2025-03-23 2009, Volume 29 Issue 1

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  • Zhiguo Liu , He Li , Yinghong Li , Yan Wang , Yiqiang Zong , Youmei Feng , Zongchen Feng , Yaozu Deng , Shen Qu

    Very low density lipoprotein receptor (VLDLR) is thought to participate in the pathogenesis of atherosclerosis induced by VLDL and β-VLDL. The present study was undertaken to elucidate the effects of VLDL and β-VLDL on VLDLR expression and its signaling pathway. RAW264.7 cells were incubated with VLDL and β-VLDL. The expression of VLDLR mRNA was detected by RT-PCR. The transcriptional activity of VLDLR gene was detected in recombinant plasmid pGL4.2VR-luciferase transfected RAW264.7. Western blot assay was used to detect the changes of phosphorylated ERK1/2 protein. Inhibitors or activators were used to observe the signal pathway involving VLDLR expression regulation. The results showed that VLDL and β-VLDL stimulated ERK1/2 activity in a PKC-dependent manner. VLDL or β-VLDL-induced VLDLR expression on macrophages was extremely abolished by inhibitors ERK1/2 or PKC. Our findings revealed that VLDL or β-VLDL-induced VLDLR expression via PKC/ERK cascades and the effect was linked to the transcriptional activation of VLDLR gene promoter.

  • Yaohui Wang , Haiyan Zheng , Shengyuan Liu

    This study investigated the effects of ATP-sensitive potassium channels on the expression of P21, P27 and leptin. The expression of receptor of ATP-sensitive potassium channels (sulphonylurea receptor, SUR) mRNA in the preadipocytes and leptin mRNA was detected by PCR after rat preadipocytes were treated with the opener (diazoxide) or inhibitor (glibenclamide) of ATP-sensitive potassium channels during the process of inducing differentiation. The expression of P21 and P27 in preadipocytes treated with diazoxide or glibenclamide was assayed by Western blot. The results showed that the expression of SUR2, not SUR1 was detected in adipose tissue, preadipocytes and adipocytes. After treatment of preadipocytes with diazoxide, the expression levels of P21 and P27 were obviously higher than those in control group, but the expression levels of P21 and P27 in glibenclamide-treated group were lower than those in control group. During the process of inducing differentiation, the expression of leptin mRNA in preadipocytes treated with diazoxide was increased greatly, but the expression of leptin mRNA in glibenclamide-treated group decreased obviously. It was concluded that ATP-sensitive potassium channels might be involved in the proliferation and differentiation of rat preadipocytes by changing the expression of P21, P27 and leptin.

  • Hui Chen , Jian Qiao , Qian Li , Jungang Deng , Zhirong Tan , Tao Guo , Weiyong Li

    Batifiban, a synthetic cyclic peptide, is a potent platelet glycoprotein GPIIb/IIIa antagonist which may be useful in the treatment and prevention of acute coronary syndromes. The pharmacokinetics and pharmacodymanic (inhibition of platelet aggregation) effects, and tolerability of batifiban were investigated in healthy subjects following single bolus injection with doses of 55, 110, or 220 μg/kg, or multiple doses of an bolus followed intravenous infusion for 24 h (180 μg/kg plus 2.0 μg/min·kg, and 220 μg/kg plus 2.5 μg/min·kg) in this phase I clinical trial. Plasma levels of batifiban and areas under the curve were found to be proportional to doses. Batifiban was rapidly eliminated with a half-life of approximately 2.5 h. Significant differences were noted for plasma levels of batifiban and areas under the curve between males and females. No significant differences in the terminal half-life were found between males and females. Batifiban reversibly inhibited ex vivo platelet aggregation in a dose- and concentration-dependent manner, consistent with its mechanism as a GPIIb/IIIa antagonist. Single and multiple intravenous doses of batifiban were found to be safe and well tolerated in healthy subjects. These results support a bolus injection plus intravenous infusion regimen of batifiban for the treatment and prevention of acute coronary syndromes.

  • Changqing Zhou , Xiangning Fu , Jiangtao Yan , Qiao Fan , Zhuoya Li , Katherine Cianflone , Daowen Wang

