Cancer, also known as malignant tumor, is the second largest disease after heart disease, which is characterized by genomic instability and mutagenicity. Ataxia telangiectasia and RAD3-related kinase (ATR) are members of phosphatidylinositol 3-kinase (PIKK) family, belonging to serine/threonine kinase, one of the key kinases in DNA damage response (DDR) and DNA repair pathway. This paper reviews the latest progress in the ATR inhibitor field including mechanism of action (MOA), therapeutic applications, and the combination therapy from the perspective of medicinal chemistry. It also discusses the possible challenges and future directions of developing ATR inhibitor antitumor drugs, which could provide the scientists in this field the convenience for access the information and application guidance for clinical studies.

A binary plant expression vector, pCM12-slm, carrying the aroAM12 mutant gene encoding bacterial 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) and the Bts1m recombinant gene consisting of 331 N-terminal amino acids of CryIAc and 284 C-terminal amino acids of CryIAb has been constructed. The truncated Bts1 gene was fused with the PR1b signal peptide sequence and expressed in tobacco plants under the control of 2E-CaMV35S promoter and the omega (omega) translation enhancer sequence from tobacco mosaic virus. The mutant aroAM12 was fused with the transit sequence of tobacco EPSPS and expressed in tobacco plants under the control of the CaMV35S promoter. Tobacco leaves were transformed with Agrobacterium tumefaciens LBA4404 harboring the pCM12-slm plasmid, and the transgenic plants were selected directly on medium containing the herbicide. Forty glyphosate resistant plants were regenerated, with a transformation frequency of 27%. Transgenic plants were initially assessed for glyphosate resistance by placing leaf discs on shoot induction media containing the herbicide. Rooted plantlets, propagated from selected transgenic tobacco, were transferred to soil in a greenhouse and tested for glyphosate resistance by spraying them with Roundup at a commercial recommended dose. The glyphosate resistance assay indicated that all the transgenic plants showed highly resistant to the herbicide. The PCR assay showed that the aroAM12 gene was present in all of the 40 T0 transfer plants, and Bts1m genes present in 28 of 40 of the transgenic plants. Southern blot analysis further confirmed that the copy number of the transgenes varied from one to three copies in different transgenic plants. Northern blot and immunodot blot showed that the aroAM12 and Bts1m genes were expressed at the transcription and translation levels. Transgenic plants containing both the aroA M12 and Bts1m genes were further assessed for insect resistance. Tobacco leaves of T0 transgenic plants were infested with tobacco bollworm H. assulta larvae for 6 days. The result (table 1) showed that the survival rate of insect larvae was between 0-10%, and the growth of insect larvae was seriously inhibited, suggesting pCM12-slm as a dual functional vector with potential application in breeding of glyphosate and insect resistance transgenic plants.
Agrobacterium tumefaciens ; genetics ; Animals ; Blotting, Northern ; Blotting, Southern ; Genetic Vectors ; genetics ; physiology ; Herbicides ; pharmacology ; Moths ; pathogenicity ; Plant Diseases ; parasitology ; Plant Leaves ; drug effects ; genetics ; parasitology ; Plants, Genetically Modified ; drug effects ; genetics ; parasitology ; Polymerase Chain Reaction ; Tobacco ; drug effects ; genetics ; parasitology

OBJECTIVETo investigate the relationship between differential expression of VEGF and its receptors and clinical characteristics of childhood acute lymphoblastic leukemia (ALL).

METHODSExpressions of VEGF and its receptors (Flt-1, KDR) were assayed by ELISA and RT-PCR in healthy donors(20 cases), ALL patients in remission (20 cases), with low risk (29 cases) and with high risk (10 cases). The clinical data of all the patients and volunteers enrolled in this study were collected and analyzed according to the expression of VEGF and its receptors.

RESULTSThe expressions of VEGF were (574.37 +/- 208.45) ng/L, (387.93 +/- 175.86) ng/L, (135.80 +/- 111.28) ng/L and (91.16 +/- 41.34) ng/L in patients with high risk, standard risk, in remission and healthy donors, respectively. Expression levels of VEGF receptors were downwards with risk grades. The clinical manifestations were also in accord with the expression levels of VEGF and its receptors.

