The molecular mechanism of autophagy is complicated and it always affects the occurrence and development of tumor in many different degrees,so it is the new hot spot in pathological study area.The angiogenesis includes physiological vascularization and pathological vascularization,and the pathological new vessels which provide nutrition for the continuous growth of tumors have a close relationship to the development and metastasis of tumor.The process of angiogenesis can be affected by regulating the autophagy of vascular endothelial cells,which provides a new method for tumor therapy.

Protein is involved in a variety of biological activities and associated with disease development,such as cancer.With the development of mass spectrometry-based proteomic technologies,qualitative and quantitative analysis of protein in clinical specimen using mass spectrometry is rapidly progressing in cancer biomarker study.Novel cancer biomarker for early diagnosis and prognosis evaluation is the key to reducing death rate and improving survival rate.This review summarizes mass spectrometry technologies and proteomic studies for cancer biomarker discovery.

Brain ; Humans ; Neoplasms ; Neurogenesis

Objective To study the expression of Gli1 protein and vascular endothelial growth factor (VEGF)-C in papillary thyroid carcinoma and their relationship with cervical lymph node metastasis.Methods The expression of Gli1 protein and VEGF-C were investigated by immunohistochemical EnVision method in 67 cases of papillary thyroid carcinoma and 32 cases of nodular goiter pathological specimen tissues.Nascent microvascular and micro-lymphatic of all pathological speimen tissues were examined by immunohistochemical monoclonal antibody CD34 and D2-40 staining assay respectively,and the microvascular density (MVD) and micro-lymphatic vessel density (MLVD) were calculated.The relationship between the positive expression of Gli1 protein and MVD,between the positive expression of VEGF-C and MLVD,and their relationship with cervical lymph node metastasis were analyzed.Results The positive rates of Gli1 protein,VEGF-C and MVD,MLVD were significantly higher in papillary thyroid carcinoma than those in nodular goiter [67.16％(45/67) vs.43.75％(14/32),70.15％(47/67) vs.31.25％(10/32),23.14 ± 2.06 vs.2.14 ± 0.31,13.36 ± 1.32 vs.3.53 ± 0.65,P ＜ 0.05].The positive expression of Gli1 protein was related to the patient' s age and the tumor size (P ＜ 0.05),while the positive expression of VEGF-C was not related to the patient' s age and the tumor size (P ＞ 0.05).The positive expression of Gli1 protein and VEGFC were significantly higher in TNM stage Ⅲ-Ⅳ of papillary thyroid carcinoma than those in TNM stage Ⅰ-Ⅱ (P ＜ 0.01) and also was significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.01).MVD and MLVD were significantly higher with cervical lymph node metastasis than without cervical lymph node metastasis (P ＜ 0.05).The positive expression of Gli1 protein was positively correlated with MVD (r =0.784,P＜ 0.01),the positive expression of VEGF-C was positively correlated with MLVD (r =0.529,P ＜ 0.01),the positive expression of Gli 1 protein was positively correlated with the positive expression of VEGF-C (r =0.586,P ＜0.01).Conclusions Gli1 protein which may participate in the formation of nascent microvascular is abnormally activated by the Hedgehog signaling pathway to express,VEGF-C which may be participate in the formation of nascent micro-lymphatic is mediated and started by the MAPK signaling pathways to express.Expressions of Gli1 protein is positively correlated with expressions of VEGF-C,therefore the Hedgehog signaling pathway may be associated with the MAPK signal pathway.Suppressing nascent microvascular and micro-lymphatic may become new target to blockingup papillary thyroid carcinoma cervical lymph node metastasis.

Objective To analyze the policy support of rural doctors for the essential medicine system,and explore the main influence factors and mechanism affecting the support degree.Methods 621 rural doctors from five cities in Shandong province were subject to a stratified random sampling for a quantitative questionnaire,to build a hypothesis model of policy support of rural doctors,which is verified using factor analysis and path analysis.Results 59.1% of rural doctors support the essential drugs policy.It was found that the essential medicine factor,the factor to influence the practitioner,and the supporting policy factor had a direct impact on policy support of rural doctors,with the effect of 0.305, 0.1 59 and 0.135 respectively;Essential medicine factor and the supporting policy factor indirectly influence the policy support of rural doctors through the factor to influence practitioners,with the effect of 0.065 and 0.032 respectively.Conclusion The essential medicine system needs to emphasize such rationality as the quality,list and pricing of essential medicines;it is imperative for the government to better oversee the purchase and supply of essential drugs,to cater to the interests of rural doctors,to encourage the enforcement of the system,and promote the development of village clinics.

Objective To investigate the relationship between diabetes and bone metabolism and the potential epigenetic mechanisms. Methods BMSCs were cultured for 7 and 15 days in cell culture medium with different concentrations of glucose. The mRNA and protein expression of HDACs and osteogenesis-related genes were detected by RT-PCR and Western blot assay ,respectively. Moreover ,the combination of HDAC to the promoter region of Runx2 was tested by the chromatin immunoprecipitation (ChIP) assay. Results ThemRNA expression of osteogenesis-related genes ,incuding OCN(P < 0.05)and Col1(P < 0.05),in the bone marrow of diabetic mice was significantly reduced compared with the control mice. The mRNA and protein expression of ALP ,OCN ,Runx2 and OSX was gradually reduced with the increasing concentration of glucose ,while HDAC2 mRNA and protein expression was increased. The binding activity of HDAC2 to the upstream and downstream of Runx2 promoter region in 25mM glucose-treated BMSCs was higher than the control group(P < 0.05). Conclusoins Diabetes might repress osteogenesis of BMSCs via inhibiting the activity of Runx2 through upregu-lating the expression of HDAC2.

