Objective To explore the Cep70 by adjusting the stability of acetylated alpha tubulin,participate in breast cancer drug resistance mechanisms.Methods (1) In order to induce taxol drug resistance cell line Michigan cancer foundation-7 (MCF-7)/pac,high-dose shock treatments taxol MCF-7 was used for 6 months,until the cells can grow in 3.5 μmol/L of paclitaxel.(2) The 3-(4,5-dimenthylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) method was used to detect inhibition rate by taxol to MCF-7 and MCF-7/pac cell.(3) Immunofluorescence and Western blot were used to test acetylated alpha-tubulin and Cep70 expression levels in MCF-7 and MCF-7/pac cells.(4) Chemical intervention was used to acetylate apha-tubulin expression,Western blot and polymerase chain reaction (PCR) were used to detect the change of acetylated alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac groups.Flow cytometry and Western blot were used to detect the change of cell cycle.Results (1) IC50 of MCF-7 and MCF-7/pac was 22.47 μ mol/L and 31.38 μmol/L,respectively.(2) Immunofluorescence and Western blot results showed that the expression of acetylation of alpha-tubulin in resistant MCF-7 cell/pac was obviously decreased.(3) Real time polymerase chain reaction (RT-PCR) and Western blot showed Cep70 expression was consistent of acetylation of alpha-tubulin.(4) After incubation with paclitaxel for 24 hours,the expressions of acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac were increased,but the extent of MCF-7 cell was much higher.Instead,incubation with nocodazole after 24 hours,the acetylation of alpha-tubulin and Cep70 in MCF-7 and MCF-7/pac cells were obviously lowered.(5) After paclitaxel intervention,compared to the same group MCF-7 cells,the G2 phase ratio in MCF-7/pac cells was lower.In addition,given nocodazole after the intervention,compared to the same group MCF-7 cells,the ratio of G2 phase in MCF-7 cell/pac was significantly decreased.Conclusions Cep70 decreased the expression of the acetylated alpha-tubulin,reduced the stability of microtubules,which could be an important mechanism of taxol drug resistance.

Objective To reveal acetylated a-tubulin acts as pressure sensors,sensing the changes in extracellular matrix to impact on the invasion and metastasis of breast cancer.Methods The acetylated alpha microtubule protein expressions were detected,and its relationship with invasion and metastasis in breast cancer clinical specimens and different molecular subtypes of breast cancer cell lines were analyzed.The expression of acetylated a-tubulin was interfered through chemical methods,and cell morphology and the change of the invasion and metastasis ability were detected under the condition of different matrix hardness.Results The expression of acetylated a-tubulin was highest in basal-like breast cancer tissue and cell lines,and was lowest expressed in the luminal B breast cancer tissue and the breast cancer cell lines.The expression of acetylated a-microtubule was positively correlated with the occurrence of breast cancer.Under the condition of soft substrate cultivation,cell polarization was declined,becoming the circular or oval shape.The acetylated a-tubulin caused reduction in cell polarity,accompanied with less invasion and metastasis ability.Conclusions The acetylated a-tubulin acts as pressure sensors,sensing the sclerosis of extracellular matrix in the process of tumorigenesis,promoting invasion and metastasis of breast cancer.

The poly ester-amide (PEA) nanofiber membrane was prepared by electrospinning and the surface topography of nanofiber membrane was observed by scanning electron microscope (SEM). It was found that with the increasing of spinning fluid concentration from 10% to 20%, the diameter of prepared fibers was increased from 180 nm to 305 nm. The chemical structure of PEA was detected by infrared spectrum, which showed that electrospinning had no significant effect on PEA. X-ray diffraction spectrogram and differential scanning calorimetry revealed that the crystallinity of PEA was decreased after electrospinning. Mechanical property test demonstrated that the mean breaking strength and elongation at break of PEA nanofiber membrane with (0.50±0.05) mm thickness were (1.00±0.18) MPa, and (18.20±2.86)%, respectively. MTT results showed that the endothelial cells proliferated actively on the PEA nanofiber membrane, and presented good adhesive and growth status.

OBJECTIVE:To study the percutaneous permeability of methoxypsoralen liposomal gel METHODS:The preparations were applied on the rat skin in vitro and in vivo with common gel and Mingbailing tincture as control,and the drug concentrations in the receiver,skin,blood and other organs were determined by RP-HPLC RESULTS:In vitro experiment,liposomal gel showed the lowest percutaneous permeation rate and the highest retention in skin In vivo studies illuminated that the skin depositions of drug encapsulated in liposome was(4 37?0 91) and (3 36?0 58) times those of the common gel and tincture respectively CONCLUSION:Methoxypsoralen liposomal gel showed an evident targeting effect on skin

Objective To explore the MRI features of Langerhans cell histiocytosis (LCH) in central nervous system (CNS) in children. Methods Clinical and MRI data of 25 cases with LCH in CNS from three children's hospitals between January 2009 and December 2014 were retrospectively studied. All cases were confirmed by surgery or biopsy. All cases underwent non?contrast and contrast pituitary and/or cranial MRI examinations. The location, morphology, MRI signal and enhanced patterns of LCH lesions were observed and analyzed. Result A total of 17 cases had hypothalamic-pituitary LCH, with 2 of them complicated with pineal cyst, 2 complicated with brain parenchymal lesions, and one complicated with both pineal cyst and brain parenchymal lesions. MR images showed that neurohypophysis lost its original hyper?intensity on T1WI, and nodular or homogeneous thickening was seen in the pituitary stalks. Dura matter was involved in 3 cases, 2 of them had single lesion, and the other one got multiple lesions. Neoplasm in choroid plexus was seen in trigone of left lateral ventricles in one case. Three cases with pineal gland involved demonstrated cystic change. Four cases had gray matter involved, with cerebellar dentate nuclei involvement in 2 cases, and both thalamus and basal ganglia involvement in the other two cases. Three cases showed white matter involvement without obvious Virchow-Robin space enlargement and brain atrophy. Conclusions MR imaging of LCH in CNS shows certain specific characteristics. The diagnosis can be made comprehensively based on both clinic features and other imaging findings.