    Dizziness, chest discomfort, chest depression and dyspnea are a group of symptoms that are common complaints in clinical practice. Patients with these symptoms are usually informed that while neurosis consequent to coronary heart disease is excluded nonetheless they remain unhealthy with no rational explanation or treatment. 165 cases of these symptoms and 85 control subjects were reviewed and underwent further medical history inquiry, routine EKG test and cardiac ultrasound examination. Thirty-five patients received coronary artery angiography to exclude coronary heart disease. Serum myocardial autoantibodies against beta1-adrenoceptor, alpha-myosin heavy chain, M2-muscarinic receptor and adenine-nucleotide translocator were tested, and inflammatory cytokines and high sensitivity C-reaction protein were measured and lymphocyte subclass was assayed by flow cytometry. All patients had a complex of four symptoms or tetralogy: (1) persistent throat or upper respiratory tract infection, (2) neck pain, (3) chest pain and (4) chest depression or dyspnea, some of them with anxiety. Anti-myocardial autoantibodies (AMCAs) were present in all patients vs. 8% in controls. TNF-α, IL-1 and IL-6 were significantly higher in patients than in controls (P<0.01). CD3+ and CD4-CD8+ lymphocytes were significantly higher and CD56+ lymphocytes lower in patients than those in controls (P<0.01). The ratio of serum pathogen antibodies positive against Coxsackie virus-B, cytomegalovirus, Mycoplasma pneumoniae and Chlamydia pneumoniae were all markedly higher in patients. These data led to identification of a persistent respiratory infection-related clinical syndrome, including persistent throat infection, neck spinal lesion, rib cartilage inflammation, symptoms of cardiac depression and dyspnea with or without anxiety.

  • Junbin Hu , Yan Wang , Yan Chen

    This study investigated the inhibitory effects of curcumin on proliferation of hematological malignant cells in vitro and the anti-tumor mechanism at histone acetylation/histone deacetylation levels. The effects of curcumin and histone deacetylase inhibitor trichostatin A (TSA) on the growth of Raji cells were tested by MTT assay. The expression of acetylated histone-3 (H3) in Raji, HL60 and K562 cells, and peripheral blood mononuclear cells (PBMCs) treated with curcumin or TSA was detected by immunohistochemistry and FACS. The results showed curcumin inhibited proliferation of Raji cells significantly in a time- and dose-dependent fashion, while exhibited low toxicity in PBMCs. Curcumin induced up-regulation of the expression of acetylated H3 dose-dependently in all malignant cell lines tested. In conclusion, curcumin inhibited proliferation of Raji cells selectively, enhanced the level of acetylated (H3) in Raji, HL60, and K562 cells, which acted as a histone deacetylase inhibitor like TSA. Furthermore, up-regulation of H3 acetylation may play an important role in regulating the proliferation of Raji cells.

  • Tiejun Zhang , Xiaoming Zhou , Yue Chen , Weiming Gu , Tao Zhang , Qingwu Jiang

    In order to study the resistance of Neisseria (N.) gonorrhoeae to the fluoroquinolone and detect mutation patterns of quinolone resistance-determining regions (QRDRs) of clinical isolates in Shanghai, China, a total of 80 clinical isolates of N. gonorrhoeae were consecutively collected from Shanghai. The MIC of fluoroquinolone for the isolates was examined by using the agar dilution method and the mutation profiles of the QRDRs of gyrA and parC were analyzed by sequencing and restriction fragment length polymorphism (RFLP). Chi-square test was used for comparison of the mutation patterns. The results showed that: (1) High percentages of the 8 isolates were resistant to ciprofloxacin (95.0%), ofloxacin (95.0%) and lomefloxacin (97.5%), only one strain was susceptible to the ciprofloxacin. (2) Sensitive strains had a substitute of Asp95→Ala in the gyrA, and all isolates that were resistant or intermediated to the ciprofloxacin, had a double mutation in the gyrA (Ser91, Ala 92 and Asp95). Some strains also had a mutation in the parC. (3) The MICs of these isolates were significantly associated with the mutation patterns in the gyrA and parC. A double mutation of gyrA combined with parC87 mutation was a predominant pattern in Shanghai and could mediate high level resistance to ciprofloxacin. It suggests that mutations in the QRDRs of gyrA and parC may be responsible for the fluoroquinolone resistance. And fluoroquinolone could not be used as the first line antibiotics for gonorrhea treatment any more in Shanghai, China.

  • Jiongxin Xiong , Shikai Zhu , Yu Zhou , Heshui Wu , Chunyou Wang

    The aim of this study was to explore the effects of parenteral supplementation with ω-3 fish oil emulsion (Omegaven®) on systemic inflammatory response syndrome (SIRS) during the initial stage of severe acute pancreatitis (SAP). In a prospective, randomized and controlled trial, 60 patients with SAP were randomized either to treat with conventional therapy (Con group, n=30) or conventional therapy plus intravenous supplementation with ω-3 fish oil emulsion 0.2 g/kg every day (FO group, n=30). The effects were analyzed by the SIRS-related indexes. The results showed that APACHE-II scores in FO group were significantly lower, and the gap increased much farther after the 4th day than those in Con group (P<0.05). Fluid equilibrium time became shorter markedly in FO group than in Con group (5.1±2.2 days vs 8.4±2.3 days). In FO group, SIRS scores were markedly decreased and the SIRS state vanished after the 4th day; Plasma level of TNF-α was significantly reduced, while IL-10 decreased markedly, most prominently between the 4th and 7th day, and the ratio of IL-10/TNF-α raised as compared with Con group (P<0.05). During the initial stage of SAP, parenteral supplementation with ω-3 fish oil emulsion could efficiently lower the magnitude and persistence time of the SIRS, markedly retrieve the unbalance of the pro-/anti-inflammatory cytokines, improve severe condition of illness and may provide a new way to regulate the SIRS.

