Objective To study the pattern of cerebral gray matter and white matter volume changes among smokers with differ-ent level of nicotine dependence (addition)using voxel-based morphometry (VBM).Methods The current case-control study recrui-ted 53 healthy male smokers and 53 healthy non-smokers from outpatients of our hospital during January 2013 to May 2014.Personal information (including for example age,sex and addition dependence of subjects)was collected using a questionnaire.3D-T1 images of whole brain structure were collected and were analyzed using DARTEL toolbox of SPM8.Smokers were divided into mild to mod-erate nicotine dependence group (n=23)and severe nicotine dependence group (n=30)based on Fagerstr?m Test for Nicotine De-pendence (FTND)score.Independent sample t-test analyses were performed to compare the volumes of gray matter and white mat-ter between smokers with different levels of nicotine dependence and non-smokers.Results Compared with non-smokers,gray and white matter volumes of smokers were smaller in multiple brain areas,mainly in the middle occipital gyrus,posterior cingulate,cer-ebellum anterior lobe,precuneus,caudate body and insula,which however,had larger number and scope of focal areas with gray and white matter atrophy in the mild to moderate nicotine dependence group than that in the severe nicotine dependence group.Conclusion Smokers with mild to moderate nicotine dependence have more pronounced gray and white matter atrophy than that smokers with severe nicotine dependence have.

BACKGROUND:Silk fibroin, chitosan, and nano hydroxyapatite are natural materials, and they al have good biological activity and physical or chemical properties. As tissue engineering materials, they have been already widely used in clinic or research work, but there are some defects in the application of these three kinds of materials. OBJECTIVE:To discuss the preparation and characteristics of silk fibroin/chitosan/nano hydroxyapatite complicated scaffolds which could be used in bone tissue engineering. METHODS:Silk fibroin, chitosan, and nano hydroxyapatite were separately prepared into 2%solution, and then mixed at the ratio of 1:1:0.5, 1:1:1, 1:1:1.5 respectively. The three-dimensional complicated scaffolds were prepared by those mixed liquids through repeated freeze drying and chemical crosslinking technology. Scanning electron microscope was used to detect the pore size of the scaffolds. Porosity, water absorption rate, and hot-water loss rate were determined. Mechanical tester was used to measure the tensile and compressive modulus of dried three-dimensional scaffolds. RESULTS AND CONCLUSION:The silk fibroin/chitosan/nano hydroxyapatite complicated scaffold in the dry state had no special smel , appeared to be a stabilized solid cylinder, and exhibited clear resiliency and flexibility with a touch. With the increased content of nano hydroxyapatite, the porosity, water absorption rate and average pore size of the scaffolds appeared to be decreased, while the hot-water loss rate and compressive strength were increased. The scaffold prepared at 1:1:1 was better for bone tissue engineering, and the average pore size, water absorption rate and hot-water loss rate were 85.67 μm, (135.65±4.56)%and (22.84±1.06)%, respectively, closer to the needs of the bone tissue engineering. Uniform pores were found within the scaffold at 1:1:1, showing the network structure, developed transport among pores, and the network structure was approximately 10μm.

