To determine the expressions of Notch1, Jagged1 and vascular endothelial growth factor (VEGF) in human non-small cell lung cancer (NSCLC) and to explore its clinical pathological significance. Immunohistochemical SP method was used to detect the expressions of Notch1, Jagged1 and VEGF in 65 patients with NSCLC and 15 normal epithelial tissues of the lung, and the relationship between them and clinic-pathological parameters were analyzed. The positive rates of Notch1, Jagged1 and VEGF in NSCLC were 81.5%, 83.1% and 93.8%, respectively, higher than those in normal epithelial tissues of the lung (P<0.05). The positive expression levels of Notch1 and VEGF were closely associated with the tumor stage and the lymph node metastasis (P<0.05). The positive expression levels of Jagged1 was positively correlated with the pathological type and lymph node metastasis. There was a positive correlation between Notch1, Jagged1 and VEGF. Notch1, Jagged1 and VEGF protein may play an important role in the pathway of carcinogenesis and progression of NSCLC. The up-regulation of Notch1, Jagged1 and VEGF protein expression probably predict NSCLC carrying relatively strong permeation and metastasis.
ABCG2 is a half ATP-binding cassette (ABC) transporter that facilitates efflux of a wide variety of substrates ranging from natural products to synthetic anticancer drugs. Over-expression of ABCG2 has been shown to cause multidrug resistance in both laboratory model cell lines and in clinical settings. The polymorphism in ABCG2 has also been suggested to affect the function and clinical outcome in cancer patients. More recently, ABCG2 has been suggested to play a protective role for cancer and normal stem cells. Thus, ABCG2 is an ideal target for therapeutic development to chemo-sensitize drug resistant cancers. In this paper, the recent progress on understanding the structure, function, and pharmacology of ABCG2 and its role in drug resistance and cancer stem cells will be reviewed.
To transfect the recombinant plasmid enhancement type green fluorescent protein C2- neurotrophic factor-3 (pEGFPC2-NT3) into the spinal ganglion cells(SGCs) of guinea pigs' cochlea injured by the excitotoxicity of hydroxyapatite particle (HAT), to inject the recombinant plasmid pEGFPC2-NT3 into the guinea pigs' cochlea, and to observe the expression of pEGFPC2-NT3 and the protective effect of pEGFPC2-NT3 on SGCs of the cochlea in guinea pigs. The recombinant plasmid pEGFPC2-NT3 with gene-green fluorescent protein was established. Kanic acid (KA) was injected into guinea pigs'cochleae and the excitotoxicity model was established. After a week the recombinant plasmid was transferred into SGCs of guinea pigs'cochlea treated with HAT. The following week the expression of NT-3 was examined by the immunohistochemical method, and the morphology of SGNs was observed under the electronic microscope after 4 weeks, in the mean time the changes of auditory brain-stem response (ABR) were examined. The excitotoxicity models were established successfully. NT-3 expression in the intracytoplasm of SGNs was observed by the immunohistochemical method 1 week after the injection, the morphologic damages of SGNs lessened under the electronic microscope after 4 weeks. ABR was partly restored, compared with ABR after the injury of the excitotoxicity. On the 7th day, NT3 gene transferred by HAT through the scala tympani can lessen the excitotoxicity of SGCs after KA was injected into the guinea pigs cochlea.
To explore the effects of NGX6 on the transcriptional activation of beta-catenin/TCF/LEF in Wnt/beta-catenin signal pathway, and to identify the role of NGX6 in Wnt signal pathway. The eukaryotic expression vector pcDNA3.1(+)-beta-catenin (WT) was constructed. pcDNA3.1(+)-beta-catenin (WT) and pCMV-myc-NGX6 were cotransfected to COS-7 and the transcriptional activity of TCF/LEF was detected by TCF-4 luciferase report system. Without extro-genous beta-catenin, pCMV-myc-NGX6 was transfected alone to COS-7 and colon cancer cell line SW620, and the transcriptional activity of TCF/LEF was detected by TCF-4 luciferase report system, and then the expression of nucleus beta-catenin and TCF-4 was detected by Western blot. The eukaryotic expression vector pcDNA3.1(+)-beta-catenin (WT) was successfully constructed. The activation of TCF-4 luciferase report gene in the cotransfection group in COS-7 was less than that in NGX6 alone transfection group (P<0.05). The activation of TCF-4 luciferase report gene in NGX6 alone transfection group without extro-genous beta-catenin was less than that in pCMV-myc transfection group in COS-7 and SW620. The expression of beta-catenin and TCF-4 was decreased after the NGX6 transfection in COS-7 and SW620 cells. NGX6 can inhibit the transcriptional activation of beta-catenin/TCF/LEF in Wnt signal pathway by its negative regulation in the nuclear translocation of beta-catenin.
