BACKGROUND:Many in vivo and in vitro experiments indicate that implantation of exogenous basic fibroblast growth factor can significantly promote the process of bone formation, but the in vivo degradation of exogenous basic fibroblast growth factor affects the therapeutic efficacy.
OBJECTIVE:To observe the effects of basic fibroblast growth factor-transfected mesenchymal stem cells which transfected using molecular biology techniques combined with al ogeneic bone in the repair of critical-size bone defects in sheep.
METHODS:Al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells, bone marrow mesenchymal stem cells combined with al ogeneic bone material stents, al ograft bone material,β-tricalcium calcium material were respectively implanted into critical-size bone defects in sheep. After 4, 8 and 12 weeks, histological and immunohistochemical staining was performed.
RESULTS AND CONCLUSION:At 12 weeks after implantation of al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells as tissue engineering bone, there were many cartilage-like structures in the operative binding region and a large amount of osteoblast-like cells in the center of operative region, and there was more material degradation in the entire operative area as compared with other groups;there were fibrous connective tissues ful of the pores, and osteoclast-like cells were commonly seen around the implant material;bone sialoprotein and col agen type Ⅰ expression were strongly positive. In the other three groups, although the cartilage-like structure appeared in the binding region, dead bone structure was found in the central area, and bone sialoprotein and type Ⅰ col agen expression was weak. These findings indicate that al ogeneic bone with basic fibroblast growth factor-transfected bone marrow mesenchymal stem cells can basical y repair critical-size bone defects in sheep.

BACKGROUND:Xenogeneic bone has a natural porous structure that is similar to human bone. In the treatment of bone defects, the porous structure is helpful to guide bone regeneration, but different degrees of immune responses wil be caused during the implantation process. OBJECTIVE:To prepare a freeze-dried antigen-extracted sheep cancelous bone scaffold and to evaluate its biocompatibility. METHODS: The sheep vertebral cancelous bone was colected to prepare two kinds of antigen-extracted heterologous bone scaffolds that were treated with chemical methods as chemical group and treated with chemical methods+cryopreservation at a-80℃ refrigerator for 4 weeks+drying in vacuum apparatus+60 RESULTS AND CONCLUSION:Freeze-dried bone had no cytotoxicity, no acute toxicity and heat reaction, and was negative for the intracutaneous stimulation test. The scaffold in the chemical group had cytotoxicity and mild irradiation as freeze-dried bone group. (1)Cytotoxicity test: Bone marrow mesenchymal stem cels isolated from sheep were cultured in extracts of the chemical group, free-dried bone group and Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12. (2) Heat reaction and acute toxicity tests: Extracts from the chemical group, freeze-dried bone group and normal saline were respectively injected into the ear vein of rabbits. (3) Intracutaneous stimulation test: Extracts from the chemical group, freeze-dried bone group and normal saline were respectively injected subcutaneously into the back of rabbits. Co acute toxicity reaction, sent heat source and had mild irritation. Results show that after freeze drying processing, the sheep vertebral cancelous bone has good biocompatibility, can meet the requirements of bone tissue engineering, but the bone that through chemical processing exhibits a relatively poor biocompatibility that cannot achieve the safety standard of biological scaffold materials.

BACKGROUND:Xenogeneic bone structure and biological characteristics are similar to human bone tissue, and the xenogeneic bone has a decreased antigenicity after physicochemical treatment, with a natural porous structure and rich source, and can be kept for a long time, which is considered to be an effective way to solve the shortage of the autogenous bone and al ograft bone.
OBJECTIVE:To compare the physical and chemical properties of xenogenic bone materials prepared by two different methods.
METHODS:Sheep cancellous bone treated with chemical method was placed into the muffle furnace at 1 000 ℃for 2 hours to prepare calcined bone. Another cancellous bone was placed into an 80 ℃ refrigerator for 4 weeks and then placed into a vacuum instrument to prepare freeze-dried bone. Cancellous bone rinsed with ultra-pure water served as controls.
RESULTS AND CONCLUSION: Three groups of samples retained three-dimensional porous structure which similar with human bone tissue under microscopic observation. The framework was stil intact, with a smal pore of 55-650μm and high porosity of 65%-80%. For the calcined bone, the toughness was decreased and the brittleness increased significantly, but the freeze-dried bone had a little changes in the mechanical properties. Through diffraction analysis, hydroxyapatite was the main composition of the three groups. However, there was a smal amount ofβ-tricalcium phosphate in the calcined bone. Spectrum analysis confirmed that calcium and phosphorus content in these three groups were al close to the human body. The results suggest the cancellous bone treated with these two methods is similar to human bone structure, and the major elements are close to the body. In addition, the cancellous bone after processing has enough smal pore and higher porosity. However, calcination process has a more influence on the mechanical property of scaffold materials, and the freeze-dried bone has a little change but the antigen cannot be completely removed that can reach the basic requirements.