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human tissue factor( hTF) gene,in order to achieve high level secretory expression in extracellular.Methods Expression plasmid, pGAPZaA-hTF, was constructed by inserting the synthesized sequence encoding human extracellular tissue factor into yeast expression vector pGAPZaA and transformed into Pichia pastoris SMD1168H with electroporation.Having been selected by Zeocin, transformants containing hTF cDNA were expressed in YPD, extracellular proteins were detected by SDS-PAGE and confirmed by Western bolt.Results Successfully constructed the recombinant pGAPZaA-hTF expression system in Pichia pastoris.SDS-PAGE showed that the molecular weight of the expression product was about 37 -40 kDa.Western-blot indicated that it was human extracellular tissue factor.The crude yield of total protein in medium was up to 1 g/L, more than 80% of which was hTF.Conclusion Truncated rTF gene is expressed in Pichia pastoris and the active products are secreted into the medium which have the same activation as the native TF.

Objective To investigate the expressions of transforming growth factor-?1(TGF-?1)and ?-smooth muscle actin(?-SMA)in the lungs of mice with intra-amniotic endotoxin priming and exposed to 60% hyperoxia after born in order to elucidate the possible relationship with bronchopulmonary dysplasia(BPD).MethodsFifty C57 pregnant mice were divided into 2 groups:lipopolysaccharide(LPS,40 ?g/L)group and saline solution group,and then received an intra-amniotic injection of corresponding solution on E15.The neonatal mice of each group were randomized to be set in 60% oxygen exposure or in room air.So there were 4 subgroups,LPS+air,LPS+hyperoxia,saline+air and saline+hyperoxia groups.On days 1,3,7,10 and 14 after birth(8 rats each time point),the lung histological changes was assessed with hematoxylin and eosin(HE)staining for radial alveolar counting(RAC).The expressions of TGF-?1 and ?-SMA proteins were detected by immunohistochemical and immunofluorescence staining,and the expressions of TGF-?1 and ?-SMA mRNA by real-time polymerase chain reaction(RT-PCR).ResultsIn the LPS+hyperoxia group and saline+hyperoxia group,RAC began to decrease on day 3,and then further declined in a time-dependent manner.Compared with saline+hyperoxia group,LPS+hyperoxia group had significantly lower RAC(P

Objective To investigate the different distribution and expression of mammalian target of rapamycin complex (mTORC) in the kidney of diabetic nephropathy (DN) mice.Methods Fourteen eight-week-old male C57BL/6 mice were assigned to 2 groups: the control group ( n=7 ) and the streptozotocin ( STZ )-induced DN group ( n=7 ) . Blood and urinary variables including glucose , albumin, creatinine and albumin/creatinine ratio were assessed 2 weeks after STZ injection.Hematoxylin-eosin staining was performed for renal pathological analyses .The distributions of mTOR , phosph-ser2448-mTOR(p-mTOR), mTORC1(Raptor), mTORC2(Rictor) and phosph-ser240/244-S6K1 (p-S6K1) were determined by immunofluorescence.The expression levels of mTOR, p-mTOR, mTORC1(Raptor), mTORC2(Rictor), S6K1 and p-S6K1 were detected by Western blotting .Results Two weeks after STZ injection , the diabetic mice developed albuminuria (P<0.01) and renal hypertrophy (P<0.05).The immunofluorescence positive staining for mTOR , Raptor, and Rictor was distributed in the epithelial cells of proximal tubules , glomerular mesangium and capillary loops as well as the medullary collecting ducts of the control mouse kidney .These positive signals increased in the DN mouse kidney ( P<0.05).However, pS6K1 was not detected in the inner medulla of control mouse and p-mTOR was not found in the glomeruli of both control and DN mice .Conclusion mTORC is widely expessed in the mouse kidney and participates in the development of DN , whereas the 2448 serine phosphorylation of mTOR may be not implicated in the hyperglycemia mediated glomerular injury .

ObjectivesTo study the effect of community interventions and management on asthma in children and its impact on knowledge, attitude, and practice (KAP)of parents and the home environment.MethodsAn asthma health management profile of 0 to 14-year-old asthmatic children (n =93 ) in Sanlitun and Liulitun communities in Chaoyang District in Beijing was established.The children were randomly divided into a management group ( n =49) and control group ( n =44 ) based on community.Community-integrated management,such as regular follow-up,condition monitoring and health education,was implemented in the asthmatic children in the management group but not in the control group.The parents' KAP and the household environment in the two groups were compared after 1 year based on the changes shown in the health management profile.ResultsThe asthma relapse rate decreased to 27.9％ (12/43) in the management group.Compared with the control group,the rates of hospitalization (x2 =8.174,P =0.004) and school absences ( x2 =4.962,P =0.026) significantly decreased.The KAP level of parents increased to 67.4％ ( 29/43 ) in the management group and 20.4％ (9/44) in the control group,and the difference was statistically significant (x2 =19.517,P ＜0.01 ).Knowledge improved the most and showed a significant difference from the control group ( x2 =19.517,P ＜0.01 ).Home environment in the management group improved to 76.7％ (33/43).The number of indoor pets ( x2 =3.906,P =0.048) and indoor cockroaches ( x2 =4.962,P =0.026 ) reduced and showed significant differences between the two groups.In addition,children's allergy-related symptoms decreased to 30.2％ ( 13/43 ) in the management group compared with 9.1％ (4/44) in the control group,which was a significant difference ( x2 =6.183,P =0.013).ConclusionsParents' knowledge of asthma,compliance behaviors,and home environmentwere effectively improved through community-integrated management.This management technique can reduce the allergy-related symptoms of asthmatic children,improve asthma severity,and reduce the influence of asthma on children's daily lives.