Table of Content

05 August 2013, Volume 33 Issue 8

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Characteristics of the whole-genome microRNA expression profiles and chromosome distribution in Down syndrome fetus

2013, 33(8):  935-940. 

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Objective To investigate the expression profiles and chromosome distribution characteristics of the whole-genome miRNAs in Down syndrome (DS) fetus. Methods Illumina deep sequencing technology was conducted to evaluate the miRNAs expression in 6 pairs of fetal cord blood mononuclear cells (CBMCs) samples from DS fetus (DS group) and normal fetus (control group), the characteristics of miRNA expression profiles and chromosome distribution were confirmed by comparing with human genome. Results Of 395 identified miRNAs encoded by 316 miRNA genes, 149 miRNAs were significantly differentially expressed (6 miRNAs up-regulated and 143 miRNAs down-regulated in DS group)，51 miRNAs were specifically expressed in the control fetal CBMCs. For the14 human chromosome 21 (Hsa21) derived miRNA genes, one miRNA (miR-802) was up-regulated in the DS group, and 4 miRNAs (let-7c, miR-99a, miR-125b and miR-155) were down-regulated in the DS group, and other Hsa21-derived miRNA genes were not expressed in both group. Although the chromosome distribution of expressed miRNA genes was similar in both samples, the chromosome distribution of miRNA expression abundance was different. DS group was major scattered in 8、16、17 and 21 chromosome, while the normal group was major scattered in 3、8、14、16、17 and 21 chromosome. Conclusion The DS fetal CBMCs have its own specific miRNA expression profiles and chromosome distribution characteristics. The miRNAs encoded by the chromosome with differentially distribution of expression abundance may play an important role in the development of the DS clinical symptoms.

The expression of FGF1 and its receptors during the osteogenic and adipogenic differentiation of human bone marrow stromal cells

2013, 33(8):  941-946. 

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Objective To study the dynamic expression profile of fibroblast growth factor 1 (FGF1) and its receptors (FGFRs) during osteogenic and adipogenic differentiation of human bone marrow stromal cells (hBMSCs), and further exploring the role of FGF1 during hBMSCs differentiation. Methods Cells were isolated from human bone marrow by density gradient centrifugation method. hBMSCs in the 3rd passage were induced into osteoblasts and adipocytes, respectively, and followed by staining experiments for identification. The expression of FGF1, FGFRs, and marker genes during osteogenic and adipogenic differentiation, were detected by real-time PCR. Immunofluorescent staining was applied for FGF1 localization of hBMSCs after induced differentiation. Meanwhile, the influence of exogenous FGF1 on osteogenic and adipogenic differentiation was detected by real-time PCR and Oil Red O stainning. Results Osteogenic and adipogenic induced hBMSCs were positively stained by Alizarin Red S and Oil Red O in vitro. The expression status of FGF1 and its receptors showed dynamic variation. Immunofluorescent staining results illustrated that FGF1 was mainly distributed in cytoplasm, and its expression amount increased in nucleus after osteogenic induction for 3 days. The expression of osteopontin increased during osteogenic differentiation in the presence of exogenous FGF1 for 3 days (P＜0.05), and the amount of lipid droplet decreased and the expression of adipogenic marker genes were suppressed during adipogenic differentiation after the introduction of exogenous FGF1 for 7 days (P＜0.01). Conclusions It has been demonstrated that FGF1 could inhibit adipogenic differentiation of hBMSCs, and promotes the expression of OPN, which may further hasten the maturity of osteogenesis.

Effects on the proliferation and apoptosis of ovary cancer cells SKOV3 co-cultrued with and apoptosis with lymphocytes infected by recombinant adenovirus Ad-IL-12

2013, 33(8):  947-954. 

