Table of Content

05 June 2013, Volume 33 Issue 6

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Considerations of the tension between doctors and patients

2013, 33(6):  643-647. 

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The increasing tension between doctors and patients in recent years has attracted more and more attention. The problem troubling previously only healthcare industry has gradually become a social issue. In the process of healthcare reform, this issue affects not only the healthy development of healthcare industry, but also social harmony and stability. In this paper, the authors analyzed relevant data and examples to reveal the factors causing the tension between doctors and patients and proposed solutions for those factors, aiming to help with easing the growing tension in doctor-patient relationship.

DJ-1 protect against rotenone-induced cell injuries via enhancing autophaty in MN9D cells

2013, 33(6):  648-654. 

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Objective To explore the protective effect of DJ-1 on MN9D cells and its mechanisms. Methods Transiently transfected wild type DJ-1 into MN9D cells. The MN9D cell viability and cell apoptosis level were observed by MTT assay and flow cytometry, respectively. The numbers of LC3-positive puncta were observed by confocal images. The ratio of LC3II to LC3I was detected via Western blot. Results DJ-1 overexpressing could attenuate decreased cell viability induced by rotenone treatment (P<0.05). The increased of apoptosis caused by rotenone was diminished by overexpression of DJ-1 (P<0.05). Overexpressing of DJ-1 could increase the numbers of LC3 positive puncta(P<0.001) and increase the ratio of LC3II to LC3I (P<0.001) in MN9D cells treated with rotenone. Giving 3-MA could make the numbers of LC3 positive puncta down to the basal level and reduce the ratio of LC3-II/I (P<0.05). Meanwhile, giving autophagy inhibitor 3-MA, the protective effects of DJ-1 on MN9D cells were disappeared. Conclusion DJ-1 upregulated the level of autophagy can relieve the toxin caused by rotenone in MN9D cells.

Recombinant human apolipoprotein(a) carboxyl terminal kringles inhibited angiogenesis

2013, 33(6):  655-660. 

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Objectives To characterize some purified recombinant Apo(a) kringles expressed by Pichia pastoris and to illustrate their antiangiogenic and antitumorogenic capacities. Methods Two recombinant proteins RHAKA (kringle V) and RHAKB (kringle IV type 10 and kringle V) were expressed by Pichia pastoris. Both RHAKA and RHAKB, recombined into pPICZ?A, were secreted by Pichia pastoris X-33. Recombinant proteins were concentrated and dialyzed before His?Tag affinity chromatography. Six amido terminal amino acids of RHAKB were analyzed through sequencing the purified protein from reverse-phase high performance liquid chromatography. We’ve also illustrated several important characters of recombinant proteins, such as glycosylation and disulfide bonds formation. Finally, recombinant proteins’ influence on in vitro cellular proliferation and in vivo angiogenesis of chick embryo chorioallantoic membrane (CAM) were tested. Results We observed that Pichia pastoris as an expression host could not only express recombinant proteins at a high level but modify them well. Both RHAKA and RHAKB could inhibit angiogenesis in vitro or in vivo, but no such inhibitory effect on cultured carcinoma cells. Conclusion Recombinant Apo(a) carboxyl terminal kringles expressed by Pichia pastoris could inhibit angiogenesis significantly.

Identification of a novel DNA aptamer against IFN-γ

2013, 33(6):  661-665. 

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Objective: To develop IFN-γ DNA aptamer that may potentially serve as a binding ligand in novel aptamer-based IFN-γ detection. Methods: A single-stranded 59nt DNA library containing 21 random oligonucleotides was synthesized in vitro. A new aptamer B3-8 was developed by SELEX technique with IFN-γ protein as target. Flow cytometry was performed to monitor the enrichment of aptamer pool and to evaluate the binding properties of B3-8. The structure of B3-8 was predicted by MFold software. Results: The aptamer B3-8 bound to the IFN-γ with a Kd of nmol/L, and had minimal cross reactivity with BSA. Conclusions: B3-8 may have application potentials in IFN-γ assay.

Effect of miR-23a on LPS-induced pro-inflammatory cytokines expression in mouse macrophage

2013, 33(6):  666-670. 

