Roles of NG2 Glia in Cerebral Small Vessel Disease / 神经科学通报·英文版

Cerebral small vessel disease (CSVD) is one of the most prevalent pathologic processes affecting 5% of people over 50 years of age and contributing to 45% of dementia cases. Increasing evidence has demonstrated the pathological roles of chronic hypoperfusion, impaired cerebral vascular reactivity, and leakage of the blood-brain barrier in CSVD. However, the pathogenesis of CSVD remains elusive thus far, and no radical treatment has been developed. NG2 glia, also known as oligodendrocyte precursor cells, are the fourth type of glial cell in addition to astrocytes, microglia, and oligodendrocytes in the mammalian central nervous system. Many novel functions for NG2 glia in physiological and pathological states have recently been revealed. In this review, we discuss the role of NG2 glia in CSVD and the underlying mechanisms.

Structure, content and psychometric properties of physical activity measurement/reporting tools applicable to children and adolecents with disabilities: a systematic review / 中国康复理论与实践

ObjectiveTo synthesize the structure, content and psychometric properties of physical activity measurement/reporting tools applied in children and adolescents with disabilities. MethodsLiterature on physical activity measurement/reporting tools for children with disabilities were retrieved from PubMed, Medline, EBSCO, Web of Science and CNKI, from establishment to June, 2023. The main structure and content characteristics of the tools were analyzed using the International Classification of Functioning, Disability and Health-Children and Youth Version (ICF-CY). The psychometric properties of the tools was assessed using the COnsensus-based Standards for the selection of health Measurement INstruments (COSMIN). ResultsSix tools were selected, and nine papers were finally included. Two tools were globalized standard tools, namely Global Matrix 4.0 Physical Activity Report Card for Children and Youth (Global Matrix 4.0) and the Global Matrix of Para Report Card on Physical Activity of Children and Adolescents with Disabilities (Para Report Card), measuring b455 functions of motor tolerance function, b7 neuromusculoskeletal and motor-related functions; d4 mobility, d450 walking, d470 using transportation, d820 school education, d910 community life, d920 recreation and leisure; e310 immediate family, e320 friends, e460 societal attitudes, and e575general social support services, systems and policies, etc. These tools showed a complete structure, high reliability, validity and feasibility, and could be performed in a self-reported manner or completed by the custodian or school teacher. The International Physical Activity Questionnaire (IPAQ) might measure d4 mobility, d450 walking, d640 doing housework, d820 school education, d920 recreation and leisure. IPAQ was good in terms of internal consistency, content and construct validity, and cross-cultural validity, with different test-retest reliability and hypothesis testing results reported in different applied studies. Children and Adolescents Physical Activity and Sedentary-Questionnaire (CAPAS-Q) might measure the d4 mobility, d450 walking, d470 using transportation, d820 school education, and d920 recreation and leisure. CAPAS-Q was good in internal consistency, retest reliability, content and construct validity, but hypothesis testing results and cross-cultural validity were unknown. Physical Activity Questionnaire for Older Children (PAQ-C) and Physical Activity Questionnaire for Adolescents (PAQ-A) might measure d4 mobility, d450 walking, d470 using transportation, d820 school education, and d920 recreation and leisure. PAQ-C and PAQ-A were strong in internal consistency, retest reliability, content and construct validity, and cross-cultural validity, without hypothesis testing results. ConclusionIn terms of content, Para Report Card is the most comprehensive and extended physical activity measurement/reporting tool for children and adolescents with disabilities. In terms of the psychometric properties of the instruments, the multidimensional, structured physical activity measures show high internal consistency, retest reliability, and content and construct validity, with varying retest reliability and correlational validity with objective measures across studies.