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Objective To investigate the effects of lymphocytes infected by recombinant adenovirus Ad-IL-12 on the proliferation and apoptosis of ovary cancer cell SKOV3. Methods The peripheral blood lymphocytes were infected by recombinant adenovirus Ad-IL-12 and Ad-GFP for 48 h respectively.The SKOV3 cells was co-cultured with lymphocytes infected with Ad-IL-12 (SKOV3/Ad-IL-12) and lymphocytes infected with Ad-GFP (SKOV3/Ad-GFP) separately, and the untreated cells (SKOV3) were set as another control. IL-12 double-subunits (P35, P40) mRNA expressions were tested by RT-PCR and the levels of IL-12P70 protein in the cell culture supernatant were detected with ELISA. The proliferation potential of SKOV3 was assessed by CCK8 assay, flow cytometry was used to detect the cell cycle and cellular apoptosis, RT-PCR and Western blot were used to detect the expression of BCL-2 and Survivin. Results The human recombinant adenovirus Ad-IL-12 could successfully infect the human peripheral blood lymphocytes and effectively secret IL-12P70 proteins; growth ability of SKOV3 cells (SKOV3/Ad-IL-12) was significantly inhibited (P<0.05). In addition, the percentages of SKOV3 cells in the G1 phase and the apoptosis (SKOV3/Ad-IL-12) were increased Significantly(P<0.05), the expression of BCL-2 and Survivin were down-regulated（P<0.05）. Conclusions The human recombinant adenovirus Ad-IL-12 could infect the human peripheral blood lymphocytes and express the IL-12 P70 protein successfully, which could inhibit the proliferation of SKOV3 cells by blocking cell cycle and inducing apoptosis.

Beraprosat Promoting the Apoptosis of Pulmonary Artery in the Pulmonary Artery Hypertension Rats Induced by Monocrotaline

2013, 33(8):  955-959. 

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Objective To investigate the effect of prostacyclin on apoptosis of pulmonary artery. Methods The pulmonary artery hypertension rats were induced by monocrotaline (MCT) intraperitoneal injection. In MCT and beraprosat group, each rat was only given an intraperitoneal injection of 60 mg/kg of MCT. In control group, each rat was only given an intraperitoneal injection of equivalent volume saline. The rats in beraprosat group were given an intragastric administration of 200mg /kg of beraprosat daily from the second day. The hemodynamics, right ventricular hypertrophy, pulmonary artery remodeling, pulmonary artery apoptosis and the molecules involved in apoptosis of PAH were observed. Results In beraprosat group, rats were with lower hemodynamics and right ventricular hypertrophy than MCT group. Beraprosat significantly improved the pulmonary artery remodeling and increased the apoptosis in pulmonary artery. Beraprosat also increased the Bax, decreased the Bcl-2 and PDK in both mRNA and protein level.Conclusions Beraprosat can improve pulmonary artery remodeling by inducing the apoptosis of pulmonary artery.

Placental growth factor induces brain microvascular endothelial cell tight junctions’opening by activation of Rho/ROCK signaling pathway

2013, 33(8):  960-965. 

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Objective To study the molecular mechanism of placental growth factor (PLGF) inducing brain vascular endothelial cell tight junctions’ opening. Methods Using human brain microvascular endothelial cells (HBMECs) to construct blood-brain barrier (BBB) modal in vitro, with which to study human recombinant PLGF protein inducing the change of electrical resistance (TEER) and permeability of the BBB. Moreover, the distribution and change of tight junction protein ZO-1 and occludin were observed by immumofluorescence and Western blotting methods to confirm the effects of PLGF on tight junction of BBB. Finally, detected the cell signals induced by PLGF on tight junction of BBB using protein inhibitors and Western blotting method.Results PLGF induced a decrease of TEER and an increased of HRP flux of BBB modal in vitro. Besides, PLGF increased soluble occludin and changed ZO-1 distribution, the inhibitor of ROCK (Y27632) blocked the HRP flux induced by PLGF, and RhoA-GTP was decteced in HBMEC after PLGF tratement. Conclusion PLGF induces tight junction’s opening of BBB in virto by activation Rho/ROCK signaling.

Effect of 5-Aza-CdR on DLK1 expression and the growth of the human hepatocarcinoma cell line HepG2

2013, 33(8):  966-970. 

