Loading...

Table of Content

    05 May 2011, Volume 31 Issue 5
    Mutation analysis of AGL gene in 7 Chinese patients with glycogen storage disease type III
    QIU Zheng-qing WANG Wei WEI Min QIU Jia-jing ZHANG Han-bing WU Xiao-yan
    2011, 31(5):  471-474. 
    Asbtract ( 2200 )   PDF (776KB) ( 1258 )  
    Related Articles | Metrics
    Objective To study the AGL gene mutation of patients with glycogen storage disease type Ⅲ in order to know whether there is common mutation in Chinese patients. Methods All 7 patients had hepatomegaly, fasting hypoglycemia, variable hyperlactacidemia and hyperlipidemia. Total genomic DNA was extracted from peripheral blood leukocytes of the patients. The coding exons 1-35 of the AGL gene were amplified by polymerase chain reaction (PCR) and sequenced directly. Results Total of 8 mutations was detected in all 14 alleles of 7 patients, including 4 previously reported mutations, c.1735+1G>T, c.665-1G>A, c.958+1G>T and c.4284T>G; 4 novel mutations, c.202delG, c.1017delT, c.1251dupA and c.2546+1G>T. Splicing mutation c.1735+1G>T is the most common mutation in our group, accounting 42.9%. Conclusion 8 mutations has been identified in this study, including 4 splicing mutations, 2 deletion mutations, 1 nonsence mutation and 1 duplication mutation. Splicing mutation c.1735+1G>T may be the most common AGL gene mutation in Chinese patients.
    Expression changes of Nicastrin in the neuronal cells and its relationship with Aβ generation after proteasomal inhibitor treatment.
    LONG Zhi-min ZHAO Lei LUO Shi-fang WANG Ke-jian HE Gui-qiong
    2011, 31(5):  475-479. 
    Asbtract ( 2071 )   PDF (842KB) ( 890 )  
    Related Articles | Metrics
    Objective To explore whether Nicastrin undergoes ubiquitination before proteasomal degradation, as well as the relationship between Nicastrin protein level and Aβ generation. Methods Cell fractionation, western blot, immunoprecipitation as well as ELISA were used to check the expression of NCT and Aβ level. Results NCT distributes primarily in ER and Golgi apparatus, less NCT in lysosome, and increased NCT accumulates in the ER and Golgi apparatus after proteasomal inhibition. NCT and ubiquitin colocalize and interact with each other in cells. The degradation of NCT was not affected by PS. Overexpression of NCT by transient NCT plasmid transfection or inhibition of NCT proteasomal degradation can decrease substrate of γ-secretase, C99 and C83, and increase production of γ-secretase, Aβ40 and Aβ42(p<0.01). Conclusion The degradation of NCT is by the way of the ubiquitin-proteasome pathway. The expression of NCT is increased following proteasomal inhibition, and over-expression of NCT can facilitate APP processing and Aβ generation.
    PGC1α synergizes PPARγ to regulate transcription of mouse CIDEC in mouse adipocyte
    2011, 31(5):  480-484. 
    Asbtract ( 3581 )   PDF (808KB) ( 1645 )  
    Related Articles | Metrics
    Objective To identify the regulation of PPARγ and PGC1α on CIDEC transcription. And to determine the Function of CIDEC in adipose metabolism. Methods Reconstructing CIDEC promoter reporters including a series of 5’-deletions and cis-element mutations. Dual-luciferase assay was performed to screen PPARγ binding sites in HepG2 cells. Pioglitazone, which is the PPARγ agonist, was used in 3T3-L1 cells to detect the CIDEC mRNA level. Adenoviral CIDEC was constructed and over-expressed CIDEC gene through adenovirus in primary hepatocytes to examine the effect of CIDEC on liver adipose metabolism. Results PPARγ and PGC1α can stimulate CIDEC transcription obviously. Over-expressing CIDEC stimulate gene FAS 3.47±0.17 (p<0.05) and ACC 3.95±0.57 (p<0.05), and adipose synthesis in hepatocytes. Conclusion Transcription of CIDEC can be activated by PPARγ and its coactivitor PGC1α. This activation result in accumulation of fat in cells and liver.
    The adenovirus construction and identification of expression and interference of PNPLA3 gene
    Ai-jun QIAO Fu-de FANG, Yong-sheng CHANG
    2011, 31(5):  485-489. 