Adaptive physical activity and its health benefits for patients with spinal cord injury based on ICF: a scoping review / 中国康复理论与实践

ObjectiveTo analyze adaptive physical activity interventions and their health benefits for patients with spinal cord injury (SCI) based on International Classification of Functioning, Disability and Health (ICF). MethodsA combination of subject headings and free words was employed to search for relevant literature on physical activity in patients with SCI in PubMed, Web of Science, EBSCO and CNKI, from January, 2017 to July, 2022. A scoping review was conducted. ResultsEight English articles were included, from Australia, the United States, Estonia, Canada, Netherlands and Brazil. The study involved 150 patients with SCI and included seven randomized controlled trials and one non-randomized controlled trial. These literatures were primarily from journals in the fields of SCI, neurology, neuromuscular medicine and physical activity, with publication dates concentrated between 2017 and 2021. The included studies involved participants with complete or incomplete SCI, presenting with paraplegia, tetraplegia, and various levels of injury severity (grades A to D). The main functional disorders related to ICF included b710 mobility of joint functions, b715 stability of joint functions, b720 mobility of bone functions, b730 muscle power functions, b735 muscle tone functions, b750 motor reflex functions, b760 control of voluntary movement functions and b770 gait pattern functions; the functions involved in activities and participation included d410 changing basic body position, d415 maintaining a body position, d420 transferring oneself, d445 hand and arm use, d450 walking, d455 moving around, d570 looking after one's health, d610 acquiring a place to live, d910 community life, d920 recreation and leisure; the environmental factors involved were e1151 assistive products and technology for personal use in daily living, and e1401 assistive products and technology for culture, recreation and sport. Physical activity intervention sites mainly included home, community, medical or rehabilitation institutions. The physical activity could be classified into prevention, health promotion, treatment and rehabilitation. Patients with SCI had completed acute and subacute rehabilitation in medical and rehabilitation institutions, and returned to their families and communities, and some of them received services in medical and rehabilitation institutions from time to time. The forms of physical activity included strength training combined with routine nursing, short-term resistance training, hydrotherapy and robot treadmill activities, functional electrical stimulation and therapeutic exercise, progressive exercise training based on exoskeleton of power machine, leg bicycle assisted by electrical stimulation combined with manual bicycle, and acute exercise with different intensity. The activity frequency was twelve to 60 minutes a time, two to three times a week, lasting for three to 16 weeks, and the activity intensity was mainly medium to high. The health benefits of physical activity on patients with SCI were mainly reflected in four aspects: physical and mental health, activity and behavior health, environmental factors, quality of life and well-being. In terms of physical and mental health, it helped to improve muscle function (muscle strength, strength perception), walking function (walking speed, walking distance), respiratory and circulatory system functions (peak oxygen uptake, aerobic endurance, cardiopulmonary function, reducing the risk of cardiopulmonary diseases, etc.), immune system related functions, and improving psychosocial function (fatigue degree). In terms of activity and behavioral health, it was helpful to improve activity skills and abilities. In terms of environmental factors, the availability and effectiveness of some auxiliary equipment were confirmed. In terms of quality of life and well-being, it could improve self-living ability and quality of life. ConclusionThis study established a PICO framework for adaptive physical activity and its health effects in patients with SCI based on ICF. Physical activity for patients with SCI can be conducted in various settings, including home, community, or medical and rehabilitation institutions. The physical activities of patients with SCI are characterized by wheelchair-based adaptive physical activities, which are mainly divided into two categories: various aerobic exercises and resistance exercises based on wheelchairs, and physical activities based on auxiliary exercise equipment or intervention methods (such as robot treadmill, power machine exoskeleton, functional electrical stimulation, etc.). The frequency of physical activity in patients with SCI is twelve to 60 minutes a time, two to three times a week, lasting for three to 16 weeks, and the activity intensity is maily medium to high. Therapists and rehabilitation professionals can provide guidance and support through various means, such as online or offline supervision and one-on-one coaching, to promote the health benefits of physical activity for patients with SCI, including improved physical and psychological function, enhanced activity levels, reduced sedentary behavior, and increased self-care abilities and quality of life.

