Objective To observe the distributive characteristics of glial cell line-derived neurotrophic factor(GDNF)in brain tissue during cerebral ischemia-reperfusion in rats,and its role in ischemic brain damage.Methods The model of focal cerebral ischemia was made by occluding middle cerebral artery(MCA) for 2 h and reperfusion for 0.5～48 h, HE Staining was used to investigate the histological features of ischemic cerebral damage,the immunohistochemical method was used to observe the distributive characteristics of GDNF in brain tissue in rats. Results The focal ischemic areas (including preoptic area, striatum and cortex) presented at 0.5 h of reperfusion and peaked at 24 h. The neurons presented irreversible degeneration at 6 h of reperfusion. At 24 h, the ischemic area in the preoptic area developed into infarct form. At 0.5 h of reperfusion, neurons in ischemic cortex showed GDNF weak positive, neurons in peri-ischemic regions showed GDNF moderate positive. During reperfusion 3～48 h, neurons in ischemic regions showed GDNF negative. Up to 48 h of reperfusion, the active microglias or macroglias in periinfart area strongly expressed GDNF.The number of GDNF positive cells in all groups decreased (P

OBJECTIVE:To review the selection process and methods of WHO Essential Drug List and to analyze the problem and challenge of the selection and evaluation of National Essential Drug List in order to provide suggestion for the selection and evaluation of essential drug list in China. METHODS:The selection process of previous National Essential Drug Lists were compared with that of National Essential Drug Lists(2009 edition)and WHO Essential Drug List. The problem and limit of National Essential Drug List were found out to put forward countermeasure. RESULTS:The prominent problem of the selection and evaluation of National Essential Drug List were as follows:the categories of drugs were changed significantly and including of some drugs was not supported by scientific evidence. There was no indirection and standard for the evidencebased selection and evaluation of essential drug. CONCLUSION:Aiming at the problem of the selection and evaluation of National Essential Drug List,evidencebased evaluation research is conducted to establish indirection and standard for the selection of essential drug list and database. Professionals and institutions for evidencebased evaluation and selection of essential drug list should be established to promote evidencebased evaluation and selection of National Essential Drug List and make more scientific decision.

OBJECTIVE To investigate the contribution of the GJB6 gene [encoding connexin 30 (C?30)] mutations in Chinese population with sporadic non-syndromic hearing impairment. METHODS PCR reactions were performed with two pair of primers for the coding sequence of GJB6 gene and for the deletion of GJB6. PCR products bidirectional sequencing was subsequently applied in 214 patients with hearing loss and 86 normal controls. RESULTS A novel heterozygous mutation-233(C→A) was found, which results in amino acid change, A78D. This mutation wasn't detected in the control subjects. The altered valine residue lies within the second conserved transmembrane domain. The large deletion△(GJB6/ D13S1830)] of GJB6 was not found in this group. CONCLUSION The large deletion of GJB6 was not found in the Chinese deafness population. A novel heterozygous mutation of GJB6 was found. These results indicated GJB6 mutations are not a major cause of hearing loss in the Chinese population.

Objective To explore the in vitro isolation of ?2m-/Thy-1+ bone marrow-derived liver stem cells(BDLSCs) which bear double features of stem and liver cells from bone marrow stem cells(BMSCs)as so to provide suitable donor cells for the treatment of liver diseases by cellular transplant. Methods ?2m-/Thy-1+ BDLSCs were isolated by magnetic bead cell sorting(MACS) method from cholestatic rats in vitro,and cell purity was detected using flow cytometry.Liver associated phenotype markers were characterized by RT-PCR and immunofluorescence staining. Results BDLSCs isolated by MACS were purified and viable,and possessed hepatocyte-like features at gene and protein levels. Conclusion ?2m-/Thy-1+ BDLSCs are special subsets of BMSCs which may have promising potentials in the stem cell-based treatment of liver diseases.

