p53 Gene and proliferating cell nuclear antigen (PCNA) are both related closely to the ra-diotherapy of non-small cell lung cancer(NSCLC). The life span of NSCLC patients with both p53 and PCNA positive is shorter than that of NSCLC patients with both p53 and PCNA negative. Studying the expression of p53 and PCNA in NSCLC, and their relationship with the radiosensitivity and prognosis of NSCLC can play a positive role in individualized treatment and increasing curative effect.

Objective:To investigate the effects of Chuanxikang decoction(a Chinese herbal medicine)on the amount of IFN-? and IL-5 in bronchoalveolar lavage fluid(BALF) and expression of MMP-9 and TIMP-1 in lung tissue of asthmatic model in guinea pig to explore its mechanism in treatment of bronchial asthma.Methods:A total of 40 guinea pigs were randomly divided into 4 groups:normal control group(A group),asthmatic model group(B group),treatment with dexamethasone(C group) or with Chuanxikang decoction groups(D group),10 guinea pigs in each group.The guinea pig models of asthma were established through injecting ovalbumin(OVA) into the abdominal cavity and inhalation of atomization.The guinea pigs of asthma underwent with different therapy.Amount of IFN-? and IL-5 in BALF and expression of MMP-9 and TIMP-1 in lung tissue were measured by immunotisssuchemical technology.Results:Compared with the normal control group(A group),the concentrations of IFN-? in the BALF was decreased significantly(P

Objective To analyze the resenilin-1 (PS-1) gene mutations in Alzheimer' s disease (AD) patients and investigate the influence of the initiation codon mutation on the mRNA expression of PS-1 and amyloid precursor protein (APP) genes and the expression of PS-1 proteins.Methods (1) All 111 AD patients were enrolled by the Department of Neurology,Second Affiliated Hospital,College of Medicine,Zhejiang University from July 2004 to June 2010.Mutations in the 13 exons and flanking regions of PS-1 gene were examined by direct sequencing.(2) cDNAs encoding full-length wild-type and mutant (c.1A ＞G) PS-1 were subcloned into enhanced green fluorescent protein.Levels of the mRNA expression of PS-1 and APP genes and PS-1 proteins expression in the transfected cells were detected by quantitative real-time PCR and Western blot,respectively.Results A new heterozygous initiation codon mutation changing from ATG to GTG in one individual was identified.Compared to the control groups,the mRNA expression of the mutant PS-1 gene in HEK293 and N2a was significantly lower than the normal PS-1 gene(116.8 ± 3.9 vs 49.5 ±3.3,t =13.27,P ＜0.01 ;69.0 ± 1.9 vs 29.5 ± 1.3,t =17.20,P ＜0.01) and the APP gene was not obviously altered.The proteins were detected by Western blot analysis in HEK293 cells but not in N2a cells.Conclusions Since we only identified one novel heterozygous initiation codon mutation (from ATG to GTG),mutations in PS-1 are likely to be rare in AD patients.Initiation codon mutation would reduce the expression of PS-1 proteins.Inactivation of some of the PS-1 proteins could be insufficient to lead to AD and could be more likelv to act as a risk factor.

Objective To investigate the role of the expressions of APC,bcl-2 and c-met gene in the progress of gastric cancer and their significances in the diagnosis of early gastric cancer.Methods The immunohistochemical technique was used to detect the expressions of APC,bcl-2 and c-met gene in 30 cases of human gastric carcinoma(GC),30 cases of intestinal metaplasia(IM),30 dysplasia(Dys) gastric mucosa,10 gastric adenoma(GA) and 20 normal gastric mucosa.Results ①The positive expression rates of APC in GC,IM,Dys and GA(53.3%,67.7% and 53.3%,respectively) were significantly lower than those in normal gastric mucosa(90.0%,P

Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P＜0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.