Objective To investigate the effects of cold preservation on the expression of GATA in intrahepatic bile duct.Methods The intrahepatic bile duct tissues of SD rats were obtained by collagenase perfusion combined with mechanical separation.After being cut into fragments,the intrahepatic bile duct tissues were cultured in rat tail collagen gel for 48 hours before experiment.All the rats were divided into the control group,cold preservation 1 hour (CP1 h) group and cold preservation 12 hours (CP12 h) group.There were 5 rats in each group.The mRNA and protein expressions of GATA were detected by Real-Time polymerase chain reaction and Western blot.Measurement data with normal distribution were presented as (x) ± s.Comparison among 3 groups was done by ANOVA and pairwise comparisons were done by LSD test.Results The mRNA expressions of GATA3,GATA4,GATA6 were detected,while the mRNA expressions of GATA1,GATA2 and GATA5 were undetectable in intrahepatic bile duct tissue of the control group.The mRNA expressions of GATA4 in the CP1 h group,CP12 h group and the control group were 0.72 ± 0.08,0.56 ± 0.07 and 0.96 ± 0.06,with significant difference among the 3 groups (F =38.981,P ＜0.05).The mRNA expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group,and the mRNA expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The mRNA expression of GATA6 in the CP1 h group,CP12 h group and the control group were 0.83 ± 0.07,0.68 ± 0.12 and 0.98 ± 0.12,with significant difference among the 3 groups (F =10.175,P ＜ 0.05).The mRNA expression of GATA6 in the CP12 group was significantly lower than that in the CP1 h group and the control group,and the mRNA expression of GATA6 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The mRNA expressions of GATA3 in the CP1 h group,CP12 h group and the control group were 0.92 ± 0.06,0.89 ± 0.05 and 0.98 ± 0.11,with no significant difference among the 3 groups (F =1.674,P ＞ 0.05).The protein expressions of GATA4 in the CP1 h group,CP12 h group and the control group were 0.78 ± 0.07,0.64 ± 0.06 and 0.99 ± 0.10,with significant difference among the 3 groups (F =24.211,P ＜ 0.05).The protein expression of GATA4 in the CP12 h group was significantly lower than that in the CP1 h group and the control group,and the protein expression of GATA4 in the CP1 h group was significantly lower than that in the control group (P ＜ 0.05).The protein expressions of GATA6 in the CP1 h group,CP12 h group and the control group were 0.90 ± 0.04,0.75 ±0.06 and 0.98 ±0.11,with significant difference among the 3 groups (F=11.651,P＜0.05).The protein expression of GATA6 in the CP12 h group was significantly lower than that in the CP1 h group and the control group (P ＜ 0.05).Conclusion The expressions of GATA4 and GATA6 in the intrahepatic bile duct tissues are decreased significantly after cold preservation,which indicate that GATA4 and GATA6 might be involved in the pathophysiological process of the bile duct after cold preservation.

Objective To investigate cognitive functions in first-episode schizophrenia and unaffected first-degree relatives of different patients.Methods A total of 52 patients with first-episode schizophrenia (patient group),48 unaffected first-degree relatives of different patients (relatives group) and 50 healthy normal controls (control group) was recruited in the study.The attention,memory,and executive functions of all the subjects were assessed by a battery of neuropsychological tests,including cancellation test (CT),trail making test (TMT),digit span (DS),visual reproduction (VR),verbal fluency task (VFK),Wisconsin card sorting test (WCST) and HANOI tower.Results There were statistically significant difference among three groups in net scores of CT,TMTA,TMT-B,backward and total scores of DS,scores of VR,VFK,total errors and perseverate errors of WCST and all indicators of HANOI Tower (P ＜0.05 or P ＜ 0.01).Post Hoc multiple comparisons showed that,compared with the control group,the patient group had worse scores in all tests above except TMT-A in the relatives group (P ＜0.05 orP ＜0.01).However,compared with the patient group,the relatives group had better scores in net scores of CT,TMT-A,TMT-B,scores of VR,VFK,total errors and perseverate errors of WCST (P ＜ 0.05 or P ＜ 0.01).Conclusions Patients with first-episode schizophrenia have global cognitive functions deficits.Unaffected first-degree relatives of different patients also have cognitive deficits to a certain extent.Our study indicates that cognitive functions deficits may be viewed as a biomarker for candidates reflecting genetic liability for schizophrenia.

BACKGROUND:Poly lactic acid as an excellent delivery has good biocompatibility.
OBJECTIVE:To prepare recombinant human bone morphogenetic protein-2 (rhBMP-2)/poly lactic acid (PLA) sustained release microspheres, and to study its physical and chemical properties.
METHODS:The rhBMP-2/PLA sustained release microspheres were prepared using w/o/w solvent evaporation method. Scanning electron microscopy, laser particle size, zeta potential, and swel ing properties were detected. ELISA kit was utilized for measurement of encapsulation efficiency, drug-loading rate and in vitro drug release rate.
RESULTS AND CONCLUSION:Under the scanning electron microscope, rhBMP-2/PLA sustained release microspheres were approximately circle with excellent dispersion. The uniform spheres were visible with a mean particle size of 839.6 nm. The zeta potential were (-32.93±3.74) mV. The swel ing coefficient was 1.157±0.059. The drug-loading rate and encapsulation efficiency of rhBMP-2/PLA sustained release microspheres were (88.943±2.878)%and (0.026±0.001)%respectively. The drug release rate at 1 day was about 10.199%, then the drug release was relatively constant, and til 19 days, the cumulative drug release rate was 54.643%. These findings indicate that the constructed rhBMP-2/PLA sustained release microspheres meet the requirement of the Chinese Pharmacopoeia (10th edition) that the encapsulation efficiency is not less than 80%and the microspheres have a good slow-release function in vitro.