Objective To examine the mRNA and protein expression of peroxisome proliferator-activated receptors (PPAR) and fatty acid binding protein-4 (FABP-4) in placenta, and to investigate their correlation with the prognosis of pre- eclampsia. Methods The data of 177 women who delivered from January 2013 to December 2015 in Chinese People′ s Liberation Army No.94 Hospital were collected. Among them, 60 cases were term pregnancy and not in labor (TN); 46 cases were term pregnancy with preeclampsia and not in labor (TPE); 42 cases were preterm pregnancy and not in labor (PN); 29 cases were preterm pregnancy with pre-eclampsia and not in labor (PPE). Real-time PCR and western blot were used to examine the PPAR and FABP-4 mRNA expression and protein expression in placentas. And linear correlation was used to analyze the relationship between PPAR and FABP-4 protein expression and the prognosis of pre-eclampsia. Results (1)Real-time PCR showed that:① PPAR-α, PPAR-β mRNA expression were not statistically different between placentas from TN and TPE (P>0.05), but PPAR-γ mRNA level in TPE (0.59±0.17) was significantly lower than that in TN (0.81±0.19, P<0.01). However, FABP-4 mRNA level in TPE (0.70±0.23) was significantly higher than that in TN (0.43±0.18, P<0.01). ②There were no statistical differences in PPAR-α, PPAR-β mRNA expression level between PN and PPE (P>0.05). However,PPAR-γ mRNA in the PPE group (0.33±0.14) was significantly lower than that in PN (0.52±0.16, P<0.01), and FABP-4 mRNA level in the PPE group (0.80±0.29) was significantly higher than in the PN group (0.63±0.22, P<0.01).(2)Western-blot showed the same tendency as the mRNA results. ①There were no statistical differences in the protein expression of PPAR-α, PPAR-β, not in term groups(TN and TPE), nor in premature groups (PN and PPE). PPAR-γ protein level in the TPE group (0.46 ± 0.17) was significantly lower than that in TN (0.65±0.20, P<0.01) and FABP-4 protein level in the TPE group (0.60± 0.19) was significantly higher than that in the TN group (0.50±0.21, P<0.05). ②The PPAR-γ protein level in the PPE group (0.30±0.16) was significantly lower than that in the PN group (0.61±0.16, P<0.05), while the FABP-4 protein expression in the PPE group (0.58±0.19) was significantly higher than that in the PN group (0.40±0.19, P<0.05).(3)Linear correlation showed that:①PPAR-γ protein expression correlated negatively with FABP-4 protein expresssion [P<0.01, R2=0.12 in the term groups(TN+TPE), R2=0.15 in the premature groups(PN+PPE)]. ②PPAR-γ protein expression correlated negatively with blood pressure recovery time, 24-hour urinary protein quantification and blood lipid recovery time (P<0.01, R2=0.37,0.35,0.18). FABP-4 protein expression correlated positively with lipid recovery time and blood cholesterol concentrations (P<0.01, R2=0.13,0.25). Conclusions The expression of PPAR-γ decreases in placentas from patients with pre-eclampsia, while the expression of FABP-4 increases. The expression of FABP-4 correlates negatively with the expression of PPAR-γ, and both are associated with the prognosis of pre-eclampsia.

Objective:To study the radiation-sensitizing effects of nimotuzumab and X-ray radiotherapy on human esophageal carcino-ma KYSE450 cells. Methods:Human esophageal carcinoma cells KYSE450 were treated with nimotuzumab, irradiation, and the combi-nation of both. Cell growth inhibition was evaluated by MTT assay, and cell cycle distribution and apoptosis were analyzed by flow cy-tometry assay. Cell radiosensitivity was tested by clonogenic assay, and the survival curve was fitted using multi-target single-hit mod-el. The combination and accelerated radiation groups were tested by microarray technology, and the differentially expressed genes were screened among the two groups. Results:The growth of KYSE450 cells was inhibited in three groups, namely, the group treated with nimotuzumab, the group treated with irradiation, and the group treated with both. The group treated with both nimotuzumab and irradiation resulted in the highest inhibition rate (35.25%±5.62%) compared with that of the nimotuzumab (16.12%±8.73%) and ir-radiation groups (27.64%± 6.66%) (F=10.953, P<0.001). The highest rates of G2 phase arrest and cell apoptosis were observed in the group treated with the combination of nimotuzumab (29.37%±7.29%) (F=17.299, P<0.001) and irradiation (18.80%±2.03%) (F=85.691, P<0.001). Multi-target single-hit model showed that the values of SF2, Do, and Dq in the group with both treatments were smaller than those of the irradiation group with sensitization enhancement ratio of 1.63, which confirmed the radiosensitization effect of ni-motuzumab on KYSE450 cells. Microarray technology analysis found that nimotuzumab can enhance the radiosensitivity of esophageal squamous cell carcinoma by cutting the genes of EGF/PDGF signaling pathways. Conclusion:This experiment shows that nimotuzumab can effectively inhibit the growth of human esophageal cancer cell KYSE450. Nimotuzumab can also promote apoptosis and G2 phase arrest when combined with X-ray radiotherapy, thereby enhancing the radiosensitivity of KYSE450 cells. This effect is associated with cutting the genes of EGFR signaling pathways.

