Objective To investigate the radiosensitivity effects of poly ADP-ribose polymerase-1 (PARP-1) inhibitor 3-amion benzamide (3-AB) on the BRCA non-mutant and BRCA mutant breast cancer cells,and to explore the regulatory mechanism of PARP-1 and BRCA in radiation-induced DNA damage repair.Methods MDA-MB-436 cells and MDA-MB-231 cells were divided into four groups respectively as the control (CTRL),ionizing radiation alone (IR),3-AB alone (3-AB),and ionizing radiation combined with 3-AB(IR + 3-AB)group.γ-H2AX foci were detected by immunofluorescence assay.The radiosensitivity of breast cancer cells was evaluated by clonogenic survival assay.The percentage of apoptotic cells was assessed by flow cytometry.Results Compared with MDA-MB-231 cells,MDA-MB-436 cells had a higher radiosensitivity and produced more γ-H2AX foci(t =4.57,P ＜ 0.05),which was further increased by 3-AB.The DNA damage of MDA-MB-436 cells in the IR + 3-AB group was the most remarkable (t =3.26,P ＜ 0.05).Flow cytometry showed that the cells in the IR + 3-AB group had the highest rate of apoptosis (t=3.81,P ＜ 0.05),and the apoptosis rate of MDA-MB-436 cells was significantly higher than MDA-MB-231 cells (t =2.96,P ＜ 0.05).Conclusions The radiosensetivity of BRCA mutant cells MDA-MB-436 is significantly higher than that of non-BRCA mutant cells MDA-MB-231.Inhibition of PARP-1 can further increase the apoptosis and radiosensitivity of BRCA-mutant cells by further blocking the repair of DNA single-strand break induced by ionizing radiation.

AIM(+/-) Ibuprofen sugar derivatives were prepared in order to decrease side-effects and increase bioavailability of (+/-) ibuprofen.

METHODSThe synthesis of derivatives were performed using 1,2:3, 4-di-O-isopropylidene-beta-D-galactopyranose, 1,3,4,5-tetra-O-acetyl-2-deoxy-2-amino-beta-D-gluctopyranose, 3,4 6-tri-O-acetyl-2-deoxy-2-N-acetyl-beta-D-gluctosylamine and 2,3,6,2',3',4',6'-hepta-O-acetyl-beta-D-lactosylamine as glycosyl donors, respectively. Target products (4, 7, 12a, 12b, 13) were obtained after deprotection.

RESULTSFive compounds (4, 7, 12a, 12b, 13) were synthesized as new compounds. The structures of all objective compounds were confirmed by 1HNMR, 13CNMR, HMQC, COSY, IR and MS.

CONCLUSIONIt was found that 12a showed better anti-inflammatory activity than (+/-) ibuprofen.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; chemical synthesis ; therapeutic use ; Ibuprofen ; analogs & derivatives ; chemical synthesis ; therapeutic use ; Inflammation ; drug therapy ; Male ; Mice ; Molecular Conformation ; Molecular Structure ; Random Allocation

OBJECTIVETo investigate the effect and its possible mechanisms of Danhong Injection (DHI) on cerebral injury during coronary artery bypass graft (CABG) operation with hypothermic cardiopulmonary bypass (CPB).

METHODSFifty patients went to CABG with CPB were randomly assigned equally to two groups, the control group and the tested group. DHI 1.5 mL/kg was pumped to where the tested group at the times of aortic pre-charging and unclamping respectively, but to the control group, equal volume of normal saline was given instead. Blood samples were taken from jugular bulb at different time points, i. e. before operation (T1, baseline), re-warming to 36 degrees C (T2 ), 30 min (T3 ) and 6 h (T4) after terminating CPB, for determination levels of superoxide dismutase (SOD) activity using xanthine oxidase method, malondialdehyde (MDA) concentration using thiobarbituric method, tumor necrosis factor alpha (TNF-alpha), as well as interleukin-6, -8 using radioimmunoassay and -10 (IL-6, IL-8 and IL-10) using ELISA.

