Objective To assess the genetic toxicity and effects of cooking oil fume on oxidation-antioxidation level, immunity function of the workers occupationally exposed to cooking oil fume. Methods 108 subjects engaged in catering trade in Dongshan district of Guangzhou city were enrolled into this study by questionnaire investigation, serum MDA, SOD, activity of Lysozyme, micronucleus formation of periphery lymphocytes, blood lipids, blood routine indexes and vital capacity were determined. Results Compared with the control, serum MDA, micronucleus formation of periphery lymphocytes, CHOL, apoB LDL,WBC and mononuclear cells in the exposed group were significantly higher and a positive correlation was found between the levels of serum MDA, CHOL, apob, LDL, micronucleus formation of periphery lymphocyfes and length of service. Conclusion Cooking oil fume may injure oxidation-antioxidation system and have genetic toxity to human. The effect on lung function needs further study.

Objective To evaluate the effect of the intermittent deferomamine(DF) therapy on relieving iron overload caused by transfusion in children with ? thalassemia.Methods Sixteen children who were finally diagnosed as ? thalassemia major were treated with deferomamine for 124 times totally to low the iron overload. The serum iron(SI), serum ferritin(SF) and urine ferritin were detected each time with radio-immunity technique and difference was compared before and after treatment. Meanwhile, weather DF involved children′s liver and renal function was observed in whole procedure.Results Iron overload exists in 16 cases of ? thalassemia major children by a long- term hypertransfusion therapy, with average level SI 33.69?6.72 mmol/L,SF 441.19? 54.70 ?g/L,urine ferritin 8.64?6.79 ?g/L. The difference was significant (paired-samples t test,t =6.173 P 0.05).Conclusion The study suggest that intermittent low-dose DF therapy is effective for iron overload caused by transfusion in ? thalassemia children, without apparent side effects.

OBJECTIVETo investigate the chemical constituents from the aerial parts of Breynia fruticosa.

METHODVarious chromatographic techniques were employed for isolation and purification of the constituents. The structures were elucidated by chemical evidence and spectral methods.

RESULTSeven compounds were obtained and identified by spectroscopic methods and compared with authentic samples as aviculin [(+)-isolariciresinol-9'-rhamno-pyranoside], friedelan-3beta-ol, friedelin, arborinone, isoarborinol, 5-hydroxy-7,8,4'-trimethoxy flavone, 2,4-dihydroxy-6-methoxy-3-methyl-acetophenone.

CONCLUSIONAll compounds were firstly isolated from B. genus, furthermore, aviculin was isolated from Euphorbiaceae for the first time.

Euphorbiaceae ; chemistry ; Glycosides ; chemistry ; isolation & purification ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification

BACKGROUNDThe delivery of glucose from the blood to the brain involves its passage across the endothelial cells of the blood-brain barrier (BBB), which is mediated by the facilitative glucose transporter protein 1 (GLUT(1)), and then across the neural cell membranes, which is mediated by GLUT(3). This study aimed to evaluate the dynamic influence of hyperglycemia on the expression of these GLUTs by measuring their expression in the brain at different blood glucose levels in a rat model of diabetes. This might help to determine the proper blood glucose threshold level in the treatment of diabetic apoplexy.

METHODSDiabetes mellitus was induced with streptozotocin (STZ) in 30 rats. The rats were randomly divided into 3 groups: diabetic group without blood glucose control (group DM1), diabetic rats treated with low dose insulin (group DM2), and diabetic rats treated with high dose insulin (group DM3). The mRNA and protein levels of GLUT(1) and GLUT(3) were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively.

RESULTSCompared with normal control rats, the GLUT(1) mRNA was reduced by 46.08%, 29.80%, 19.22% (P < 0.01) in DM1, DM2, and DM3 group, respectively; and the GLUT(3) mRNA was reduced by 75.00%, 46.75%, and 17.89% (P < 0.01) in DM1, DM2, and DM3 group, respectively. The abundance of GLUT(1) and GLUT(3) proteins had negative correlation with the blood glucose level (P < 0.01). The density of microvessels in the brain of diabetic rats did not change significantly compared with normal rats.

CONCLUSIONSChronic hyperglycemia downregulates GLUT(1) and GLUT(3) expression at both mRNA and protein levels in the rat brain, which is not due to the decrease of the density of microvessels. The downregulation of GLUT(1) and GLUT(3) expression might be the adaptive reaction of the body to prevent excessive glucose entering the cell that may lead to cell damage.