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Objective To investigate if demethylation drug 5-aza-2'-deoxycytidine (5-Aza-CdR) could inhibite the expression of DLK1 and cell proliferation,invasion in hepatoma cell line HepG2. Methods After different concentrations of 5-Aza-CdR was treated on HepG2 cells, RT-PCR、Western blot was used for detection of DLK1 mRNA and protein level change; MTT, Transwell and flow cytometry were used for detection of cell growth, cycle and invasiveness. Results After treatment with 5μmol/L 5-Aza-CdR, the expression of DLK1 mRNA and protein decreased,the cell growth rate slowed, the number of G1 phase cells decreased where the S phase cells increased, Transwell confirmed invasive ability decreased significantly ( P < 0.05 ). Conclusion 5-Aza-CdR can effectively inhibit DLK1 gene expression, thereby inhibiting tumor cell growth, proliferation, invasion of HepG2.

Delay of skeletal muscle atrophy after transplantation of mesenchymal progenitor cell into transected position

2013, 33(8):  971-975. 

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Objective To study the delaying denervated skeletal muscle atrophy after transplantation of mesenchymal progenitor cell (MPC) into the transected position. Methods MPC were isolated from bones of hind limbs of GFP transgenic C57 mice for cultivation and identification. 36 C57 mice were divided into 3 groups evenly in random, MPC transplantation group, the transected group and the control group. 5μL of MPC suspension and 5μL of Phosphate buffered saline (PBS) were injected into the sciatic nerve transected position in the MPC transplantation group and the transected group respectively while nothing was injected in the control group. The activity ability of hind limbs of mice were observed. At the time point of 2 and 4 weeks after the operation, the retain ratio of wet weight of gastrocnemius muscle and cross sectional area of muscle fiber was measured and the ultrastructural organization was observed. The expression of α-actin, myoglobulin (MHC) were detected by western blot and the expression of Myogenin and MyoD were detected by RT-PCR. Results At the time point of 2 and 4 weeks after the operation, the wet weight of gastrocnemius muscle and the retain ratio of cross sectional area of muscle fiber of mice of the MPC transplantation group was higher than that of the transected group significantly (P < 0.05). At the time point of 4 weeks after the operation，compared with the degeneration of myocyte, mitochondria and sarcoplasmic reticulum and the extent of musculus fibrosis of the transected group, that of the MPC transplantation group were lower significantly while compared with the the expression of α-actin, MHC, Myogenin and MyoD of the transected group, that of the MPC transplantation group were higher significantly(P < 0.05). Conclusion The transplantation in vivo of allogenic mesenchymal progenitor cells is effective for delaying denervated muscle atrophy.

Effect of 5-aza-2’-deoxycytidine on CHD5 gene expression and proliferation of Human Colon Carcinoma cell

2013, 33(8):  976-980. 

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Objective To study the effect of 5-Aza-2-deoxycytidine (AZA) on re-expression of the hypermethylated and silenced CHD5(Chromodomain, helicase, DNA-binding 5) gene in human colon carcinoma cell and cell proliferation. Methods Lovo and SW480 cells were exposed to 5μmol/L AZA for 72h, and then CHD5 expression was determined by quantitative RT-PCR (qPCR), methylation status of CHD5 genes was determined by bisulfite sequencing PCR (BSP), and the proliferation of the cells was evaluated by MTT assay. Results After treatment with 5μmol/L AZA, the promoter region of the CHD5 gene exhibited a demethylation status, and CHD5 mRNA was re-expressed．Meanwhile, the growth of Lovo and SW480 cells were reduced. Conclusions AZA could reverse methylation of CHD5 gene to regulate the expression of CHD5 gene, and could effectively inhibit the cellular proliferation of human colon carcinoma cell.

Effects of shRNA targeting COL1A1 gene on the invasion and migration of human breast cancer MDA-MB-231 cell line in vitro

2013, 33(8):  981-985. 

