Objective To evaluate recurrence risk factors in postoperative breast cancer patients after 5-year adjuvant endocrine therapy.Methods From Jan 2006 to Dec 2011,a total of 327 patients were enrolled for this study.Kaplan-Meier curves were applied to estimate survival rates and COX's proportional hazards model to identify prognostic variables.Results Among these 327 eligible patients,42 (12.8％) patients suffered from distant metastasis and 34 (10.4％) patients experienced locoregional recurrence after 5-year adjuvant endocrine therapy.Survival analysis showed that patients with histologic grade 3 disease,lymph node metastasis,Ki-67 high expression,high TNM stage and radiotherapy were statistically significant with poorer relapse-free survival (RFS,P =0.000,0.003,0.000,0.003,0.034)and poorer distant metastasis-free survival (DMFS,P =0.000,0.002,0.000,0.002,0.023),respectively.In multivariate analysis,patients with histological grade 3 disease (P =0.002) and more than 3 positive nodes (P =0.032) were risk factors for lower RFS.However,only histological grade 3 (P =0.015) was risk factor for DMFS.Conclusions Late relapse after completion of 5-year adjuvant endocrine therapy was still common,patients with grade 3 disease and more than 3 positive nodes may benefit from extended endocrine therapy.

Objective To discuss the clinical effect of facial contour remodeling using transplantation of the autologous granular fat grafting combined with botulinum toxin A injection.Methods Negative-pressure liposuction was carried out in the inner thigh or abdomen using liposuction needle connected with an injector,to let stand for layered,discharge lower layer water,rinse,extract the fat particles into a 2 ml syringe,inject into facial depression area with multipoint,multiple-tunnel,and multilayer manner,over injection of about 20％-30％ every time.89 patients received the injection;some accepted two injections;13 cases were injected in temple (14.6％),and 16 in forehead (17.98％).30 of them received subcutaneous injection of botulinum toxin A,each side dose was less than 20 U.Results Patients had been followed up for 3-36 months,and obvious improvements were observed in facial contour in the frontal and temporal region as well as the skin texture;the survival rate of fat granule reached to 60％-80％.No hematoma,nodes and infection been observed.Patients combined with botulinum toxin A injection had more ideal facial contour and more satisfaction.Conclusions Autologous fat granules transplantation in the face has stable clinical effect,and combining with botulinum toxin A injection can improve facial contour.It is a safe,ideal treatment in facial rejuvenation and facial contour remodeling and it therefore can be widely recommended in clinical treatment.

Objective To investigate the correlations between P53 expression and clinicopathologic factors and prognosis of Luminal breast cancer.Methods From January 2009 to December 2012,a total of 283 patients with Luminal breast cancer in the Shanghai General Hospital Affiliated to Shanghai Jiaotong Univer-sity were included.P53 expressions of them were assayed by immunohistochemistry.Survival analysis was applied using Kaplan-Meier curve and Log-rank test.Single factor analysis and mutiple-factor analysis were applied using Cox proportional hazard regression model.Results The rate of P53 expression was associated with tumor size (χ2 =6.285,P =0.043),lymph node metastasis (χ2 =15.881,P =0.000),histological grade (χ2 =8.132,P =0.043)and Ki-67 (χ2 =6.476,P =0.039),but it was not associated with age (χ2 =0.955,P =0.328),menopausal status (χ2 =3.808,P =0.051),pathological pattern (χ2 =0.847,P =0.655),estrogen receptor (χ2 =1.867,P =0.172),progesterone receptor (χ2 =0.937,P =0.333)and human epidermal growth factor receptor-2 (χ2 =0.110,P =0.741 ).In all the 283 patients,the 5-year relapse-free survival rates for P53-positive group and P53-negative group were 66.7% and 90.7% respectively (χ2 =12.609,P =0.000),while the 5-year overall survival rates were 84.4% and 93.4% respectively (χ2 =4.153,P =0.042),with significant differences.In Cox mutiple-factor analysis,lymph node metastasis (HR =2.484,95%CI:1.393-4.431,χ2 =9.497,P =0.002)and P53 over-expression (HR =3.627,95%CI:1.061-12.401,χ2 =4.220,P =0.040)were independent prognostic factors for the relapse-free survival of patients with Luminal breast cancer,and lymph node metastasis (HR =3.451,95%CI:1.891-6.297,χ2 =16.290,P =0.000)and higher histological grade (HR =2.806,95%CI:1.091-7.219,χ2 =4.582,P =0.032)were independent prognostic factors for overall survival.Conclusion P53 over-expression of patients with Luminal breast cancer is associated with prognostic factors such as lymph node metastasis and histological grade,which indicates the worse prognosis.

