Endogenous erythroid colony assay in patients with polycythemia vera and its clinical significance

Background Polycythemia vera (PV) is a malignant disorder of hemaopoietic stem cells which is characterized by clonal hyperproliferation and a low rate of apoptosis. This study was to assess endogenous erythroid colony (EEC) formation in the bone marrow of PV patients and determine its clinical significance.Methods The bone marrow mononuclear cells of 26 patients with PV, 2 patients with secondary erythrocytosis (SE), and 19 normal controls were cultured by Marsh‘ s method for EEC evaluation,and the clinical significance was evaluated.Results EECs appeared in 25 patients with PV but not in 2 patients with SE and 19 normal controls.The number of EECs and the EEC ratio [ EEC/erythropoietin ( EPO)-dependent colony forming uniterythroid (CFU-E)] in PV patients positively correlated with hemoglobin (Hb) levels. Their EEC number did not correlate with white blood cell (WBC) counts, platelet (PLT) counts, or leukocyte alkaline phosphatase (LAP) scores. Their EEC did not correlate with serum EPO levels. Fifteen patients with PV were treated with hydroxyurea ( Hu ) and/or interferon-alpha (IFN-α). Their EEC ratio before treatment positively correlated with the treatment time required for complete remission (CR) and negatively correlated with the time before relapse. The EEC numbers of 7 PV patients treated with Hu/IFN-α decreased after the blood cell counts dropped to normal levels. There was a positive correlation between the EEC ratio and the incidence of attacks of vascular thrombosis in PV patients. The numbers of apoptosised bone marrow mononuclear cells in PV patients were lower than those in normal controls. The EEC numbers of PV patients negatively correlated with the rate of apoptosis of bone marrow mononuclear cells.Conclusions EEC formation is characteristic in PV patients. EEC number in PV patients positively correlates with Hb levels, the time required for CR, and the incidence of attacks of vascular thrombosis. EEC number negatively correlates with the time before relapse. Bone marrow suppressive treatment might decrease EEC number. Thus, EEC number is a sensitive and specific parameter reflecting the abnormal hematopoietic clone burden induced by polycythemia vera. EEC number is an important diagnostic parameter for PV patients.     

Ad/CMV-hTGF-β1 treats rabbit intervertebral discs degeneration<Emphasis Type="Italic">in vivo</Emphasis>

Summary  To investigate therapeutic efficiency of Ad/CMV-hTGF-β1 gene for rabbit intervertebral dise degeneration model. 60 Japanese white rabbits were selected to form the L5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral dise degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 μl (10×10⁶ pfu) of Ad CMV-hTGF-β1 gene, constituted the therapy group, 12 were injected with 20 μl (10×10⁶ pfu) of Ad CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group, 3 weeks after injection, examples were taken for investigation of HE staining, MRI. Western Blotting and immunohistochemical research TGF-β1. Wide distribution of TGF-β1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-β1 content in intervertebral dises treated with TGF-β1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral dise pathological degree improved. Ad CMV-hTGF-β1 gene transfection is a potential method to increase TGF-β1 content and reverse intervertebral dise degeneration. ZHAN Zirui, male, born in 1970 Associate Professor The Project Sponsored by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, Ministry of Education P.R.C. (No. [2001]345).     

Childhood asthma hospitalizations and ambient air sulfur dioxide concentrations in Bronx County, New York

The association between asthma hospitalizations and ambient sulfur dioxide (SO2) concentrations was examined in a case-control study in Bronx County, New York. Cases comprised 2629 children aged 0-14 yr who were admitted to hospitals for asthma. There were 2236 controls who were admitted for reasons other than asthma. Daily ambient SO2 concentrations were categorized into quartiles of both average and maximum levels and various exposure windows (i.e., day of admission and 1-, 2-, and 3-d lags). Cases were exposed to higher daily average concentrations of SO2 than controls. The authors compared the highest exposure quartile with the lowest, and the odds ratios were 1.66, 1.90, 2.05, and 2.21 (all p < 0.01 for same-day, 1-, 2-, and 3-d lags, respectively), with a similar finding for daily SO2 maximum exposure. The results suggest a consistent positive association between SO2 exposure and hospitalizations for childhood asthma.     