    Asbtract ( 2901 )   PDF (922KB) ( 1411 )  
    Related Articles | Metrics
    Objective To construct a adenovirus vector of PNPLA3 expression and interference, a gene related to metabolism of glucose and lipid and liver fatty disease. Methods The expressional and interferce primers were designed according to PNPLA3 gene sequence,the sequences were cloned into pAdTrack-CMV or pAdTrack-U6 vector with BglII and xhoI restriction site. The E.coli BJ5183 sensitive bacterias were cotransfected with lined vector cutted by PmeI enzyme and adenovirus vector pAdEasy-1. The obtained recombinant adenovirus vector was cutted with PacI enzyme, then adenovirus was obtained in 293A cells transfected with lined recombinant adenovirus plasmids. The titer of virus was measured based on the expression level of green fluorescent protein. The transfection efficiency of green fluorescent protein into mouse primary hepatocytes was calculated. Results DNA sequencing and digestion identification demonstrated that the expression and interference adenovirus vectors of PNPLA3 gene were constructed. The titer of concentrated virus was 1.5×1011VP/ml.The infection efficiency reached above 90% and interference efficiency decreased endogenous PNPLA3 mRNA in hepatocytes by more than 80% when mouse primary hepatocytes were stabled transfected with 120 infection multiplicity. Conclusion The expressional and interference adenovirus vector of PNPLA3 gene is successfully constructed.
    Human adipose-derived mesenchymal stem cells differentiated into endothelial cells on poly-ε-caprolactone
    CHEN Jia ZHAI Yu-Jia ZHANG Hong-gang ZHANG Jing ZHANG Qiu-ju Rui-juan XIU
    2011, 31(5):  490-495. 
    Asbtract ( 1801 )   PDF (1432KB) ( 862 )  
    Related Articles | Metrics
    Objective to study on adhesion ability, growth ability and differentiation into endothelial cells of human adipose-derived mesenchymal stem cells (hADSCs) planted on poly-ε-caprolactone material. Methods we choose the poly-ε-caprolactone with pore diameters of 60~80μm(called “small pore-size material” for short) and 180~200μm(called “large pore-size material” for short). Absorption and degradation rate were calculated on the 14th days. After hADSCs implantation in the materials with different diameters, comparison of seeding rate and growth were carried out by immunofluorescence with DAPI and scanning electron microscope (SEM). CCK-8 method was used to draw the growth curve. Endothelial differentiation with 40ng/ml VEGF and 10ng/ml bFGF after 50 days, vWF and VE-cadherin were detected by flow cytometers and Flk-1 was observed by immunofluorescence way. Results Absorption rates of small and large pore-size material on the 14th day were respectively calculated as 200% and 216.7%, while degradation rates were 13.3% and 21.7%. After implantation of hADSCs, cells looked long fusiform attaching on the material by SEM. Seeding rate of small pore- size material was 98.3%±0.3%, which was comparatively more than seeding rate as 90.3%±1.5% of large pore-size material, and growth curve observed the same. Hence, endothelial differentiation was carried out with small pore-size material. Differentiating after 50 days, positive ration of vWF was 80.9%±0.9% and VE-cadherin was 84.3%±1.1%. Flk-1 expression was observed in most of cells via immuofluorescence. Conclusion Poly-ε-caprolactone material with pore diameter of 60~80 μm is suitable for the growth of hADSCs which are also successfully differentiated into endothelial cells on it.
    Effects and immune mechanism of different adjuvants on polyepitope chimeric vaccine M.RCAg-1&M.RCAg-3
    MA Rui-sen LIN Ya-hui ZHANG Wen-wen MA Ming WANG Heng
    2011, 31(5):  496-502. 
    Asbtract ( 1897 )   PDF (950KB) ( 830 )  
    Related Articles | Metrics
    Objective To investigate the formulations of Plasmodium falciparum polyepitope chimeric vaccine M.RCAg-1 & M.RCAg-3 with different adjuvants in order to increase their immunogenicities and to find out their basic immune mechanisms. Methods The polyepitope chimeric vaccines formulated with different adjuvants (AD, ADL, CpG, CpG&Al) were injected subcutaneously into mice for three times. The immunization was performed on week 0, 2, 4. The concentration of anti-M.RCAg-1 and anti-M.RCAg-3 IgG, the IgG isotype and the recognition level with M.RCAg-1/M.RCAg-3 single epitopes were detected at ten days after week 4. The recognition of antibodies in serum was tested with indirect immunofluorescence assay (IFA). Results We showed here that ADL and AD can enhance the immunogenicities of M.RCAg-1 and M.RCAg-3 respectively. Compared with another groups, ADL and AD formulated with M.RCAg-1 and M.RCAg-3 respectively can enhance their antibodies recognition with natural parasites greatly. Their antibodies can recognize the single epitopes generally. Conclusion ADL and AD can greatly increase the immunization antibody level of M.RCAg-1 and M.RCAg-3 and their recognition with natural parasites. In conclusion, ADL and AD, as potential adjuvants, paved the way for the clinical use of M.RCAg-1 and M.RCAg-3 respectively.