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Objective To investigate the effect of miR-23a on the pro-inflammatory cytokine expression in mouse peritoneal macrophage stimulated with LPS. Methods Real-time PCR was used to analyze the expression of miR-23a in mouse macrophage treated with LPS and the expression of pro-inflammatory cytokines including IL-1?、IL-6 and TNF-??in mouse peritoneal macrophage transfected with miR-23a mimics or inhibitors；ELISA was used to measure the IL-6 secretion. Results After LPS stimulation, the expression of miR-23a in the mouse peritoneal macrophage was remarkably decreased. While the cells transfected with miR-23a mimics, the expression and secretion of IL-6 was significantly increased(P <0.05), with no notable influence on IL-1??and TNF-??expression??And after transfected with miR-23a inhibitors, the expression of IL-6 was decreased. Conclusions LPS stimulation can suppress the expression of miR-23a. MiR-23a can promote the expression of pro-inflammatory cytokine IL-6, but have no notable influence on IL-1? and TNF-??

Value of positive labial salivary glands pathology results in diagnosing Sj?gren’s syndrome

2013, 33(6):  671-674. 

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Objective To detect the effect of positive labial salivary glands pathology results derived from different methods for diagnosis of Sj?gren’s syndrome(SS). Methods From January 2009 to October 2010, 21 patients with SS and positive labial salivary glands pathology were studied. 10 of the biopsies were focal lymphocytic sialadenitis (FLS), with focus score (FS) ≥1 (group A). The presence of focal infiltrates of lymphocytes in labial gland interstitial tissue was defined as positive in the other 11 biopsies (group B). Clinical data were collected, and student's t-test, chi-square test were used to evaluate the effect of positive labial glands pathology results derived from different methods for diagnosis of SS. Results 1) The positive rate of arthralgia was significantly higher in group A than group B (p<0.01). 2) Group A’s left eye BUT was lower than group B, and the number of corneal fluorescein staining spot was more than group B, while the positive rate of left corneal staining was higher than group B (p<0.05). 3) There was no difference between two groups in positive rates of high titer ANA and anti-SSA/SSB antibodies. Conclusion The labial salivary glands pathology derived from the two different methods had no obvious difference in diagnosing SS.

The role of iNOS in cardiomyocyte apoptosis of rat trauma model

2013, 33(6):  675-679. 

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Objective To observe the role of iNOS in cardiomyocyte apoptosis of rat trauma model. Methods Trauma rats model was established by Noble-Collip drum. Cardiomyocyte apoptosis was determined by Caspase-3 activity and DNA ladder detection. The protein expression if iNOS was investigated by Western blot. Nitric oxide (NO) content in myocardial tissue was detected by chemiluminescene method. Myocardial nitrotyrosine (NT) content was measured by ELISA. Results Cardiomyocyte apoptosis was increased in mechanical trauma (P<0.01). Myocardial iNOS expression obviously increased after trauma (P<0.01). In addition, the product of NO2-/NO3- and NT, was elevated significantly in myocardial tissue after trauma (P<0.01). 1400W, a selective inhibition of iNOS, significantly decreased cardiomyocyte apoptosis(P<0.05), and the product of NO2-/NO3- and NT (NO2-/NO3-: P<0.01; NT: P<0.05). Conclusion iNOS caused cardiomyocyte apoptosis by increasing the levels of NO and peroxynitrite acid in myocardial tissue.

The effect of long non-coding RNA-TP53TG1 on glucose deprivation stress response in glioma cells

2013, 33(6):  680-684. 

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Objective To construct full length clone of TP53TG1 into eukaryotic expression vector, and to explore its overexpression effect on glucose deprivation stress response. Methods TP53TG1 was amplified from human glioma cell by RT-PCR, and the eukaryotic expression clone of TP53TG1 was constructed. Then, its expression in U87MG cells was detected by real time PCR. Furthermore, we overexpressed TP53TG1 and meanwhile treated with low glucose (0.3g/L, 8h) in U87MG cells, and measured the expression of GRP78, IDH1 and PKM2 mRNA by real time PCR. Results TP53TG1 eukaryotic expression clone was successfully constructed. After the clone was transfected into U87MG cells for 36 hours, green fluorescence was seen. The expression of TP53TG1 was increased by 2.9x106 times in U87MG cells (P<0.05). As a result of over expression of TP53TG1 and low glucose treatment simultaneously in U87MG cells, GRP78 and IDH1 mRNA expression were significantly increased (P<0.05), while PKM2 mRNA significantly reduced (P<0.05). Conclusion Results suggest that TP53TG1 may be involved in the stress response of U87MG cells under glucose deprivation through influencing the expression of GRP78, IDH1 and PKM2 mRNA.