Simulation research on magnetoacoustic B-scan imaging of magnetic nanoparticles / 生物医学工程学杂志

As drug carriers, magnetic nanoparticles can specifically bind to tumors and have the potential for targeted therapy. It is of great significance to explore non-invasive imaging methods that can detect the distribution of magnetic nanoparticles. Based on the mechanism that magnetic nanoparticles can generate ultrasonic waves through the pulsed magnetic field excitation, the sound pressure wave equation containing the concentration information of magnetic nanoparticles was derived. Using the finite element method and the analytical solution, the consistent transient pulsed magnetic field was obtained. A three-dimensional simulation model was constructed for the coupling calculation of electromagnetic field and sound field. The simulation results verified that the sound pressure waveform at the detection point reflected the position of magnetic nanoparticles in biological tissue. Using the sound pressure data detected by the ultrasonic transducer, the B-scan imaging of the magnetic nanoparticles was achieved. The maximum error of the target area position was 1.56%, and the magnetic nanoparticles regions with different concentrations were distinguished by comparing the amplitude of the boundary signals in the image. Studies in this paper indicate that B-scan imaging can quickly and accurately obtain the dimensional and positional information of the target region and is expected to be used for the detection of magnetic nanoparticles in targeted therapy.

Preparation and evaluation of long-acting injectable formulations of memantine / 中国药科大学学报

@#:In order to improve the compliance of patients with Alzheimer′s disease and maintain the continuity of treatment by reducing administration frequency of memantine hydrochloride, a series of memantine long-acting nanosuspension-based injectable formulations were prepared using a hydrophobic salt formation method. Four hydrophobic salt forms of memantine were prepared, including memantine oleate(Mem-Ole), memantine stearate(Mem-Ste), memantine palmitate(Mem-Pal)and memantine pamoate(Mem-Pam). The salt forms of memantine were characterized using fourier transform infrared(FTIR)spectroscopy, proton nuclear magnetic resonance(1H NMR)spectroscopy and powder X-ray diffraction(PXRD)analysis. The equilibrium solubilities of different salt forms of memantine and the in vitro drug release of long-acting injectable formulations were investigated. In comparison with memantine alone, the equilibrium solubilities of Mem-Ole, Mem-Ste, Mem-Pal and Mem-Pam in simulated body fluid were decreased by 95. 1%、96. 2%、96. 7% and 99. 6%, respectively. Meanwhile, the equilibrium solubilities of Mem-Pam in simulated body fluid with pH ranging from 5 to 8 were all lower than 0. 07 mg/mL. The order of the in vitro drug release rate of the four long-acting injectable formulations with nanosuspensions of memantine was Mem-Ste> Mem-Pal≈Mem-Ole> Mem-Pam> Memantine. The Mem-Pam nanosuspensions could sustain drug release for seven days and exhibited a zero-order drug release profile(y=0. 549 9x+7. 594 2, r=0. 988 3). In conclusion, injectable Mem-Pam nanosuspensions showed desired drug release behavior and might potentially be applied in vivo for a week with a steady plasma drug concentration-time profile.

Effect of miR-126 on proliferation and apoptosis in colon cancer cells via targeting regulation of SOX2 expression / 中国比较医学杂志

Objective To explore the effect of miR-126 on proliferation and apoptosis in colon cancer cells via targeting regulation of SOX2 expression. Methods miR-126 mimics and miR-126 NC were transfected into SW480 cells by liposome LipofectamineTM2000. The expression of miR-126 was detected by RT-PCR. Cell viability was determined by MTT staining. Cell apoptosis and cell cycle were detected by flow cytometry. The expression of SOX2 protein and mRNA was measured by western blot and RT-PCR. Luciferase reporter analysis was performed. Results Compared with miR-126 NC, the expression of miR-126 was upregulated(P< 0.01),cell viability was reduced(P< 0.01),early cell apoptosis rate and late apoptosis rate were increased(P< 0.01), cell cycle was arrested at G1 phase(P< 0.01), meanwhile, miR-126 mimics targeted downregulation of the expression of SOX2 protein and mRNA(P< 0.01). Conclusions miR-126 mimics can inhibit SW480 cell proliferation and induce cell apoptosis by targeting downregulation of expression of SOX2.