The prevalent rates of overweight and obesity are steadily increasing all over the world. Previous studies of some candidate genes have indicated that most of the genes are associated with obesity in human adipose tissue. As much as 40% of the variations in body mass could be attributed to genetic difference. The beta-adrenergic receptor (beta-AR) plays a major role in the regulation of energy balance in humans. A high sympathetic nervous system activity has been shown to be associated with obesity and is believed to have pathogenetic relevance. Several common single nucleotide polymorphisms (SNPs) including Gly389Arg in beta(1)-AR, Gln27Glu in beta(2)-AR, and Trp64Arg in beta(3)-AR in humans could alter receptor function and these variations of beta-ARs were shown to have certain association with obesity. Here we summarize the genetic polymorphisms of human beta-ARs and their potential impacts to obesity.
To determine the effect of CagA(+) Helicobacter pylori(H.pylori)strain and anti-H.pylori drugs on the expression of connexin 43(Cx43) and cell proliferation of BGC-823 cells in vitro,and to investigate the relation between the changes of Cx43 expression, cell proliferation of BGC-823 cells and CagA(+)H.pylori. BGC-823 cells were co-cultured with CagA(+) H.pylori strain(NCTC J99) or CagA(-) H.pylori strain(NCTC 12908)at bacteria/cells ratio of 20:1,100:1 and 500:1 for 24 hours and 48 hours respectively. anti-H.pylori drugs was given in the group co-cultured at bacteria/cells ratio of 100:1 after 16 hours. In the control group, BGC-823 cells were cultured for 24 hours and 48 hours respectively,but without H.pylori or antij H.pylori drugs. Immunocytochemical SABC method and the image analysis of the computer were applied to detect the changes of Cx43 expression in BGC-823 cells. The cell proliferation was examined by methyl tetrazolium (MTT) method. (1)The expression of Cx43 in the control group after cultivation for 48 hours was higher than that for 24 hours (P0.05). Cx43 expression in the groups co-cultured with CagA(-) H.pylori strain at the ratio of 100:1 and 500:1 was lower than that in the control group, and Cx43 expression at the ratio of 500:1 was lower than that at the ratio of 20:1 for 24 hours and 48 hours. Cx43 expression increased after the intervention with anti-H.pylori drugs for 48 hours. (2) In the groups co-cultured with CagA(+)H.pylori strain, the optical density value of MTT indicated that the cell proliferation at the bacteria/cells ratio of 100:1 was higher than that in the control group, but no significant difference was found in other two groups co-cultured for 24 hours. After co-culturing for 48 hours, the cell proliferation at the bacteria/cells ratio of 20:1 and 100:1 was significantly accelerated, while the cell proliferation at 500:1 was inhibited. In the groups co-cultured with CagA(-) H.pylori strain,there was no change in the cell proliferation. Intervention with anti-H.pylori drugs could suppress the cell proliferation. CagA(+) H.pylori can down-regulate the expression of Cx43 in BGC-823 cells,which is related to the reaction time and the density of H.pylori. Low density of CagA(+)H.pylori suspensions can accelerate the proliferation of BGC-823 cells, while high density can suppress the cell proliferation. The CagA(-) H.pylori has no effect on the cell proliferation. Intervention with anti-H.pylori drugs can up-regulate the expression of Cx43,and suppress the cell proliferation of BGC-823 cells.
