Objective To observe the expression of vascular endothelial growth factor receptor-1 (VEGFR-1) and VEGFR-2 in hypoxic chorioretinal endothelial cells of monkeys (RF/6A),and to evaluate the effect of minocycline.Methods RF/6A was cultured and divided into four groups:control group,hypoxia group,hypoxia and low dose of minocycline group (0.5 μmol/L),hypoxia and medium dose of minocycline group (5 μmol/L),and hypoxia and high dose of minocycline group (50 μmol/L).Real-time reverse transcription-polymerase chain reaction (RT-PCR) and immunohistopathological staining were used to measure the mRNA and protein expression of VEGFR-1 and VEGFR-2,respectively.Results RT-PCR showed that the expression of VEGFR-1 mRNA did not vary significantly between groups (F24 h =0.17,F48 h =1.53,F72 h =2.04; P＞0.05).Compared with hypoxia group,the expression of VEGFR-2 mRNA in all minocycline treated groups were significantly down-regulated (low minocycline,medium minocycline,high minocycline:t=4.69,20.16,17.12; P＜0.001).The immunohistopathological study showed the cells with positive staining of VEGFR-1 can be observed in all groups,and the staining was relatively weak and mainly located in cell membrane and cytoplasm.The optical density value analysis showed that the protein expression of VEGFR-1 did not vary significantly between groups at all time points(F24 h =0.251,F48 h=0.340,F72 h =0.589; P＞0.05).The VEGFR-2 positive staining cells were also observed in all groups,and the staining was relatively high.Brown staining particles of VEGFR-2 were observed in the cell membrane with minor staining particles in cytoplasm.The staining density of VEGFR-2 was significantly higher in hypoxia group than control group.Compared with the hypoxia group,the protein expression of VEGFR 2 in minocycline treated groups was significantly lower (F24 h =19.147,F48 h =14.893,F72 h =11.984; P＜0.05).Conclusion The expression of VEGFR-2 is up-regulated in RF/6A,and minocycline somewhat shows an inhibition effect.

Objective To investigate the three-dimensional spatial relationships of the sphenopalatine foramen ( SPF) , vidian canal ( VC ) , and foramen rotundum ( FR ) with regard to an endoscopic endonasal approach to the pterygopalatine fossa(PPF)using three-dimensional reconstruction of high resolution computed tomography (HRCT) scans. Methods The HRCT scans of 17 patients and a cadaver specimen were retrospectively evaluated .The morphology of the SPF, VC, and FR as well as the spatial relationship between the SPF and VC were measured on the image of three -dimensional CT reconstruction.Results The mean diameters of the SPF, VC, and FR were(6.26 ±1.59)mm,(2.35 ± 0.77)mm and(2.75 ±0.77)mm, respectively.The mean distance between the VC and posteroinferior margin of SPF was (4.03 ±1.15) mm.The mean vertical and horizontal distances on the image of three-dimensional CT reconstruction were between the VC and FR were(4.94 ±1.35)mm and(9.22 ±3.07)mm, respectively.The whole or partial margin of the VC was above the inferior margin of the SPF in 92%(33/36) of the VC and lateral to the posterior margin of the SPF in 97%(35/36) of the VC.Conclusion The endoscopic endonasal approach to the PPF is performed with greater safety through comprehension of the spatial relationships between the SPF , VC, and FR.