Expression of transcription factors Sp1 and Survivin in breast cancer and their correlation / 实用肿瘤学杂志

Objective The objective of this study was to investigate the expression and clinical significance of transcription factors Sp1 and Survivin in breast cancer and to further analyze their correlation.Methods Seventy cases of breast cancer patients were diagnosed as breast cancer from June 2013 to June 2015.The expression of Sp1 and Survivin in 70 cases of breast cancer and 20 cases of adjacent normal tissues were detected by immunohistochemical staining.The relationship was detected between the protein expression and the clinicopathological parameters of breast cancer.Results The positive rates of Sp1 and Survivin in breast cancer were 71.43% and 78.57%,respectively.The positive rates of Sp1 and Survivin were 30% and 10% in adjacent normal breast tissues.They showed a significant difference(P0.05).There were also no differences in the age,tumor size and histological grade of patients with breast cancer(P>0.05).In breast cancer tissues,there was a significant positive correlation between Sp1 and Survivin expression(r=0.517,P<0.01).Conclusion The transcription factors Sp1 and Survivin are closely related to the pathogenesis of breast cancer,which may serve as an adjunctive index for the diagnosis of breast cancer.It has certain guiding value for clinical diagnosis,treatment and prognosis of breast cancer.

Effects of Four-week Intermittent Fasting on Skeletal Muscle Mass and Autophagy in Rats / 中国运动医学杂志

Objective To observe the changes of body weight,fat mass and skeletal muscle mass of rats after 4 weeks of intermittent fasting,and explore relationship with autophagy in skeletal muscle,so as to provide theoretical basis for intermittent fasting.Method Twenty male Sprague Dawley rats were randomly divided into a control group (Con) and an intermittent fasting group (IF),each of 10.The rats of IF group were forbidden to eat food every Wednesday and Friday,and the body weight of both groups was recorded weekly.After 4 weeks,Dual-Energy X-Ray Absorption (DEXA) was used to analyze the body fat mass,then the bilateral soleus was separated to record the wet weight and measure the cross-sectional area of the soleus fibers by testing laminin with immunofluorescence confocal laser scanning microscope.The form of autophagic vacuole of soleus was observed using a transmission electron microscopy.The expression of autophagy-related protein LC3,p62 and regulating protein AMPK,pAMPK and ULK1 were measured using Western blotting.Result After 4 weeks of intermittent fasting,the weight and fat mass of IF were significantly lower than those of Con (P＜0.01),but there were no significant differences between them in wet weight and cross-sectional area of soleus (P＞0.05).The expressions of AMPK,p-AMPK,ULK1 in IF were significantly higher than those in Con (P＜0.01).Compared with Con,the expression of LC3-Ⅱ/LC3-Ⅰ of IF increased significantly,while the expression of p62 decreased significantly (P＜0.01).Conclusion Four weeks of intermittent fasting decreases the fat mass significantly,and control the weight efficiently.Intermittent fasting can maintain the skeletal muscle mass by promoting moderate autophagy through the AMPK-ULK1 pathway.It should be a potential lose weight method for further research.

Effects of IL-13 on SDF-1 and EGF expression in fibroblasts co-cultured with breast cancer cells / 中国免疫学杂志