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Objective To investigate the effects of shRNA-mediated human collagen type 1 alpha 1 （COL1A1）gene silence on the migration and invasion of Breast Cancer Cell Line MDA-MB-231. Methods Two shRNA vectors and one negative control vector were designed and stably transfected into MDA-MB-231 cells. Western blot analysis was used to screen for the group which had the highest inhibitory rate. Monolayer colony formation assay was performed to assess a single cell proliferation ability. The cell adhesion, migration and invasion potencies were observed by cell-matrix adhesion assay, and Transwell assay. Results Cells transfected with pshRNA-COL1A1-2 vector had the stronger inhibition of COL1A1 protein, with the inhibition rate being 66.98%±2.08%, and cells in this group were used for the subsequent experiments. Compared with control group and pshRNA-scramble group, the monolayer colony formation rate decreased obviously(P<0.05). The optical absorbance value of adherent cells and the number of invading and migrating cells which moved across the matrix barrier for pshRNA-COL1A1 group were less that than that of control group and pshRNA-scramble group(P<0.001). Conclusion COL1A1 gene silencing of human MDA-MB-231 cells could inhibit cell matrix adhesion, migration and invasion potencies in vitro.

The intracellular antiviral activities of a novel engineered M1GS ribozyme that targets to 5' untranslated region of hepatitis C virus genome

2013, 33(8):  986-991. 

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Objective Hepatitis C virus (HCV) is one of the major pathogens that lead to viral hepatitis. At present, Interferon treatment in combination with ribavirin is the first line clinical therapeutic approach. However, the responses are usually poor and the viral infection reoccurs. Therefore, exploring new antiviral agents and therapies is under urgent needs. Methods The sequence and structure of the 5' untranslated region of HCV genome were analyzed through the two computer software, DNAMAN and RNA Structure. The cytosine 67 nt downstream of the first base of HCV genome RNA was identified as the optimal target cleavage site. Based on the flanking sequence of this assumed cleavage site, a guide sequence (GS) was designed and covalently linked to the 3 prime terminus of the M1 RNA, which is catalytic subunit of the RNase P derived from Escherichia coli using PCR. We named this new targeting ribozyme M1GS-HCV/C67 and it antiviral activities were analyzed in cultured cells. Results In the in vitro cleavage assay, The M1GS-HCV/C67 ribozyme could effectively cleave the HCV target RNA into two fragments at the specific cleavage site. Moreover, comparing to the blank control, this engineered M1GS ribozyme could reduce the core protein expression of more than 75% in the HCV-infected host cell and lead to a 800-fold reduction of HCV RNA copies in the culture supernatant. An another M1GS ribozyme, M1GS-HCV/C67*, which has the same guide sequence but does not contain the bridge sequence, did not exhibit apparent inhibition for the expression of HCV core gene and viral proliferation in our paralleled assay . Conclusion We successfully constructed an M1GS ribozyme showing affective and specific cleavage of target viral RNA. Further results showed that the engineered ribozyme had notably antiviral activity in cultured cells, thus provided a new promising approach for clinical anti-HCV therapeutic strategy.

The effect of lidamycin against mouse myeloma in vivo and in vitro

2013, 33(8):  992-997. 

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Objective This study is to investigate the effect of Lidamycin (LDM) against mouse myeloma in vivo and in vitro and provide the basis for its use in therapy. Methods MTS assay was used to detect SP2/0 cell proliferation. Cell cycle distribution and cell apoptosis were measured by flow cytometry combined with propidium iodide (PI) staining. The expression of proteins was determined by Western blot analysis. In vivo antitumor activity was performed using a murine myeloma model in BALB/c mice. Results There was a significant reduction of proliferation in the cells treated with LDM. The overall growth inhibition correlated with apoptotic cell death. LDM induced cell apoptosis was associated with decrease of NF-κB, Bcl-2, and Survivin. LDM induced G2/M phase arrest through increase the expression of p27 protein in SP2/0 cells. LDM markedly suppressed tumor growth in murine myeloma model. Conclusion LDM demonstrates a significant antitumor efficacy against myeloma SP2/0 in vivo and in vitro. Taken together, our data provided some clues for further research of the effects of LDM on human multiple myeloma.

Hepatic stellate cell through SDF-1/CXCR4 axis induces epithelial–mesenchymal transition in hepatocellular carcinoma invasion

2013, 33(8):  998-1003. 