0.05),but these positive rates were notable decreased comparing with nPCR and immunohistochemistry techniques(P

OBJECTIVE:To study the pharmacokinetics of emodin(an active component of Qingyi-Ⅱgranula) in rats after i.g administration of Qingyi-Ⅱgranula.METHODS:Rats were i.g administered with Qingyi -Ⅱgranula(2.5 g?kg~(-1) and 5.0 g?kg~(-1)).Serum concentrations of emodin were determined at different time points by HPLC,and the pharmacokinetic parameters were computed.RESULTS:The pharmacokinetic parameters of emodin in rats after administration of Qingyi-Ⅱgranula(2.5g?kg~(-1) and5.0g?kg~(-1)) were asfollows:t_(1/2?):(9.468?8.46) hand(21.68?17.867) h;t_(1/2?):(15.388?5.46) h and.(39.63?24.39) h;t_(max):(2.500?3.479) h and(5.333?3.266) h;C_(max):(0.058?0.004) mg?L~(-1)and(0.101?0.007) mg?L~(-1);CL:(33.027?9.365) L?h~(-1)?kg~(-1) and(9.405?5.846) L?h~(-1)?kg~(-1);AUC_(0-∞):(0.652?0.201) mg?h~(-1)?L~(-1)and (1.364?0.267) mg?h~(-1)?L~(-1).The pharmacokinetic process was in line with two-compartment model.CONCLUSION:The method for the detection of serum concentration of emodin in rats and the related pharmacokinetic parameters had been established in our study,which serves as a theoretic basis for the pharmacokinetic study of Qingyi-Ⅱgranula.

Objective To evaluate the diagnostic value of urine-based prostate cancer antigen 3 ( PCA3 ) score in detecting prostate cancer during initial prostatic biopsy .Methods Urine was collected after digital rectal examination ( DRE) ( three strokes per lobe ) from 248 men before prostate biopsy .The specimens were collected between January 2010 and December 2012.The expression of PCA3 mRNA and prostate specifc antigen (PSA) mRNA was determined by quanti-tative real time polymerase chain reaction ( qRT-PCR ) .PCA3 scores were calculated by PCA 3 mRNA/PSA mRNA × 1000 .The ability of the PCA3 score to predict the biopsy outcome was assessed with AUC-ROC analysis and compared with the serum PSA levels.Results The rate of positive prostate biopsy was 32.3%(80 patients with positive prostatic biopsy versus 168 patients with negative prostate biopsy ) .PCA3 scores were significantly higher in patients with positive biopsy than in those with negative biopsy results (P<0.001).The ROC curve analysis demonstrated that the area under the ROC curve (AUC) of serum total PSA (tPSA), PCA3 score and the duplex model combining tPSA and PCA 3 score was 0.620, 0.693 and 0.724, respectively.Further analysis of the diagnostic performance of PCA3 score revealed that at a cut-off of 90.2456, the sensitivity was 67.5%and the specificity was 61.9%for discriminating positive biopsy from negative biopsy. The duplex model combining tPSA and PCA 3 score represented a better approach than tPSA alone in PCa diagnosis by pros-tatic biopsy (P=0.011), but there was no statistically significant difference between tPSA and PCA 3 score (P=0.160). In addition , a comprehensive diagnostic model based on multiple risk factors of prostate cancer combined with PCA 3 score could further improve the predictive accuracy of prostate cancer .Conclusion PCA3 could be a good predictor of prostate cancer in initial prostate biopsy in Chinese population .The comprehensive diagnostic model can improve the diagnostic potency .Further large-scale multicenter studies in China are needed to confirm our findings .

Objective To systematically evaluate the association between lactotransferrin(LTF) gene rs1126478 polymor‐phism and caries susceptibility .Methods The published literatures on the association between LTF gene rs1126478 polymorphism and caries susceptibility were retrived from the electronic databases of Pubmed ,Web of science ,Embase ,EBSCO ,Springerlink , CNKI ,VIP ,Wanfang and CBM by computer .The pooled odds ratios (ORs) and its 95% confidence intervals (95% CIs) were used as the effect indicators .The statistical analysis was performed by using the RevMan 5 .2 and Stata 12 .0 softwares .The bias evalua‐tion and sensitivity analysis were also performed .Results A total of 5 case‐control studies involving 720 cases and 412 controls were included .The meta analysis results showed that no statistical significance in the association between LTF gene rs1126478 pol‐ymorphism and caries susceptibility was revealed in all four genetic models (GG+ AG vs .AA :P= 0 .75 ,OR= 0 .93 ,95% CI=0 .59-1 .46 ;G vs .A :P=0 .88 ,OR=0 .97 ,95% CI=0 .68 -1 .38 ;GG vs .AA :P=0 .84 ,OR=1 .07 ,95% CI =0 .57 -1 .99 ;GG vs .AG+AA :P=0 .52 ,OR=1 .12 ,95% CI = 0 .79-1 .58) .Similarly ,the subgroup analysis by ethnicity showed no statistical sig‐nificance in the onset risk between Caucasian and Asian populations as well (both P>0 .05) .Conclusion The LTF gene rs1126478 polymorphism may have no relation with the susceptibility to caries .