To screen the differentially expressed gene profile from the smooth muscles in the fundus uterus at the active stage of labor, and to provide candidate genes for picking out the drug targets related to uterine contraction. Differentially expressed genes of uterine smooth muscles in the corpus from pro and post spontaneous parturition and those induced by oxytocin,as well as those from the corpus and the lower portion spontaneous parturition,were scanned respectively by human full-length genetic cDNA microarray with 8064 probe sets. Semi-quantitative RT-PCR was applied to testify the expression of voltage dependent calcium channel-L subtype (CACNA). The differentially expressed genes in the structure and function of the drug targets were picked out by bio-informatics to serve as candidate drug targets related to uterine contraction. The expressions of 29 genes were upregulated in fundus smooth muscles from the pro and post natural parturition, the pro and post inductive parturition of oxytocin, and the natural parturition. The expression of CACNA gene in RT-PCR was in accordance with that in the microarray. Among the 29 genes, neuromedin B receptor (NMBR) gene and neuropeptide Y (NPY) gene were the genes which not only had the targets of uterine contracted medicine, but also could contract the uterine. The differential expression ratios of NMBR in the above 3 types of uterine myometrium were 6.9,11.3, and 9.0, respectively while those of NPY were 6.0,29.8, and 2.9 respectively. NMBR, whose expression in the uterine smooth muscles is always up-regulated at different parturition conditions, is likely to be an ideal candidate target of uterotonic drugs.
Hepatic stellate cells (HSCs) play an important role in hepatic fibrogenesis. In response to liver injury, HSCs undergo a process called activation, which involves 2 steps jonit nation from quiescent phenotype to myofibroblast-like phenotype, and perpetuation that maintains the activated phenotype of HSCs. The fate of the activated HSCs depends on the apoptotic and survival signals that they receive. The apoptosis of HSCs results from a series of complex and interrelated signaling events. Apoptotic signals for the activated HSCs include proteins from membrane receptors, such as death receptors, nerve growth factor receptor and peripheral-type benzodiazepine receptor, as well as proteins from cytoplasm such as Bcl-2 family members. The survival signals for the activated HSCs are induced by some kinases and cytokines including tissue inhibitors of metalloproteinase-1, Rho/Rho kinase, platelet-derived growth factor, transforming growth factor beta-1, and insulin-like growth factor-1. Approaches that specifically initiate HSC apoptosis are promising to be direct and effective strategies to treat liver fibrosis. Although it remains unclear whether the activated HSCs could be reversed back to the quiescent phenotype, the different expression and sensitivity of pro-apoptotic and survival molecules between quiescent and activated HSCs provide a prospect to develop therapeutic approaches that specifically targets apoptosis of the activated HSCs. These therapeutic strategies to induce HSC apoptosis are current research hotspot and the future for the patients with liver fibrosis and cirrhosis.
To analyze the evaluation and treatment of blood loss during total hip arthroplasty (THA) and total knee arthroplasty (TKA) in the aged . We retrospectively surveyed the blood loss and the rehabilitation of 46 cases of TKA and 146 cases of THA older than 60. In the group younger than 70, the mean total blood loss of THA was 1425 mL and the hidden hemorrhage 729 mL (51%); following the TKA, the mean total loss was 1386 mL and the hidden hemorrhage was 890 mL (64%). In the group 70 and older, the mean total blood loss of THA was 1435 mL and the hidden hemorrhage was 769 mL (53%)û following the TKA, the mean total loss was 1380 mL and the hidden hemorrhage was 910 mL (65%). The difference of hidden hemorrhage between the THA and the TKA was both significant by different (p 0.05). The aged have poor resistance to blood loss. Hidden hemorrhage in the TKA or THA perhaps is the primary part of the blood loss. Prompt treatment is helpful for the rehabilitation.