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Objective Recent studies have shown that hepatic stellate cell in tumor microenviroment is an important regulator for hepatocellular carcinoma cell behavior. The aim of this study is to explore the impact of hepatic stellate cell in hepatocellular carcinoma invasion through SDF-1/CXCR4 axis. Methods The expressions of SDF-1 and CXCR4 were examined in hepatic stellate cell LX02, four hepatocellular carcinoma cell lines by Western blotting at protein levels and Real-time RT-PCR at mRNA level respectively. In addition, Transwell invasion assay was carried out to analyze the influence of hepatic stellate cell LX02 and SDF-1 on invasion of hepatocellular carcinoma cell HepG2 under normal condition or CXCR4 gene silence condition. Then, Western blotting was performed to evaluate the expression of epithelial mark E-Cadherin and mesenchymal mark vimentin. Results The expression of SDF-1 was high in hepatic stellate cell LX02, and increased levels of expression of CXCR4 were found in all hepatocellular carcinoma cells. Co-culture with hepatic stellate cell LX02 or treatment with SDF-1 both induced the epithelial–mesenchymal transition and increased the invasion of hepatocellular carcinoma cell HepG2. Furthermore, inhibition of CXCR4 by gene silence in HepG2 suppressed the enhanced invasion and epithelial–mesenchymal transition of HepG2 cells which induced by stellate cells or SDF-1. Conclusion Hepatic stellate cells promote hepatocellular carcinoma cell invasion through chemokine SDF-1/CXCR4 axis, the mechanism may involve the epithelial-mesenchymal transition of carcinoma cell.

Association of paired box4 gene Arg121Trg polymorphism with type 2 diabetes mellitus

2013, 33(8):  1004-1008. 

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Objective To investigate the association of paired box4 (Pax4) gene Arg121Trg single nucleotide polymorphism with impaired glucose tolerance in Kunming, Yunnan province. Methods 430 patients with type 2 diabetes mellitus(T2DM) , 200 people with impaired glucose tolerance (IGT) and 446 normal volunteers(NGT) were recruited in the study．Pax4 gene Arg121Trg single nucleotide polymorphism was identified by high resolution melting and DNA direct sequencing. The distribution of genotype and allele in three groups were investigated. We also observed the association of clinic phenotype with two subsets in type 2 diabetes mellitus group. Results 1) This study indicated that GG genotype and A allele frequencies of Pax4 gene Arg121Trg variant were 0.909, 0.047 in T2DM group, 0.935 and 0.035 in IGT group , 0.939 and 0.03 in NGT group respectively. 2) Among 430 people with type 2 diabetes mellitus, rate of insulin users was higher in GA/AA genotype compared with the patients with GG genotype (P=0.001). No significant difference of other clinical data was observed in T2DM patients with different genotype. Conclusions A allele of Pax4 gene Arg121Trg polymorphism might be a molecular marker for β cell function progressively deterioration in Kunming T2DM patient, Yunnan province.

High concentration of glucose increases apoptosis of Schwann cell by downregulation of autophagy

2013, 33(8):  1009-1013. 

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Objective To observe the effect of the high concentration of glucose on autophagy of rat’s Schwann cells in vitro and the effect of autophagy on the proliferation and apoptosis of Schwann cells cultured in high glucose medium. Method RSC96 cells were cultured. The expression of beclin1 and caspase-3 were detected by immunofluorescence and Western blot. The proliferation was detected by MTT. Result The expression of beclin1 in RSC96 cells cultured in high glucose medium was reduced (P<0.05), as well as the proliferation was decreased (P<0.05, P<0.01). Inhibition of autophagy resulted in aggravating the decrease of the proliferation of RSC96 cells cultured in high glucose medium (P<0.01) and upregulating the activity of caspase-3 (P<0.01). Conclusion The impairment of the high concentration of glucose increased apoptosis by downregulation of autophagy.

Bone morphogenetic protein 4 affects angiogenesis in vitro of human umbilical cord vein endothelial cells through ERK/MAPK pathway

Qin WANG, ZHANG Hong-gang ZHANG Qiu-ju Rui-juan XIU

2013, 33(8):  1014-1018. 