Objective:The effects of IL-13 ( Interleukin-13 ) on SDF-1 ( Stromal cell derived factor 1 ) and EGF ( Epidermal growth factor) expression in fibroblasts co-cultured with breast cancer cells were investigated to explore the mechanism for IL-13 in the development of breast cancer.Methods:The co-culture of human breast cancer cell line MDA-MB-231 with the human skin fibroblast line CCC-ESF-1 ( ESF) was used in vitro and in tumor-burdened nude mice.The effects of IL-13 on SDF-1 and EGF expression in the co-cultured fibroblasts in vitro were analyzed using reverse transcription quantitative polymerase chain reaction ( RT-qPCR ) , flow cytometry and Western blot assay.The proliferation of the co-cultured human breast cancer cells in vitro was detected by Cell counting kit-8(CCK-8).The effects of IL-13 on SDF-1 and EGF expression in the fibroblasts of tumor tissue of tumor-burdened nude mice were analyzed using immunofluorescence and laser confocal microscope, and the tumor volumes were examined.Results: IL-13 could up-regulate SDF-1 and EGF expression in the fibroblasts co-cultured with breast cancer cells in vitro,and promoted the proliferation of the co-cultured breast cancer cells.In tumor-burdened nude mice,IL-13 enhanced SDF-1 and EGF expression of fibroblasts in tumor tissue, and accelerated tumor growth.Conclusion:IL-13 up-regulates SDF-1 and EGF expression of fibroblasts co-cultured with breast cancer cells.The molecular mechanism of promoting effect of IL-13 on breast cancer relates to SDF-1 and EGF of fibroblasts in breast cancer stroma.

Study of the efficacy and toxicity of gemcitabine combined with nedaplatin in anthracycline and/or taxane -pretreated patients with metastatic triple -negative breast cancer / 中国基层医药

Objective To evaluate the short -term efficacy and adverse reactions of gemcitabin combined with nedaplatin to patients with metastatic triple -negative breast cancer after taxane and/or anthracycline treatment. Methods Thirty -three patients with anthracycline and/or taxane -resistant metastatic triple -negative breast cancer received gemcitabin and nedaplatin regimen:gemcitabin 1 000 mg/m2,d1,8 days,intravenous;nedaplatin 80mg/m2,d1,intravenous.Treatments were repeated every 21 days and response rate was evaluated every two cycles. Results Of 33 patients, complete remission ( CR) in 1 case ( 3.03%), partial remission ( PR) in 13 cases (39.39%),stable disease(SD) in 11 cases(33.33%),progressive disease(PD) in 8 cases(24.24%),objective response rate was 42.42%.The main adverse reaction was bone marrow suppression and gastrointestinal reaction. Conclusion Combination therapy of gemcitabin and nedaplatin is an effective and well tolerated rescue regimen in anthracycline and/or taxane -resistant metastatic triple -negative breast cancer patients.

NGX6 gene combined with cisplatin in treatment of lung cancer in vitro and in vivo / 中国生化药物杂志

Objective To evaluate the effect of NGX6 combined with cisplatin on the inhibition rate of A549 cells and NCI-H1975 cells and antitumor effects in vivo.Methods The NGX6 was loaded in the LPD, and prepare Liposome protamine DNA complexes.A549 cells and NCI-H1975 cells were seperately divided into NGX6 group ( 30 μg/mL NGX6 concentration ) , cisplatin group, NGX6 +cisplatin group, PBS as negative control group.The effect of cytotoxicity of four group on A549 cells and NCI-H1975 cell were evaluated by MTT assay.the clone forming rate and the inhibition rate were determined by Cell colony count.lung transplantation tumor model were successfully established, then nude mice were divided into four groups as abeve, each group of 10, tumor size and survival period were determined tumor cell apoptosis were observed.Results The cell viability of A549 cells and NCI-H1975 cells of (NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The cloning efficiency of A549 cells and NCI-H1975 cells of ( NGX6 +cisplatin) were lower than that of NGX6 group, cisplatin group and saline group, respectively(P<0.01).The tumor inhibitory rate was in vivo for (NGX6 +cisplatin) was higher than other group(P<0.01).The median survival of nude mice in (NGX6 +cisplatin), NGX6, cisplatin and saline group were 43,31,29 and 15 days.Conclusion NGX6 combination with cisplatin can inhibit the cell proliferate of lung cancer cells and inhibit the tumor growth and the combination of NGX6 and cisplatin may be a potentially effective treatment for lung cancer.