  • Kai Xu , Fengjing Guo , Shuwei Zhang , Cheng Liu , Feixiong Wang , Zhiguo Zhou , Anmin Chen

    The roles of Indian hedgehog (Ihh) signaling pathway in the proliferation and apoptosis of precartilaginous stem cells (PSCs) were investigated. PSCs, labeled with fibroblast growth factor receptor 3 (FGFR-3), were isolated from neonatal rats by immunomagnetic separation. After identification with FGFR-3 and Col II, the cells were incubated with different concentrations of cyclopamine (cyclo), the specific inhibitor of Ihh signaling pathway. The morphologic changes of the cells were observed under the inverted phase contrast microscope. The mRNA expression levels of Ihh, parathyroid hormonerelated peptide (PTHrP), protein Patched (Ptch), Bcl-2 and p21 were detected by RT-PCR. The protein expression levels of Ihh and Ptch were measured by Western blot. MTT assay was used to examine the effects of cyclo on proliferation of PSCs. Apoptosis rate of PSCs was examined by Annexin V/PI assay of flow cytometric analyses. After PSCs were incubated with cyclo, obvious morphologic changes were observed as compared with the control group. The mRNA expression levels of PTHrP, Ptch and Bcl-2 were decreased to varying degrees in a cyclo dose-dependent manner. However, the expression levels of Ihh and p21 mRNA were increased. The protein expression of Ptch and Ihh had the same change as the mRNA expression. Meanwhile, cyclo could obviously inhibit the proliferation and promote the apoptosis of PSCs. The results indicated that Ihh signaling pathway plays an important role in regulating the proliferation and apoptosis of PSCs, which is probably mediated by Bcl-2 and p21.

  • Ming Cai , Guobin Wang , Kaixiong Tao , Changxue Cai

    This study constructed siRNA recombinant expression vector targeting survivin gene and observe the apoptosis induction effect of it in human colon cancer cells. siRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cells. The effect of siRNA recombinant expression vector was detected by RT-PCR, Western blot, MTT reduction assay and flow cytometry. It was confirmed by restriction endonuclease and sequence analysis that siRNA recombinant expression vector targeting survivin gene was constructed successfully. Inhibition rate of survivin siRNA at mRNA and protein levels was 36.33% and 44.65% respectively. Growth of cancer cells was inhibited and the apoptosis rate was (17.24±2.13)%. The siRNA recombinant expression vector targeting survivin gene has been constructed successfully. It not only can inhibit the expression of survivin gene, but also can induce apoptosis in human colon cancer cells remarkably.

  • Chunping Liu , Huaxiong Pan , Zhi Li , Lan Shi , Tao Huang

    In order to investigate the relationship between the extent of tumor invasion and the tumor size, axillary lymph nodes metastasis, Her-2 gene overexpression, and histologic grading in breast invasive ductal carcinoma as well as the optimal extent of excision during the breast-serving surgery, the clinical data of 104 patients with breast invasive ductal carcinoma who had received modified radical mastectomy were analyzed. The correlation analysis on invasive extent, which was evaluated by serial sections at an interval of 0.5 cm from 4 different directions taking the focus as the centre, and the tumor size, axillary lymph nodes metastasis, Her-2 gene overexpression, and histologic grading was processed. There was a significant correlation between invasive extent and tumor size (r=0.766, P<0.01), and lymph nodes metastases (r=0.574, P<0.01), but there was no significant correlation between invasive extent and Her-2 expression (r=0.106, P>0.05), and histologic grading (r=0.228, P>0.05). The 100% negative rate of infiltration in patients without nipple discharge with tumor size <2, 2–3 and >3 cm was obtained at 1.5, 2.0 and 2.5 cm away from the tumor respectively. It is concluded that the performance of breast-serving surgery in patients with breast invasive ductal carcinoma should be evaluated by tumor size in combination with axillary lymph nodes involvement to decide the possibility of breast-serving and the secure excision extent.

  • Jiaquan Huang , Haiyan Huang , Yuntao Jiao , Guo Ai , Tiejun Huang , Lan Li , Haijing Yu , Ke Ma , Fei Xiao