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Objective To observe the effects of BMP-4 on HUVECs proliferation, migration and capillary sprouting, and signal pathway involved in these biological behaviors. Methods HUVECs were treated with BMP-4. HUVEC proliferation was determined by BrdU incorporation, wound-healing was applied to observe HUVEC migration. Cell lysates were analysed by Western blotting for ERK1/2 phosphorylation. Results Treatment with BMP-4 results in dose dependent inhibition of HUVEC proliferation, migration and capillary sprouting at lower concentrations(5、10 and 20 ng/mL), but at higher dose of BMP-4(50 and 100 ng/mL), this inhibition disappeared. Conclusions HUVEC proliferation , migration and capillary sprouting induced by BMP-4 might be dependent on ERK phosphorylation.

Observation of the spinal microcirculation in rat through a closed spinal window by fluorescence microscopy

2013, 33(8):  1019-1023. 

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Objective To investigate the feasibility and efficacy of improved closed spinal window applied to assess the rat dorsal pial microcirculation by fluorescence microscopy. Methods On the 8-week-old SD rats, we made a precise improved closed spinal window with a laminectomy at the T10 level using a dental acrylic resin and a cover glass. The window was placed over the back of the rat with intact dura. Pasted the dental acrylic resin onto the spinous processes at the T9 and T11 as fixed aids. Collect 100μL hematocrit red blood cells (BRC) by centrifugation (2000×g). The RBC labelled by Dil (5 mg/L) in vitro were injected in jugular vein. Leukocytes labelled with rhodamine 6G were injected intravenously (0.3 g/L). The spine was fixed with vertebral fixation device. Local microcirculation is recorded in real time through the window. Results Microvascular morphology, distribution and blood vessel density can be observed. Dil-labeled RBC velocity and adhesion were observed. Leukocyte adherence to pial vessels and assessment of perfusion were observed with the help of rhodamine 6G. Conclusion By improved closed spinal window and vertebral fixation device, we observed the microcirculation in the rat pia.

Reticulocyte Hemoglobin Content in screening for iron deficiency in women

2013, 33(8):  1024-1027. 

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Objective To examine the utility of the reticulocyte hemoglobin content (CHr) in screening for iron deficiency in women. Methods 506 female subjects in department of medical examination were selected, and all the subjects were divided into iron deficiency group (iron deficiency without anemia group and iron deficiency anemia group, 121 cases), and healthy controls group(394 cases). The levels of red blood cell counting(RBC), hemoglobin(HGB), hematocrit(HCT), red blood cell volume distribution width(RDW), serum ferritin (SF), serum iron(SI), and CHr were determined. Results The levels of CHr(27.2 ± 3.0) in the iron deficiency group were significant lower than that (30.6 ±1.3) in the control group, P<0.001. Receiver operator characteristic curve analysis demonstrated that the area under the curve of CHr was 0.814 and significantly greater than RBC and HGB and HCT and MCV and MCH and MCHC and RDW in the diagnosis of iron deficiency in women, P<0.05. Conclusions CHr could be used for screening for and early diagnosis of iron deficiency in women.

Reduced expression of Toll-like receptors 3 in peripheral blood mononuclear cells of chronic hepatitis B patients

2013, 33(8):  1028-1031. 

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Objective To investigate the Toll-like receptor 3 (TLR3) expressions in peripheral blood mononuclear cells of chronic hepatitis B patients and explore its clinical significance. Methods The peripheral blood were collected from chronic hepatitis B patients and healthy volunteers, serum HBV DNA replication levels were detected by quantitative PCR; TLR3 mRNA and protein expression levels in peripheral blood mononuclear cells were detected by RT-PCR, flow cytometry and Western blot respectively; the serum levels of TNF-α and IFN-β were measured by ELISA. Results The TLR3 expression in peripheral blood mononuclear cells of chronic hepatitis B patients was significantly lower than that in healthy volunteers and related to the level of serum HBV DNA replication; the serum TNF-α，IFN-β concentrations of chronic hepatitis B patients were significantly lower than that in healthy volunteers and related to the level of serum HBV DNA replication level. Conclusion The TLR3 expression in peripheral blood mononuclear cells of chronic hepatitis B patients was associated with the proliferation of hepatitis B virus.

The inhibition effect of Juglone on proliferation of lung cancer A549 cells through lowering AKT activity

2013, 33(8):  1032-1037. 

