity or slightly high SI with relatively higher ADC value and lesion-liver ADC ratio than those of HCCs on DWI,which is characteristic for its diagnosis and differentiation.

Objective To investigate the effects of human leptin ( hLEP) gene transfection on rat bone marrow stro-mal cells ( rBMSCs) .Methods rBMSCs were cultured and transfected with adenoviruses encoding hLEP ( Ad5-hLEP-EGFP) in vitro as experimental group while rBMSCs transfected with Ad 5-EGFP and non-transfected were control groups.The proliferation was detected by MTT and the expression of collagen type Ⅰ(Col-Ⅰ) and alkaline phosphatase ( ALP) were assessed by real-time PCR.The ability of mineralized nodule forming was also examined by Alizarin red staining .The combination of transfected rBMSCs and β-tricalcium phosphate (β-TCP ) was con-structed and the osteogenic ability of the construction was evaluated in nude mice .Results hLEP could be trans-fected into rBMSCs successfully by adenovirous .After transfection , the proliferation was not affected while Col-Ⅰand ALP expressions were more pronounced in rBMSCs transfected with Ad 5-hLEP-EGFP ( P<0.05 ) .Alizarin red staining showed the ability of mineralized nodule forming was also up-regulated in Ad5-hLEP-EGFP group (P<0.05).In addition, the transfected rBMSCs adhered to β-TCP and survived well and the combination showed more new bone like tissue formation in nude mice compared to control groups .Conclusions rBMSCs transfected with hLEP might be potently used in bone or periodontal tissue regeneration .

Objective To explore the quantitative characteristics of pancreatic ductal adenocarcinoma(PDAC) in single-source dual energy spectral CT imaging. Methods From January 2013 to December 2014, 113 patients underwent dual phase contrast-enhanced gemstone spectral imaging(GSI) on Discovery CT 750 HD. All diagnoses were pathologically confirmed by surgery or biopsy. The spectral HU curves of PDAC were observed, the monochromatic CT values, the effective atomic number(Zef ), the iodine concentration(IC), water concentration(WC), and the corresponding normalized values(normalized monochromatic CT values, normalized Zef , normalized IC, normalized WC)of the lesion and the pancreatic parenchyma in late arterial phase(AP) and portal venous phase(PP) were recorded . The measurements were performed three times repeatedly. Paired t test (normal distribution) or Wilcoxon test (non-normal distribution) were used for analyzing the differences between the two phases and between PDAC and pancreatic parenchyma. Results The monochromatic CT values of PDAC in AP were lower than in PP at each energy level and the difference was more marked at lower energy. The normalized monochromatic CT values increased with the increase of energy level in both AP and PP and the difference was more distinct at lower energy. The Zef , IC and normalized IC of PDAC all had significant differences(P<0.05), while the WC, normalized Zef , and normalized WC had no difference between AP and PP. The Zef and IC of pancreatic parenchyma had significant differences(P<0.05), while the WC had no difference between AP and PP. The differences of Zef , IC, and WC between PDAC and pancreatic parenchyma were significant in both two phases (P<0.05). Conclusions Dual phase CT spectral imaging showed characteristic quantitative parameters of pancreatic ductal adenocarcinoma. The monochromatic CT values, Zef , and iodine concentration of PDAC were lower than those of pancreatic parenchyma in both AP and PP. The monochromatic CT values, Zef , and iodine concentration of PDAC in late arterial phase were lower than those in portal venous phase. The differences were all more distinct at lower energy.

Aim To investigate the expression of stro-mal interaction molecule 1 (STIM1) in rat pulmonary arterial hypertension ( PAH ) tissues and effects of STIM1 on arterial muscle cells proliferation. Methods PAH was induced by a single intraperitoneal injec-tion of MCT at a dose of 60 mg·kg - 1 . The mRNA or protein expressions of STIM1 in monocrotaline-induced pulmonary hypertensive rats were measured by real-time PCR or Western blot, respectively. The arterial smooth muscle cells A7R5 were transiently transfected with STIM1 plasmids to prepare STIM1 overexpressed cells. Cell proliferations were detected by using CCK-8 kits. The expressions of Akt/ mTOR pathway molecules of A7R5 were measured by Western blot. Results The right ventricular systolic blood pressure ( RVSP) and right ventricular mass index ( RVMI ) were markedly elevated in MCT-treated rats (P < 0. 01) in comparison to control rats. The mRNA and protein ex-pression levels of STIM1 in monocrotaline-induced pul-monary hypertensive rats were 2. 19 and 1. 66 folds of control rats, respectively. STIM1 were transiently over-expressed in cultured A7R5. Cells transfected with STIM1 grew more quickly than non-transfected control. Overexpression of STIM1 significantly increased the phosphorylation of Akt, mTOR, p70-S6K, and 4E-BP1, but did not change their protein expression lev-els. Conclusion STIM1 are over-expressed in rat PAH tissues. Overexpression of STIM1 can promote ar-terial smooth muscle cells proliferation by regulating Akt/ mTOR pathway.

