Objective To prepare the composite of Polycaprolactone(PCL) and Bone powder(BP) for bone repairing,and evaluate the biocompatibility of the material.Methods The materials were prepared in college of chemical engineering and materials science,Shanghai Jiaotong University,and the animal tests were fulfilled in Animal Experimental Center of the Second Military Medical University of PLA from June 2010 to March 2011.The PCL and BP were blended together to get the copolymer by haake ( Rheocord 900 ) torque rheometer.Then the extracts of the material in DMEM for 24 hours were obtained.Firstly biological safety detection of the materials was performed:①cytotoxicity test:the extracts were mixed with DMEM to seed rabbit osteoblasts for 24 hours.The cell proliferation was detected by MTT method.The cell proliferation rate and toxicity grate were also calculated ( cytotoxicity of grade 0 or 1 was qualified).②hemolysis test:the extracts were mixed with fresh rabbit blood,hemolysis rate were calculated ( hemolysis mte less than 5％ was qualified).③pyrogenic test:the extracts were injected into the rabbit ear vein with the dose of 10 ml/kg.The anus temperature was measured per hour for three times.The elevator degree was calculated by subtracting the natural temperature from the highest one.Secondly,cellular adhesion test of the material composite was performed.The composite was casted on the cover glasses.The samples were put in 6-well tissue culture plates,totally 6 × 104 osteoblasts were seeded each well and cultured for 3 days.The cells were observed at 24 and 72 hours of culture,and photography was performed using inverted phase contrast microscope.④general toxioity test:the extracts were injected into mouse vein with the dose of 50 ml/kg.The toxic symptoms were observed within 72 hours.Results The novel copolymer of PCL/BP was prepared.①Biological safety:the cell viability of the composite was 87.8％ and cytotoxicity was grade 1.The hemolysis rate was 2.4％,less than 5％.The maximal variety of body temperature in rabbits were 0.4℃,which accorded with the national standard ( ＜0.6℃ ).The general toxicity test showed that there were no adverse effects in the animals,such as death,convulsion,paralysis,respiratory depression,diarrhea and decreasing weight.②Cell adhesion:adhesive cells manifold obviously and assembled 24 hours later,inosculated gradually 3 days later,and the cells were in good fettle.Conclusion The composite of PCL/BP accords with cytotoxicity demand of the biological materials,presenting no toxicity,pyrogenicity or hemolysis.It possesses good biocompatibility and cellular adhesion.

Objective To study the relationship between Xsp Ⅰ restriction fregment length polymorphism (RFLP) in exon 4 of low-density lipoprotein receptor (LDLR) gene and hypercholesterolemia. Methods PCR-RFLP method was applied to determine the polymorphism of LDLR gene in 446 cases of hypercholesterolemia, 284 of borderline hypercholesterolemia and 187 normal control subjects. Results Three genotypes, X+X+, X+X- and X-X-, were found in the population. (1) The frequencies of the X+X+ genotype and X+ allele in the group with hypercholesterolemia were higher than those of the other 2 groups (P<0.05). (2) From X-X- to X+X+ genotype, cholesterol and low-density lipoprotein-cholesterol increased gradually, while high-density lipoprotein-cholesterol decreased (all P < 0.05). (3) With logistic regression analysis, LDLR genotype was associated with hypercholesterolemia. Conclusion Polymorphism in Xsp I restriction site at the exon 4 of LDLR (X+ X+ genotype and X+ allele) may be a risk factor of hypercholesterolemia in Chinese population.