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Objective To explore the effect and mechanism of PIN1 inhibitor Juglone on cell proliferation of human lung cancer cells A549 in vitro. Methods A549 cells were treated by different concentrations of Juglone. Cell proliferation was measured by MTT assay respectively. The apoptosis rate and cell cycle of A549 cells were detected by Flow cytometry. Real-time PCR was carried to detect the mRNA expression of PIN1 and AKT. Western blot were used to observe the protein expression of PIN1 and AKT-pS473 in cells. Results With the increasing of Juglone’s concentration, the inhibition effect on proliferation and the apoptosis rate of A549 cells enhanced in a dose-depended manner, and A549 cells cycle were blocked in G2/M and S period. Real-time PCR result showed the mRNA level of PIN1 in A549 cells treated by Juglone significantly decreased compared with control group. Western blot showed that the protein expression of PIN1 in Juglone groups were 1.032±0.056, 0.892±0.024, 0.596±0.023, 0.396±0.021, significant lower than control group 1.280±0.046 (P＜0.05).the protein expression of AKT-pS473 in Juglone groups were 0.554±0.023, 0.464±0.018, 0.362±0.015, 0.228±0.020,significant lower than control group 0.626±0.015(P＜0.05). the protein expression of PIN1 and AKT-pS473 in cells treated by Juglone for 48h were 0.575±0.036 and 0.338±0.014, significant lower than 0.764±0.032 and 0.436±0.023 of 24h group(P＜0.05). Conclusion Inhibiting the expression of PIN1 with Juglone can significantly depress the proliferation activity of A549 through weakening the phosphorylation level of AKT.

The Hans，Choi immunophenotyping and c-myc genetic features between pediatric and adult diffuse large B-cell lymphoma

2013, 33(8):  1038-1042. 

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Objective To explore the difference of Hans, Choi immunophenotyping and c-myc gene characteristics between pediatric and adult diffuse large B-cell lymphoma. Methods To Collect 60 cases of DLBCL clinicopathological data and follow-up, 17 cases of children group, adult group of 43 patients. To observe CD10, bcl-6, MUM1, FOXP1, GCET1 and CD5 protein expression by immunohistochemical SP method and accord to Hans and Choi immunophenotyping standard genotyping. To detect c-myc gene circumstances Interphase fluorescence insituhybridization (FISH) . Results 1) Hans typing：the Children DLBCL group of GCB were 11 cases, non-GCB type were 6 cases； the GCB type in the adult DLBCL group were nine cases, non-GCB type were 34 cases. 2) Choi typing：the Children DLBCL group of GCB type were 13 cases, ABC type were 4 cases； the adult DLBCL group of GCB type were 13 cases, ABC type were 30 cases (P = 0.008). 3) c-myc gene：The c-myc gene disruption of children DLBCL were 5 cases; c-myc gene in 43 adult cases were normal(P=0.006). Conclusions Child DLBCL are mainly GCB-type with better prognosis；but non-GCB-type are more common type in adult with worse prognosis.The c-myc gene disruption in Children DLBCL was significantly higher than that of adult ones.

HGF and G-CSF induce the differentiation of rat bone marrow mesenchymal stem cells into hepatocyte-like Cells

2013, 33(8):  1043-1049. 

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Objective To investigate if the hepatocyte growth factor（HGF）and granlocyte-colony stimulating factor (G-CSF) could induce differentiation of rat bone marrow mesenchymal stem cells（BM-MSCs）into hepatocyte-like cells .Methods The BM-MSCs were isolated and cultured in- vitro by adherent method，then identified by flow cytometry.The BM-MSCs of the third passage were used as four groups - control group（10%FBS）and intervation groups（0.1μmol/L G-CSF;20ng/mLHGF;0.1μmol/L G-CSF+20ng/mLHGF).The mRNA and protein were extracted on 7,14,21 days，then detected by RT-PCR and Western-blot. Results.The third passage BM-MSCs of intervation groups (20ng/mLHGF and 0.1μmol/L G-CSF+20ng/mLHGF) got hepatocyte like feafure after induction.The results tested by Flow cytometry analysis were as follows: CD34-PE0.3%,CD45-FITC0.1%,CD90-FITC99.6%,CD105-PE99.8%. The mRNA and protein of ALB and AFP were detected respetively by RT-PCR and Western-blot on day 7,14,and 21 from inervation groups (10%FBS+20ng/mLHGF;10%FBS+0.1μmol/L G-CSF+20ng/mLHGF), however ,the amount of ALB increased as time went by ,while AFP decreased to extremely low on day 21.Meaningfully,the difference between the two groups is significant（P<0.05）. Conclusion HGF can induce the differentiation of BM-MSCs into hepatocyte-like cells, which can be significantly enhanced by G-CSF.