Objective To observe the dynamic changes of vascular endothelial growth factor (VEGF) levels in the patients with acute intracerebral hemorrhage during different phases and study the relationships between the VEGF levels and the volume of hemorrhage or stroke severity. Methods Fifty-six patients with acute intracerebral hemorrhage were divided into three groups by the volume of hemorrhage:group A(＜ 20 ml,23 caees),group B(20-30 ml, 18 cases),group C(＞ 30 ml, 15 cases) and light group (1-15 scores,22 cases),midrange group (16-30 scores,20 cases),severe group (31-45 scores, 14 cases)by China stroke scale (CSS) score in 24 hours of hospitalization. The serum VEGF level was determined at 24 h,48 h,72 h,7 d and 14 d after the onset of disease by enzyme linked immunosorbent assay(ELISA).Results The serum levels of VEGF during different phases (within 24 h,48 h,72 h,7 d,14 d) had statistical differences between group A and group C (P ＜0.01). Within 24 h,48 h,72 h,7 d,there was statistical difference between group B and group C (P ＜0.01).Within 48 h,72 h,7 d,14 d,there was statistical significance between group A and group B (P＜ 0.05). The serum levels of VEGF during different phases(24 h,48 h,72 h,7 d, 14 d) were higher in severe group than those in light group(P＜ 0.01). Within24 h,48 h,72 h,7 d,there was statistical difference between severe group and midrange group (P＜ 0.01).Within 48 h, 72 h, 7 d, 14 d, there was significant difference between midrange group and light group (P ＜0.05). The volume of hemorrhage had positive correlation with the CSS scores of neurologic impairment on admission (r = 0.916, P ＜ 0.05). Conclusion The serum VEGF levels might be related to the volume of hemorrhage and the severity of the disease.

Objective To investigate the effect of salmeterol/fluticasone (SM/FP) on serum inflammatory factors in the treatment of bronchial asthma in children.Methods 80 children with bronchial asthma from January 2015 to October 2015 in department of pediatrics of first affiliated hospital of Zhejiang Chinese medicine university were selected and randomly divided into two groups.The control group were given routine clinical treatment, the experimental group were treated on the basis of the control group with salmeterol/fluticasone (SM/FP), for 4 weeks.The serum IL-2, IL-4, IFN-α, T-lymphocyte subsets and clinical efficacy between the two groups were compared.Results Compared with control group, the serum levels of IL-2 and IFN-γin experimental group were higher, IL-4 in experimental group was lower ( P <0.05 ); the serum CD3 +T, CD4 +T and CD4 +T /CD8 +T levels in experimental group were higher, the serum CD8 +T was lower than those in control group (P<0.05); the total efficiency of the experimental group (92.5%) was significantly higher than that of control group (75.0%) (P<0.05).Conclusion The salmeterol/fluticasone (SM/FP) has the good efficacy in the treatment of bronchial asthma in children, which could effectively regulate T lymphocyte subsets proportion and the level of cytokines.

Objective To explore the clinical feature of acquired pure amegakdryocytic thrombocytopenic purpura(AATP).Methods 18 patients were analyzed retrospectively.And 112 patients with ITP in the same term were to be control group.Each group's megakaryocytic (MK),platelet (PLT) count,mean platelet volume (MPV) and platelet associated immunoglobulin G(PAIgG) were rested.Results The MK total number of AATP group was obvious decreased or even absent,while the ITP group was increased[(0.8?1.5) vs (195.0?47.3),P0.05).About the PAIgG posi- tive rate,AATP group was 55.6 % while ITP group was 83% (P

As the major pathway mediating specific protein degradation in eukaryotes, ubiquitin-proteasome system (UPS) is involved in various physiological and pathological processes such as cell cycle regulation, immune response, signal transduction and DNA-repair. Deubiquitinases (DUB) maintain the balance of UPS and related physiological processes via reversibly removing ubiquitin from the covalently modified protein substrates, which have been implicated in various disease processes in case of their imbalance expression. Because DUB plays critical regulating roles in the UPS pathway, they may be also the ideal drug targets for severe and intractable human diseases, such as cancer and neurodegenerative disease. With the rapid development of proteomic technology, systematical investigation of specific substrates and interacting proteins of varied DUB via mass spectrometry approach may shed light on these DUB's biological function and regulating roles in the physiological and pathogenic states. In this review, we briefly introduce the characteristics of DUB and summarize the recent application and progresses of proteomics in DUB research.
Humans ; Mass Spectrometry ; Proteasome Endopeptidase Complex ; metabolism ; Proteomics ; Signal Transduction ; Ubiquitin ; metabolism ; Ubiquitin-Specific Proteases ; metabolism