Objective To investigate the correlation between hypoglycemia and brain injury of newborns.Methods Medical records and follow-up data of 110 newborns with hypoglycemia (blood glucose level≤2.2 mmol/L) who admitted into neonatal department of Peking University First Hospital from December 2006 to December 2009 were studied.All patients were divided into 3 groups:no brain injury group,mild and severe brain injury group according to their clinical manifestation,cerebral radiological characteristics and cerebral functional tests.By using receiver operating characteristic (ROC) curve and x2 test,the potential optimal blood glucose level and duration of hypoglycemia for predicting brain injury were confirmed.Multivariate Logistic regression was taken to determine independent predictors for brain injury.The analyzed factors included gender,preterm/small for gestational age,hyperbillirubinemia,fetal distress,asphyxia,infection,seizures and maternal hypertensive disorder complicating pregnancy and hyperglycemia.Results Among the 110 hypoglycemia newborns,33 (30.0％) infants suffered from brain injury,of which 23 were mild and 10 were severe.Blood glucose ≤1.7 mmol/L had high specificity (73％) and sensitivity (60％)for predicting brain injury.When blood glucose≤ 1.7 mmol/L,the incidence of brain injury and severe brain injury was 43.6％ (24/55) and 18.2％ (10/55),which was higher than those [16.4％(9/55) and 0.0％ (0/55)] of patients whose glucose level ＞1.7 mmol/L(x2 =9.74 and 11.00,P＜0.01 respectively).Blood glucose ≤ 1.2 mmol/L had high specificity (100％) and sensitivity (81％) for predicting severe brain injury.When blood glucose ≤1.2 mmol/L,the incidence of severe brain injury was higher than that of the patients whose glucose level was higher than 1.2 mmol/L [34.5％ (10/29) vs 0.0％ (0/81),x2 =30.72,P＜0.01].Duration of hypoglycemia ≥12 h had specificity (100％) and sensitivity (36 ％) for predicting brain injury.When duration of hypoglycemia ＜12 h,the incidence of brain injury was lower than that of the patients whose duration of hypoglycemia≥12 h [21.4％ (21/98) vs 6/6,x2 =27.69,P＜0.01].Multivariate Logistic regression showed that fetal distress (OR=4.69,95％CI:1.47-14.97,P=0.009),glucose level≤1.2 mmol/L (OR =5.16,95％CI:1.56-17.03,P=0.007),duration of hypoglycemia≥12 h (OR=8 885 220 297.12,95％CI:0.00-∞,P =0.000) and maternal hyperglycemia (OR =3.34,95％CI:1.01-11.02,P=0.048) were independent risk factors for neonatal brain injury.Conclusions Low blood glucose level and prolonged hypoglycemia might induce injury of neurol system.Fetal distress and maternal hyperglycemia might increase the incidence of brain injury in newborns with hypoglycemia.

Objective To analyze the effects of fertilization methods and sperm sources in intra‐cytoplasmic sperm injection (ICSI) on the developmental capacity of surplus embryos .Methods A retrospective study was carried out to compare the blasto‐cyst rate of the surplus embryos from 2 697 patients .According to the fertilization methods ,the embryos were divided into IVF group and ICSI group .According to sperm sources ,the ICSI group was divided into ejaculated group and testicular sperm group . The rates of blastocyst formation and good quality blastocysts were compared between different fertilization methods and sperm sources .Results There were 8 426 embryo developed in 2 697 patients .The blastocyst formation rate of surplus embryos was high‐er in the IVF group(n=1 048 ,53 .18% ) than that in the ICSI group(n=1 378 ,49 .27% ) ,but with no statistically significant differ‐ent(P> 0 .05) .The rates of blastocyst were not statistically significant different between the IVF group and in the rescue ICSI group(P>0 .05) .The rates of blastocyst were not statistically significant different between the ejaculated group and the testicular sperm group(P>0 .05) .Conclusion There were not statistically significant different of the rate of blastocyst between different fer‐tilization methods and sperm sources in ICSI .