Oxidized low density lipoprotein down-regulates expression of Rictor in vascular endothelial cells

2013, 33(6):  685-689. 

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Objective To investigate the effect of oxidized low density lipoprotein on the expression of Rictor in cultured vascular endothelial cells. Methods Human umbilical vein endothelial cells were isolated and cultured and incubated with oxidized low density lipoprotein (10, 20, 40, 80mg/L) for 24h. RT-PCR and Western blot were employed to detect the expression of Rictor mRNA and protein. The phosphorylation of Akt and eNOS were analyzed with Western blot. The contents of NO2-/NO3- in the culture medium were measured by nitrate reductase assay. Results Oxidized low density lipoprotein significantly down-regulated expression of Rictor mRNA and protein in endothelial cells (P<0.01) and decreased the amount of rictor-mTOR complex. In endothelial cells transfected with Rictor, increased expression of Rictor enhanced the phosphorylation of Akt and eNOS, and restored endothelial NO production. Conclusions The inhibition of Rictor expression by oxidized low density lipoprotein may play an important role in promoting endothelial dysfunction.

Screening and potential functional analysis of substrates of neuronal adapter protein Dok6

2013, 33(6):  690-693. 

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Objective To screen the protein which can interact with Dok6 and explore the potential function . Methods Immunoprecipitation, western blot, and sliver staining were employed to screen the candidate proteins interact with Dok6. liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to identify the binding proteins of Dok6. Finally, we analyzed its potential physiological functions by bioinformatics prediction. Results After screening the binding proteins of Dok6 with IP and LC-MS/MS, 26 candidate binding proteins were identified. These proteins are involved in many physiological processes in the body. In order to narrow the scope of the study, we selected nine proteins associated with nervous system development and intracellular signal transduction as object of study for following study of the function. Conclusions The potential Dok6 downstream proteins identified by mass spectrometry and bioinformatics will provide some clues to determine physiological functions and related molecular mechanisms of Dok6 .

Screening and identification of long noncoding RNAs in PCBP2 gene locus

2013, 33(6):  694-698. 

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Objective Screening and identification of long noncoding RNAs in PCBP2 gene locus based on ESTs. Methods Screening ESTs with introns transcription, with the analysis of CPC and statistical comparison, four ESTs were recognized as potential lncRNAs（two of the four are human ESTs, the other two are mouse ESTs）, those ESTs were identified with polymerase chain reaction( PCR ) and then cloned in pGEM-T Vector-System for sequencing. We test the express of these ESTs in human cell lines or mouse tissue. Results The four ESTs are potential long noncoding RNAs, three out of four ESTs were identified, their expression were different. Conclusions With a new method for searching lncRNAs based on ESTs, we recognized some potential long noncoding RNAs, one of which may be a functional lncRNA influence mouse brain development.

ERK1/2 and JNK involved in the inhibitory effect of DL0805 on vasoconstriction induced by angiotensin II in the rat thoracic aortic rings

2013, 33(6):  699-703. 

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Objective To observe the effect of DL0805 (a Rho kinase inhibitor) on angiotensin II (Ang Ⅱ) -induced vascular contractions in the rat thoracic aortic rings and to investigate the possible mechanism. Methods The isometric vascular tone was measured in both endothelium-intact and endothelium-denuded rat thoracic aortic rings. Phosphorylations of p44/42 extracellular signal-regulated kinase (ERK1/2) and c-Jun amino-terminal kinase (JNK), and protein level of angiotensin type 1 receptor (AT1R) in rat aortic rings were detected by Western blot. Results DL0805 (10、25 and 50 μmol/L) inhibited Ang Ⅱ（100 nmol/L）-induced vascular contractions in both endothelium-intact and endothelium-denuded rat aortic rings in a dose dependent way （P < 0.01, P < 0.001）. Furthermore, activation of ERK1/2 and JNK by Ang Ⅱ（100 nmol/L）stimulation was significantly inhibited by DL0805 (25 and 50 μmol/L) in both endothelium-intact and -denuded rat aortic rings （P < 0.05, P < 0.01 and P < 0.001）. However, the protein level of AT1R in response to Ang Ⅱ was not affected by DL0805 (5、25 and 50 μmol/L) in rat aortic rings. Conclusion DL0805 inhibits Ang Ⅱ-induced rat aortic rings contraction and the mechanism involves the inhibition of Ang II-induced ERK1/2 and JNK activation.