Animals ; Blood Glucose ; analysis ; Brain ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; Glucose Transporter Type 1 ; analysis ; genetics ; Glucose Transporter Type 3 ; analysis ; genetics ; Glycated Hemoglobin A ; analysis ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Streptozocin

Mammalian cells are prone to apoptosis when cultured in large scale for production of biopharmaceuticals. And this will reduce production duration and result in high cost of production. Apoptosis is triggered by various factors, and delicately regulated by a set of genes. Bcl-2, a component integrated in mitochondria membrane, is an important member of these genes. By maintaining the integrity of mitochondria membrane, Bcl-2 keeps cytochrome C from releasing into cytoplasm, and thus blocks the activation of caspases, and subsequent onset of apoptosis. Over-expression of Bcl-2 has proven to be useful in blocking apoptosis in various cell lines, including CHO, hybridoma, myeloma, lymphoma and insect cells. Ammonia, a metabolite of cultured cells, however, showed apparent pro-apoptosis activity. In living cells, ammonia can be utilized by glutamine synthetase (GS) to synthesize glutamine, and thus lower the concentration of ammonia in medium, and its negative effects. Glutamine is essential to living cells. If not added into medium, glutamine can only be synthesized by GS, which makes GS a qualified selection marker. This marker can be used for gene amplification by adding into medium increased concentration of MSX, an inhibitor of GS. In this study, we over-expressed Bcl-2 using GS amplification in a recombinant CHO cell line stably expressing human interferon-beta. The modified cell line, with higher expression of Bcl-2 and lower production of ammonia, exhibited good anti-apoptosis quality and higher interferon-beta production in continuous culture.
Animals ; Apoptosis ; genetics ; physiology ; Biopharmaceutics ; CHO Cells ; cytology ; metabolism ; Cricetinae ; Cricetulus ; Glutamate-Ammonia Ligase ; genetics ; metabolism ; Interferon-beta ; metabolism ; Models, Genetic ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism

Serum used widely in mammalian cell culture is also a potential source of bacterial, mycoplasmal and viral contaminations. In addition, the complex biological components in serum make harder the subsequent product recovery process. High cost, high batch variation and potential source limitation are among the other shortcomings. So serum-free or even protein-free medium are preferable for recombinant protein production. However, without serum to provide essential components such as hormones, growth factors and binding proteins, cells are easy to die. In this study, CHO-dhfr- cells were genetically engineered to make them adapted to IMEM, a protein-free medium, and resistant to apoptosis. The genes in choice are insulin-like factor (Igf-1), Bcl-2 and cyclin E. Bcl-2 is a mitochondrial membrane-integrated protein. It can block the release of cytochrome c by maintaining the integrity of mitochondrial membrane, and thus inhibit apoptosis. Igf-1 is similar both in structure and function to insulin, a growth factor added to serum-free medium to promote cell growth and is the only protein component in many currently used serum-free media. cyclin E is a cell cycle protein expressed continuously in G1 phase. When cyclin E accumulates to certain amount, cell cycle was driven to S phase. So cyclin E is a proliferation-promoting protein. By co-express Igf-1/Bcl-2 or Bcl-2/ cyclin E in CHO-dhfr- cells with a dicistronic expression vector, we constructed two cell lines: CHO-IB and CHO-BC. The high expression of each protein was confirmed by Western blot and flow cytometry. Apoptosis was analyzed by flow cytometry and DNA ladder detection, and the two cell lines were both found much more resistant to apoptosis induced by withdrawal of serum or addition of actinomycin D than the CHO-dhfr- parent cell. Cell proliferation assay by MTT method showed that the two cell lines proliferated much faster than CHO-dhfr- in IMDM medium without serum. Continuously culture assay proved that the two cell lines grow very well in IMEM protein-free medium supplemented with fibronectin and vitronectin to ease adherence. When compared to CHO-dhfr-, the two cell lines exhibited much more viable cell numbers and faster growth rate.
Animals ; Apoptosis ; CHO Cells ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Culture Media ; Cyclin E ; genetics ; Genes, bcl-2 ; Insulin-Like Growth Factor I ; genetics

3a and 7a are nonstructural proteins of SARSCoV, which are encoded separately by ORF 3a and ORF 7a in SARSCoV genome. The expression of 3a has been founded in cells infected by virus in vivo or in vitro. Firstly, the pGL3Control vector was reconstructed , the pGL3Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN? promoter gene was cloned into the pGL3Enhancer vector and pGLIP21, the Luciferase reporter plasmid with IFN? promoter was established. The availability of pGLIP21 was verified by NDV ,the inductor of IFN?, the Luciferase activity was assayed in cells transfected with pGLIP21 by Luminometer. In order to see the function of 3a and 7a protein of SARSCoV,CHO cells expressing 3a or 7a protein were transfected with pGLIP21, the intensity of luciferase activity was analyzed . By analysis, in vitro, 3a and 7a protein of SARSCoV had the similar ability in triggering the expression of Luceferase gene, i.e 3a and 7a protein of SARSCoV could effectively activate the promoter fragment of IFN? gene. This result will help studying the function of 3a and 7a protein and provide a method to study the nosogenesis mechanism of SARSCoV.