Objective To investigate the correlation between anemia and outcome in a large cohort of unselected patients with acute cerebral infarction.Methods Consecutive acute cerebral infarction patients who were hospitalized were prospectively recruited from August 2010 to November 2013.Eight hundred and fifty-eight patients were enrolled,and the baseline data including age,sex,National Institute of Health Stroke Scale(NIHSS) scores,type of Oxfordshire Community Stroke Project(OCSP:total anterior circulation infarct,partial anterior circulation infarct,posterior circulation infarct and lacunar infarct),serum creatinine,initial hemoglobin level,initial hematocrit level,etc,were recorded.Hemoglobin level and hematocrit level during hospitalization were also recorded.Domestic criteria were used to define if the patient had anemia on admission.Recovery was assessed by modified Rankin Scale (mRS) 180 days after stroke by telephone interview (mRS scores ≤ 2 reflected good prognosis,and mRS scores ＞ 2 reflected unfavorable prognosis).The influence on outcome by anemia on admission,initial hemoglobin level,nadir hemoglobin level,nadir hematocrit level was analyzed by multinomial Logistic regression analysis.Results Odds ratio of initial hemoglobin level for poor outcome was 1.013 (95％ CI 1.001-1.024,P =0.027) with each decrease in hemoglobin of 1 g/dl.Initial anemia(OR =2.417,95％ CI 1.202-4.859,P =0.013) was a independent prognostic factor for mortality;odds ratio of nadir hemoglobin level for mortality was 1.016(95％ CI 1.002-1.030,P =0.026) with each decrease in hemoglobin of 1 g/dl;odds ratio of nadir hematocrit level for mortality was 1.047(95％ CI 1.003-1.093,P =0.037) with decrease in hematocrit of one percentage point.Conclusions Initial hemoglobin level was a independent prognostic factor for poor outcome in patients with acute cerebral infarction.Anemia on admission,nadir hemoglobin level,nadir hematocrit level were independent prognostic factors for mortality in patients with acute cerebral infarction.

Objective Folate receptors ( FRs) , overexpressed on the surface of a variety of tumor cells , are potential targets for tumor targeting therapy .This study aimed to prepare an FR-mediated drug nanocarrier with folate conjugated pullulan acetate ( FPA) to target chemotherapeutic agents to FR-overexpressed tumor cells and investigate its tumor-suppressing effect in vitro. Methods The cytotoxicity of epirubicin-loaded FPA nanoparticles ( FPA/EPI NP) against HepG2 and Hela cells was evaluated by MTS assay.The HepG2 and Hela cells were divided into five groups to be treated with NPs (NP control), chlorpromazine, chloro-quine, amiloride, and folate, respectively, followed by detection of the fluorescence intensity by flow cytometry . Results FPA/EPI NP was successfully formulated into NPs , with the mean particle size of (268.5 ±12.0) nm, by dialysis with an almost spherical shape . The drug-loading rate and entrapment rate of FPA/EPI NP were (6.45 ±1.04) and (72.45 ±11.50) %, respectively.The survival rates of the HepG2 and Hela cells were both >95%after 24 hours of incubation with FPA NP at 5, 40, 200, 400 and 1000μg/mL and 90.0%after 72 hours.The survival rates of the HepG2 cells treated with 5, 40, 200, 400 and 1000μg/mL FPA/EPI NP for 24 hours were (92.3 ±5.2), (70.4 ±4.6), (50.0 ±4.0), (41.1 ±4.1) and (27.0 ±3.6) %, respectively.Compared with the NP control group, the Hela cells of the chlorpromazine , amiloride and folate groups showed a significantly lower rate of NP uptake (P<0.05), and so did the HepG2 cells pretreated with chlorpromazine or amilo-ride (P<0.05).At 72 hours, the half maximal inhibitory concentrations (IC50) of FPA/EPI NP against HepG2 and Hela cells were 168 and 105μg/mL, respectively . Conclusion Clathrin-mediated endocytosis and macropinocytosis are involved in the internaliza-tion of FPA/EPI NP in HepG2 cells, while clathrin-and FR-mediated endocytosis in that of Hela cells .FPA NP may serve as a new drug carrier for tumor-targeted therapy .