Objective:To explore and compare the value of radiomic features based on 18F-fluorodeoxyglucose (FDG) PET and CT in distinguishing epidermal growth factor receptor (EGFR) mutation status in patients with lung adenocarcinoma. Methods:Pretreatment 18F-FDG PET/CT images and EGFR gene status of 114 patients (64 males and 50 females, aged range: 35-84 (average age: 61) years) with primary lung adenocarcinoma between January 2017 and December 2017 were retrospectively collected. The volume of interest was drawn manually slice by slice, then the features were extracted by the LIFEx software. The parameters were screened by least absolute shrinkage and selection operator (LASSO) method for 200 times, and ten-fold cross-validation was used to select the best tuning parameter λ. Three models, namely M PET, M CT, M PET+ CT, were constructed by binary logistic stepwise regression. The receiver operating characteristic (ROC) curve was generated and the corresponding area under the curve (AUC), sensitivity, specificity and accuracy were calculated. The AUCs of three models were compared by Delong test. Results:Totally, 53.51%(61/114) patients were with wild type EGFR and 46.49%(53/114) patients had EGFR mutation. There were 3, 3, 7 parameters selected to form M PET, M CT, M PET+ CT, respectively. The AUCs for M PET, M CT, M PET+ CT were 0.730, 0.752 and 0.866 respectively. When the cut-off values were 0.427, 0.522, 0.378 for M PET, M CT and M PET+ CT, the Youden index were up to the maximum as 0.420, 0.405, 0.630, with sensitivities of 83.0%(44/53), 58.5%(31/53), 92.5%(49/53), specificities of 59.0%(36/61), 82.0%(50/61), 70.5%(43/61) and accuracies of 70.2%(80/114), 71.1%(81/114), 80.7%(92/114), respectively. There was no significant difference between AUC of M PET and M CT ( z=-0.320, P>0.05). The differences of AUCs between M PET+ CT and M PET, M PET+ CT and M CT were statistically significant ( z values: 2.963, 2.523, both P<0.05). Conclusions:PET, CT and PET+ CT radiomic features are all associated with EGFR gene expression in lung adenocarcinoma. M PET+ CT has the highest predictive efficiency.

Objective To investigate the applied perineal anatomy of the pudendal nerve of female and its direction to perineal operation.Methods The gross anatomy were done on 20 female cadaver(ensuring the pelvic,the hip and the perineum were intact).In each cadaver,the dissection proceeded from intrapelvic,the pudendal canal(Alcock'canal) and the ischiorectal fossa.The composition,track,abut,ramus and domination area of female pudendal nerve were examined.Results The gross anatomy showed that the pudendal nerve was composed of the ventro-branches of sacral nerve 2~4(S_2~S_4).The pudendal nerve sends off anal branch at level of the ischial spine.And the trunk courses in the internal pudendal canal forwardly and inwardly send off the perineal nerves and dorsal nerve of clitoris.The perineal nerves sends off branches to back lunar labium,muscle and urethra.The dorsal nerve of clitoris sends off the nerve to skin of clitoris,and some to the urethra simultaneously.The pudendal nerve has different abut in different anatomical position.Conclusions The perineal anatomy of the pudendal nerve has the meaningful direction to the perineal operations.

OBJECTIVE To establish a new cell line that can stably express humanα2A-adrenoceptor (α2A- AR). METHODS Recombinant plasmid of α2A- AR with hygromycin B (Hygro) resistance (pcDNA3.1/Hygro-HA-α2A-AR)was stably transfected into Chinese hamster ovary(CHO)cells which had expressed protein kinase A catalytic subunits(PKAcat) with labeling of enhanced green fluorescent protein(EGFP)by a Lipofectamine based method. A single positive clone expressingα2A-AR was selected through cultivation in the presence of 200 mg · L-1 hygromycin B followed by PKA redistribution assay. The transcriptional expression ofα2A-AR was detected by quantitative real-time PCR(qRT-PCR). Time-resolved fluorescence resonance energy transfer immunoassay was used to identify the function of inhibiting cAMP accumulation of α2A-AR. RESULTS The CHO-PKAcat-α2A-AR cell line No.7 exhibited stable response in PKA redistribution assay. qRT-PCR analysis demonstrated that the high expression ofα2A-AR in the cell line remained stable after a few generations compared with CHO-PKAcat-EGFP cells (P<0.01). The cAMP accumulation caused by forskolin was significantly inhibited by α2A-AR agonist in CHO-PKAcat-α2A-AR cells(P<0.01). CONCLUSION CHO-PKAcat-α2A-AR cell line is constructed successfully, which provides an effective model for drug screening and studies of mechanisms.