The rice Rim2/Hipa is a stress-induced transposon superfamily recently identified in Oryza genomes. Genomic variation was found in the Rim2 core region among rice genetic resources/genomes, indicative of high genomic divergence accumulated during the Rim2 evolution. Based on the divergence and quiescent state of the Rim2 elements, a Rim2 paralog display-based fingerprinting approach was developed to effectively identify rice genetic resources and explore their genetic relationships within a set of rice germplasm including 45 accessions ofO. sativa and 8 accessions of its wild relatives O. rufipogon. A dendrogram showed not only clear genetic diversity of rice germplasm, but also considerable genetic differentiation among wild rice resources. The wild rice relatives were either clustered as an independent group, or among the japonica varieties. This Rim2-based fingerprinting approach could also serve as a sensitive tool to identify rice hybrids from their parents, and variety stability, demonstrating its great potential in evolution study ofrice genomes and in rice breeding and seed production.

Objective　To evaluate the correlation between vascular endothelial growth factor(VEGF) expression and microvessel density(MVD) in gastric carcinoma (GC) and the relationship of VEGF and MVD with clinicopathologic characteristics and prognosis of GC. Methods　The expression of VEGF and intratumoral microvessel density (MVD) in one hundred and sixteen resected specimens of the patients with GC were observed and counted, and the relationship of VEGF and MVD with clinicopathologic factors and prognosis of GC were analysed. Results　The expression of VEGF was found in 60.34% of the specimens. The MVD value was much higher in VEGF(+) group than that in VEGF(-) group (26.16±8.50 and 19.22±8.20, respectively, P<0.01). Expression of VEGF was significantly higher in patients with lymph node metastasis than that in patients without lymph node metastasis (p<0.05).Expression of VEGF was highly correlated with the stage of tumor (P<0.05). MVD correlated with lymph node metastasis (P<0.01) and abdomen metastasis (P<0.01) and increased with the TNM stage (P<0.01). The total five-year survival rate in VEGF(-) group and low MVD group were significantly higher than that in VEGF(+) group and high MVD group respectively(both P<0.01). Multivariate analysis indicated that the expression of VEGF and MVD were independent prognostic factors of GC. Conclusions　The expression of VEGF and MVD can reflect the malignant degree of GC. They may serve as the prognostic factors and guide the decisions on the therapy.

Objective To investigate the impact of synthetic urine (synurine solution) at differentpH values on skin surface.Methods Sixty-four healthy volunteers were enrolled into this study.Based on the results of lactic acid test and questionaires,these subjects were enrolled as sensitive skin group and normalskin group.The 4-,20-,and 24-hour occlusive patch tests were successively performed with synthetic urine at various pH values (2.0,3.5,5.0,6.5,8.0 and 10.0)on the two groups of volunteers.The distilled waterand 0.25%sodium lauryl sulphate (SLS)served as control.The skin surface pH values were measured bypotable pH meter before,and at 24 h,48 h,72 h after the first patch.Half an hour after the latter three detections of pH values,the skin response was evaluated clinically,as well as by transepidermal water loss (TEWL)and pH values,which were measured by pH-900 pH Meter.Results The responses to these solutions were similar between sensitive and normal skin groups.The order of skin surface pH values measured by the two pH meters was consistent with that of the original solutions.The skin surface pH value was altered by the synthetic urine,and this change was still present at 24 hours after the patches were removed.The TEWL values of skin challenged by synthetic urine were not higher than those by distilled water.The skin responses at 72 hours were more intense to the synthetic urines with pH values of.5,8.0,10.0than to distilled water.CondusionsSynthetic urines with different pH values can alter skin surface pH values,and skin responses to synthetic urines with a pH range 2.0-10.0 are similar to those to distilled water.

