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Objective:To characterize clinical and neuroimaging features, etiologies, and mechanisms of bilateral middle cerebellar peduncle (MCP) infarctions.Methods:Consecutive patients with bilateral MCP infarctions treated in the Beijing Tiantan Hospital, Capital Medical University between January 1, 2020 and April 30, 2022 were enrolled in this retrospective study. The demographic data, vascular risk factors, clincial manifestations and the National Institutes of Health Stroke Scale (NIHSS) scores were collected. Brain diffusion-weighted imaging was used to assess the regions of cerebral infarction, and the extracranial and intracranial segments of the vertebrobasilar artery were evaluated using magnetic resonance angiography, or computed tomography angiography. The stroke etiology and underlying mechanism were evaluated according to the Chinese Ischemic Stroke Subclassification.Results:Ten patients with bilateral MCP infarctions (8 men and 2 women) were analyzed ultimately. The onset age were 51.0-86.0 (64.8±11.4) years. NIHSS scores were 2.0-12.0 (4.9±2.9) points at admission. All patients had vascular risk factors, most of which were hypertension (10 cases) and dyslipoproteinemia (8 cases). The most common clinical manifestations were vertigo (10 cases), followed by ataxia (9 cases) and dysarthria (8 cases). Four cases were isolated bilateral MCP infarctions, while 6 patients were combined with other vertebrobasilar artery infarctions, 4 of which were combined with cerebellar hemisphere infarctions, consistent with the clinical symptoms. The etiology in all patients was large atherosclerosis (severe stenosis or occlusion of V4 segment of vertebral artery and anterior inferior cerebellar artery; 9 cases). Five patients were classified as hypoperfusion/impaired emboli clearance, while 4 patients were considered as artery-to-artery embolism, and 1 was considered as the parent artery (plaque or thrombosis) occluding penetrating artery.Conclusions:Bilateral MCP infarctions are an extremely rare cerebrovascular disease characterized by vertigo, ataxia, and dysarthria. Cerebral infarction can be isolated or often combined with cerebellar hemisphere infarction. The etiology was mostly stenosis or occlusion of V4 segment of vertebral artery and anterior inferior cerebellar artery.
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Objective:To explore the factors on malnutrition or risk of malnutrition in patients with Alzheimer′s disease (AD)-related cognitive impairment,and to further analyze the association between the severity of behavioral and psychological symptoms in dementia (BPSD) and nutritional status.Methods:The clinical data of 247 patients with AD-related cognitive impairment were collected continuously from the Chinese Imaging, Biomarkers and Lifestyle Study of Alzheimer′s Disease (CIBL) cohort between June 1, 2021 and August 31, 2022. The patients were divided into well-nourished group ( n=128) and malnourished group ( n=119) according to the scores of Mini-Nutritional Assessment scale (MNA). The sociodemographic data (sex, age, body mass index, waist-to-hip ratio, education level), the medical history of olfactory dysfunction, combination with more than two chronic diseases, and gastrointestinal diseases, presenting BPSD, and the scores of the Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment (MoCA), Neuropsychiatric Inventory (NPI), Activity of Daily Living (ADL), Caregiver Burden Inventory (CBI) and Dietary Diversity Score (DDS) were compared between the two groups. The factors with statistically significant differences in hypothesis test and univariate Logistic regression analysis were enrolled in multivariate Logistic regression analysis to further identify independent factors associated with malnutrition in patients with AD-related cognitive impairment. Furthermore, the association between NPI scores and MNA scores was analyzed by Spearman′s rank correlation test. Results:Compared with those in the well-nourished group, patients in the malnourished group had higher age [(66.70±7.01) years vs (69.14±8.87) years, t=-2.39, P=0.018], lower body mass index [(24.68±2.84) kg/m 2vs (22.69±3.63) kg/m 2, t=4.78, P<0.001], and higher proportion of presenting BPSD [22.66% (29/128) vs 76.47% (91/119), χ 2=71.49, P<0.001]; lower scores of MMSE, MoCA, and DDS [24.27±4.69 vs 18.95±8.40, t=6.09; 20.29±5.18 vs 14.55±8.12, t=6.56; 8.00 (8.00, 9.00) vs 8.00 (7.00, 8.00), Z=-4.66; all P<0.001], and higher scores of NPI, ADL and CBI [1.00 (0, 6.00) vs 10.00 (2.00, 25.00), Z=-6.50; 20.00 (20.00, 22.00) vs 27.00 (20.00, 40.00), Z=-7.08; 1.00 (0, 14.75) vs 12.00 (2.00, 35.00), Z=-5.13; all P<0.001]. There were no statistically significant differences in the sex, waist-to-hip ratio, education level, and the medical history of olfactory dysfunction, combination with more than two chronic diseases, and gastrointestinal diseases between the two groups. The multiple Logistic regression analysis demonstrated that the decreased body mass index ( OR=0.79, 95% CI 0.70-0.89, P<0.001), presenting BPSD ( OR=7.84, 95% CI 3.67-16.73, P<0.001), elevated ADL scores ( OR=1.15, 95% CI 1.06-1.24, P<0.001) and CBI scores ( OR=0.98, 95% CI 0.97-1.00, P=0.026), and decreased scores of DDS ( OR=0.66, 95% CI 0.51-0.84, P=0.001) were independently associated with malnutrition in patients with AD-related cognitive impairment. The MNA scores were significantly negatively associated with NPI scores ( r=-0.483,95% CI -0.58--0.38, P<0.001). Conclusions:The decreased body mass index, dietary diversity, and ability of daily living, and presenting BPSD and heavy burden of caregivers can independently contribute to the malnutrition in patients with AD-related cognitive impairment. The more serious the BPSD, the worse the nutritional status.