Stability of haemoglobin A1c in different Storage temperature and time

2013, 33(6):  704-707. 

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Objective To investigate the stability time of HbA1c samples in 4 different storage temperature conditions. Methods 20 samples with different HbA1c levels were collected. Each sample is thoroughly mixed and distributed into 45 parts. In each level group, a random selected sample was measured immediately, and the remaining samples were immediately grouped and stored at room temperature (22 ~ 25℃), refrigerated (2 ~ 8℃), frozen (≤-20℃), cryopreservation (≤-70℃), which were measured at 1,2,3,7,15,20 day and 1,2,3,6 month and 1y respectively. HbA1c levels were detected by HPLC method. The changes of average level of HbA1c versus time with 4 different temperatures were observed and the stability of HbA1c was evaluated by different method or judgment. Results Judge by different less than 6%, HbA1c was stable for 1 year at -70℃; HbA1c in low concentration levels was stable for 7 days at -20℃, while in high concentration levels could be stable for 20 days with 11.2% average annual decline rate. HbA1c at room temperature and refrigerated storage conditions could be stable for 3 days, till 7 days, HbA1c level dropped by average -9.1% and -7.1% respectively. Conclusions This study provides a basis to formulate and improve HbA1c specimens preservation conditions. The Cryopreservation is more favorable for large-scale epidemiological studies of HbA1c.

Risk factors of postoperative cardiac events in elderly CAD patients undergoing non-cardiac surgery

2013, 33(6):  708-712. 

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Objective To determine the preoperative and intraoperative predictors of postoperative cardiac events (PCE) in elderly patients with coronary artery disease (CAD) undergoing non-cardiac surgery. Methods 1510 elderly CAD patients, aged 60 years and older, who underwent intermediate- to high-risk non-cardiac surgery at five academic medical centers in China, were enrolled in this clinical prospective observational study from Jan. 2008 to Dec. 2009. Clinical variables were evaluated, The PCE were followed by the examinations of 12-lead ECG and Troponin I(cTnI) levels. Results PCE occurred in 104 patients (6.9%). Cardiac death occurred in 9 patients (0.6%). The independent risk factors were age ≥ 75 years old, history of MI, high-risk surgery, intraoperative hypoxemia, and intraoperative hypotension. Conclusions Age ≥ 75 years old, or history of MI, or high-risk surgery could increase the risk of PCE in CAD patients individually. Preventing intraoperative hypoxemia and hypotension may reduce the occurrence of PCE in these high risk patients.

Over-expression of estrogen receptor-alpha 36 promotes the invasion and metastasis ability of human breast cancer cell MCF-7

2013, 33(6):  713-717. 

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Objective To explore the effect of ER-α36, a novel variant of estrogen receptor α, on the invasion and metastasis potential in the human estrogen receptor-positive cell and its mechanism. Methods Wild type breast cancer cell line MCF-7 (MCF-7/W), MCF-7 cells transfected with pcDNA3.1 empty vector cells (the MCF-7/pcDNA3), MCF-7 cells transfected with the recombinant vector pcDNA3.1/ER-α36 cells (the MCF-7/ ER-α36) were the research objects. The adhesion between tumor cells and extracellular matrix was observed by cell adhesion test. The cell invasion and metastasis was detected by Transwell chamber experiments. The expression levels of NF-κB P65, MMP-2, MMP-9, and TIMP-1 were analysed by Western blot. MCF-7 / ER-α36 cells. Results Compared with MCF-7 / W group, the adhesion rate after growing 2h and the number of penetrating cells after growing 24h were increased had significant differences (P <0.05). The Western blot analysis results showed that the relative expression level of NF-κ P65, MMP-2, MMP-9, TIMP-1 and MMP- 2/TIMP-1, MMP-9/TIMP-1 in MCF-7/ERα36 group had a significant difference (P <0.05). Conclusions ER-α36 can promote the ability of the cell invasion and metastasis potential in the ER-α66-positive breast cancer cells. The mechanism possibly is ralated with increasing the expression level of MMP-9 and MMP-2 through NF-κB pathway, and breaking the balance between this and TIMP-1.