IL-13 induces STAT6 phosphorylation and promotes fibrosis in human hepatic stellate cells / 中华微生物学和免疫学杂志

Objective To investigate the effects of IL-13 on fibrosis in hepatic stellate cells and its molecular mechanism .Methods The effects of IL-13 on the proliferation of hepatic stellate cell line LX-2 were measured by MTT assay .The transcription level of collagen typeⅠ( COLⅠ) in LX-2 cells was detec-ted by RT-PCR.The secretion of COLⅠin LX-2 cells was measured by ELISA assay and hydroxyproline as-say.Western blot assay was used to analyze the effects of IL-13 on the phosphorylation of signal transducer and activator of transcription(STAT6).Results Compared with control group, IL-13 (10 ng/ml, 20 ng/ml and 50 ng/ml ) significantly stimulated the proliferation of LX-2 cells in a dose-dependent manner ( P0.05).The expression of phosphorylated STAT6 protein in LX-2 cells was significantly enhanced upon the stimulation with 50 ng/ml of IL-13 ( P<0.05 ) for60 min or 120 min.C onclusion IL-13 promoted the proliferation of human hepatic stellate cells and up-regulated the expression of COLⅠat mRNA and protein levels .IL-13 might promote the fibrosis in human hepatic stellate cells through activating STAT 6 phosphorylation .

Clinical significance of CD+4CDHi25CDLo127 regulatory T cell in the peripheral blood of patients with lung cancer / 肿瘤研究与临床

Objective To evaluate the changes and clinical significance of CD+4CDHi25CDLo127 regulatory T cell(Treg) in peripheral blood of patients with lung cancer. Methods 30 patients with lung cancer and 20 heathy volunteers were included in this study. The proportion of Treg population in CD4+ T cells stained with three colors was analysed by flow cytometry. The serum level of IL-10 and TGF-β were measured by ELISA. Results The proportion of Treg in patients with squamous cell careinoma(n=20), adenocarcinoma (n=10) were all significantly higher than that of healthy controls (P <0.05), but there was not obvious difference between the two groups with different pathological types(P0.05). Increased serum level of IL-10 and TGF-β was also detected in lung cancer patients. Conclusion The proportion of Treg is increased in lung cancer patients, which may result in the inhibition of host anti-cancer immune response by excreting IL-10 and TGF-β.

Tyrosine kinase inhibitor A77-1726 inhibiting STAT6 phosphorylation and cfos expression induced by IL-13 / 中国药理学通报

Aim To investigate the effects of tyrosine kinase inhibitor A77-1726 on IL-13 induced STAT6 phosphorylation and c-fos expression in Dami cell and to provide novel experimental basis to the clinical application of A77-1726 and the study of IL-13 pathway.Methods Total RNA was extracted from Dami cells incubated with or without IL-13 and A77-1726 respectively for various time points.RT-PCR and agar gel electrophoresis were used to examine the expression of c-fos mRNA.The expression of STAT6 and c-fos proteins was detected by Western blot.A densitometric rel-ative quantitation of PCR and Western blot products was quantitated using Image Quant software.Results STAT6 was phosphorylated by treatment of 100 ?g?L-1 IL-13 in Dami cells.Phosphorylation of STAT6 induced by IL-13 was inhibited by treatment of 50?mol?L-1 A77-1726.The expression of c-fos mRNA was significantly induced by IL-13 treatment in Dami cells(P