Down-expression of cystathionine-?-synthase in the brain of spontaneous hypertensive rat

2013, 33(8):  1050-1051. 

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Metformin can decrease the expression of FASN in MCF-7 cultured in high glucose and insulin

2013, 33(8):  1052-1053. 

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RAG gene and B cell lymphoma

2013, 33(8):  1054-1058. 

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Recombination Activating Gene(RAG) plays an important role in diverse and complex DNA recombination of B cell antigen-receptor determining regions. The possible mechanisms of RAG genes inducing B cell lymphoma is explained in four aspects as follows: 1)RAG complex can recognize and cut the main breaking region of Bcl-2 gene which will lead the translocation of t(14,18) and promote the lymphoma; 2)the synergy effect between RAG-1/2 genes and AID also induce the translocation of chromosome; 3)extraordinary of V(D)J recombination because of the abnormal degradation of RAG2 protein; 4)EBNA-1 expressing in EBV infected mice may induce the re-expression of RAG1/2 genes.

Estrogen and matrix metalloproteinase(MMPs)

2013, 33(8):  1059-1062. 

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Many research results have shown that the effect of estrogen on matrix metalloproteinase is very complex. For example, the influence on different matrix metalloproteinases under different circumstances in different histiocytes is different. In addition, the effects can be observed on the levels of messenger RNA and the expression or the activation of proteins.

Recent advances of antimicrobial peptides in dermatology

2013, 33(8):  1063-1066. 

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Antimicrobial peptides have been shown to serve as immediate first-line defense against potential pathogens in the skin. The antibacterial mechanism of cationic antimicrobial peptides in skin disease process has been extensively studied，but anionic antimicrobial peptides is relatively unknown. The role of antimicrobial peptides in epidermal immunity and low predilection for developing bacterial resistance has been studied extensively thus far, the potential health benefits of the use of synthetic bioactive peptides is promising in dermatological disease. But some technical obstacles must be overcome before they can be successfully translated into therapeutics.

Progress of inducing stem cells into dopamine neurons in vitro

2013, 33(8):  1067-1070. 

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Given limits of traditional medical and surgical treatments in the progress of Parkinson’s disease, the therapeutic strategy of cell transplantation to replace with loss dopamine neurons has received more attention gradually. Many researches have domenstrated stem cells could be induced into dopamine neurons in vitro through different methods. Here, recent data and findings are reviewed. By evaluation, better methods and cell sources could be found and further investigated in future, which may facilitate translation of stem cell transplantation techniques into a clinical treatment for Parkinson’s disease.

The progress of researches on the role of dendritic cells in gastric cancer

2013, 33(8):  1071-1074. 

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The dendritic cell is the most powerful antigen-presenting cell, which is important in the process of antigen capture and antigen presenting. Dendritic cells are not only associated with gastric carcinogenesis, progression and prognosis, but also have application in the treatment of gastric cancer which has been confirmed.

A Clinically Oriented Anatomy Lecture Program

2013, 33(8):  1075-1078. 

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In order to cooperate with Anatomy teaching reform, the department proposed a new teaching reform plan to combine the teaching of anatomy and clinical medical education. The reform includes re-arrangement of curriculum, application of multiple teaching methods, exchange and collaboration with foreign medical schools. A novel ‘Clinically-Oriented Anatomy Lecture’ program is launched. These programs not only strengthened the anatomy knowledge, but also emphasized the application of the knowledge in clinical application, and preliminarily achieved a good effect.