Objective: To observe the effect of acupuncture on serum peptide YY (PYY) and nesfatin-1 in obese patients with insulin resistance. Methods: Ninety-eight obese patients with insulin resistance were divided into a control group and an observation group by the random number table method, with 49 cases in each group. The control group received exercise and dietary interventions, and the observation group received additional acupuncture treatment to the exercise and dietary interventions. The body mass index (BMI), body fat percentage, fasting insulin (FINS), fasting plasma glucose (FPG), homeostasis model assessment for insulin resistance (HOMA-IR), triglyceride (TG), total cholesterol (TC), and serum PYY and nesfatin-1 levels were compared before and after treatment. The efficacy was evaluated after treatment. Results: The total effective rate was significantly higher in the observation group than in the control group (P<0.05). After treatment, the BMI, body fat percentage, and serum TG and TC levels decreased significantly in both groups (P<0.05), and were significantly lower in the observation group than those in the control group (P<0.05). The FINS, FPG, HOMA-IR, and serum PYY and nesfatin-1 levels of the control group were not significantly changed after treatment (P>0.05). The FINS, FPG and HOMA-IR of the observation group decreased significantly after treatment (P<0.05), and were lower than those in the control group (P<0.05). The serum PYY and nesfatin-1 levels of the observation group increased significantly after treatment (P<0.05), and were higher than those in the control group (P<0.05). Conclusion: Based on exercise and dietary interventions, acupuncture is effective for obese patients with insulin resistance. It can reduce the BMI, body fat percentage, blood lipids, blood glucose, and serum insulin levels and improve insulin resistance. The action may be associated with the up-regulation of serum PYY and nesfatin-1.

Tezepelumab(AMG 157/MEDI9929)is a human monoclonal antibody against the epithelial cell-derived cytokine thymic stromal lymphopoietin(TSLP).It is primarily used to treat moderate to severe asthma,particularly in patients with a non-eosinophilic inflammatory phenotype,whose asthma remains uncontrolled despite the use of long-acting beta-agonists and moderate to high doses of inhaled glucocorticoids.This article will summarise the mechanism of action,clinical trial efficacy and safety and tolerability of Tezepelumab in order to provide a comprehensive understanding of the drug and inform clinical work.

Objective To evaluate the dose distribution of target volume and normal tissues in forward intensity modulated radiotherapy (fIMRT) and inverse intensity modulated radiotherapy (iIMRT) modes for breast cancer after radical mastectomy.Methods Both fIMRT and iIMRT plans were developed for 10 patients with breast cancer after radical mastectomy.On each patient's CT images the supraclavicular area, chest wall, and internal mammary area were delineated.The prescription dose was 50 Gyin 25fractions.In the fIMRT plan X-ray irradiation at the dose of 6 MV was adopted for the supraclavicular and the chest wall areas and electron irradiation at the dose of 9 - 12 MeV was adopted for the internal mammary area, and the doses of cold and hot spots were adjusted according to the fitting doses of these 3 regions.In the iIMRT plan the supraclavicular area, chest wall, and internal mammary area were taken asa whole target, 6 MV X-rays was used, and inverse optimal design was performed.The dose distribution oftarget volume and normal tissues, conformal index (CI) , and heterogeneous index (HI) , and acceleratormonitor unit (MU) were analyzed using dose-volume histogram (DVH)for the two intensity modulated modes.Results The maximum dose of PTV of the iIMRT plan was significantly lower than that of the fIMRT plan(t = -3.23,P ＜0.05), the minimum dose and V95% of PTV of the iIMRT were significantly higher than those of the fIMRT plan(t = 4.08, -2.69, both P ＜0.05).The CI level of the iIMRT plan was significantly higher than that of the fIMRT plan and the HI level of the iIMRT plan was significantly lower than that of the fIMRT plan (t = -3.13, 2.74, both P ＜0.05).There were not significant differences in V10, V20, V25, V30, and Dmean of the ipsilateral lung between these 2 groups.However, the V15 of ipsilateral lung of the iIMRT group was significantly lower by 4.2% than that of the fIMRT group (t= 3.2, P ＜ 0.05).There were not significant differences in the mean dose (Dmean) and V30 of heart, and Dmean of contralateral lung and contralateral breast between these 2 groups.Conclusions Compared with fIMRT, the iIMRT plan results in more PTV coverage, higher conformity index, and more homogeneous dose distribution, with lower dose upon the lung at the affected side, and better protection of the contralateral lung, heart, and breast.