    Ischemia postconditioning regulating TLR4 expression in Tree shrews’s cortex of thrombotic cerebral ischemia
    Rui FENG Shu-qing LI
    2011, 31(5):  503-508. 
    Asbtract ( 1785 )   PDF (1002KB) ( 735 )  
    Related Articles | Metrics
    Objective To investigate the effect of postconditioning after brain ischemia on the expresssion of MPO(myeloperoxidase)and TLR4(Toll-like receptor 4)and inflammation in cortex. Methods The thrombotic focal cerebral ischemia was induced by photochemical reaction in tree shrews, and ischemic postconditioning was established by 3 repeated cycles of 5 min of temporary right carotid artery clipped at 4h after the onset of photochemical reaction and 5 min reperfusion. The changes of histology of cortex (by Hematoxylin-and-eosin), MPO expression (by immunohistochemistry), TLR4 expression (by Western Blot Analysis) and TLR4 mRNA (by RT-PCR) were observed. Resulsts Our study found extensive neuronal injury and inflammation in cortex at 4, 24 and 72h, peak at 24h after the cerebral ischemia, while significantly attenuated after the postconditionng with less inflammation cells’ infiltration. Immunohistochemistry analysis showed that cerebral ischemia caused an increase in MPO expression in cortex, and ischemic postcondionting decreased the expression at 24h and 72h(P <0.05). Cerebral ischemia caused an increase of the TLR4 expression in cortex (P <0.05),peak at 24h. Postcondionting caused a decrease in the TLR4 expression in cortex at 4 hours and 24 hours(P <0.05), but an increase at 72 hours(P <0.05). The levels of TLR4 mRNA in cortex showed the similar variation trend in accordance with the protein expression. Conclusion The protection mechanisms of the prostconditioning may be associated with inhibitting inflammation and regulating TLR4 expression.
    Ketamine aggravates Tau hyperphosphorylation by up-regulating activity of GSK-3? in A?25-35 induced PC12 cells
    XU Jie ZHANG Rui YUE Yun ZUO Ping-ping YANG Nan JI Chao
    2011, 31(5):  509-514. 
    Asbtract ( 2111 )   PDF (900KB) ( 911 )  
    Related Articles | Metrics
    Objective: The pathological characteristics of AD are both extracellular A? senile plaques and intracellular hyperphosphorylated tau neurofibrillary tangles formed. Increasing evidences indicate that hyperphosphorylated Tau plays a key role in the pathogenic events that occur in AD. We set out to assess whether ketamine can induce Tau hyperphosphorylation by increasing activity of glycogen synthase kinase-3? (GSK-3?) in A?25-35 induced PC12 cells. Methods: We performed these studies in cultured rat pheochromocytoma cells(PC12) and randomly allocated PC12 cells into four groups: control group(C); 10μmol/LA?25-35 (A); 1mmol/L ketamine (K); A?25-35 and ketamine (AK), above all groups were incubated for 6 hours. The cell viability was determined by MTT assay. Western blotting was performed to observe the protein expression of Tau phosphorylation of different sites, GSK-3? and p-GSK-3?ser9. In addition, Lithium Chloride, inhibitor of GSK-3?, was administered to all above grounps to investgate the changes of protein expression. Results: The cell survival rate was significantly decreased in AK group compared with the others groups. The level of Tau phosphorylation in the sites of Ser396、Ser404 and Thr231 increased after intubation with ketamine and A?25-35. And these increases were accompanied by up-regulation of activity of GSK-3?. Lithium Chloride attenuates these changes above all. Conclusion: Ketamine increases Tau hyperphosphorylation by up-regulating activity of GSK-3? in A?25-35 induced PC12 cells.
    Application of two types of RNA in setting up DJ-1 knockdown cell model
    2011, 31(5):  515-519. 
    Asbtract ( 1999 )   PDF (863KB) ( 905 )  
    Related Articles | Metrics
    Objective To explore the application of artificial microRNA in setting up gene knockdown Cell model, and compared with siRNA. Methods First of all, we constructed vectors, and prepared siRNA fragments targeting on DJ-1. Then we transcient transfected the artificial miRNA and siRNA into MN9D cells by lipofectamine2000 reagent, the mRNA and protein expression level of DJ-1 gene were detected by real-time PCR and Western blots,respectively. Results Compared with control group, DJ-1 expression level was decreased significantly in both artificial miRNA and siRNA groups. DJ-1 was knockdowned by different level. DJ-1 was decreased 90%(p<0.05)at mRNA expression level, and decreased 70%~85% (p<0.05) at protein level in MN9D cells transfected with the artificial miRNA. While DJ-1 was decreased 50%~70%(p<0.05)at mRNA expression level, and decreased 20%~50%(p<0.05)at protein level in MN9D cells transfected with siRNA. Comparing with siRNA, miRNA was more effective in silencing DJ-1. Conclusion The artificial miRNA and siRNA are both effective in silencing gene. miRNA has more significant function in knocking down DJ-1 than siRNA.