Synergistic effect of Scutellaria baicalensis and grape seed proanthocyanidins on scavenging reactive oxygen species in vitro

Scutellaria baicalensis (SbE) is a commonly used Chinese herb medicine and grape seed proanthocyanidins is a popular herbal supplement in the United States. Both herbs have been shown to possess potent antioxidant effects. Using an in vitro model to produce the reactive oxygen species (ROS) generation (H2O2/FeSO4 for hydroxyl radicals, xanthine/xanthine oxidase for suproxide), we observed that Scutellaria baicalensis and grape seed proanthocyanidins acted synergistically to scavenge ROS. Our data suggest that a combination of these two herbs can potentially enhance their antioxidant efficacy, allowing lower dosages of each drug to be used. This has the advantage of avoiding possible side effects that may arise when higher doses of a single herb are used in an attempt to achieve a maximum degree of antioxidant activity.     

葛根素对急性心肌梗死患者梗死面积的影响及其机制探讨

目的探讨葛根素对急性心肌梗死患者梗死面积及脂肪酸代谢、炎症反应及斑块稳定性的影响。方法61例急性心肌梗死患者随机分为对照组(30例)和葛根素组(31例),两组均予常规治疗,葛根素组加用葛根素500mg／d,疗程2周。测定治疗前后血浆游离脂肪酸、基质金属蛋白酶-9(matrix metalloproteinases-9,MMP-9)、C-反应蛋白水平及用Ideker QRS记分法测定梗死面积。结果治疗前血浆自由脂肪酸、MMP-9、C-反应蛋白与梗死面积呈正相关(r=0．43、0．42、0．39,P<0．01);与治疗前比较,葛根素组自由脂肪酸、MMP-9及C-反应蛋白分别降低30％、41％、23％,梗死面积明显缩小(P<0．01),对照组无明显变化(P>0．05)。结论葛根素治疗可明显缩小急性心肌梗死面积,可能与降低血浆自由脂肪酸、抑制炎症及稳定动脉粥样硬化斑块有关。     

Comparison of Epstein-Barr virus DNA level in plasma, peripheral blood cell and tumor tissue in nasopharyngeal carcinoma

BACKGROUND: The plasma Epstein-Barr virus DNA (EBV-DNA) level has been found to be an indicator for staging and prognosis of nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: The EBV-DNA level in plasma, peripheral blood cells (PBC) and neoplastic tissues was quantitatively analyzed and potential associations with clinical parameters of NPC were investigated. RESULTS: The plasma EBV-DNA detecting rate and level in NPC (92%, 82,500 copies/ml) was significantly higher than that in NPC after treatment (19%, 0 copy/ml) and in controls (12%, 0 copy/ml) (p < 0.001); while there was no significance of the PBC EBV-DNA detecting rate and EBV-DNA load in NPC before (24%, 0 copy/actin) and after treatment (14%, 0 copy/actin), and in controls (16%, 0 copy/actin). The plasma EBV-DNA level was not correlated to the PBC EBV-DNA load in NPC before (p = 0.92) and after treatment (p = 0.267), and in controls (p = 0.735). The EBV-DNA level in NPC tumor (27.8 copies/actin) was significantly higher than that in nasopharyngitis and was positively correlated to the ratio of EBER1-positive cells on the NPC section (p = 0.001). The plasma EBV-DNA level was significantly increased in TNM stages I, II, III and IV NPC, whereas there was no significant difference of PBC EBV-DNA load in different stage NPC. CONCLUSION: Our results indicate that plasma EBV-DNA is a more sensitive and reliable biomarker than PBC EBV-DNA for diagnosis, staging and therapeutic effect evaluation at a molecular level in NPC clinical practice. Plasma EBV-DNA may derive from the cancer cells and PBC EBV-DNA from circulating mononuclear cells in NPC patients.     