Life quality of patients after cholecystectomy / 中华外科杂志

<p><b>OBJECTIVE</b>To evaluate the quality of life in patients who underwent laparoscopic and open cholecystectomy for chronic cholecystolithiasis.</p><p><b>METHODS</b>A prospective survey was made on 25 patients receiving laparoscopic cholecystectomy (LC group) and 26 patients receiving open cholecystectomy (OC group). The quality of life was measured with the gastrointestinal quality of life index (GLQI) preoperatively, and at 2, 5, 10 and 16 weeks after the operation.</p><p><b>RESULTS</b>The mean preoperative GLQI scores of all dimensions of the quality of life were 112.5 and 110.3 in the LC and OC groups respectively. In the LC group, the quality of life was not considerably reduced at 2 weeks after operation, with a mean GLQI score of 110. There was a significant improvement both in total mean score and in the aspects of symptomatology, emotional and physiological status from 5 to 16 weeks after LC operation. In the OC group, the GLQI score reduced to 102.0 at 2 weeks after surgery (P < 0.05). There were significant reductions in the aspects of symptomatology, physiological and social status as well. The GLQI scores reached to the preoperative level of 115.6 at 10 weeks after the operation (P > 0.05). The patients experienced significant improvements of GLQI at 16 weeks after OC operation (P < 0.01 or P < 0.05). The LC group showed better GLQI scores than did the OC group for up to 10 weeks postoperatively (P < 0.05).</p><p><b>CONCLUSIONS</b>LC is betler or more rapidly than OC is improving the quality of life postoperatively. The assessment of the quality of life is valuable for measuring the outcome of surgical treatment.</p>

Effect on the expression of proto-onogene c-mpl in megakaryocytes line-HEL by interleukin-13 / 中国病理生理杂志

AIM:To investigate the relevance of the proliferation of megakaryocytic cell line-HEL stimulated by the recombinant human interleukin-13 (IL-13) to the expression of pro-oncogene c-mpl in HEL cells. METHODS: MTT colorimetric assay and reverse transcrition polymerase chain reaction (RT-PCR) are separately used in this study to observe the effect on the proliferation of HEL cells and the expression of c-mpl mRNA in HEL cells by rhIL-13. RESULTS: RhIL-13 stimulated the proliferation of HEL cells and upregulated the expression of c-mpl mRNA in HEL cells. CONCLUSION: Our results suggest that rhIL-13 stimulated the proliferation of HEL cells and provide the evidence that its mechanism is partly because of increasing the pro-oncogene c-mpl expression in HEL cells.

Upregulation of proto-oncogene c-mpl by interleukin-13 in Dami cells / 中国病理生理杂志

AIM: To investigate the effects of recombinant human interleukin-13 (rhIL-13) on the expression of proto-oncogene c-mpl in Dami cells, a human megakaryobiastic leukemia cell line. METHODS: The expression of c-mpl mRNA in Dami cells was investigated with RT-PCR. The expression of membrane-bound protein c-mpl on Dami cells was investigated with ligand combination experiment. RESULTS: In RT-PCR experiment, we found the quantitis of expression of c-mpl mRNA in 25 ?g/L rhIL-13 group increased by 24.8%. In ligand combination experiment, we found quantitis of expression of membrane-bound protein c-mpl in 100 ?g/L rhIL-13 group increased by 28.5% ( P

Cloning,transfer and anti-human breast cancer cell growth of the extracellular domain of cadherin 5 / 中国病理生理杂志

AIM:In order to study the effect of the extracellular domain of cadherin 5 on the growth of a human breast cancer cell line MDA-MB435.METHODS:Cadherin extracellular domain repeats 1 to 4(CED1-4)was cloned by using RT-PCR technique,and inserted into the plasmid vector pMSCV.pMSCV-CED1-4 was propagated in XL-blue strain of Escherichia coli,extracted and purified.CED1-4 was cut by restriction endonuclease,examined by using agar gel electrophoresis,and finally sequenced.CED1-4 gene was transferred into MDA-MB435 cell line.The expression of CED1-4 gene in MDA-MB435 cell was analyzed by methods of RT-PCR and Western blotting.The effect of CED1-4 on the growth of MDA-MB435 cell was observed by the methods of proliferation experiments in vitro and the experiments in nude mice in vivo.RESULTS:The recombinant vector pMSCV-CED1-4 was successfully constructed.CED1-4 band appeared between the 1 636 bp and 1 018 bp in agar gel electrophoresis.The sequence result showed that CED1-4 had 1 452 bp and codes 484 amino acids.PCR and Western blotting identified that CED1-4 mRNA and protein were expressed in the transfected MDA-MB435 cells.Cell proliferation experiments showed that the proliferation rate of MDA-MB435 cells was lower in the experimental group than that in the experimental control group and the blank control group.The mean volume and weight of tumors in nude mice were lower in the experimental group than those in the experimental control group and the blank control group.CONCLUSION:The growth of a human breast cancer cell line MDA-MB435 is inhibited in vitro and in vivo by cadherin 5 extracellular domain CED1-4.