    The Difference of Blood Pressure and Related Factors identified between Han and Korean-Chinese Adults in Heilongjiang China
    PAN Yang-xing QI Bao-shen ZHOU Xiao-mei HAN Shao-mei LI Zi-tong ZHU Guang-jin
    2011, 31(5):  520-524. 
    Asbtract ( 1936 )   PDF (564KB) ( 821 )  
    Related Articles | Metrics
    Objective To Compare the difference of blood pressure between Korean-Chinese ethnic and Han ethnic in Heilongjiang province and analyze the relationship of the difference of blood pressure between two races with the related factors. Method Systemic vascular resistance (SVR), blood pressure, and so on were determined by BIOZ.com Cardio Dynamics Monitor in 1247 Hans and 676 Korean-Chinese aged from 19 to 80 living in the same area of China. The lipids and glucose of blood were measured by related biochemistry methods. Data were analyzed by SPSS 16.0 software. Results Ethnic(Han=1,Korean-Chinese=2)was positive correlated with systolic blood pressure(SBP), diastolic blood pressure(DBP), triglyceride (TG), low density lipoprotein (LDL), glucose (GLU), Calcium ( Ca2+). After adjustment for gender, age and BMI, the average systolic/diastolic blood pressures in Korean-Chinese cohort was 3.73/3.54mmHg higher than those in Han cohort. TG, LDL, GLU and Ca2+ produced effect of -0.5%/0.5%, -36.1%/-61.7%, -74.6%/-83.0% and -71.4/-81.6% on SBP/DBP respectively by single factor regression analysis. Conclusion The difference of blood lipids, glucose and Ca2+ play an important role on difference of the blood pressure between two ethnic.
    Insulin Receptor Exon 8 NsiI Polymorphism is not associated with Type 2 Diabetes Mellitus in the Hans living in Liangshan, Sichuan Province
    Wei-wei XU WEI Da-ying WANG Chun-xiu ZHANG Jian-hua HE Hui-jing MA Ming-ju WANG Bin Guang-liang SHAN
    2011, 31(5):  525-528. 
    Asbtract ( 1911 )   PDF (523KB) ( 759 )  
    Related Articles | Metrics
    Objective To investigate the relationship between Insulin Receptor Gene Exon 8 NsiI Polymorphism and type 2 diabetes mellitus Methods A cross-sectional survey was conducted among Han residents living in 6 counties, Sichuan province. Questionnaire survey, anthropometric measurement and blood test were carried out. We selected type 2 diabetes cases as case group(n=162) and healthy control individuals as control group(n=162) with frequency matching(1:1) of sex and age. Polymerase chain reaction and restriction fragment length polymorphism (PCR–RFLP)were adopted to examine the individual genotype. Results There were no significant differences in the genotype and gene frequency between cases and controls. Conclusion NsiI polymorphisms may be not associated with the type 2 diabetes in Han people of Liangshan ,Sichuan province
    Clinical and molecular genetic analysis for a patient with glycogen storage disease Ⅰb
    SUN Xiao-fang CHEN Shi Lin LU, QIU Zheng-qing Xin-hua XIAO,
    2011, 31(5):  529-533. 
    Asbtract ( 2006 )   PDF (1003KB) ( 906 )  
    Related Articles | Metrics
    Objective To analyze the clinical and molecular genetic characteristics of a patient with glycogen storage disease Ⅰb (GSDⅠb). Methods Clinical profile and laboratory data were collected. Genomic DNA was extracted from leukocytes of peripheral blood of the patient. All nine exons and the exon-intron boundaries of the SLC37A4 gene were amplified by PCR. The mutations were identified by bidirectional sequencing of the PCR products. Results The patient was diagnosed as GSDⅠb according to the clinical presentations and laboratory examinations, SLC37A4 mutation was identified as a homozygous C>T transition at nucleotide position 527 of the cDNA. The missense mutation is located in exon 3, at codon 191, changing proline to leucine, which changes the conformation of the glucose-6-phosphate translocase (G6PT). Conclusion Through SLC37A4 gene mutation analysis, the clinical diagnosis of GSDⅠb was confirmed. The alternation of G6PT structure caused by SLC37A4 gene mutation is the molecular genetic mechanism to explain clinical manifestation of the patient. To our knowledge, this is the first report of P191L as a cause of GSD1b in Han China mainland. In the future, GSDⅠb could be diagnosed through genomic DNA mutation analysis.