BACKGROUND:The cardiomyocytes obtained by traditional culture methods exhibit poor purity and viability, cells grow in clusters, the connection between cells is less, and most of cells beat separately. OBJECTIVE: To obtain high-purity cardiomyocytes presenting with a sheet-like growth by modified culture method. METHODS: Myocardial tissues were digested using 0.1% trypsin at 4℃ overnight, and then digested repeatedly through the addition of type Ⅱ collagenase, and finally were isolated and purified by differential adhesion method combined with chemical inhibitors. The morphology of cardiomyocytes was observed under inverted phase contrast microscope. The purity of cardiomyocytes was identified by cardic troponin T antibody after cultured for 48 hours. The beating frequency of cardiomyocytes was recorded every two days for 10 consecutive days. RESULTS AND CONCLUSION:The cardiomyocytes adhered within 24 hours of culture, distributed in clusters at 72 hours and grew in sheets at 5-7 days of culture. The survival rate of cardiomyocytes was 96% and the purity identified by immunofluorescence was over 97%. There was no significant difference in the beating frequency at different time. These results suggest that the high-purity and high-viability cardiomyocytes are obtained by the modified culture method, which is an effective method for culturing neonatal cardiomyocytes.

BACKGROUND:Bone morphogenetic protein-2 (BMP-2) is a key to bone formation and repair.However,it has some disadvantages such as easy to lose and degrade and difficult to sustain continuous effect.OBJECTIVE:To study the preparation and properties of silk fibroin/chitosan/nano-hydroxyapatite (SF/CS/nHA) scaffold loading BMP-2.METHODS:After silk degumming,dissolution and purification,2％ SF solution was obtained.BMP-2 was dissolved in 2％ CS solution,and then fully mixed with equal volume of SF solution and proper amount of nHA.At last,the SF/CS/nHA scaffold loading BMP-2 was prepared using freeze-drying method as experimental group.The SF/CS/nHA scaffold was soaked in the BMP-2 solution as control group.The scaffold porosity was measured by Archimedes method,the surface morphology of the scaffold was observed by scanning electron microscope,the compressive strength was measured by universal testing machine.Scaffolds in the two groups were soaked in PBS,and the release of BMP-2 was measured by ELISA method at different time points.RESULTS AND CONCLUSION:(1) The scaffolds in the two groups had irregular porous structure,interconnected pores and uneven pore wall.There was no significant difference between the two groups in mean pore diameter,porosity and maximum compressive strength.(2) On the 1st day,the release rate of BMP-2 was 4.63％ in the experimental group,and the release curve increased slowly.After 28 days,the release curve of BMP-2 was transferred to the plateau stage.But in the control group,the release rate of BMP-2 on the 1stday was 58.84％,and it was a significant initial burst release.The release curve increased rapidly,and was transferred to the platform stage on the 10th day.The release rate of BMP-2 release was significantly different between the two groups at days 1,2,4,10 (P ＜ 0.05).These results show that the SF/CS/nHA scaffold loading BMP-2 could sustainably and slowly release BMP-2.