Experimental study of endothelin-1 gene expression in uterine tube tissue / 中国病理生理杂志

AIM: The distribution of endothelin-1(ET-1) in uterine tube and its source were investigated in order to find out relation between endothelin-1 and function of uterine tube.METHODS: The distribution of endothelin-1 and the expression of ET-1 mRNA in the rabbits uterine tubes were studied using SABC immunohistochemistry and in situ hybridization histochemistry with digoxigenin-labelled rat ET-1 cRNA probe. RESULTS: ET-1 granules with red color were seen in the uterine tube epithelial cells and they were located mainly above the cell nucleus and under the cell surface membrane. Less ET-1 granules were seen in the myometrium of uterine tube. ET-1 mRNA positive hybridization signals with deep blue were distributed in the uterine tube epithelial cells. These signals were strong and dense. They were distributed mainly above the nucleus and near the cell surface membrane. ET-1 mRNA positive hybridization signal was not seen in the myometrium of uterine tube.CONCLUSION: Our results indicate that epithelial cells of rabbit's uterine tube epithelial cells secrete ET-1.

High-efficient genetic transfection of CD41~+,UT7,U937 and MDA-MB-435 cells with a recombined murine stem cell retroviral vector / 中国病理生理杂志

AIM: Gene transduction with a recombined murine stem cell retroviral vector has been investigated to find an effective way of gene transduction and to offer theory and experimental basis for the recombined murine stem cell retroviral vector used for gene transduction. METHODS: 1. Construction of retrovirus vector: EC1-4 gene (repeats 1-4 of cadherin-5 extracellular domain) and mutant (Ser 222A) MEK1 gene were cloned into retrovirus vector pMSCV after cut by Bgl Ⅱ and EcoR 1 restriction endonuclease. 2. Obtaining CD41 + cells and cell culture: Cells expressing CD34 + from cord blood were isolated. The inducement of cells expressing CD41 from CD34 + cells was performed by using TPO and cells CD41 + were selected by FACS. NIH 3T3 cells were cultured in high sugar DMEM medium and U937 in RPMI 1640 medium. UT7 cells which is cytokine-dependent cell line were grown in Iscove's modified Dulbeco's medium supplemented by GM-CSF. 3. Determination of viral titers: Retroviral vectors were transferred to packaging cell line 293. Retroviral containing supenatant was collected after transfection. The viral titers was tested on infection of NIH 3T3 cells by FACS analysis. 4. Western blot: Transduced CD41 +,UT7,U937 and MDA-MB-435 cells were analyzed by western blot to examine expression of transduced genes. RESULTS: A packaging cell line 293 produces high-titer MEK1 pMSCV retroviruses (3.1?10 7) and EC1-4 pMSCV retroviruses (1.0?10 8). With 8-folds dilution retroviruses,60.73% GFP positive cells have been obtained in MEK1 pMSCV transduced UT7 cells,72.56% in U937 cells and 30.57% in CD41 + cells,respectively. GFP positive cells have reached up 97.54 % in EC1-4 pMSCV transducted MDA-MB-435 cells. Phosphorylated MEK1 has been decreased in experiment group when TPO has stimulated CD41 + and UT7 cells or serum has stimulated U973 cells. This indicates that is a dominant negative effect of mutation MEK gene. EC1-4 gene transduced MDA-MB-435 cells have expressed EC1-4. CONCLUSION: The recombined murine stem cell retrovirus can effectively mediate gene transduction of CD41 +,UT7,U937 and MDA-MB-435 cells,and transduced genes can be stably expressed.