RESULTSLevel of SOD activity significantly decreased during (T2) and after CPB (T3 and T4) in the control group, as compared to the T1 (P < 0.01), but it was unchanged in the tested group; level of MDA increased during and after CPB in both groups (P < 0.01), but more significantly in the control group (P < 0.05), so comparison after CPB between the two groups showed a higher SOD and lower MDA level in the tested group. Plasma levels of TNF-alpha, IL-6, IL-8 and IL-10 significantly increased in both groups after CPB (T3 and T4, P < 0.01) as compared to the T1, but the comparison between groups showed lower plasma TNF-alpha, IL-6 and IL-8 levels at T3 and T4, and higher IL-10 level at T4 in the test group (P < 0.01). All patients had stable life signs with no occurrence of adverse reaction.

CONCLUSIONDHI has obvious protective effect on cerebral injury in patients undergoing CABG with CPB, the mechanisms may be associated with the actions of anti-oxidation, anti-inflammation and regulation on immune factors.

Aged ; Brain Injuries ; blood ; drug therapy ; surgery ; Coronary Artery Bypass ; Drugs, Chinese Herbal ; administration & dosage ; Extracorporeal Circulation ; Female ; Humans ; Injections ; Interleukin-10 ; blood ; Interleukin-6 ; blood ; Interleukin-8 ; blood ; Male ; Middle Aged

We used metabolomics to investigate the ability of a traditional Mongolian medicine called modified Tabusen-2 (MT-2) to improve kidney yang deficiency (KYD) in rats. All animal experiments were conducted under the guidance and standards of the Medical Ethics Committee of Inner Mongolia Medical University. SD rats were divided into 6 groups of six rats: a normal group, a model group, Jinkuishenqi pill administration group (1.26 g·kg^-1), and MT-2 administration in high-, medium- and low-dose groups (1.512, 0.756, and 0.378 g·kg^-1). KYD was established by intramuscular injection of hydrocortisone (HC) and biochemical indicators and clinical characterization was used to confirm that KYD was established. All groups received intragastrically administered drug (Jinkuishenqi pill or MT-2) or saline. Serum from each group was collected after 8 weeks and analyzed by UPLC-Q-exactive-MS to measure various biochemical indicators. The biomarkers affected by MT-2 were identified and the metabolic pathways of KYD regulated by MT-2 were analyzed by metabolomic analysis. The results show that MT-2 can decrease serum creatinine (Cr) in KYD rats and significantly increase (P<0.05) the content of thyroid stimulating hormone (TSH) and luteinizing hormone (LH). In serum samples, 38 biomarkers such as corticosterone, L-phenylalanine, and DL-tryptophan were measured as possible indicators for disease development in KYD rats. MT-2 lowered 18 biomarkers of KYD, including corticosterone, deoxycorticosterone, and L-phenylalanine, and altered 13 related metabolic pathways including phenylalanine, tyrosine and tryptophan biosynthesis, phenylalanine metabolism and steroid hormone biosynthesis, resulting in an overall improvement in KYD. MT-2 appears to be important in improving KYD in rats mainly by regulating metabolites such as amino acids, steroids and lipids.

The adhesion properties of tumor cells with extracellular matrix(ECM) are closely associated with their invasion and metastasis.Our work reported here was intended reveal the relevant biomechanical and biorheological manifestations of human lung cancer. Using micropipette aspiration technique, we investigated quantitatively the adhesive mechanics properties of high metastatic human giant cell carcinoma(PG) cells as well as low metastatic adenocarcinoma(PAa) cells of lung based on cell culture in vitro. The results showed that the adhesion forces of PAa and PG cells to collagen Ⅳ were significantly higher than those to glass surfaces, but at the lower concentrations(1.00μg/ml and 2.00μg/ml) of collagen Ⅳ, the amplitude for the increase of adhesion forces of PG cells were less than the amplitude for that of PAa cells, and most of the adhesion force values of PAa cells to the coated surfaces of incorporation of laminin along with 2 μg/ml collagen Ⅳ were significantly greater than those of PG cells. At the lower concentrations(0.625μg/ml for PAa cells,and 0.625 μg/ml, 1. 25 μg/ml for PG cells) of laminin tested,the adhesion force values of PAa and PG cells all decreased, but the amplitude and level for the decreased values of adhesion forces of PG cells were greater than those for the PAa cells. In conclusion, the adhesive and proteolytic behaviour of cancer cells to extracellular matrix might be mediated mainly by tumor cell membrane receptors such as integrin receptors and laminin receptors, it might affect the biological characteristics and the metastasis of the tumor cells. The results may benefit to explain some questions in biomechanical views about how the highly metastatic PG cells are prone to migration and invasion.