Berberine inhibits myocardial inflammatory in rat with chronic heart failure

2013, 33(6):  718-721. 

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Objiective To detect the expression of Toll Like Receptors4 (TLR4) in experimental model rat with chronic heart failure (CHF), and investigate the possible mechanism of berberine treating inflammation response of CHF. Methods Rat model of CHF was established by intraperitoneal injection of adriamycin with 2.8 mg/kg per week for 10 weeks. The rat model of CHF were randomly divided into berberine group,placebo group and normal group.Each group contained 10 rats. Berberine group was given berberine with 21 mg/kg per day for 4 weeks while other groups were given distilled saline water. Serum BNP, TNF-α,IL-1and IL-6 were assessed by ELISA,iNOS mRNA was measured by qRT-PCR,TLR4 protein expression and distribution were detected by immunohistochemistry. HE stain was performed to observe morphological features. Results Serum BNP,TNF-α,IL-1,IL-6 and iNOS mRNA level in berberine group were significantly attenuated compared with in placebo group (P<0.05).TLR4 Absorbance value in berberine and placebo group were enhanced compared with normal group (P<0.05), while in berberine group significant decreased compared with in placebo group(P<0.05). Conclusion Berberine can inhibit the inflammatory response in CHF may through down-regulation signal transduction of TLR4．

Hepatitis B virus(HBV) preS1 gene-specific anti-gene locked nucleic acid(LNA) significantly inhibits preS1 gene expression in vitro

2013, 33(6):  722-725. 

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Objective To investigate the inhibitory effects of hepatitis B virus(HBV) preS1 gene-specific anti-gene locked nucleic acid(LNA) on HBV replication and expression in HepG2.2.15 cells. Methods The anti-gene LNA which were complementary to the purine rich region of HBV preS1 gene were designed, synthesized, and transfected by cationic liposomes into HepG2 2.2.15 cells. The HBsAg,preS1-Ag and HBV DNA of supernatant was tested by time-resolved fluorescence immune assay(TRFIA) ,real-time fluorescent quantitative PCR(FQ-PCR) and enzyme linked immunosorbent assays(ELISA) at 1, 3, 5 and 7 d after treatment. LNA’s cyto-toxicity on cell was evaluated by MTT method. Results The anti-gene LNA that targeting on the purine rich region of HBV preS1 gene showed strong inhibitory effects on replication of HBV DNA and the expression of HBsAg and preS1-Ag with the inhibition rates of 65.99%,67.49% and 63.88% respectively after 7 days.There’s no obvious toxicity on cell. Conclusion anti-gene locked nucleic acid(LNA) that targeting on the purine rich region of HBV preS1 gene has show strong inhibition on HBV in vitro.It has a therapeutic potential in the treatment of patients infected with HBV.

Effects of insulin to the apoptosis and MAPK pathway of rat colonic smooth muscle cells

2013, 33(6):  726-730. 

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Objective To study the effects of insulin to the apoptosis and mitogen-activated protein kinase( MAPK ) signal transfer pathway of rat colonic smooth muscle cells ( SMCs ). Methods Enzymatic method for the isolation and culture of SD rat colonic SMCs; α-actin for immunohistochemical identification; division of the rat colonic SMCs into normal group, insulin group, and insulin+PD98059 group; MTT method for the detection of SMCs proliferation; flow cytometric Annexin V-FITC/ PI for the detection of SMCs apoptosis; and western blot for the detection of p-ERK, ERK, p-P38MAPK, P38MAPK and p-JNK, JNK expressions. Results Compared to the normal group, cells proliferation of insulin group is much more significant, apoptosis rate is reduced, p-ERK expression is enhanced, the ratio of p-ERK/ERK is increased( 110.36 ± 9.5 vs 50.92 ± 6.01 ) ( P < 0.01 ); and no differences for p-P38MAPK, P38MAPK, p-JNK and JNK expressions; while for the PD98059 group, cells proliferation is decreased significantly, apoptosis rate is increased, p-ERK expression is weakened, and the ratio of p-ERK/ERK is decreased( 15.69 ± 2.11 vs 50.92 ± 6.01 ) ( P < 0.01). Conclusion Insulin can promote proliferation and inhibit apoptosis in colonic SMCs through activation of the ERK route of MAPK pathway, P38MAPK and JNK routes may not be involved in this process.