Objective To analyze the MR manifestations of the focal nodular hyperplasia (FNH)in liver and observe the MRI changes after 30-36 months.Methods 12 (14 lesions)of the 13 patients (1 5 lesions)were firstly diagnosed as FNH by MRI with plain scan and dynamic Gd-DTPA enhancement,and then were confirmed by the follow-up survey with similar MRI after 30 - 36 months.The last one patient was also scanned with Gd-EOB-DTPA enhancement but with similar plain scans.The size,signal,en-hancement patterns of the lesions were analyzed and compared with those in the follow-up survey.Results Typical MR manifesta-tions were found in 93.33%(14/1 5)of the lesions with hypo-or iso-intensity on T1 WI,hyper-or iso-intensity on T2 WI,marked en-hancement in arterial phase and attenuation but still equal to or higher than normal liver parenchyma in portal and delayed phases af-ter Gd-DTPA administration.In addition,the central scar showed hypointensity on T1 WI,hyperintensity on T2 WI,and enhance-ment in delayed phase.The enhanced small vessels inside lesions and the pesudocapsule enhancement could be seen in delayed phase. The only one lesion with Gd-EOB-DTPA enhancement showed marked enhancement in arterial and portal phases,and slight hyperin-tensity in hepatocellular phase with hypo-intense central scar in any phases.As for the only one lesion with atypical manifestation,it showed isointensity on T1 WI,slightly hyperintensity on T2 WI,nonenhancement in arterial phase,slightly enhancement in portal phase,and small vascular enhancement inside the lesion in delayed phase.After the follow-up survey of 30-36 months 92.86%(13/14)lesions did not change,however one lesion (10mm)disappeared.Conclusion Typical FNH shows similar MRI characteristics, iso-intense or slightly hypo-intense signal on T1 WI,iso-intense or slightly hyper-intense signal on T2 WI,marked enhancement in ar-terial phase,the central scar enhancement and the inside small vascular enhancement of lesions in delayed phase.It is valuable in dif-ferential diagnosis.

Objective To explore the significance of the suppressor of cytokine signaling-1 (SOCS-1)gene methylation in the genesis, development and prognosis of diffuse large B-cell lymphoma (DLBCL).Methods The methylation state of CpG island in SOCS-1 gene were detected by methylation-specific polymerase chain reaction (PCR) and the level of SOCS-1 gene was measured by the real-time PCR. The clinical data of 30 patients with DLBCL were collected, and according to the international prognosis index (IPI), they were divided into low risk group and high risk group. Results Aberrant methylation of SOCS-1 in 17 DLBCL patients (56.7 %) were positive, however, in control group aberrant methylation was negative(P ＜0.01).The methylation level of DLBCL patients with positive SOCS-1 methylation was higher than that of patients with negative (P ＜0.05). Combined with the clinical data, the positive rate of methylation in patients with high level of serum LDH or the numbers of extra-nodal lesions＞l were significantly higher than that in patients with normal LDH level or the numbers of extra-nodal lesions ≤ 1, respectively. Hence, the positive rate of methylation in the high risk group of DLBCL was higher than that in the low risk group (P ＜0.05). Conclusion There were aberrant methylations of the SOCS-1 gene in the patients with DLBCL. The methylations of SOCS-1 can silence the gene expression, which indicates that SOCS-1 and its methylations play some role on genesis and development of DLBCL and can evaluate the prognosis of the patients with DLBCL.

Objective To investigate the protective effect of Danxiong Recipe (DR) on brain in hypoxia mice.Methods Sixty mice were equally randomized to 6 groups:normal group,model group,propranolol group and low-,model- anti high-dose DR groups.Except the normal group,the mice in other groups were given the corresponding drug intra- gastrically according to the experimental design,once a day,for 7 days.After treatment,the survival time under nor- mobaric hypoxia condition in different groups was calculated.Meanwhile,the activities of superoxide dismutase(SOD) and nitric oxide synthase (NOS),and the contents of malondiadehyde (MDA) and lactic acid (LD) in the brain ho- mogenate were detected with test kit.Results The results showed that DR could prolong the survival time of the mice, inhibit obviously the abnormal increase of MDA and LD,reduce NOS activity,and enhance SOD activity in mice brain. Conclusion DR exerts protective effect on brain in hypoxia mice under normal pressure.

Twenty healthy subjects in each of 3 groups were fed with monounsaturated fatty-acid diet,polyunsaturated fatty-acid diet, or saturated fatty-acid diet separately for 3 days. It suggested that monounsaturated fatty-acids may ameliorate the oxidative stress and improve insulin sensitivity.