    Possible signal pathway of Resveratrol-induced apoptosis in human skin cancer cell line
    Dong-zhi ZHAO Xiao-shuang LI Hai-mei SUN Guo-quan ZHANG Ji-hong YAN De-shan ZHOU
    2011, 31(5):  534-539. 
    Asbtract ( 2278 )   PDF (1312KB) ( 950 )  
    Related Articles | Metrics
    Objective To investigate the potential signal molecule of Resveratrol (Res)-induced apoptosis of human skin cancer cell line, A431 cells. Methods A431 cells were divided into 4 groups, including Group A (controls), Group B (treated with Res), Group C (pre-treated by SP600125, Group D (inhibitor control, SP600125 only), respectively. Phase contrast microscope, Western blot and immunofluorescence staining were used to observe morphology and the expression of Caspase-3, Bcl-2, Bax, JNK and P-JNK in A431 cells. Result Apoptosis were often observed after Res treatment in the A431 cells. Meanwhile, the expression of Caspase-3 and rate of Bax/Bcl-2 increased, P-JNK expression increased significantly and aggregated in the nucleus(increased to 289469.2±3729 from 225463±672,P <0.05). Pre-treated with SP600125, the above changes decreased significantly (248198.4±4366.9,P <0.05), and the growth of A431 cells was as the same as controls. Conclusion The phosphorylated JNK entering into the nucleus to regulate the expression of Bcl-2, Bax and Caspase-3 were involved in the signal molecule of Res-induced apoptosis in A431 cells.
    Expression of TRAIL and TRAIL receptors in interstitial lung disease of experimental autoimmune myositis
    Hua ZHAO Guo-chun WANG
    2011, 31(5):  540-545. 
    Asbtract ( 1547 )   PDF (1330KB) ( 858 )  
    Related Articles | Metrics
    Objective To investigate the pathologies and pathogenesis of interstitial lung disease (ILD) in experimental autoimmune myositis (EAM) models. Methods Established EAM mode by injecting the homogenate of rabbit skeletal muscle with Freund's complete adjuvant. The histopathological features of the muscle and lung from the EAM model were investigated by staining with hematoxylin and eosin and Masson's trichrome. The expressions of TRAIL and TRAIL receptors (TRAIL-R1, TRAIL-R2, TRAIL-R3, and TRAIL-R4) in lung tissues were measured by immunohistochemical method and computer image analysis. Result 16 of 45 rats with autoimmune inflammatory myopathies were found varying degrees of interstitial lung disease (P <0.05); TRAIL expression was increased in lung tissue (39265±27595) and that of DCR1 was decreased (37377±14126) (P <0.05); Inflammatory cells infiltrated locally were found with high expression of DCR2; Alveolitis was related with CD8 + T lymphocytes (P <0.001) and pulmonary fibrosis was related with TRAIL expression in lung tissues (P <0.05). Conclusion The study firstly demonstrates that the features of ILD shown in EAMs were related with the local immune inflammatory response. The changes in the expression of TRAIL and its receptors between EAMs and controls suggest TRAIL-system probably play a role in the pathogenesis and the pathology of IIM with ILD.
    VEGFsiRNA enhance drug sensitivity of leukemia cells to Ara-C
    Chun-xia JING Huan ZHANG Guang YANG
    2011, 31(5):  546-550. 
    Asbtract ( 1701 )   PDF (804KB) ( 923 )  
    Related Articles | Metrics
    Objectives To explore the effects of VEGFsiRNA on chemosensitivity of leukemia cells to Ara-C. Methods VEGFsiRNA was constructed in vitro and then was transferred to HL60 cells. The proliferation was detected by CCK8 method. The level of VEGFmRNA was detected by reverse transcription and quantitative polymerase chain reaction (RT-PCR). The expression level of VEGF protein was determined by ELISA in cell culture medium. The cell apoptosis was detected by flow cytometry. Results VEGFsiRNA could increase the sensitivity of leukemia cells to Ara-C, and VEGFsiRNA combined with Ara-C significantly inhibited the survival of HL60 cells in which the inhibition rate was 82.9 percentages (F=121.63, P<0.01). VEGFsiRNA decreased significantly expression of VEGFmRNA121(F=17.17, P<0.01) and VEGF protein (F=3290.49, P<0.01). However, VEGFsiRNA didn’t promote the level of apoptosis induced by Ara-C. Conclusion VEGFsiRNA increases the sensitivity of leukemia cells to Ara-C by inhibiting proliferation.