Introduction to oxidized α1-antitrypsin quantitative detection kit and its use in testing Chinese male smokers

2013, 33(6):  731-735. 

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Objective To introduce a serum oxidized α1-antitrypsin (Ox-AT) quantitative detection kit, and analyze serum Ox-AT in Chinese male smokers. Method Set up a sandwich ELISA method with Ox-AT and a specific anti-Ox-AT monoclonal antibody. Analyze serum Ox-AT level of 20 pairs of smoking and non-smoking male. Results Ox-AT sELISA has a good linear in range of 0-500μg/L. The limit detection is 2.58μg/L, CV among batches is 11.27%, the accuracy is about 88.19%, the shelf life under 4℃ is more than 6 months. Detecting with this sELISA, serum Ox-AT level in smoking male is 55.43 ± 49.94μg/L, non-smoking control group is 11.01 ± 12.15μg/L, p <0.001. Conclusion The Ox-AT detection kit can be used for quantitative testing serum Ox-AT. Ox-AT might be more useful than α1-AT in evaluating Chinese COPD.

Drug-containing serum of Liu Wei Di Huang Wan reduces the damage of PC12 cells induced by KCl

2013, 33(6):  736-739. 

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Object To investigate the effect of the drug-containing serum of Liu Wei Di Huang Wan on PC12 cells which were damaged by KCl. Method We established the PC12 cell models induced by KCl which were divided into three groups: control group, model group, Liu Wei Di Huang Wan containing serum group. The morphology of cells in each group were observed with inverted phase contrast microscope. The proliferation of PC12 cells was detected by MTT assay and the concentration of LDH in PC12 cells were analysed. The level of CREB mRNA was analysed by RT-PCR. The expression of CREB was analysed by SABC-immunohistochemistry stain. Result The activity of LDH released from cells was increased and the level of CREB was decreased in the model group （P<0.01）compared with the control group. It significantly improved the activity of cells and the level of CREB mRNA in Liu Wei Di Huang Wan serum group compared with the model group （P<0.01）. Conclusion It can reduce the damage of PC12 cells induced by KCl by the drug-containing serum with Liu Wei Di Huang Wan.

Clinicopathological features of renal cancer in different ages

2013, 33(6):  740-743. 

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Objective To investigate the clinical pathological characteristics of different age patients with renal malignant tumor. Methods We analyzed the clinical and pathological data of 2,268 patients with renal malignant tumor admitted in PUMC hospital between 2002 and 2012. Patients were grouped according to age and pathological type. Results Among the 2,268 RCC patients, there were 260（11.5%），1178（51.9%），and 830（36.6%） patients aged <40, 40~59 and over 60 years old respectively. There were no significant differences in tumor size, and TNM stage by age group. However, patients <40 years old were significantly more likely to present with symptomatic tumors. There were no significant differences in histology by age; The surgical treatment of kidney cancer patients of different age groups was in different ways and the percentage of patients treated with partial nephrectomy declined with age. Conclusions the health examination of the youth crowd should be strengthened. the main form of treatment for kidney cancer is radical surgery, but we should pay attention to nephron sparing surgery.

Aristolochic acid induces apoptosis of human umbilical vein endothelial cells by suppressing the ERK1/2 signaling pathway

2013, 33(6):  744-748. 