Normal flora: living vehicles for non-invasive protein drug delivery

Feasibility to use probiotic bacteria as a living protein delivery system through oral route was assessed in vitro. Lactococcus lactis transformed with a plasmid to express and secret beta-lactamase was used to deliver beta-lactamase through Caco-2 monolayer, an intestine epithelium. Transport of beta-lactamase through Caco-2 monolayer was carried out in the transwells. The viability and integrity of the cell monolayers co-cultured with L. lactis was examined by trypan blue exclusion method and by measuring the transport of mannitol and propranolol as well as the transepithelial electrical resistance (TEER). Results show that it is feasible to use cell culture technique to evaluate the drug delivery by normal flora. The transport rate of beta-lactamase when delivered by L. lactis was 2.0 +/- 0.1 x 10(-2)h(-1) (n = 9) and through free solution form was 1.0 +/- 0.1 x 10(-2)h-1. When co-cultured with L. lactis, Caco-2 cell viability decreased to 98, 96, and 94% at 6, 8, and 10h, respectively. Transport of mannitol through Caco-2 cell monolayer was significantly increased and the transport of propranolol through Caco-2 cell monolayer was significantly decreased in the presence of L. lactis. Increase in the amount of protein delivered is probably due to the concentrate of the protein by L. lactis on the monolayer (absorption surface) and the opening of the tight junction of Caco-2 monolayer by L. lactis.     

Quality evaluation of commercial extracts of Scutellaria baicalensis

Botanical products have been widely used for various illnesses and general well-being. However, quality control of botanical products is often not performed due to lack of standardization, resulting in inconsistent efficacies and sometimes serious toxicity. The goals of this study were to determine the correlation between chemical composition and biological activities and to establish a method to measure authenticity, chemical consistency, and biological potency of botanical products. A high-performance liquid chromatography method was used to analyze the authenticity and chemical composition of 10 different commercial extracts. The cell viability assay and prostaglandin E2 (PGE2) enzyme immunoassay were used to analyze biological potency and consistency. Our results showed all extracts contained marker components (baicalein and/or baicalin), confirming their authenticity. However, significant product-to-product and batch-to-batch variation of these marker components was observed with 4 products containing no baicalin at all and baicalein concentration ranging from 0 to 52.3 g/mg. The 50% growth inhibition concentration of the extracts ranged from 0.18 to 2.0 mg/ml, more than an 11-fold variation. PGE2 levels varied from 19.5 to 111.1 pg/106 cells, more than a 5.7-fold difference. These results demonstrated significant variation in chemical composition and biological activities of the commercial extracts and that the amount of marker components may not reflect biological activity levels. Therefore, chemical analysis alone is inadequate for quality control, and biological assays must be included for botanical products to ensure chemical authenticity as well as pharmacological/biological potency and consistency.     

A novel tumor cell line cloned from mutated human embryonic bone marrow mesenchymal stem cells

A novel tumor cell line, denominated F6, was established from mutated human embryonic bone marrow mesenchymal stem cells (MSCs) which were induced by the GM-CSF and IL-4 in vitro. The characteristics of the F6 cell line, such as surface antigens, cell cycle, growth curve, gene expression, morphology, cytogenetics and tumor model were analyzed. The F6 cells were round and grew suspended in a plastic dish. The cell line has a strong self-renewal capability, was positive for CD13, CD29, CD44, but negative for CD1alpha, CD3, CD10, CD14, CD23, CD33, CD34, CD38, CD41, CD45, CD54 and HLA-DR. The surface antigens were lower than those of human embryonic MSCs. The karyotype of F6 cells was abnormal. The cell cycle included: G0/G1 phase, 52.24%; G2/M phase, 8.00%; S phase, 41.76%. After the cells had been passaged serially for more than 17 months (62 passages), their characteristics were still retained. The F6 cells resulted in tumors in SCID nude mice in vivo (8/8) and caused metastasis (3/8). The pathologic examination revealed that the tumor cells extensively invaded surrounding normal tissues such as dermis, muscular tissue, nerve tissue, adipose tissue and lymphoid tissue. F6 cell line, tumor tissues derived from F6 cells and the MSCs expressed different levels of the nucleostemin gene. These findings suggested that F6 may be a novel tumor cell line. It may provide evidence for the theory that cancer originates from stem cells, and may be useful for the investigation on safety of human MSCs in the clinical application.