Objective To investigate the effect and current situation of the standardized construction of laboratories of schis?tosomiasis control institutions in Hubei Province,so as to provide the evidence for establishing and improving the quality control system of diagnosis of schistosomiasis after the transmission of schistosomiasis was under control. Methods According to the procedures of self?examination,field operation,and laboratory on?site,five laboratories were assessed,and all the results were analyzed comparatively. Results The average number of staffs were(7.00 ± 1.58)persons,and the staffs of the laboratories of the schistosomiasis control institutions with senior professional titles in the city level were more than that in the county level(t=5.563,P<0.05). The average space was(3.20±1.64)rooms,and the average area was(117.00±88.29)m2. The average score of field operation was(96.40 ± 4.49)points. The average score of laboratory on?site assessment was(106.6 ± 6.15)points. The highest and lowest of the laboratory on?site assessment scores were environment and facilities(19.60 ± 0.55)points and manage?ment system of laboratory quality control(15.70 ± 2.39)points(F=2.869,P<0.05),respectively. Conclusion The cultiva?tion of laboratory staff should be strengthened,and the diagnostic capacity should be maintained and improved. The laboratory quality control system should be paid more attention to,and the construction and management of schistosomiasis laboratories should be standardized.

Objective To investigate the relationship between the nerve growth factor ( NGF ) induced hippocampal neuroregeneration and homeobox gene Lhx 8.Methods Seventy-two SD rats were divided into control group , transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection . Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group.The choline acetyltransferase ( ChAT)/Lhx8 double labeled cells in subgranular zone ( SGZ) of hippocampus in each group were detected by immunofluorescence .Results The expression of Lhx8 gene and protein in the transected , NGF group and especially in the transected combined with NGF group was obviously higher than in the control group .The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group . Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

Objective To investigate the effect of Jagged1 on hippocampal radial glial cells (RGCs) proliferation and neuronal differentiation in vitro.Methods Hippocampal RGCs were cultured in vitro, the agonist Jagged1 and(or) inhibitor DAPT of Notch signaling were added into the culture medium , and then the cells were divided into control group , Jagged1 group, Jagged1 combined with DAPT group and DAPT group .CCK-8 regent was used to detect cells ’ vitality;immunofluorescent was used to detect the number of BLBP /Ki67 double labeled cells and differentiated microtubule associated protein-2(MAP-2) positive cells.Results Cell vitality in Jagged1 group was obviously higher than that of the other groups .The number of BLBP/Ki67 double labeled cells and differentiated MAP-2 positive cells were more than other groups.Conclusion Jagged1 promotes the proliferation and neuronal differentiation of hippocampal RGCs in vitro.

Objective The aim is to observe the role and mechanism of estradiol ( E2 ) on the proliferation of rat hippocampal neural stem cells ( NSCs ) .Methods Twenty hippocampi from embryonic 17-day ( E17 ) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol .The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay .The expression of estrogen receptors ( ERαand ERβ) was measured by immunofluorescence staining combined with Nestin double labeling . Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10 -10 to 10 -8 mol/L.The number of cell proliferation and the viability of cells were best at the concentration of 10 -8 mol/L compared to the other groups .However, when the estradiol concentration was increased from 10-8 to 10 -6 mol/L, the cell proliferative capacity declined gradually .Double immunofluorescence labeling showed that the two types of estrogen receptors ( ERαand ERβ) were expressed in the cultured hippocampal NSCs .Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range , and ERαand ERβmay be involved in the estradiol-induced proliferation .