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Objective To make sure whether the ERK1/2 signaling pathway plays an important role in aristolochic acid (AA) - induced the apoptosis of human umbilical vein endothelial cells (HUVECs). Methods Cell viabilities were determined by the MTT assays. Apoptotic morphologic changes of the treated cells were observed under fluorescence microscope. The proportions of apoptosis were assessed by flow cytometry. Western blotting was used to evaluate ERK1/2 phosphorylation. Results The MTT assays showed that AA significantly decreased the viabilities of HUVECs in a dose- and time-dependent manner. The flow cytometry showed that AA significantly increased the apoptosis of HUVECs in a dose-dependent manner. Microscopic apoptotic changes were also observed in these cells. Furthermore, AA inhibited ERK1/2 activation, which, however, was attenuated when the cells were pretreated with PD98095. A increase of apoptosis induced by AA was inhibited when the cells were pretreated with PD98095. Conclusion AA induces apoptosis of HUVECs via the ERK1/2 signaling pathway.

Optimization of the conditions for inducing mouse reticulocyte by hytrazinobenzene hydrochloride

2013, 33(6):  749-751. 

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Objective To choose the optimum conditions for inducing mouse reticulocytes by using hytrazinobenzene hydrochloride. Methods One percent hytrazinobenzene hydrochloride was injected to back subcutaneous tissue of the ICR and BALB/c mouse everyday. The dose was100 μl, 200 μl, 300 μl and 400 μl, respectively for 9 days. Then tail vein blood was taken every day, and observed. After staining the reticulocyte induced efficiency was statistical analyzed by using the bright tar blue staining. Results The morphology and proportion of reticulocytes were the best when the mice were injected 200 μl hytrazinobenzene hydrochloride for 4 days. The ratio of induced reticulocyte was about 83.16% ±3.41%. The cell shape of reticulocytes was irregular, the state of cells was poor and animals began to die as well when mice were injected 300 μl or 400 μl hytrazinobenzene hydrochloride. Conclusions The best conditions for inducing reticulocyte are as follows: weight 25 to 35 g mice daily injection of 200 μl 1% hytrazinobenzene hydrochloride for 4 days.

Association between polymorphisms of Kir4.1 and AQP4 and temporal lobe epilepsy in a Chinese Han Population

2013, 33(6):  752-756. 

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Objective To investigate the relationship between the genetic polymorphisms of Kir4.1 and AQP4 and the susceptibility to temporal lobe epilepsy (TLE). Methods A real time polymerase chain reaction based on TaqMan-BHQ probes was used to identify the SNPs in the two genes in 438 TLE patients and 695 matched controls from Chinese Han origins. Results Five SNPs in Kir4.1 and seven SNPs in AQP4 were selected. There was no statistically significant difference between TLE and control groups. Haplotype analysis was also performed and no significant difference was found. Whether the single SNP was associated with TLE phenotypes including hippocampus sclerosis and febrile seizure was also investigated, but no significant association was detected. Conclusions These results suggested that polymorphisms of Kir4.1 and AQP4 might have no important roles in TLE occurrence and development.

Wnt2 expression in Hepatocellular Carcinoma and its inhibitory effect on the proliferation of HepG2

2013, 33(6):  757-762. 

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Objective To explore the expressions of Wnt2 in human hepatocellular carcinoma(HCC) and the suppressive effect of knocking down Wnt2 expression by RNAi on Hepatocellular cell line HepG2. Methods Real-time quantitative PCR (RT-qPCR) method and immunohistochemistry methods was used to detect the expression of Wnt2 in HCC, paired Paracancerous tissue and normal liver tissue;After RNAi was used to knock down the expression of Wnt2 in HepG2,testing the change of Wnt2 and cell proliferation level and cycle change. Result 1).Wnt2 expressed in the highest in cancer, paired Paracancerous tissue is the second smallest,and normal liver minimum. 2). Using RNAi technology,The expression of Wnt2 was downregulated.HepG2 proliferation was inhibition and the cell growth mainly detained in G0/G1 and the percentage of S decreased. Conclusion Wnt2 was over expression in Hepatocellular Carcinoma;siRNA can inhibit expression of Wnt2 as well as the growth of HepG2.Wnt2 may be a novel target in gene regulation of Hepatocellular Carcinoma.

Protective effects of DJ-1 via antagonizing ROS-induced Beclin-1 up-regulation in hypoxia-reoxygenated HL-1 cardiomyocytes

2013, 33(6):  763-768. 