To explore a simple and effective method to determinate the volume of CD34(+) cells in the peripheral blood of donors received drug mobilization for stem cell transplantation by using flow cytometry, the mobilized peripheral blood from donors and 100 micro l fresh whole blood were labeled with monoclonal antibodies Anti-CD34-PE and Anti-CD45-FITC, after lying the red blood cells, and assessed with flow cytometer FL2 (log) vs SSC (log) and FL1 (log) vs SSC (log) were mainly used for analysis windows. The results showed that a level of CD34(+) cells in whole nucleated cells as low as 0.05% - 0.1% can be detected effectively using this method when 10(5) nucleated cells were counted. At day 5 or day 6, the level of CD34(+) cells in most samples of patients reached a peak volume, some of samples and the levels were more than one percent in. It was concluded that CD34(+) cells can be effectively determined by using this method. According to the relative rate of CD34(+) cells, the time to harvest the stem cells in blood can be determined.
Antigens, CD34 ; blood ; Blood Donors ; Flow Cytometry ; methods ; Hematopoietic Stem Cells ; cytology ; Humans

BACKGROUNDMutations in fumarylacetoacetate hydrolase (FAH) gene can lead to tyrosinemia type 1 (HT1), a relatively rare autosomal recessive disorder. To date, no molecular genetic defects of HT1 in China have been described. We investigated a Chinese family with a HT1 child to identify mutations in FAH.

METHODSDNA sequencing was used for mutations screening in FAH gene. Real-time polymerase chain reaction (PCR) was performed to determine the FAH gene expression level. To confirm the presence of degradation by the nonsense-mediated mRNA decay pathway (NMD), the fragments containing R237X mutations were analyzed by primer introduced restriction analysis-polymerase chain reaction (PIRA-PCR) and cDNA sequencing. Finally, the effects of the mutations reported in this study were predicted by online softwares.

RESULTSA boy aged 3 years and 8 months was diagnosed clinically with HT1 based on his manifestations and biochemical abnormalities. Screening of FAH gene revealed two heterozygous mutations R237X and L375P transmitted from his mother and father respectively. In this pedigree, the amount of FAH mRNA relative to a healthy control was 0.44 for the patient, 0.77 for his mother and 1.07 for his father. Moreover, both PIRA-PCR and cDNA sequencing showed significant reduction of the FAH mRNA with R237X nonsense mutation. The missense mutation of L375P was not reported previously and prediction software showed that this mutation decreased the stability of protein structure and affected protein function.

CONCLUSIONSThis is the first case of HT1 analyzed by molecular genetics in China. The R237X mutation in FAH down- regulates the FAH gene expression, and the L375P mutation perhaps interrupts the secondary structure of FAH protein.

Child, Preschool ; China ; Humans ; Hydrolases ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Mutation, Missense ; genetics ; Nonsense Mediated mRNA Decay ; genetics ; Real-Time Polymerase Chain Reaction ; Tyrosinemias ; genetics

Multi-stage countercurrent extraction technology, integrating solvent extraction, repercolation with dynamic and countercurrent extraction, is a novel extraction technology for the traditional Chinese medicine. This solvent-saving, energy-saving and high-extraction-efficiency technology can at the most drive active compounds to diffuse from the herbal materials into the solvent stage by stage by creating concentration differences between the herbal materials and the solvents. This paper reviewed the basic principle, the influence factors and the research progress and trends of the equipments and the application of the multi-stage countercurrent extraction.
Countercurrent Distribution ; methods ; Drugs, Chinese Herbal ; isolation & purification ; Particle Size ; Plants, Medicinal ; chemistry ; Solvents ; chemistry ; Technology, Pharmaceutical ; instrumentation ; methods ; Temperature ; Time Factors