BACKGROUND:The researches about the effect of retinoic acid on the proliferation of adipose-derived stem cells are rare, and the researches on the testosterone are mainly on the inhibition of cellaging. OBJECTIVE: To study the effects of retinoic acid and testosterone or combination on the cellcycle of adipose derived stem cells. METHODS:Adipose derived stem cells were isolated from adult female Sprague Dawley rats with 2 months age and cultured in vitro til passage 3 adipose derived stem cells, and then the 3rd passage adipose-derived stem cells were performed with adipogenic induction, osteogenic induction and surface marker identification. The cells were divided into six groups:(1) Control group;(2) 10-5 mol/L retinoic acid group;(3) Retinoic acid group;(4) 10-5 mol/L retinoic acid+testosterone group;(5) 10-6 mol/L retinoic acid+testosterone group;(6) Testosterone group. The adipose-derived stem cells in the control group were cultured with Dulbecco’s modified Eagle’s medium+10%fetal bovine serum culture medium, and the adipose-derived stem cells in the other five groups were induced with corresponding dose of retinoic acid and testosterone on the basis of control group. After cultured for 36 hours, the flow cytometry was used to detect the changes of cellcycle. RESULTS AND CONCLUSION:Compared with the control group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group were increased significantly, and the cellproportions in phase S were decreased. Compared with control group, the cellproportion in phase G 1 of testosterone group was significantly reduced, and the cellproportion in phase S was increased. Compared with 10-5 mol/L retinoic acid group and 10-6 mol/L retinoic acid group, cellproportions in phase G 1 of 10-5 mol/L retinoic acid+testosterone group and 10-6 mol/L retinoic acid+testosterone group were reduced significantly and the cellproportions in phase S were increased. Retinoic acid can inhibit the cellcycle of adipose-derived stem cells in phase G 1 , and delay the process of the cellcycle from phase G1 to phase S;while testosterone can promote the cellcycle of adipose-derived stem cells from phase G1 to phase S;the combination induction of retinoic acid and testosterone can accelerate the process of the cellcycle of adipose-derived stem cells from phase G 1 to phase S.

Objective To investigate the expression of survivin/Human leukocyte antigen class I ( HLA-Ⅰ) proteins and its physiological significance in clear cell renal cell carcinoma ( CCRCC ) . Methods Immunohistochemistry was used to analyze survivin/HLA-Ⅰ protein expression in 90 cases of CCRCC and 10 normal tissues to study relationships with clinical symptoms and disease prognosis . Resutl s The level of survivin protein expression was found to be significantly higher in CCRCC tissues 82.2%( 74/90) than in normal tissues( 0/10).However, the relative amount of HLA-Ⅰprotein in colorectal cancer tis-sue was also found to be significantly lower 67.8%(61/90) than in normal tissues 90%(9/10).Survivin expression was associated with tumor grade , stage,and lymph node metastasis ( P=0.000, P=0.016, and P=0.001, respectively ) .Conversely , lost HLA-Ⅰexpression did not have any associations with clinicopath-ological data (P>0.05).Survivin negative patients (25.0%, 4/16) had a higher tumor-free survival rate than patients (52.7%, 39/74)with survivin expression (P=0.037).Patients (27.6%, 8/29) with normal HLA-Ⅰlevels had a higher tumor-free survival rate than those ( 60.7%, 37/61) with reduced HLA-Ⅰlev-els (P=0.020).The univariate and multivariate analyses indicated that expression of survivin and HLA indi -vidually and in combination were independent predictors for CCRCC patient survival . Conclusions Over-expression of survivin but reduced HLA-Ⅰ expression is associated with CCRCC development and progres-sion.

Objective To assess the value of computed tomography (CT) and magnetic resonance imaging (MRI) in the diagnosis of pancreatic neuroendocrine neoplasms (PNEN) and to analyze the factors influencing thepreoperative imaging diagnosis of PNEN.Methods From January 2016 to November 2016, patients with PNEN diagnosed by surgery and biopsy were collected. CT and MRI data of them were analyzed. The CT values or signal intensity of the lesions and the pancreatic parenchyma were measured and the contrast-to-noise ratio (CNR) of the lesion was calculated. Detecting sensitivity and diagnosis accuracy of CT and MRI were compared. Detecting sensitivity of different MRI sequences was also analyzed. Diagnosis accuracy of non-functional PNEN and functional PNEN was compared and analyzed. Lesion CNR was compared between arterial phase and portal venous phase of the contrast enhanced CT. The sensitivity, accuracy and constituent ratio were compared by nonparametric analysis. Independent sample t test and one-way analysis of variancewere performed for the quantitative parameters comparison. Results A total of 54 patients with 56 lesions of PNEN were included for two of whom had two lesions each. CT and MRI were both performed in 44 patients (46 lesions).Detecting sensitivity and diagnosis accuracy of CT were 97.8% (45/46) and87.0% (40/46), respectively. Detecting sensitivity of MRI were 97.8% (45/46) and89.1% (41/46), respectively. There was no significant difference in detecting sensitivity and diagnosis accuracy between CT and MRI (both P>0.05). The CNR of lesion in arterial phase was higher than that of portal venous phase(4.7±3.8 vs 3.4±2.5), and the difference was statistically significant (t=2.949, P<0.05). Detecting rates of T1 weighted imaging with fat suppression (T1WI-FS) image, T2 weighted imaging with fat suppression (T2WI-FS) image, diffusion weighted imagingand dynamic contrast enhanced T1WI-FS image were 90.0% (45/50), 88.0%(44/50), 86.0%(43/50), and 91.7% (44/48), respectively. There was no significant difference in detecting rate among these images sequences (Q=2.526, P=0.510). Tumor diameter in non-functional PNEN was significantly larger than that in functional PNEN ((2.9±1.6) cm vs (1.7±0.7) cm)(t=3.479,P<0.05). The overall diagnosis rate of non-functional PNEN with CT and MRI before operation was 70.8% (17/24), which was significantly lower than that of functional PNEN (100.0%, 31/31) (χ2=10.360，P=0.002).Conclusions CT and MRI are both sensitive in detectingPNEN, and they were two complementary modalities. CT image in arterial phase delineated the lesion better than that in portal venous phase. MRI images with different sequences can becomplementary and there is no significant difference in detecting sensitivity for PNEN among different sequences. CT and MRI play an equal rolein the diagnosis of PNEN before operation. Because of atypical CT and MRI findings, the diagnosis of non-functional PNEN is more difficult thanfunctional PNEN.