1.Cep70 drives taxol drug resistance through regulating the acetylation α-tubulin to reduce microtubule stability
Yufang HE ; Ni QIU ; Hongsheng LI
Journal of Chinese Physician 2017;19(2):195-200
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.
2.Acetylated a-tubulin sense the change of extracellular matrix to regulate the invasion and metastasis of breast cancer
Ni QIU ; Yufang HE ; Hongsheng LI
Journal of Chinese Physician 2017;19(2):186-190
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.
3.Preparation and biocompatibility of poly ester-amide electrospinning nanofiber membrane
Xiaofeng LIU ; Xiumei MO ; Changlong HE ; Hongsheng WANG
Chinese Journal of Tissue Engineering Research 2009;13(51):10045-10048
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.
4.Characteristics of Molecular Recognition of Norfloxacin Template Polymer
Hongsheng GUO ; Xiwen HE ; Jie ZHOU ; Hong LIANG
Chinese Journal of Analytical Chemistry 2001;29(2):128-132
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.
5.MRI features of Langerhans cell histiocytosis in central nervous system in children
Hongwu ZENG ; Yungen GAN ; Wenxian HUANG ; Ling HE ; Hongsheng LIU
Chinese Journal of Radiology 2016;50(4):252-255
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.
6.Relationship between expression of EGFR and E-cadherin and metastasis of esophageal cancer
Hongsheng LI ; Yuandong WANG ; Xiaojun TAN ; Weixing HE ; Guangri ZHAO
Cancer Research and Clinic 1997;0(03):-
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
7.The Percutaneous Permeability of Methoxypsoralen Liposomal Gel
Hongxia WU ; Wen HE ; Hongsheng CAI ; Shunde LUO ; Xianzhou ZHANG
China Pharmacy 2001;0(11):-
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
8.Construction of a eukaryotic expressing vector containing CP15 gene of C.parvum and its expression in Hela cells
Hongxuan HE ; Xichen ZHANG ; Hongsheng OUYANG ; Al ET
Chinese Journal of Immunology 1986;0(04):-
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
9.Four cases of hemophilia complicated with hematorrhachis in infants
Rujuan LING ; Hongsheng LIU ; Yun ZHENG ; Liya HE
Chinese Journal of Applied Clinical Pediatrics 2021;36(2):140-142
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.
10.The molecular mechanism of inhibition of murine Lewis lung carcinoma metastasis by weimaining in vivo
Jinli LOU ; Hongsheng LIN ; Quanying QIU ; Yingxia PEI ; Xin QI ; Xiujuan HE
Chinese Journal of Pathophysiology 1986;0(04):-
AIM: To investigate the anti-metastasis effect of weimaining, extracted from fragopyrum cymosum meissn, a Chinese medicine, on murine Lewis lung carcinoma (3LL). METHODS: The anti-metastasis effect of weimaining in vivo was detected in the grafting lung metastasis model of murine Lewis lung carcinoma. The effects of the drug on the expression of CD34 and E-cadherin were investigated by immunohistochemical staining and RT-PCR. RESULTS: Weimaining effectively inhibited the lung metastasis of 3LL at a concentration of 250 mg?kg -1?d -1, significantly suppressed the expression of CD34 and increased the expression levels of E-cadherin protein and mRNA in 3LL cells. CONCLUSIONS: Weimaining inhibits the metastasis of murine Lewis lung carcinoma (3LL) in vivo via increasing the expression of E-cadherin and decreasing microvessel density of tumor tissue.