CONCLUSIONALL patients with highly expressed VEGF and its receptors are usually with higher tumor burden, and refractory treatment. Detection of VEGF and its receptors might be one of prognostic marker for ALL treatment.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; RNA, Messenger ; genetics ; Receptors, Vascular Endothelial Growth Factor ; genetics ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVE: To investigate the effects of fenofibrate on the proliferation and apoptosis and endothelial nitric oxide synthase (eNOS) mRNA expression of cultured human umbilical vein endothelial cells (HUVECs) induced by lysophosphatidylcholine (LPC). METHODS: HUVECs were cultured in vitro. The study was designated to 5 groups according to fenofibrate concentration: control group, LPC group, LPC + low-concentration fenofibrate (10 micromol/L), LPC + middle-concentration fenofibrate (50 micromol/L), and LPC + high-concentration fenofibrate (100 micromol/L). The study was designated to 6 groups according to the intervention time: control group, LPC group, LPC + fenofibrate (50 micromol/L) 6 h, LPC + fenofibrate 12 h, LPC + fenofibrate 24 h, and LPC + fenofibrate 48 h. The proliferation and apoptosis of HUVECs were evaluated by MTT assay, flow cytometry and fluorescence microscopy, respectively. eNOS mRNA were assayed by real time-PCR. RESULTS: Compared with the control group, LPC could inhibit the proliferation and induce apoptosis, and downregulate eNOS mRNA expression and decrease NO production of HUVECs. Fenofibrate could increase the proliferation and decrease the apoptosis, and up-regulate eNOS mRNA expression and enhance NO production in HUVECs. CONCLUSION Fenofibrate could improve the proliferation and inhibit the apoptosis, and up-regulate eNOS mRNA expression of HUVECs induced by LPC, which may be responsible for fenofibrate to prevent and treat atherosclerosis.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelium, Vascular ; cytology ; Fenofibrate ; pharmacology ; Humans ; Hypolipidemic Agents ; pharmacology ; Lysophosphatidylcholines ; pharmacology ; Nitric Oxide Synthase Type III ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Umbilical Veins ; cytology

OBJECTIVETo establish the inner blood-retinal barrier (BRB) model in vitro by co-culturing RF/6A cells and C6 cells and to investigate the effects of EMP (200 kV/m, 200 pulses) exposure on the permeability of the inner BRB model in vitro.

METHODSRF/6A cells and C6 cells were co-cultured on transwell, and the characteristic of the inner BRB model was assessed by detecting transendothelial electrical resistance (TEER) and the permeability of horseradish peroxidase (HRP). The co-cultured model was exposed or sham exposed to the EMP (200 kV/m 200 pulses) for 0.5, 3, 6, 12, 24 h in vitro, then TEER and the permeability of HRP were measured for studying the effects of EMP on the permeability of inner BRB model in vitro.

RESULTSTEER value (145 Ωcm(2)) of the co-culturing inner BRB model significantly increased, as compared to that of RF/6A cells alone model (P < 0.05) on the 6th day after inoculation. There was significant difference of permeability of HRP between the co-culturing inner BRB model and RF/6A cells alone model (P < 0.05). The ability of inhibiting large molecular materials in the co-culturing inner BRB model enhanced. The TEER value decreased and the permeability of HRP increased as compared to the sham group at 0.5, 3, 6 h after the exposure.

CONCLUSIONThe inner BRB model by co-culturing RF/6A cells and C6 cells in vitro is efficient and suitable to study the alterations of the restricted permeability function of the inner BRB. EMP (200 kV/m for 200 pulses) could induce the enhanced permeability of the inner BRB model in vitro.

Animals ; Blood-Retinal Barrier ; physiology ; Cell Line, Tumor ; Coculture Techniques ; Electric Impedance ; Electromagnetic Fields ; Endothelial Cells ; physiology ; Macaca mulatta ; Permeability ; Rats ; Retina ; cytology

Objective To detect the dynamic visual acuity ( DVA) before and after vestibular habituation of subjects in order to optimize the DVA assessment criteria .Methods The vestibular function examination system was applied to the detection of static and dynamic visual function in 16 healthy subjects .Results When the speed of left or right swinging was fast enough , DVA before and after vestibular habituation was different .Conclusion Subjects with vestibular habituation can reduce their sensitivity to the vestibular system , the changes in DVA are better than before habituation , and the vestib-ular function adaptability training may have effect on DVA .

BACKGROUND:Epidermal growth factor is an auxiliary growth factor,but its effect on the growth of bone marrow mesenchymal stem cells (BMSCs) is uncertain.OBJECTIVE:To establish a mature method for isolation,extraction and identification of rat BMSCs,to investigate the effects of epidermal growth factor (EGF) on the proliferation and migration ability of BMSCs and to explore its potential mechanisms at the same time.METHODS:Rat BMSCs were isolated and purified using the improved whole bone marrow adherence method.After the cells were subcultured to the third generation,we detected the expression of cells surface antigens CD29,CD45 and CD90 by flow cytometry.BMSCs were further identified by osteogenic and adipogenic differentiation.Meanwhile,the effect of EGF on the proliferation of passage 3 BMSCs was measured by cell counting kit-8 and clonogenic assay.And the migration of P3 cells was verified by Transwell chamber.In addition,we detected the expression of proteins related to PI3K/Akt and nuclear factor-κB signaling pathways by western blot assay.RESULTS AND CONCLUSION:The primary BMSCs were polygonal and spindle-shaped,and then gradually appeared to be spindle-shaped.The results of flow cytometry demonstrated that the passage 3 cells were positive for CD29 and CD90,but negative for CD45.Furthermore,we successfully induced the osteogenic and adipogenic differentiation of BMSCs in vitro.Additionally,our data demonstrated that EGF promoted the proliferation and migration of passage 3 BMSCs.The relative expression levels of p-Akt and Bcl-2 of PI3K/Akt signaling pathway was up-regulated and the expression of Bax was down-regulated.At the same time,the relative expression level of phosphorylated p65 of nuclear factor-κB signaling pathway was up-regulated and the expression of phosphorylated inhibitor κB was down-regulated.Moreover,the downstream protein of matrix metalloproteinase-9 was up-regulated.Those proteins were related to the migaration of BMSCs.In summary,our results suggest that EGF could promote the proliferation and migration of BMSCs.