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Objective To explore the potential mechanism of DJ-1 via antagonizing ROS-induced Beclin-1 up-regulation on HL-1 cardiomyocytes during hypoxia-reoxygenation. Methods DJ-1 down-regulation was induced in mouse HL-1 cardiomyocytes by lentivirus transfection. The expression of DJ-1，LC3 and Beclin-1 protein expression change after hypoxia-reoxygenation and lentivirus transfection were detected by Western blotting, autophagosomes were detected by MDC, ROS levels were detected by DCFH-DA, Cardiomyocytes death was detected by AnnexinⅤ and PI. Results Compared to the control group cells，significant down-regulation of protein levels of DJ-1 was observed in cardiomyocytes with shDJ-1 virus，while protein levels of Beclin-1 and LC3-Ⅱ in cardiomyocytes with shDJ-1 virus were up-regulated after hypoxia/reoxygenation, NAC decreased protein levels of Beclin-1 and LC3-Ⅱ in cardiomyocytes with shDJ-1 virus. ROS and autophagosome levels were significantly increased after hypoxia/reoxygenation, and apoptoses of those were significantly increased after hypoxia/reoxygenation in cardiomyocytes with shDJ-1 virus, and NAC could reverse of these. Conclusion The effect of DJ-1 via antaganizing ROS induced Beclin-1 up-regulation might involve in the ischemia-reperfusion injury.

Clinical analysis of Behcet’s disease associated with malignancy：41 cases reports

2013, 33(6):  769-772. 

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Objective：To investigate the clinical characteristics of Behcet’s disease (BD) associated with malignancy. Methods Medical records at Peking Union Medical College Hospital spanning from December 1995 through June 2012 were reviewed to identify patients who were diagnosed as BD associated with malignancy. Results Of 651 patients with BD in our hospital, 41 patients (25 female and 16 male) developed malignancy including 29 in hematologic malignancy and 13 in solid cancer. Myelodysplastic syndrome (n= 20) was the most common hematologic malignancy associated with BD, followed by leukemia (n = 4), aplastic anemia (n= 2) and lymphoma (n= 1). While, in solid cancer, colorectal cancer were more commonly associated with BD (n= 3, one case was also complicated with endometrial cancer), followed by bladder carcinoma (n= 2), esophageal carcinoma, stomach cancer, pancreatic cancer, thyroid carcinoma, breast cancer, cervical cancer, renal cell carcinoma, metastatic adenocarcinoma (n= 1, each). Older age and intestinal involvement were more frequent in patients with malignancy (P < 0.05). Comparisons of clinical characteristics between patients with solid cancer and those with hematologic disease showed that older age, long disease duration and low disease activity were more commonly seen in BD patients with solid cancer. Conclusions The possibility of development of malignancy in BD patients should not be ignored. Older age and intestinal involvement is more likely to be the risk factors for the BD patients associated with malignancy. Further studies will be required to ascertain the pathogenic link between BD and malignancy.

Soluble MHC I was generated in the late endosome in HeLa cell

2013, 33(6):  773-774. 

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L-carnitine on decreases intestinal ischemia-reperfusion injury in rats

2013, 33(6):  775-776. 

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Virus supernate enhances the insulin secretion of cultured insulinoma cell line INS-1

2013, 33(6):  777-778. 

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The research progress in the role of myeloid cells in tumor angiogenesis and its clinical application

2013, 33(6):  779-782. 

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For the past few years, growing evidences have shown that bone marrow-derived myeloid cells play a key role in regulating the formation of blood vessels in tumors. Myeloid cells promote tumor angiogenesis in various manners, including VEGF secretion and VEGF independent way. The latter constitutes the mechanism of tumor refractoriness to anti-VEGF therapy. Myeloid cells also promote tumor recurrence after radiotherapy or chemotherapy. Last, we discuss its concomitant therapeutic implications.

Advances in imaging of metastasized stomach cancer lymph node

2013, 33(6):  783-786. 

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In order to perform accurate lymph node excision ,we ought to resect all metastatic lymph nodes and keep normal lymph nodes. This challenge the present examinational methods of lymph node metastasis before and during operation. In the past, the CT、MRI、PET-CT have low accuracy and specificity, poor and false negative rate , in order to achieve the goal infrared electronic endoscope, high resolution PET-CT、MRI equipments and high polymer nanoparticles molecular targeted imaging contrast agents have been developed.