Objective:To construct an eukaryotic expressing vector pCR3 1 15 containing CP15 gene of Crypstosporidium parvum(C.parvum) and express it in Hela cells Methods:CP15 gene of C parvum was obtained from pMD18 T 15 disgested by BglⅡ and was inserted into eukaryotic expressing vector pCR3 1(+) in BamHⅠ site,and then Hela cells were transfected with recombinant by liposomes The transcription and expressed products of CP15 in the transfected Hela cells were assayed by RT PCR,ELISA and indiret immunofluorescence assay after screening with G418 Results:It showed that pCR3 1 15 was constructed successfully CP15 gene was transcripted in transfectants and CP15 protein with obvious biological activity was highly expressed in Hela cells Conclusion:CP15 gene in recombinant vector is proved to be expressed in Hela cells and obvious biological activity of expression production in transfected cells was detected

A molecular template synthetic polymer highly selective for norfloxacinum was prepared by a molecular imprinting technique. The selective binding characteristics of the template polymer was evaluated by Scatchard analysis. The multiple-sites binding model was used to calculate the maximum numher of binding sites and the dissociation constant. The results showed that the template polymer using methacrylic acid (MAA) as functional monomer could form two kinds of binding sites. The dissociation constants were estimated to be Kd1 = 2.9 × 10-5mol/L and Kd2 = 3.2 × 10-3 mol/L. The selective binding experiment for substrates indicated that the polymer gave much higher affinity and selectivity for norfloxacin than for acid pipemidic and cefalexin. It is possible to be a good adsorption and binding material in the selective enrichment and determination of trace norfloxacin in complex biosamples.

Clinical data of 4 cases of infants with hemophilia complicated with hematorrhachis diagnosed in Guangzhou Women and Children′s Medical Center from January 2017 to December 2018 was retrospectively analyzed.The clinical manifestations of 4 children was atypical, only showing irritability and crying, poor spirit and weak head in the early stage, and torticollis and limb weakness in the late stage may easily contribute to a misdiagnosis.The spinal magnetic resonance imaging is an important diagnosis of hematorrhachis.The most common site of disease was cervicothoracic and thoracolumbar spinal epidural hemorrhage.Of the 4 patients in this article, 1 patient was treated with surgery combined with infusion factor Ⅷ replacement therapy.Three patients were treated with infusion factor Ⅷ replacement therapy alone to obtain clinical rehabilitation.The prognosis of early diagnosis was good, the time from onset to diagnosis in 3 cases was within 1 week, after treatment all had rehabilitation; 1 case within 2 weeks of diagnosis, partial rehabilitation.It is suggested that hemophilia complicated with hematorrhachis in infants is hidden, and it can only be irritable and crying in the early stage, and neurological symptoms appear in the late stage.Improving diagnostic awareness, early diagnosis and early infusion factor replacement therapy are critical for prognosis.

Objective To study the relationship between expression of EGFR and E- cadherin and metastasis of esophageal cancer. Methods The expression of EGFR and E- cadherin in esophageal cancer tissues of 47 patients by LSAB immunohistochemistry method was studied. Results The positive rates of EGFR in distant metastasis group and local lymph node metastasis group were 47.62 % and 56.52 % respectively. They were higher than individual corresponding control groups(27.08 % and 26.32 %)(P

Objective To study exogenous expression and subcellular localization of wild type (WT ) and mu-tant PAX3 proteins in vitro by generating their expression plasmids for further study of pathogenesis of Waarden-burg syndrome (WS) .Methods The plasmids pECE-PAX3 and pcDNA3 .0-HA were ligased after they were cut by double enzyme digestion using molecular cloning technique to generate recombinant eukaryotic expression plasmid pcDNA3 .0-PAX3-HA ,which was as a template to generate expression plasmids pcDNA 3 .0 -H80D -HA and pcDNA3 .0-H186fs-HA of novel mutations H80D and H186fs of PAX3 gene .All constructs were verified by di-rect nucleotide sequencing .NIH3T3 cells were transfected transiently with the expression plasmids of PAX3 ,H80D and H186fs respectively .The exogenous expression of WT PAX3 protein and mutant H80D ,H186fs proteins were analysed using Western blot assay ,while their subcellular distribution were observed using immunofluorescence as-say .Results The DNA sequences of expression plasmids of PAX3 and its mutant H80D ,H186fs were correct . Both WT and mutant PAX3 proteins were detected at the expected size .WT PAX3 and H80D proteins were only lo-calized in the nucleus ,whereas H186fs protein showed aberrant localization in both cytoplasm and nucleus .Conclu-sion We successfully generated the recombinant eukaryotic expression plasmids of PAX 3 gene and its mutants and drew preliminary conlusion of gene mutation having effect on subcellular distribution of WT PAX 3 proteins in vitro , which lays experimental basis for further study of the moceluar mechanism of WS caused by PAX3 gene mutations in China .