    The therapeutic effects of anluohuaxian tablet combined with γ-IFN on schistosomal liver fibrosis and its mechanism were studied in a murine model and clinical cases of schistosomal liver fibrosis. Fifty Kunming mice were randomly divided into 5 groups: normal control group, infection control group, anluohuaxian tablet-treated group, γ-IFN-treated group and combined treatment (anluohuaian tablet+γ-IFN) group. Pathologic changes in liver, including hepatic pigmentation and the size of schistosomal egg granuloma, were observed by HE staining after treatment for 8 weeks. The expression of the type I and collagen III, and TIMP-1 was detected by immunohistochemistry. TGF-β1 mRNA expression was examined by real-time fluorescent quantitative PCR. Sixty patients with schistosomal liver fibrosis were divided into treatment group and control group. The patients in treatment group were treated with anluohuaxian tablet in combination with γ-IFN for 6 months. Before and after treatment, the changes of symptoms and signs, liver function, serum liver fibrosis indexes and imaging indexes were observed. The results showed that as compared with infection control group, all forms of treatments relieved the hepatic pathological injury with apparently diminished size of schistosomal egg nodules and decreased percentage of pigmentation (P<0.05). Furthermore, the expression of collagen I and III, TIMP-1, and TGF-β1 mRNA in combined treatment group was significantly decreased as compared with anluohuaxian tablet-treated and γ-IFN-treated groups (P<0.05). In the clinical observation, the serum liver fibrosis indexes, the portal vein width as well as the spleen thickness was significantly reduced in treatment group as compared with control group (P<0.05). It was concluded that the combined use of anluohuaxian tablet with γ-IFN in schistosomal liver fibrosis could protect liver function, alleviate liver fibrosis, and could be used as a choice in treating patients with schiatosomal liver fibrosis.

  • Qinglei Gao , Fei Ye , Xi Xia , Hui Xing , Yunping Lu , Jianfeng Zhou , Ding Ma

    In order to investigate the role of the PTEN expression in carcinogenesis and development of endometrial carcinoma and clarify whether and how PTEN and PI3K/Akt pathway relate to endometrial carcinoma, the expression of PTEN and phospho-Akt was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) methods and Western-blot from 24 cases of endometrial carcinoma, 10 cases of endometrial atypical hyperplasia, 10 cases of endometrial hyperplasia, and 10 cases of normal endometrium. SP immunohistochemical methods were used to measure levels of PTEN protein expression in following 5 study groups: 31 cases of endometrium in proliferative phase, 30 cases of endometrium in secretory phase, 71 cases of endometrial hyperplasia, 25 cases of atypical hyperplasia and 73 cases of endometrial carcinoma. Immunostaining score of PTEN was 3.39±0.15 in proliferative phase, 1.90±0.21 in secretory phase, 3.34±0.29 in endometrial hyperplasia, 0.62±0.11 in atypical hyperplasia, and 0.74±0.19 in endometrial carcinoma, respectively. PTEN mRNA relative value in normal endometrium, endometrial hyperplasia, endometrial atypical hyperplasia, and endometrial carcinoma was 2.45±0.51, 2.32±0.32, 0.46±0.11, and 0.35±0.13 respectively. The expression levels of PTEN mRNA and protein in patients with endometrial carcinoma and atypical hyperplasia were significantly lower than in those of proliferative phase and with endometrial hyperplasia. The level of PTEN expression in patients with endometrial carcinoma was significantly related to tissue type (P<0.005), differentiation (P<0.05) and clinical stage (P<0.05), but not to depth of myometrium invasion (P>0.05). Western blot analysis revealed that Phospho-Akt level in PTEN negative cases was significantly higher, and there was a negative correlation between PTEN and phospho-Akt (r=−0.8973, P<0.0001). It was suggested that loss of PTEN expression was an early event in endometrial tumorigenesis. The phosphorylation of Akt induced by the loss of PTEN took part in the tumorigenesis and development of endometrial carcinoma.

  • Rui Gao , Jing Wang , Yi Shen , Ming Lei , Zehua Wang

    Voltage-gated sodium channels (VGSCs) are known to be involved in the initiation and progression of many malignancies, and the different subtypes of VGSCs play important roles in the metastasis cascade of many tumors. This study investigated the functional expression of Nav1.5 and its effect on invasion behavior of human breast cancer cell line MDA-MB-231. The mRNA and protein expression of Nav1.5 was detected by real time PCR, Western Blot and immunofluorescence. The effects of Nav1.5 on cell proliferation, migration and invasion were respectively assessed by MTT and Transwell. The effects of Nav1.5 on the secretion of matrix metalloproteases (MMPs) by MDA-MB-231 were analyzed by RT-PCR. The over-expressed Nav1.5 was present on the membrane of MDA-MB-231 cells. The invasion ability in vitro and the MMP-9 mRNA expression were respectively decreased to (47.82±0.53)% and (43.97±0.64)% (P<0.05) respectively in MDA-MB-231 cells treated with VGSCs specific inhibitor tetrodotoxin (TTX) by blocking Nav1.5 activity. It was concluded that Nav1.5 functional expression potentiated the invasive behavior of human breast cancer cell line MDA-MB-231 by increasing the secretion of MMP-9.