Aim To investigate and compare the phar-macokinetics of doxapram injection in healthy subjects of different Chinese nationalities including Han, Mon-golian, Korean, Hui and Uigur, and the influence of gender,in order to provide instruction and help for the usage of doxapram for both clinic and remedy of battle wound. Methods An HPLC-UV method was used to determine the plasma concentration of doxapram. Fifty healthy subjects ( five males and five females of each nationality) were recruited for the study. A single dose of 50 mg doxapram was administered intravenously to the healthy subjects, and blood samples were collected at various predetermined time points. The pharmacoki-netic parameters were calculated by DAS software and were compared by SPSS 13. 0 software, in order to as-sess the influence of nationality or gender on pharmaco-kinetics of doxapram. Results The results indicated that the pharmacokinetic profile of doxapram in vivo could be described as two-compartment model. The main pharmacokinetic parameters for Han, Mongolian, Korean, Hui and Uygur were as follows: Cl ( 0. 25 ± 0. 11 ) , ( 0. 33 ± 0. 11 ) , ( 0. 27 ± 0. 07 ) , ( 0. 26 ± 0. 06) and (0. 39 ± 0. 25) L·h-1 ·kg-1 , while Cmax (1. 55 ± 0. 52 ) , ( 1. 02 ± 0. 30 ) , ( 1. 31 ± 0. 47 ) , (1. 48 ± 0. 46 ) and ( 0. 99 ± 0. 35 ) mg · L-1 . The AUC0-12. 5 , AUC0-∞ and Cmax of Chinese Han were sig-nificantly higher than those of Uigur and Mongolian ( P<0. 05 ) , while there was no significant difference in other parameters ( P>0. 05 ) . There were statistically significant differences in Vc , Vd and CL between young males and females ( P < 0. 05 ) . Conclusion The large inter-individual variation in the main pharmacoki-netics suggests the dosage of doxapram should be ad-justed for different nationalities for both clinic and rem-edy of battle wound.

Purpose To investigate the dosimetric differences among RapidArc (RA) plans which were designed on different target volumes in hepatocellular carcinoma (HCC).Methods A total of 10 HCC patients underwent 3D-CT scan under free breathing ( FB),end inspiration hold ( EIH ) associated with active breath coordinator (ABC) and 4D-CT scan.The 4D-CT were sorted into 10 sets of CT images according to respiratory cycle.The gross tumor volume (GTV) was manually contoured on different CT images.The individual internal gross target volume ( IGTV1 ) was obtained from 4D-CT,and the individual margins from GTVFB to IGTV1.IGTV2 were obtained from GTVFB using individual margins.The planned target volumes (PTV-1,PTV-2,PTV-3 and PTV-4 ) were obtained from GTVFB,IGTV1,IGTV2 and GTVEIA applying different margins.The RA plans (RA1,RA2,RA3 and RA4 ) were designed from different PTVs,and for RA1,RA2 and RA3 the simple 358° arc were used,while three 135° arcs were used for RA4.The dosimetric differences were compared.Results The PTV-1 and PTV-3 were larger than PTV-2 and PTV-4; the mean values of PtV-1/PTV-2 and PTV-1/PTV-4 were 2.5 and 1.9,respectively.There were no significant differences in conformal index,homogeneity index,maximum dose,and minimum dose of PTV among 4 RA plans.The irradiation dose of normal liver of RA3 and RA4 were 8.23 Gy and 7.63 Gy respectively,both significantly lower than those of RA1 and RA2 (10.21 Gy,9.62 Gy,x2 =10.68,P ＜0.05 ),and the V30of RA3 and RA4 were 5.24％ and 5.05％ respectively,both significantly lower than those of RA1 and RA2 (7.76％,6.12％,x2 =14.76,P ＜ 0.05 ).There were no significant differences in irradiation doses of stomach and duodenum among different plans.Conclusions Using 4D-CT or ABC technology with RapidArc in HCC can define the target volume accurately and achieve prefect dose distribution sparing more normal liver volume,compared to the traditional margins.4D-CT and ABC play similar roles in sparing normal liver.