Objective:To explore the effect of calcium phosphate cement（CPC）scaffold loaded with emodin（EMO）on osteogenic activity of osteoblasts.Methods:The bone cement scaffold was prepared by mixing EMO powder and CPC powder（ratio 1∶9），adding citric acid and then was poured into polytetrafluoroethylene mold（EMO-CPC group）. A dose of 0.36 g CPC powder was mixed with citric acid and injected into the polytetrafluoroethylene mold（CPC group）. General morphology，setting time（initial setting time and final setting time），injection rate and compressive strength of stents were compared between the two groups. Primary osteoblasts were extracted and co-cultured with two sets of scaffolds. After co-culture for 3 days，their characterization was observed by scanning electron microscopy. Live/dead cell staining and 3-（4,5-dimethylthiazole-2）-2,5-diphenyltetrazolium bromide（MTT）colorimetric method were used to detect cell viability，toxicity and proliferation activity of scaffolds. Two sets of scaffolds were stained with immunofluorescence for osteopontin（OPN），and protein expression was observed under an inverted fluorescence microscope. After co-culture for 7 days，tetrazolium nitro blue/5-bromo-4-chloro- 3-indolyl-phosphate（NBT/BCIP）staining method was used for alkaline phosphatase（ALP）staining. After co-culture for 14 days，two sets of scaffolds were stained with Alizarin Red to detect their osteogenic activity.Results:Two sets of stents showed relatively smooth and flat topography under the scanning electron microscope. There were no significant differences in initial setting time，final setting time，injection rate and compressive strength of stents between two groups（ P > 0.05）. After co-culture for 3 days，the osteoblast clusters were adhered to the surface of the EMO-CPC scaffold，with good shape. Viable cell rate reached（98.2 ± 0.1）% in EMO-CPC group and（90.2% ± 0.1）% in CPC group（ P <0.05）. Cell proliferation activity in EMO-CPC group was stronger than that in CPC group（ P < 0.05）. OPN-specific staining showed that EMO-CPC group had stronger OPN protein fluorescence expression compared to CPC group. After co-culture for 7 days，expression of ALP in EMO-CPC group was higher than that in CPC group. After co-culture for 14 days，staining intensity of Alizarin Red in EMO-CPC group was more significant than that in CPC group. Conclusions:The EMO-CPC scaffold can provide a suitable environment for the growth of osteoblasts for it has better biocompatibility，cell proliferation and osteogenic activity than the CPC scaffold.

Objective To evaluate the safety and efficacy of botulinum toxin type A for injection in the treatment of post-stroke upper limb spasticity (dosage was 200 U,or 240 U if combined with thumb spasticity).Methods The study was a multi-center,stratified block randomized,double-blind,placebocontrolled trial.All the qualificd subjects were from 15 clinical centers from September 2014 to February 2016.They were randomized (2∶1) to injections of botulinum toxin type A made in China (200-240 U;n =118) or placebo (n =60) in pivotal phase after informed consent signed.The study was divided into two stages.The pivotal trial phase included a one-week screening,12-week double-blind treatment,followed by an expanded phase which included six-week open-label treatment.The tone of the wrist,finger,thumb flexors was assessed at baseline and at weeks 0,1,4,6,8,12,16 and 18 using Modified Ashworth Scale (MAS),disability in activities of daily living was rated using the Disability Assessment Scale and impaction on pain,muscle tone and deformity was assessed using the Global Assessment Scale.The primary endpoint was the score difference between botulinum toxin type A and placebo groups in the tone of the wrist flexor using MAS at six weeks compared to baseline.Results Muscle tone MAS score in the wrist flexor of botulinum toxin type A and placebo groups at six weeks changed-1.00 (-2.00,-1.00) and 0.00 (-0.50,0.00) respectively from baseline.Botulinum toxin type A was significantly superior to placebo for the primary endpoint (Z =6.618,P ＜ 0.01).The safety measurement showed 10 subjects who received botulinum toxin type A had 13 adverse reactions,with an incidence of 8.47％ (10/118),and three subjects who received placebo had three adverse reactions,with an incidence of 5.00％ (3/60) during the pivotal trial phase.All adverse reactions were mild to moderate,none serious.There was no significant difference in adverse reactions incidence between the botulinum toxin type A and the placebo groups.During the expanded phase three subjects had four adverse reactions and the incidence was 1.95％.All adverse reactions were mild,none serious.Conclusion Botulinum toxin type A was found to be safe and efficacious for the treatment of post-stroke upper limb spasticity.Clinical Trial Registration:China Drug Trials,CTR20131191