  • Guang Shan , Tian Tang , Xiaobin Zhang

    Lead is a ubiquitous environmental and industrial pollutant that may have toxic effects on the male. Vitamins may protect against toxic effects of lead in the liver and reproductive system, which is confirmed by our initial research. The aim of this study was to further investigate the protective effects of vitamins (ascorbic acid combined with thiamine) on lead acetate (Pb)-induced reproductive toxicities in mice and study the possible mechanisms underlying these effects. Forty-five male mice were randomly divided into 3 groups, 15 mice in each and received daily intragastric administration with control, Pb (20 mg/kg), and Pb+vitamins (ascorbic acid of 420 mg/kg+thiamine of 30 mg/kg) for 6 weeks, respectively. The Pb-treated animals showed significant decreases in the epididymal sperm count and motility compared to the control group, while the Pb+vitamins group had significant increases for these variables. Moreover, an increasing apoptosis of germinal cells induced by Pb was reduced by vitamin treatment. Pb induced the activation of Caspase-3, Fas/Fas-L and Bcl-2 with elevated levels, and the adaptor protein primarily regulated signaling through Fas and required for Fas-induced apoptosis. In conclusion, ascorbic acid combined with thiamine exhibited protective effect on reproductive system by inhibiting Pb-induced excessive cell apoptosis.

  • Zhiqiang Zhu , Jianguo Wen , Xiangyu Zheng , Daoxie Wang , Qingwei Wang , Changhui Fan

    The expression of octamer binding factor 4 (Oct4) gene in bladder cancer cell line T24 and its effects on the biological characteristics of the cells were investigated. RT-PCR and Western blot were employed to detect the expression of Oct4 in T24 cells. The changes of biological characteristics in T24 cells were analyzed before and after gene-silencing by Boyden chamber and MTT. The results showed that the expression of Oct4 gene was detectable in T24 cells by RT-PCR and Western blot. The expression of Oct4 gene and protein was down-regulated by siRNA, and average number of transwell cells in interference group, negative control group and blank control group was 101.40±4.56, 104.20±10.03 and 111.00±11.90, respectively. There was significant difference in the proliferation ability of the cells from 48 h, 72 h to 96 h after the interference by siRNA between interference group and negative group or blank control group (P<0.05). It was suggested that Oct4 gene was related with proliferation ability of T24 cells, but not with invasive capability.

  • Yonghua Wang , Qianyuan Zhuang , Siwei Zhou , Zhiquan Hu , Ruzhu Lan

    B7-H1, a recently described member of the B7 family of costimulatory molecules, is thought to be involved in tumor immune escape by inducing T-cell apoptosis. In order to investigate the relationship between B7-H1 and immune escape of bladder cancer, B7-H1 expression in 50 cases of bladder cancer was detected by using immunohistochemical method. Survival curves were constructed using the Kaplan-Meier method and independent prognostic factors were evaluated using the Cox regression model. Our results showed that the positive rate of B7-H1 immunostaining in normal bladder tissue and bladder cancer was 0 and 72% respectively. The expression of B7-H1 was strongly associated with the pathological grade, clinical stage and recurrence (P<0.05). The survival rate was significantly lower in patients with B7-H1 positive group than in those with B7-H1 negative group and multi-variable analysis revealed that B7-H1 could be regarded as an independent factor in evaluating the prognosis of bladder cancer. It is concluded that the expression of B7-H1 is strongly associated with neoplastic progression and prognosis of bladder cancer. The manipulation of B7-H1 may become a beneficial target for immunotherapy in human bladder cancer.

  • Hong Wang , Liang Ma , Shuhua Yang , Zengwu Shao , Chunqing Meng , Deyu Duan , Yanjun Li

    In order to investigate the effect of Arg-Gly-Asp (RGD) peptide-modified silk biomaterial on the adhesion and proliferation of bone marrow-derived mesenchymal stem cells (MSCs), MSCs of third generation were seeded onto the surface of RGD-decorated silk (silk-RGD group), silk alone (silk group) or tissue culture plate (TCP group). After incubation for 4 or 12 h, MSCs were examined quantitatively by using precipitation method for cell attachment. The cell proliferation, which was defined as cell density, was compared among the three groups after culture for 1, 2, 3, and 4 days. Cell skeleton, which was labeled fluorescently, was observed under laser confocal microscope after 24 h of culture. The results showed that cell adhesion rate in silk-RGD group was higher than in silk group (P<0.05), but similar to that in TCP group after incubation for 4 or 12 h (P>0.05). There were no significant differences in the cell proliferation among the three groups at different time points (P>0.05 for all). Laser confocal microscopy revealed that in silk-RGD group, MSCs, strongly fluorescently stained, spread fully, with stress fibers clearly seen, while in silk group, actin filaments were sparsely aligned and less stress fibers were found. It was concluded that RGD peptide could improve the adhesion of MSCs to the silk scaffold, but had no impact on the proliferation of the cells.