Objective To compare the dosimetric difference in forward intensity modulation radiotherapy (fIMRT)and inverse IMRT(iIMRT)planning for breast cancer.Methods Six patients received radiotherapy alone after left breast-conserving surgery were selected.For each patient,two treatment phns(flMRT and iIMRT)were designed with Pinnacle3 7.4f.In each plan,the volume of PIT received prescription dose was not less than 95 %.The dosimetrie parameters were assessed with dose volume histograms in planning target volume (PIT)and organ of around risk(OAR).Results Of flMRT and ilMRT plans,the PTV average canformal indexwere(0.67±0.06)and(0.66±0.06)(t = 2.423,P ＞ 0.05),average homogeneity index were (28.2±6.0)% and(26.1±6.8)%(t = 2.164,P ＞ 0.05);the volume of left lung received 20 Gy(V20)were(18.7±3.3)% and(17.0±2.8)%(t =5.087,P<0.05),and V30 of left lung were(15.5±3.0)%and(14.0±2.6)%(t =7.272,P<0.05);V30 of heart were(4.1±3.1)% and(3.5±2.5)%(t=1.916,P ＞ 0.05);the total monitor units were(262±5)MU and(308±14)MU(t = 7.515,P < 0.05).Conclusions There were no significant differences of CI,HI,and V30 of heart between flMRT and iIMRT.Because of fewer MUs,fIMRT plan could reduce the machine abrasion and treatment time,but V30 and V30 of left lung are higher significantly than iIMRT plan.

OBJECTIVETo investigate the significance of mutation and single nucleotide polymorphism (SNP) of class III receptor tyrosine kinases such as PDGFRbeta and SHIP in acute myeloid leukemia (AML) patients.

METHODSScreening of the mutation and SNP of PDGFRbeta and SHIP by genomic PCR, RT-PCR, directly sequencing and Mass-ARRAY system was carried out in 273 AML patients.

RESULTSThe mutations of PDGFRbeta R685C and SHIP Q1153L were detected for the first time in AML patients. The positivity ratio was 0.73% and 0.36% respectively.

CONCLUSIONThe mutations of PDGFRbeta R685C and SHIP Q1153L may contribute to leukemogenesis of AML.

Humans ; Inositol Phosphates ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Mass Spectrometry ; Mutation ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Receptor, Platelet-Derived Growth Factor beta ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVEAcute monocytic leukemia (AML)-M5 is the common type of acute myeloid leukemias in children. Studies have shown that there are abundant lipopolysaccharide (LPS) receptor (designated as CD14) molecules on the cell membrane of M5 cells and they play an important role in the diagnosis of M5, since they can be recognized and bound by mouse-anti-human CD14 monoclonal antibody (McAb). However, mouse-originated antibodies are largely not suitable for clinical application due to the severe side effects, thus "humanized antibody" is desired. As the first step for developing humanized antibody, the construction and expression of single chain antibody (scFv) with functional protein are necessary. The present study aimed to express ZCH-7-2F9 ScFv (scFv(2F9)) in eukaryotic cells and obtain the scFv(2F9) protein with a high biological activity.

METHODSFour primers were synthesized to construct the eukaryotic expressional vector, which included SfiI and EcoRI enzyme cleaving site, 6 x His and stop code TGA sequences. scFv(2F9) gene was amplified through splicing by overlap extension (SOE) using the high fidelity Taq polymerase. Positive recombinants (pSectag2A/scFv(2F9)) were identified through enzyme cleaving and sequenced before expression and were transformed into Chinese hamster ovary (CHO) cells for expression. Western-Blot and flow cytometry (FCM) were carried out to determine the relative molecular mass (Mr) and binding activity of scFv(2F9).

RESULTSThe cloned scFv(2F9) gene was identified to be functional by sequencing and expressing. The interested protein could be detected in the culture supernatant of transformed CHO cells with an Mr of 31 000. The blocking test showed that the positive cell percentages, the mean fluorescence intensity (MFI) and the peak of channel (peak Ch) were reduced by 90.02%, 63.30% and 63.38%, respectively after blocking with scFv(2F9), while those were 4.55%, 10.09% and 5.02% after blockage using the supernatant from the CHO cells transfected with blanked vector pSectag2A.

CONCLUSIONSThe scFv(2F9) against human CD14 antigen was successfully expressed in eukaryotic cells and showed a high biological activity, which may be useful for the further studies on its humanized antibodies.

Amino Acid Sequence ; Animals ; Antibodies, Monoclonal ; genetics ; isolation & purification ; Base Sequence ; CHO Cells ; Child ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Humans ; Lipopolysaccharide Receptors ; immunology ; Molecular Sequence Data ; Single-Chain Antibodies ; genetics ; isolation & purification