Purpose To identify the role of tyrosine kinase receptor Axl for anti-apoptosis which was induced by cisplatin (DDP) and methotrexate (MTX) chemotherapy and to analyze the relationship between P-Axl and apoptosis-related proteins in osteosarcoma.Methods Osteosarcoma cell lines MG63,143B and U2OS were used in apoptosis assays,Axl siRNA transfection,cytotoxicity assays,cell cycle analysis,etc.A total of 41 cases of osteosarcom patients were included for immunohistochemistry of EnVision two-step staining and clinico-pathological relative analysis.TUNEL assay was performed in ten cases for apoptosis detection.Results Among the osteosarcoma cell lines,Gas6/Axl could obviously protect tumor cells from apoptosis induced by DDP and MTX (P ＜ 0.05).Axl siRNA transfection enhanced cell apoptosis,whereas Gas6 prone to function upon previous knockdown by Axl siRNA.Among the 41 cases,the positive rate of P-Axl,BCL-2,and Bax was 85.4％,70.7％,and 36.6％,respectively.In contrast,the positive rate of them was 22.2％,11.1％,and 11.1％ in osteofibrous dysplasia,respectively.The expression levels of these apoptosisrelated factors were significantly higher in osteosarcoma than in osteofibrous dysplasia (P ＜ 0.05).Through clinico-pathological analysis,there were significant relationships between the survival status and BCL-2 or Bax expression (P ＜ 0.05).Pearson correlation analysis demonstrated that BCL-2 was positively correlated to P-Axl with statistical significance (r =0.842,P ＜ 0.0001).By Cox univariate analysis,BCL-2 or Bax was correlated with the patients' prognosis.TUNEL assay also demonstrated that P-Axl high expression inhibited apoptosis in osteosarcoma tissues.Conclusion Gas6/Axl protects osteosarcoma cells from the apoptosis induced by DDP and MTX chemotherapy and inhibits apoptosis in osteosarcoma tissue,possibly through the regulation of apoptosis-related protein BCL-2.

Objective To establish a rat model at the same time in accordance with the “hot sheng syndrome” of traditional Chinese medicine and primary immune thrombocytopenic purpura of peripheral blood platelet reduction. Methods Using back multi-point injection of 20% dry yeast suspension on SD rats and 1∶4 dilution of rabbit anti SD rats platelet serum (APS) by intraperitoneal injection to establish a primary immune thrombocytopenic purpura “heat sheng”rat model.And observing rats of TCM syndrome characteristics, hemogram, myelogram and serotonin (5-HT) level of the temperature regulating center in thalamus.Results After injection of 2 h ~6 h temperature and daily water of the model group rats increased significantly,toe purper showed in fourth day of modeling and intestinal mucosal bleeding in thirty day of modeling(P <0.05);Platelet count in peripheral blood decreased significantly, bone marrow megakaryocyte number reduced significantly((P <0.05);5-HT level of the temperature regulating center of brain increased significantly((P <0.05).Conclusions The study of the primary immune thrombocytopenic purpura heat sheng rat model of combination of disease and syndrome reflected basically the pathological characteristics of purpura caused by “heat sheng” in primary immune thrombocytopenic purpura rat mode.

BACKGROUND:Severe abdominal infection after liver transplantation is the serious perioperative complications in liver transplant recipients, and it is one of the major reasons of death or loss of liver function. OBJECTIVE:To investigate the etiology, diagnosis and treatment of severe abdominal infection after orthotopic liver transplantation. METHODS:The clinical data of 186 cases of abdominal infection that received orthotopic liver transplantation between March 2004 and November 2011 were retrospectively analyzed. RESULTS AND CONCLUSION:Among the 186 patients, 16 patients had severe abdominal infection. Among the 16 patients, five patients had the infection due to the biliary anastomotic leakage caused large effusion in the gap under liver;10 patients had infection due to the peripheral liver massive hematocele caused by liver transplant surgery wounds extensive bleeding;one patient had injection due to left subphrenic large effusion caused by lower esophagus fistula after transplantation. Twelve patients had second operation within 3 days after diagnose, and there was no death;four patients had second operation after diagnosed for 3 days, one patient dead due to multiple organ failure at 21 days after liver transplantation and 5 days after second surgery. The results show that severe abdominal infection after liver transplantation is one of the serious perioperative complications in liver transplant recipients, and active recovery, multiple organ support and removal of infected lesions with control ing surgery as wel as the adequate drainage and other comprehensive treatment measures are the key points for the treatment of severe abdominal infection after liver transplantation.