Objective To study the clinical features and curative effect of hepatitis associated aplastic anemia (HAAA) in children of China.Methods Patients' records in China Biological Medicine Database (CBM-disc 1980-2011) and Wan Fang Database were reviewed.The clinical data of the children with newly diagnosed HAAA were respectively studied,including clinical manifestations,blood routine,bone marrow examination and viral serology results,as well as the treatment and clinical effect.Results A total of 50 children were confirmed as HAAA.There were 41 boys and 9 girls.The median age was 8.2 years(range 0.8 to 15.0 years) on diagnosis.The causes of hepatitis could not be identified.The median interval between hepatitis occurrence to blood cell reduction was 10 weeks.Twenty-two cases were diagnosed as severe aplastic anemia and 13 cases as very severe aplastic anemia.Fifteen children died within 1 month after diagnosis.In the immunosuppression treatment group,the percentage of overall responders was 69.2％,which was higher than that of the non-immunosuppression treatment group (18.5％) (x2 =9.920,P ＜ 0.01).Conclusions Severe HAAA is very common in school children,especially in boys.The children with HAAA have a higher early death rate.Immunosuppression therapy is effective if combined with androgenic hormone with an earlier diagnosis.

Objective To discuss the clinical curative effect of percutaneous vertebroplasty(PVP)combined with percutaneous pedicle screw fixation for thoracolumbar fracture.Methods Retrospectively analyzed the clinical data of 43 patients with thoracolumbar fracture who underwent PVP combined with percutaneous pedicle screw fixation in our hospital from November 2015 to June 2017.Those patients included 28 males and 15 females,and the age of patients ranged from 50 to 66 years old,with an average age of(58.26 ±3.67)years old.The func-tional outcome were evaluated by VAS scores and ODI scores before and after the operation.The sagittal Cobb angle was used to evaluate the reduction of fracture.Results All these patients all successfully completed the operation,and there was no complications after operation.The operation time ranged from 60 to 126 min,with an average time of(96.07 ±15.69)min;the blood loss ranged from 60 to 180 min,with an average time of(113.26 ±24.7)min.All the patients were followed up for 4 to 23 months,with an average time of(12.07 ±4.01)months. The VAS score,ODI score and sagittal Cobb angle were significantly decreased in the last follow -up period compared with those before surgery,and the difference was statistically significant(P<0.05).Conclusion PVP combined with percutaneous pedicle screw fixation in the treatment of thoracolumbar fracture has smaller incision,less blood loss,shorter operation time and better improvement of local pain,func-tional movement and kyphosis.

OBJECTIVE: To determine the effect of different doses of atorvastatin on the serum soluble intercellular adhesion molecules-1 (sICAM-1) in patients undergoing percutaneous coronary intervention (PCI). METHODS: The study consisted of 38 patients with unstable angina and 10 patients with old infarction who underwent elected PCI for stenotic lesions of the coronary artery. Patients were randomly assigned to either aggressive group or conventional one. After PCI the patients took atorvastatin 20 mg per day or 10 mg per day. Blood lipid profile was examined before, and 3 months after the PCI. SICAM-1 was examined before the PCI, 48 hours and 3 months after the PCI. RESULTS: The total cholesterol and LDL-Cholesterol 3 months after the PCI in the 2 groups were lower than those before the PCI (P<0.01). The aggressive group showed greater reduction in concentrations of TC and LDL-C than the conventional group (P<0.01). The changes in concentrations of HDL-C between pre-PCI and 3 months after the PCI and TG were not obvious (P>0.05). sICAM-1 in the 2 groups 48 hours after the PCI significantly higher than that before the PCI (P<0.01). But sICAM-1 in the 2 groups 3 months after the PCI significantly lower than that before the PCI (P<0.01 or P<0.05). The aggressive group showed greater reduction than the conventional group (P<0.01). TC and LDL-C were positively correlated with sICAM-1(r=0.2413, r=0.2691, all P<0.05). CONCLUSION Atorvastatin 20 mg per day reduces TC, LDL-C, and sICAM-1 to a greater extent than atorvastatin 10 mg per day. The effect on sICAM-1 is partly related to reduce lipid profile.
Aged ; Atorvastatin ; Female ; Heptanoic Acids ; administration & dosage ; therapeutic use ; Humans ; Intercellular Adhesion Molecule-1 ; blood ; Male ; Middle Aged ; Percutaneous Coronary Intervention ; Pyrroles ; administration & dosage ; therapeutic use