  • Ke Jiang , Lin Cheng , Jiangjun Wang , Jinsong Li , Jun Nie

    This study investigated the expression of hemeoxygenase-1 (HO-1) in rats with acute lung rejection and its implication. A valid rat orthotopic left lung transplantation model (SD rat→Wistar rat) was established by using an improved three-cuff anastomosis technique. The rats were divided into control group, CoPP (HO-1 inducer)-treated group and ZnPP (HO-1 inhibitor)- treated group. The severity of acute rejection was graded on the basis of the morphologic changes of the lung samples stained with HE. The expression of HO-1 protein in lung tissue was detected by using immunohistochemistry and Western blot, and HO-1 mRNA activity was assayed by RT-PCR. The results showed that the expression of HO-1 protein was significantly increased with the acute rejection grading in rats (P<0.01). As compared with control and ZnPP-treated groups, the severity of acute rejection was not alleviated and the grade not reduced significantly in CoPP-treated group (P>0.05). It was concluded that HO-1 protein might be involved in the pathological process of post-graft acute rejection. The expression of HO-1 protein was increased gradually with aggravation of acute rejection, and HO-1 protein might be used as an index to monitor acute rejection after lung transplantation.

  • Mingjun Cai , Ruifan Xie , Lin Han , Rudong Chen , Baofeng Wang , Fei Ye , Dongsheng Guo , Ting Lei

    The effects of RNAi-mediated gene silencing of LRIG3 expression on cell cycle and survival of human glioma cell line GL15 and the possible mechanisms were explored. The plasmids pGenesil2-LRIG3-shRNA1 and pGenesil2-LRIG3-shRNA2 were transfected into GL15 glioma cells respectively by using Metafectine, and the transfected cells that stably suppressed LRIG3 expression were selected by G418. The control cells were transfected with negative control shRNA. The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot. The apoptosis rate and cell cycle were analyzed by flow cytometry. As compared with the negative shRNA-transfected GL15 cells, LRIG3 mRNA expression in GL15 cells transfected with pGenesil2-LRIG3-shRNA1 and pGenesil2-LRIG3-shRNA2 was silenced by 52.4%, 63.8%, and LRIG3 protein expression was reduced by 50.9% and 67.4% respectively. The LRIG3-specific siRNA transfected cells had higher proliferation rate than control cells. Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and the proliferation index significantly (P<0.01). Silencing LRIG3 could inhibit the apoptosis of GL15 cells (P<0.05). These findings suggest that the siRNA targeting LRIG3 gene shows a dramatic inhibitory effect on RNA transcription and protein expression, then promoting the proliferation of GL15 cells, arresting GL15 cells in G2/M phase, and suppressing apoptosis of GL15 cells.

  • Jian Chen , Xianping Xu , Heping Wang

    To investigate the expression of integrin-α3 mRNA in meningiomas and its correlation with proliferation and invasion, the expression of integrin-α3 subunit was detected by using in situ hybridization in patients with meningiomas (36 cases) and normal dura (2 cases) and arachnoid tissues (2 cases). SABC immunohistochemical assay was used to study the expression of Ki-67 nuclear antigen. The results showed that the expression of integrin-α3 mRNA in benign, atypical and malignant meningiomas was 2.52±0.362, 1.75±0.316 and 1.42±0.633, respectively. The expression levels of integrin-α3 mRNA was significantly lower in atypical or malignant meningiomas than those in benign meningiomas (P<0.05, P<0.01, respectively). The expression of integrin-α3 mRNA in those with invasive biological behavior was lower than that in those without invasion (1.63±0.462 vs. 2.61±0.526, P<0.01). Moreover, the expression of integrin-α3 mRNA was inversely associated with Ki-67 labeling index in all cases. It suggested that integrin-α3 subunit participated in the modulatory process of growth of meningiomas. The proliferation activity and malignant grade of meningiomas were increased with the decreased expression of integrin-α3 subunit, and the down-regulation of integrin-α3 mRNA was associated with the invasive biological behaviors in meningiomas.

  • Zhentao Zhang , Tao Wang , Xuebing Cao , Shenggang Sun , Lan Wang

    This study investigated the effect and mechanism of cell cycle reentry induced by 6-hydrodopamine (6-OHDA) in PC12 cells. By using neural differentiated PC12 cells treated with 6-OHDA, the apoptosis model of dopaminergic neurons was established. Cell viability was measured by MTT. Cell apoptosis and the distribution of cell cycle were assessed by flow cytometry. Western blot was used to detect the activation of extracellular regulator kinase1/2 (ERK1/2) pathway and the phosphorylation of retinoblastoma protein (RB). Our results showed that after PC12 cells were treated wtih 6-OHDA, the viability of PC12 cells was declined in a concentration-dependent manner. Flow cytometry revealed that 6-OHDA could increase the apoptosis ratio of PC12 cells in a time-dependent manner. The percentage of cells in G0/G1 phase of cell cycle was decreased and that in S phase and G2/M phase increased. Simultaneously, ERK1/2 pathway was activated and phosphorylated RB increased. It was concluded that 6-OHDA could induce cell cycle reentry of dopaminergic neurons through the activation of ERK1/2 pathway and RB phosphorylation. The aberrant cell cycle reentry contributes to the apoptosis of dopaminergic neurons.