Objective Coralline hydroxyapatite (CHA),in comparison with Bio-Oss bone meal,is a material with extensive resources but no immunogenicity or risk of disease-transmission. The aim of this article was to study the clinical application of CHA in ridge preservation in the maxillary anterior zone. Methods Twenty-six patients underwent extraction of maxillary anterior teeth (n=26) for chronic periodontitis or periapical periodontitis. The patients were randomly assigned into a CHA and a control group of equal number to receive ridge preservation with CHA and Bio-Oss bone meal respectively. Cone beam computed tomography (CBCT) was performed immediately and at 4 months after ridge preservation to compare the vertical and horizontal alterations of the alveolar ridge be-tween the two groups of patients. Results After ridge preservation,both the CHA and control groups showed a reduction in the width ([1.1±0.7] vs [1.3±1.9] mm) and height of the alveolar ridge ([1.3±1.6] vs [1.2±1.4] mm),but with no statistically significant differences between the two groups (P<0.05). Conclusion For ridge preservation in the maxillary anterior zone,CHA has a similar effect to that of Bio-Oss bone meal and therefore is an ideal material for bone graft.

OBJECTIVETo compare efficacy and side effects of patient-controlled epidural analgesia (PCEA) with levobupivacaine, ropivacaine and racemic bupivacaine after cesarean section.

METHODSIn this prospective, randomized double-blind study, 90 ASA I-II full-term nulliparous women (aged 25-38 years with body weight of 59-87 kg) undergoing elective cesarean section under spinal-epidural anesthesia equally allocated into 3 groups. PCEA was administered with 0.125% levobupivacaine and 20 microg/ml morphine (group L, n=30), 0.125% ropivacaine and 20 microg/ml morphine (group R, n=30), and 0.125% bupivacaine and 20 microg/ml morphine (group B, n=30), respectively. The Visual Analog Scale (VAS) score, satisfaction rate, patients' overall impression of treatment, modified Bromage motor score, and incidence of side effects were recorded at regular intervals after operation.

RESULTSThe three groups were comparable with respect to the efficacy of analgesia, patients' overall impression of treatment, motor blockade and side effects. There was significant difference in patients' satisfaction rate between group R (70%) and the other two groups (93.3% in group L and 96.6% in group B, P<0.05).

CONCLUSIONPCEA with 0.125% levobupivacaine and morphine 20 microg/ml produces better analgesic effect with little side effects after cesarean section.

Adult ; Amides ; administration & dosage ; adverse effects ; Analgesia, Epidural ; Analgesia, Obstetrical ; Analgesia, Patient-Controlled ; Bupivacaine ; administration & dosage ; adverse effects ; analogs & derivatives ; Cesarean Section ; Double-Blind Method ; Drug Therapy, Combination ; Female ; Humans ; Morphine ; administration & dosage ; Nausea ; chemically induced ; Pregnancy ; Prospective Studies ; Pruritus ; chemically induced ; Treatment Outcome ; Vomiting ; chemically induced

Without serum to provide adherent factors, CHO-dhfr- cells grow in suspension when cultured in serum-free medium. Although this offers advantages in some applications, in most production systems adherent cell growth is preferable. Gene transfection, clonal selection and amplification can be easier for adherent cells; the density of immobilized cells is often higher than those in suspension culture, which results in a higher protein productivity; washout of cells by perfused medium during continuous fermentation can be avoided; for high-throughput microplate assays, adherent cells are preferred to facilitate medium changes and cell washing. It has been proved that purified vitronectin alone was able to mediate attachment and spreading of CHO cells in serum-free medium. So we constructed a tricistronic expression vector expressing Igf-1, Vitronectin and Bel-2 at the same time. The vector was transfected into CHO-dhfr- cells and one clone, namely CHO-IVB2, expressing high level of the three proteins was screened out by Western blot. The cell line showed similar apoptosis-resistant and serum-independent properties to CHO-IB, an engineered cell line constructed before. When cultured in IMEM protein-free medium without any components supplemented, CHO-IVB can grow adherently. The viable cell numbers and growth rate of CHO-IVB were much higher than CHO-IB, making CHO-IVB an apoptosis-resistant host for production of recombinant proteins which can grow adherently in protein-free medium.
Animals ; Apoptosis ; CHO Cells ; physiology ; Cell Adhesion ; Cell Line ; Cell Proliferation ; Cricetinae ; Cricetulus ; Culture Media, Serum-Free ; Flow Cytometry ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; Vitronectin ; genetics