To investigate the expression of haptoglobin in the lesions of condyloma acuminatum (CA) at the mRNA and protein level, and to explore its role in the pathogenesis of CA. The expressions of haptoglobin protein and mRNA in the skin tissues of 30 patients with CA and 20 normal controls were detected by immunohistochemistry(IHC), Western blot, and hybridization in situ. The in situ hybridization study showed that haptoglobin mRNA was expressed in the epidermal cells in the lesions of CA. The distribution of haptoglobin mRNA expression in the lesions of CA was similar to that of the normal controls, and the expression of haptoglobin mRNA in CA was higher than that of the normal controls. There was a significant difference in the positive expression of haptoglobin mRNA between the CA group and the control group (P<0.05). The immunohistochemical study showed that haptoglobin protein was expressed in the whole layers of epidermal keratinocytes in the lesions of CA at a high level and stronger staining was seen in the stratum basale and stratum spinosum. Haptoglobin protein was expressed predominantly in the stratum basale in normal skin tissues, while weak staining was seen below the stratum spinosum.There was a significant difference in the mean gray value between the CA group and control group (P<0.05). Western blot showed that the haptoglobin expression in CA lesions significantly increased compared with the normal skins (P<0.05). The expression of haptoglobin mRNA in the CA lesions obviously increases and the epidermal cells in the CA lesions are able to synthesize haptoglobin protein. Haptoglobin in the CA lesions may involve in the local immunity escape by preventing Langerhans cell functional maturation and inhibiting the immunocompetence of keratinocyte.
Multiple myeloma (MM) is the second most common hematological malignancy and remains incurable. The marked variation in survival of patients with symptomatic myeloma ranging from few months to more than 15 years can be explained by differences in tumor mass, proliferative activity and, more recently, by cytogenetic and molecular genetic characteristics of the myeloma clone. Oligonucleotide microarray-based gene expression analysis was applied to CD138-enriched plasma cells from newly diagnosed patients with symptomatic or progressive multiple myeloma treated with melphalan-based high-dose therapy. Here we discuss recent progress made in the development of molecular-based diagnostics and prognostics for MM from Myeloma Institute for Research and Therapy of University Arkansas for Medical Sciences, where we treat more patients with myeloma than anywhere else in the world. Seven distinct entities of myeloma were elucidated by genomic profiling. Expression extremes of 70 genes from a high-risk signature profile,30% of which were derived from chromosome 1, were strongly linked to disease-related survival. CKS1B located on chromosome 1q21, responsible for promoting cell cycle progression by inducing the degradation of p27Kip1, represented a strong candidate gene related to rapid patient death and was studied in detail. The data suggest that CKS1B influences myeloma cell growth and survival through SKP2j and P27(Kip1) -dependent and independent mechanisms and that therapeutic strategies aimed at abolishing CKS1B function may hold promise for the treatment of high-risk disease for which effective therapies are currently lacking.
To investigate the effect of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, on the activation of hepatic stellate cells (HSCs) and its mechanism. Primary HSCs isolated from SD rats were cultured and treated with different concentrations (1, 3 or 10micromol/L) of ADMA for various periods (12 approximately 48h). Expression of alpha-smooth muscle actin (alpha-SMA) and synthesis of type-I collagens in HSC were determined. Messenger RNA levels of the transforming growth factor-beta1 (TGF-beta(1)) in the HSCs were determined using RT-PCR. Intracellular reactive oxidant species (ROS) production was measured using oxidant-sensitive fluorescent indicator. Activation of nuclear factor-kappaB (NF-kappaB) was detected by electrophoretic mobility shift assay (EMSA). ADMA could increase alpha-SMA-positive cells ratio and Type I collagens production of HSCs in a concentration- and time-dependent manner, concomitant with the increase of the TGF-beta(1) mRNA level. Treatment with ADMA (10micromol/L) significantly increased the intracellular ROS production and activated NF-kappaB. Such effects of ADMA on the level of TGF-beta(1) mRNA could be markedly attenuated by pretreatment with antioxidant pyrrolidine dithiocarbamate (25micromol/L). ADMA can induce the HSC activation by increasing TGF-beta(1) expression through ROS-NF-kappaB-dependent pathway. Therefore, ADMA should be a novel and endogenous activator of HSC, which may be involved in the development of liver fibrosis.