  • Xiaofang Dai , Dan Tao , Hongge Wu , Jing Cheng

    The low dose hyper-radiosensitivity (HRS) in human lung cancer cell line A549 was investigated, the changes of ATM kinase, cell cycle and apoptosis of cells at different doses of radiation were observed, and the possible mechanisms were discussed. A549 cells in logarithmic growth phase were irradiated with 60Co γ-rays at doses of 0–2 Gy. Together with flow cytometry for precise cell sorting, cell survival fraction was measured by means of conventional colony-formation assay. The expression of ATM1981Ser-P protein was examined by Western blot 1 h after radiation. Apoptosis was detected by Hoechst 33258 fluorescent staining, and Annexin V-FITC/PI staining flow cytometry 24 h after radiation. Cell cycle distribution was observed by flow cytometry 6, 12 and 24 h after radiation. The results showed that the expression of ATM1981Ser-P protein was observed at 0.2 Gy, followed by an increase at >0.2 Gy, and reached the peak at 0.5 Gy, with little further increase as the dose exceeded 0.5 Gy. Twenty-four h after radiation, partial cells presented the characteristic morphological changes of apoptosis, and the cell apoptosis curve was coincident with the survival curve. As compared with control group, the cell cycle almost had no changes after exposure to 0.1 and 0.2 Gy radiation (P>0.05). After exposure to 0.3, 0.4 and 0.5 Gy radiation, G2/M phase arrest occurred 6 and 12 h after radiation (P<0.05), and the ratio of G2/M phase cells was decreased 24 h after radiation (P<0.05). It was concluded that A549 cells displayed the phenomenon of HRS/IRR. The mode of cell death was mainly apoptosis. The activity of ATM and cell cycle change may take an important role in HRS/IRR.

  • Zheng Xue , Xuezhen Wang , Fei Liu , Shaoxian Hu , Suiqiang Zhu , Suming Zhang , Bitao Bu

    In order to better understand the clinical manifestation of systemic lupus erythematosus (SLE) with intracranial hypertension syndrome (IHS), we analyzed the clinical features and treatment of a typical SLE patient with IHS. SLE is one of the most unpredictable autoimmune diseases involving multiple organ systems that is defined clinically and associated with antibodies directed against cell nuclei. IHS is an uncommon manifestation of neuropsychiatric SLE (NPSLE) and is characterized by an elevated intracranial pressure, papilledema, and headache with occasional abducens nerve paresis, absence of a space-occupying lesion or ventricular enlargement, and normal cerebrospinal fluid chemical and hematological constituents. IHS has been reported in a few sporadic cases in patients with SLE worldwide, but rarely has been reported in China. In this study, a 34-year-old female SLE patient with IHS was reported and pertinent literature reviewed. The clinical presentation, image logical features, and investigatory findings were discussed.

  • Xuezhen Wang , Ronghua Tang , Zheng Xue , Feng Jiang , Min Zhang , Bitao Bu

    In order to explore the role of TNF-α in Niemann-Pick type C (NPC) disease, lentiviral-delivered RNA interference (RNAi) was used to silence the expression of murine TNF-α gene in vitro and in npc mice. Interference efficiency of the lentivirus expressing TNF-α-siRNA, previously constructed with the concentration of 2×108 ifu/mL, was determined by RT-PCR and ELISA in BV-2 cells and astrocytes. At the same time, the constructed Lenti-TNF-α-siRNA was intracerebroven-tricularly infused into 4-week old npc mice for a 4-week period, and the mice were divided into 3 groups: Lenti-TNF-α-siRNA (n=6), control lentivirus (n=6), and NPC mice without any intervention (n=4). By using immunohistochemistry and real-time PCR, the down-regulation of the target genes was detected. The Lenti-TNF-α-siRNA downregulated the expression of murine TNF-α gene efficiently in vitro and the interference efficiency was 66.7%. Lentivirus could be expressed stably for long-term in the npc mice brain. Immunohistochemistry and real-time PCR revealed that, as compared with non-intervention group and Lenti-control group, Lenti-TNF-α-siRNA efficiently down-regulated the expression of murine TNF-α gene with the interference efficiency being 66.9%. TNF-α-siRNA down-regulated the expression of TNF-α gene in vitro and in vivo, which provided a potential tool for studying and treating neurodegenerative diseases and TNF-α-related diseases.

  • Huicong Kang , Qi Hu , Xiaoyan Liu , Feng Xu , Lin Chen , Suiqiang Zhu

    The clinical characters, diagnosis and differential diagnosis of paroxysmal kinesigenic choreoathetosis (PKC), and efficacy of the anti-epileptic drugs (AEDs) were investigated. Thirty-one patients with PKC were collected, and the clinical characters and change of EEG were analyzed. The average age of the first attack was 16.8 years old and the pinnacle was 10 to 20 years old. There were definite causes for every attack and the sudden movement was the most common one (92%). Time for the whole attack was always less than 1 min. The attack presented with muscle tension disturbance (83.9%), movement like dancing (16.1%), abnormal movement of mouth and face and other symptoms (16.2%). The attack tended to be very frequent and 71% patients were beyond once per day. The EEG examination and image scan of primary PKC were normal in most patients. Low dosage of AEDs could control the attack of 50%–77.3% patients. It was concluded that PKC was a common disease of movement disorder. The therapy by AEDs was very effective. PKC should be differentiated from epilepsy and the relationship between PKC and epilepsy needs further research.