    IL-17 expression in patients with chronic HBV infectious diseases and hepatic fibrosis
    Zhao-min ZHENG Yan-guo WANG Hai-tao ZHANG Wen-jun DU Shi-jun CHEN
    2011, 31(5):  551-555. 
    Asbtract ( 2239 )   PDF (941KB) ( 951 )  
    Related Articles | Metrics
    Objective To explore the role of interleukin-17(IL-17) in the pathogenesis and progress of chronic hepatitis B virus(HBV) infectious diseases, especially in liver fibrosis. Method Serum IL-17 level and liver fibrosis index type IVcollagen, LN,HA,type IIIcollagen was determined by enzyme linked immunosorbent assay(ELISA). IL-17 mRNA expression in peripheral blood mononuclear cell(PBMC) was analysed by reverse transcriptase-polymerase chain reaction(RT-PCR).IL-17 expression in liver was measured by immunohistochemistry. Result Serum IL-17 protein levels and IL-17 mRNA expression of PBMC in four HBV infectious groups (P<0.01each) were higher than in normal control. IL-17 and IL-17 mRNA in LC was higher compared with other groups(p<0.001 each). Liver IL-17 expression and serum fibrosis index raised as severity of disease with LC higher than CHB,CHB higher than ASC(Liver IL-17 expression p<0. 01 each, serum fibrosis index p<0. 001 each). IL-17 mainly located in portal area and was associated with fibrosis extent closely. Lymphocyte, fibroblast and sinusoid endotheliocyte were stained positively. Liver IL-17 expression was correlated with serum fibrosis index type IVcollagen, LN,HA,type IIIcollagen positively(r=0.883,0.834,0.793,0.722,P <0.01each) Conclusion IL-17 may contribute to the development of HBV infectious disease especially to the pathophysiology of LC.
    in situ Analysis of gene expression in single cells with Padlock-RCA
    Hong-zhen DU De CHANG Li-min LI
    2011, 31(5):  556-560. 
    Asbtract ( 1982 )   PDF (998KB) ( 857 )  
    Related Articles | Metrics
    Objective To apply Padlock rolling-circle amplification (Padlock-RCA) for the analysis of in situ gene expression in individual cells during cell differentiation. Method Retinoic acid induced differentiation of mouse teratocarcinoma F9 model was used, Nanog mRNA expression in F9 cells before and after induced differentiation were analyzed with Padlock-RCA and compared with analysis using real time RT-PCR. Result Padlock-RCA can efficiently detect the level of Nanog mRNA expression in single cells . Nanog specific Padlock-RCA products were significantly decreased in F9 cell following retinoic acid induction, and confirmed with real time RT-PCR (P<0.05). Conclusion We have successfully applied Padlock-RCA to analyze differential Nanog mRNA expressions before and after induced differentiation in single cells, and provided a reliable method for molecular and functional analyses of gene expression in single cell or cell subpopulations.
    CDK2 accelerates early erythroid differentiation of K562 cells
    Jun LI Rui-hua YUE Jun-le SHEN Jun XIAO
    2011, 31(5):  561-564. 
    Asbtract ( 1802 )   PDF (535KB) ( 831 )  
    Related Articles | Metrics
    Objective To study the roles of a cell cycle regulator Cyclin-dependent kinase 2 (CDK2) in erythroid differentiation of K562 cells.Methods K562 cells were transfected with the construct expressing CDK2 and siRNAs specifically targeting CDK2, respectively. The effects of over-expression or knocking-down of CDK2 were examined by Western blot. Quantitative RT-PCR was performed to detect the levels ofγ-globin mRNA expression. The benzidine staining assay was used to access the differentiation state of K562 cells. Results CDK2 was up-regulated at the early stage of K562 erythroid differentiation. Over-expression of CDK2 in K562 cells accelerated erythroid differentiation, in contrast, inhibition of CDK2 attenuates globin accumulation in K562 cells.Conclusion CDK2 is required for early erythroid differentiation of K562 cells.
    Depressive behavior of rats induced by CUMS involved in abnormal pathomorphology and BDNF expression in hippocampus of rats
    Han WANG Na LI Wei WEN Rong JIANG Qi-xin ZHOU
    2011, 31(5):  565-569. 