BACKGROUND: At present, treatments for spinal cord injury are limited, with poor outcomes. Therefore, it is of great importance to explore new treatment methods. OBJECTIVE:To observe the therapeutic effect of bone marrow mesenchymal stem cells(BMSCs)injection via the caudal vein on spinal cord injury in rats. METHODS:BMSCs were isolated and cultured in vitro by whole bone marrow adherence method.The surface markers were identified by flow cytometry. Thirty Sprague-Dawley rats were randomly divided into control group, spinal cord injury group and BMSCs transplantation group, 10 rats in each group. A rat spinal cord injury model was established by occlusion of the 10ththoracic vertebra using an aneurysm clamp, and 2×106BMSCs were injected through the caudal vein at 10 minutes after modeling. Basso-Bettle-Bresnahan (BBB) score for motor function recovery was assessed at 0, 10, 20, 30 days after transplantation. The therapeutic effect was evaluated by hematoxylin-eosin staining and electron transmission microscopy at 30 days after implantation. RESULTS AND CONCLUSION: Under the microscope, fusiformis-shaped or fibroblast-like cells were observed. The expression rate of CD44 and CD90 was more than 90% and the expression of CD45 was less than 2%, by which, the BMSCs were identified. The BBB scores were significantly higher in the BMSCs transplantation group than the spinal cord injury group at 20 and 30 days after transplantation (P < 0.05). Hematoxylin-eosin staining showed that there was spinal cord tissue damage, vascular rupture injury, neuronal cell degeneration and inflammatory cell infiltration in the spinal cord injury group. After BMSCs transplantation, the number of spinal cord neurons was markedly increased with intact cytomembrane and clear nucleolus. Electron microscopic results showed that spinal cord axon swelling, demyelination, nerve axon deformation and necrosis were observed in the spinal cord injury group, while after BMSCs transplantation, the rat spinal cord axon structure was repaired,and partially lost myelin was recovered with uniform thickness.To conclude,BMSCs transplantation via the caudal vein has a significant therapeutic effect on spinal cord injury in rats by repairing the spinal cord structure and protecting the integrity of axon and myelin structures.

BACKGROUND: Until now, there is no effective treatment for peripheral neuropathy caused by acrylamide. Therefore, it is necessary to explore new treatment methods. OBJECTIVE: To explore the protection role and its mechanism of bone marrow mesenchymal stem cells (BMSCs) against acrylamide-induced intoxication in the spinal cords of rats. METHODS: BMSCs were cultured by the whole bone marrow adherence method and identified by morphological observation and flow cytometry detection. Thirty Sprague-Dawley rats, clean grade, were randomly divided into three groups (n=10 for each group): normal control group, acrylamide group and BMSCs transplantation group. The latter two groups received acrylamide by gavage, 50 mg/(kg?d), 5 days per week, for 2 weeks with an interval of 2 days. Then, in the BMSCs transplantation group, 3×106BMSCs were transplanted by the caudal vein, 5 days per week, for 3 consecutive weeks. Hematoxylin-eosin staining was utilized to observe the morphological changes of the spinal cord. Tunel assay was used to detect cell apoptosis. Western blot assay was adopted to detect the expression levels of Bcl-2 and Bax. RESULTS AND CONCLUSION: In the acrylamide-exposed rats, the damage to the structure was found in the spinal cords by morphological observation, which was significantly alleviated after BMSCs transplantation. The disturbed expression levels of Bax and Bcl-2 were also significantly inversed after BMSCs transplantation (P < 0.05). These results suggest that BMSCs transplantation can inhibit cell apoptosis in the spinal cords of acrylamide-intoxicated rats, probably by up-regulating expression of Bcl-2 and down-regulating expression of Bax.