  • Ming Chen , Jing Wang , Mingxing Xie , Xinfang Wang , Qing Lv , Lei Wang , Yan Li , Manli Fu

    Left ventricular remodeling index (LVRI) was assessed in patients with hypertensive heart disease (HHD) and coronary artery disease (CAD) by real-time three-dimensional echocardiography (RT3DE). RT3DE data of 18 patients with HHD, 20 patients with CAD and 22 normal controls (NC) were acquired. Left ventricular end-diastolic volume (EDV) and left ventricular end-diastolic epicardial volume (EDVepi ) were detected by RT3DE and two-dimensional echocardiography Simpson biplane method (2DE). LVRI (left ventricular mass /EDV) was calculated and compared. The results showed that LVRI measurements detected by RT3DE and 2DE showed significant differences inter-groups (P<0.01). There was no significant difference in NC group (P>0.05), but significant difference in HHD and CAD intra-group (P<0.05). There was good positive correlations between LVRI detected by RT3DE and 2DE in NC and HHD groups (r=0.69, P<0.01; r=0.68, P<0.01), but no significant correlation in CAD group (r=0.30, P>0.05). It was concluded that LVRI derived from RT3DE as a new index for evaluating left ventricular remodeling can provide more superiority to LVRI derived from 2DE.

  • Lei Wang , Jing Wang , Mingxing Xie , Xinfang Wang , Qing Lv , Ming Chen , Shaoping Zheng

    We studied the wall motion characteristics of the ascending aorta by velocity vector imaging (VVI) in primary hypertension patients. The ascending aortas both in 30 patients with primary hypertension and 30 normal controls were examined by Acuson sequoia 512 equiped with VVI. The maximum velocity (Vs, Ve) of every point on the anterior wall of ascending aorta both in systole and diastole was measured. The aortic diameter was wider in the hypertension patients than that in the healthy subjects (P<0.05). The movement amplitude of the anterior wall of the ascending aorta in long axis view in the hypertension patients was lower than that in the healthy subjects (P<0.05). The motion and time to peak in systole of each point of the ascending aorta in the healthy subjects had no significant difference (P>0.05). The velocity curves of the anterior wall of ascending aorta both in the hypertension and healthy subjects were regular, and the curve in systole was named S wave and that in diastole named E wave. The velocity of S wave and E wave was slower in the hypertension patients than that in the healthy subjects (P<0.05). The time to peak of S wave on the anterior wall of ascending aorta in systole was shorter in the hypertension patients than in the healthy subjects (P<0.05). VVI could be used to accurately and directly observe the movement character of the ascending aorta walls, which would help us understand the elasticity of great arteries in patients with hypertension.

  • Sihai Gao , Ping Li , Jinping Zhao , Yunfeng Zhang , Yu Sun

    To establish a murine carotid artery transplantation model for the study of the chronic rejection, 80 rats were divided into two groups, an allotransplant (ACI-Lewis) group and an isotransplant (Lewis-Lewis) group (control group). The donor carotid artery and the recipient carotid artery were anastomosed by using a polyethylene cuff (internal diameter: 0.7 mm, length: 3 mm).The pathological changes of carotid artery transplant were observed 14, 28 and 56 days after the transplantation. The results showed that the model was successfully established in 95% of the animals. The chronic rejection-associated arteriosclerosis was induced 28 days after the transplantation. The new chronic rejection model of carotid artery by using cuff technique caused fewer traumas and was easy to make. The pathological changes of the transplant mimicked the chronic rejection-associated arteriosclerosis found in human transplant.

  • Qian He , Yan Feng , Ying Wang , Zengzhen Wang

    In order to develop the heroin abstainers’ cue-sensitization questionnaire (HACSQ), and evaluate its reliability and validity, during March and April in 2006, the cross-sectional sample survey and cluster sampling were adopted among 939 volunteer participants who were heroin abstainers from 5 compulsive detoxification centers in China. The data were collected by the self-report questionnaires. The reliability analysis included Cronbach’s α coefficient, split reliability and test-retest reliability, while construction validity was analyzed in the light of exploratory and confirmatory factor analysis. Based upon the exploratory factor analysis, 4 factors were extracted, which were social emotion cue (SEC), direct implement cue (DIC), personnel cue (PC) and indirect implement cue (IIC), and they together explained 64.52% variance. According to the data, test-retest reliability coefficient ranged from 0.57 to 0.63, and Cronbach’s α coefficient ranged from 0.86 to 0.95 across the 4 factors and HACSQ. The adjusted goodness of fit index was 0.75. It was suggested that the HACSQ demonstrated good validities and reliabilities, and may be used as a reliable assessment tool for the cue-sensitization among the heroin abstainers.