    Asbtract ( 165 )   PDF (963KB) ( 907 )  
    Related Articles | Metrics
    Objective Establish depression model of rat through chronically unpredicted mild stress(CUMS)in order to investigate its connection with the changes of hippocampal pathomorphology and Brain-derived neurotrophic factor(BDNF). Methods Rats were randomly divided into two groups,control and model.CUMS was used to establish depression model of rats. The rat behavior, SOD activity and MDA level of cortex, hippocampal pathomorphology, and BDNF expression were tested by the methods of open-field, xanthine oxidase, TBA,HE staining,and immunohistochemisty, respectively. Results It was shown that there existed significantly decreasing open-field scores,SOD activity,and BDNF expression(A value 803±422 vs 237±96,p<0.01) with obviously increasing MDA level and karyopyknosis of hippocampal neurocytes in rats treated by CUMS,compared with control rats. Conclusions Depression behavior of rats can be induced by CUMS,the mechanism of which may be associated with damage of hippocampal neurocytes and decrease of BDNF expression from imbalance of oxidation-antioxidation system and increase of free radicals in rat brains.
    Radioactive Seed Migration After Transperineal Interstitial Prostate Brachytherapy
    Yi ZHOU Han-zhong LI Wei-gang YAN
    2011, 31(5):  570-573. 
    Asbtract ( 172 )   PDF (500KB) ( 662 )  
    Related Articles | Metrics
    Objective To study the incidence of seed migration after prostate cancer brachytherapy using a loose-seed (125I) implantation technique and to identify the predictors associated with its occurrence. Methods Between December 2003 and June 2010, 270 patients staging T1~T2 underwent permanent prostate brachytherapy at our hospital and were eligible for the study. Pelvic CT and Chest X-rays obtained at follow-up were examined for the number and location of seeds. The patient and treatment variables potentially associated with the occurrence and number of seed migrations were analyzed. Results Incidence rate of seed migration in patient is 8.9%,(24/270), 27 seeds were found migrated(0.16%,27/17010), among which 6 seeds migrated to thoracic cavity, 19 seeds migrated into retropubic space and subcutaneous tissue, 2 seeds were discharged through urine or sperm. Conclusions To Decrease Prostate volumn, total number of implanted seeds and extraprostatic placement of seeds can decrease the frequency of seed migration. With history of TURP and hyperplasia of middle lope of prostate are associated with an increased frequency of seed migration. The small proportion of migrated implanted seeds will not effect the dosimetry of target organ of prostate. No severe complications were observed in cases of seeds migration.
    Clinical characteristics and outcome of primary pulmonary non-Hodgkin’s lymphoma
    Jian LI Li JIAO Wei ZHANG Ming-hui DUAN Nong ZOU Dao-bin ZHOU Ti SHEN
    2011, 31(5):  574-577. 
    Asbtract ( 183 )   PDF (473KB) ( 661 )  
    Related Articles | Metrics
    Objective To study the clinical characteristics and outcome of patients with primary pulmonary non-Hodgkin’s lymphoma (PPL). Methods The clinical features, diagnostic methods, pathological types and outcome of 30 patients with PPL diagnosed in PUMC Hospital were analyzed retrospectively. Results Of these 30 patients, 46.7% of patients were male and the median age was 46 years. These patients didn’t have specific clinical symptoms and radiological features and 20% patients were asymptomatic. The median time from onset to diagnosis of all patients was 8 months. The positive diagnostic rate of transbronchial lung biopsy, lung biopsy guided by computer tomography, and lung biopsy by thoracoscopy was 33.3%, 50% and 100%, respectively. The mucosa-associated lymphoid tissue (MALT) lymphoma was main type in PLL, which was found in 56.7% patients. 7 of 30 patients were dead after 39 months of median follow-up time. Compared with patients with non-MALT lymphoma, patients with MALT lymphoma had lower international prognostic index (IPI) score and better outcome. Conclusions The PPL is a rare extra-nodular lymphoma, which is difficult to diagnose. The patients with non-MALT lymphoma have better outcome in PPL.
    Clinical analysis of POEMS syndrome with pulmonary arterial hypertension
    Shu-fen LIU Wei-guo ZHU Xue-jun ZENG Wen-bing XU
    2011, 31(5):  578-581. 
    Asbtract ( 146 )   PDF (495KB) ( 763 )  
    Related Articles | Metrics
    Objective To describe the clinical characteristics of POEMS syndrome associated with pulmonary arterial hypertension. Methods Medical records of patients with POEMS syndrome who were identified in Peking Union Medical College Hospital were reviewed. The clinical manifestations of patients complicated with PAH (pulmonary arterial hypertension, PAH) diagnosed by echocardiography (uCG) was compared with those without PAH and those without echocradiography results. Results One hundred and seventeen patients with POEMS syndrome were recruited. Forty-nine patients have PAH,thirty-three patients have no PAH and thirty-five patients didn’t undergo echocradiography. The prevalence of PAH was 41.9%(49/117). Chest distress is the main manifestation in group of POEMS syndrome with PAH, and the frequency is 42.9%(21/49),higher than the other two groups. These patients with PAH are more likely to have pleural effusion, ascites, or pericardial effusion. Conclusion The prevalence of PAH in POEMS syndrome patients was more than 40%, and over half of them don’t have any symptoms like chest distress. We suggest patients with POEMS syndrome should have echocradiography to screen PAH.