The flavonoid-3-O-glucosyltransferas (3GT) is one of the most important enzymes for biosynthesis of plant secondary metabolites. In this paper, the homology analysis and gene special primers design were used. With the methods of modern molecular biology, the full-length gene of 3GT (GenBank Accession No. JN092127) which was cloned from Saussurea involucrata Kar. et Kir by RT-PCR and RACE. The cDNA sequence of 3GT consisted of 1548 bp open reading frame (ORF) encoding 516 amino acid, the deduced 3GT protein shared 91% and 89% identities with that of Fragaria x ananassa GT6, Manihot esculenta anthocyanidin 3-O-glucosyltransferase. Homology analysis showed that deduced 3GT protein has a glycosyltransferase signature domain PSPG-box. The transcripts of 3GT members were found mainly in leaves and callus. 3GT gene of S. involucrata was under the control of the cauliflower mosaic virus (CaMV) 35S promoter, homologous transformation used an Agrobacterium rihizogenes-mediated transformation system. The results on UV spectrophotometry showed S. involucrata callus after suspension culture that an average of total flavonoids on transgenic callus was 2.06 times higher than non-transgenic callus.
Amino Acid Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Glucosyltransferases ; genetics ; Molecular Sequence Data ; Open Reading Frames ; genetics ; Plant Proteins ; genetics ; Saussurea ; enzymology ; genetics

Objective To investigate the effect of extracellular signal-regulated kinases(ERK) signal transduction in the process of NSCs differentiating into neurons in the fimbria-transected hippocampi's extracts. Methods Twelve Sprague-Dawley rats'right fimbrias were transected. The extracts were gained from the fimbria-transected hippocampi at the 14th day normal rat, and the extracts supernatant fluid was collected after centrifugal process, then the protein concentration in the extracts was determined. In the serum-free medium,NSCs from the fetal hippocampus were planted on 24 well culture plate, then were divided into three group and eight wells for each group as follows: the transected group contained the extracts of the fimbria-transected hippocampi;the normal group contained the extracts of the normal hippocampi;the pure control group have no extracts. After cultured for 14 days,the cells were detected by using MAP-2 and p-ERK immunofluorescence. Result The number, area, perimeter of MAP-2 positive neurons were all declined in transected group, the normal group and the control group orderly. Statistic results showed significant difference between every two groups. The number of MAP-2/p-ERK double-positive neurons were decreased in transected group, the normal group and the control group orderly, but the percentage of double-labeled neurons in total MAP-2 positive neurons were increased in turn. In these two aspect, there were also significant difference between every two group. And most of the MAP-2/p-ERK double-positive neurons were immature. Conclusion The extracts of the fimbria-transected hippocampi had obvious effects on promoting NSCs differentiating into neurons and speeding up the maturation of neurons than those of the normal hippocampi. The morphological results showed that ERK signal transduction might be related to the differentiation of NSCs into neurons.

BACKGROUND:Clinical application of neural stem cells is under exploration.Currently,the indicative differentiation of neural stem cells into specific neurons to replace lost and degenerative neurons needs to solve.OBJECTIVE:To explore the effect of 83 ku protein in rat hippocempi on inducing neural stem cell differentiation into acetylcholine esterase (ACHE) positive neurons.DESIGN,TIME AND SETTING:In vitro controlled observation of cytology was performed at the Medical College of Nantong University between October 2003 and April 2008.MATERIALS:A total of 12 SD rats,of clean grade,and SD fetal rats,aged 17 days,were provided by the Experimental Animal Center of Nantong University.METHODS:The normal hippocampi and hippocampi on the 14th day after the hippocampal fimbria transection were prepared into homogenate used for 10% native-polyacrylamide gel electrophoresis,and the differential proteins of 83 ku were electroeluted.The protein concentration was adjusted to 300 mg/L.The forebrain tissues of fetal rats were harvested and neural stem cells were isolated and in vitro cultured:blank control cells were cultured in serum-free DMEM/F12 medium;83 ku normal and 83 ku transection groups were separately cultured in serum-free DMEM/F12 medium containing 10 mg/L 83 ku protein from normal and hippocampal fimbria transection rats for 12 days.MAIN OUTCOME MEASURES:AChE histochemical staining was used to detect the differentiation of neural stem cells into AChE positive neurons.RESULTS:After 12 days of culture,there was a large amount of AChE positive neurons in 83ku transection group and their bodies were very big and the processes were abundant;The AChE positive neurons in 83ku normal group were less than 83 transection group,and their bodies were small with short processes.A few of AChE positive neurons were seen in control group.There were significant differences in number of AChE positive neurons among three groups (P＜0.05).CONCLUSIONHippocampal 83 ku protein can induce neural stem cells to differentiate into AChE positive neurons.