    Cardiovascular abnormalities in patients with Turner's syndrome
    Yan-lin ZHU Chao NI Li-lin GUO
    2011, 31(5):  582-585. 
    Asbtract ( 164 )   PDF (495KB) ( 745 )  
    Related Articles | Metrics
    Objective To analyse the associated cardiovascular abnormalities in patients with Turner's syndrome (TS). Methods Clinical features and cardiovascular abnormalities of 25 female patients with TS were analyzed according to karyotype. Results Congenital cardiovascular malformations were detected in 40% of TS patients, including bicuspid aortic valve, mitral/aortic lengthy or prolapse, coronary sinus dilation, cardiac chamber dilation, and noncompaction of the ventricular myocardium. The patients with monosomy X, more often than those with mosaicism, had a higher incidence of congenital cardiovascular malformations (40% vs 14%, p<0.01). Aortic size index, including root and assending aorta were significantly greater in TS than the healthy contro(27±5mm vs 18±2mm, 28±6mm vs 18±2mm, p<0.001). Conclusion Regular evaluation of congenital cardiovascular malformations and accompanied risk factors may help to predict the risk of cardiovascular death.
    Clinical Manifestations and Differential Diagnosis of Laryngeal Tuberculosis
    2011, 31(5):  586-590. 
    Asbtract ( 288 )   PDF (1214KB) ( 729 )  
    Related Articles | Metrics
    Objective: To analyze the clinical patterns of current laryngeal tuberculosis. Methods: Clinical analysis of 10 patients with pathologically confirmed laryngeal tuberculosis was carried out retrospectively. Analyzed 6 Wegener granuloma cases that involved the larynx and 24 supraglottic laryngeal carcinoma cases for differential diagnosis of laryngeal tuberculosis. Results: The age of patients of laryngeal tuberculosis ranged from 37 to 74 years. The male-to-female ratio was 1.5 to 1. The most frequent complaint was hoarseness. The epiglottis, arytepiglottic fold, the false vocal cord and the true vocal cord were commonly involved, 8 patients had active pulmonary tuberculosis, and 2 were proven to have normal lung status. All the patients received proper antituberculosis chemotherapy. Conclusions: The patterns of laryngeal tuberculosis have changed over the years. It can occur without pulmonary involvements. The characteristics of lesions seem to be more nonspecific. The differential diagnosis of this disease is more difficult. Prompt and proper treatment is very important in decreasing the complications of laryngeal tuberculosis.
    New insight of active vitamin D in chronic kidney disease
    Dan LIU Lin-li LV Bi-cheng LIU
    2011, 31(5):  591-593. 
    Asbtract ( 224 )   PDF (374KB) ( 1456 )  
    Related Articles | Metrics
    Chronic kidney disease is usually complicated with deficiency of vitamin D which causes severe impairment of kidney function through stimulating RAS (renin–angiotensin system)and other pathways. Both in vivo and vitro studies have demonstrated that active vitamin D and its analogues supplementation could attenuate glomerulosclerosis and renal tubulointerstitial fibrosis and the renoprotective effects could delay the progression and reduce the mortality rate of CKD.
    Efficiency of the intensive orientation for the new graduate medical students to become emergency residents
    Ji-hai LIU Xue-zhong YU Hua-dong ZHU Zhong WANG Teng-da XU Veronica Pei Jeffery Smith
    2011, 31(5):  594-596. 
    Asbtract ( 141 )   PDF (462KB) ( 732 )  
    Related Articles | Metrics
    Objective Orientation is a kind of training for the residency who just graduated from medical school. We perform different types of orientation and discuss their efficiency for the new graduate medical students to become emergency residents. Methods 15 new graduate medical students randomly divided into 2 groups. One group is intensive orientation training, and the other group is traditional group. We compare their written exam scores before and after the orientation training. We perform surveys on their self-evaluation after they finish the orientation and 3 months later。Differences between these 2 groups are analyzed. Results The intensive training group has better scores on the written exam scores than the traditional group(78.25±5.90 Vs. 59.71±3.90,P=0.02). They have more confidence 3 months after orientation training(91.20±8.76 Vs. 75.46±6.56,P=0.03). Conclusion Intensive orientation training helps the medical students to become a residency more efficiently.