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Objective:To analyze the correlation between related indexes of serum lipid and insulin resistance and cognitive impairment in middle-aged and elderly people with mild cognitive impairment (MCI).Methods:In this cross-sectional study, 262 middle-aged and elderly patients with a Montreal Cognitive Function Scale (MoCA) cognitive score greater than or equal to 18 points who underwent physical examination in the Health Management Center of Beijing Tiantan Hospital Affiliated to Capital Medical University from January 1 to July 31, 2021 were selected as subjects. According to the cognitive function and MoCA score, the patients were divided into MCI group (143 cases) and normal cognition group (119 cases). Basic data, fasting blood glucose, triglyceride (TG), total cholesterol, apolipoprotein E(ApoE) genotype and other clinical indicators were collected. Hypothesis test was used to compare the differences in basic data, related indicators of blood lipid and insulin resistance between the two groups. Spearman correlation analysis was used to analyze the correlation between related indicators of blood lipid and insulin resistance and MoCA score in the two groups.Results:The age and the proportion of patients with hypertension, coronary heart disease and diabetes in the MCI group were all significantly higher than those in normal cognition group [(54.83±8.29) vs (50.76±6.34) years, 37.76% vs 31.93%, 4.20% vs 0.84%, 16.08% vs 8.40%] (all P<0.05). The elevation of serum TG ( r=-0.50, 95% CI:-0.88--0.12), TG glucose product index (TyG) ( r=-0.75, 95% CI:-1.29--0.20) and TG to high-density lipoprotein cholesterol ratio (TG/HDL-C) ( r=-0.52, 95% CI:-0.91--0.13) were all negatively correlated with MoCA score (all P<0.05). After adjusting for age and gender, the elevation of TG ( r=-0.39, 95% CI:-0.75--0.31) and TG/HDL-C ( r=-0.43, 95% CI:-0.80--0.05) were both still negatively correlated with MoCA score (both P<0.05). There was no significant correlation between all indexes and MoCA scores in the normal cognition group (all P>0.05). The elevated TG was negatively correlated with MoCA score in the MCI group ( r=-0.70, 95% CI:-1.23-0.16, P=0.017). There was no significant correlation between elevated TG and MoCA score in patients carrying ApoE ε2 and ApoE ε3 genotypes in MCI group (all P>0.05). Conclusion:Elevated related indexes of blood lipids and insulin resistance are negatively correlated with cognitive scores in middle-aged and elderly people with MCI, and it′s more obvious in patients with ApoE ε4 genotype.
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Objective:To analyze the influencing factors of post stroke cognitive impairment (PSCI) and their correlation with cognitive scores in patients with acute ischemic stroke.Methods:In this cross-section study, 36 patients diagnosed with acute ischemic stroke and post stroke cognitive impairment (PSCI) admitted to the Department of Vascular Neurology of Beijing Tiantian Hospital Affiliated to Capital Medical University from June 1, 2022 to September 30, 2022 were selected as the PSCI group. And one to one matching was performed for patients without PSCI (PSNCI group) with an age±1 year and same gender admitted to the hospital during the same period (as control, 36 cases). Basic clinical data of the two groups were collected, the laboratory and imaging examinations were completed. Mini Mental State Examination (MMSE) and Montreal Cognitive Assessment Scale (MoCA) were used for cognitive evaluation by neuropsychologists. Hypothesis testing was used to compare the differences in basic data, laboratory tests and lesion sites between the two groups. Multi-factor conditional logistic regression was performed to analyze the influencing factors of PSCI, and Spearman correlation analysis was carried out to analyze the correlation between influencing factors of PSCI and the cognitive scores.Results:Compared with those in PSNCI group, the proportion of patients with stroke/transient ischemic attack history, hyperhomocysteinemia (HHcy), apolipoprotein E(ApoE) ε4 carriers and the ratio of temporal lobe and thalamus infarction were higher in PSCI group (41.7% vs 13.9%, 36.1% vs 2.8%, 30.6% vs 5.6%, 22.3% vs 2.8%, 25.0% vs 5.6%), the MMSE and MoCA scores were lower in PSCI group [16.50 (8.25, 19.00) vs 28.00 (27.00, 30.00), 10.00 (4.25, 14.50) vs 27.00 (25.00, 28.00)] (all P<0.05). Logistic regression analysis showed that HHcy was a positive correlation factor for PSCI ( OR=2.342, 95% CI=1.186-4.622, P=0.014). Spearman correlation analysis showed that MMSE ( r=-0.415) and MoCA ( r=-0.417) scores were negatively correlated with homocysteine (Hcy) (both P<0.05). Conclusion:HHcy is an important factor affecting the occurrence and development of PSCI in patients with acute ischemic stroke, and Hcy level is negatively correlated with cognitive scores in those patients.
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Objective:To explore the impact factors of behavioral and psychological symptoms of dementia (BPSD) in patients with Alzheimer′s disease (AD).Methods:The clinical data of 116 patients with AD admitted to the Outpatient Department of Cognitive Neurology of Beijing Tiantan Hospital, Capital Medical University from June 1, 2021 to March 1, 2022 were collected continuously. The patients were divided into BPSD group ( n=85) and control group ( n=31) according to the presence or absence of BPSD. The sociodemographic data (gender, age, body mass index, years of education), the medical history of hypertension, diabetes mellitus, hyperlipidemia, and the scores of the Mini-Mental State Examination (MMSE), the Activity of Daily Living (ADL), Mini-Nutritional Assessment scale (MNA) and Caregiver Burden Inventory (CBI) were compared between the two groups with hypothesis test and univariate logistic regression analysis. The statistically significant factors in hypothesis test and univariate logistic regression analysis were enrolled in multivariate logistic regression analysis to further identify the factors associated with BPSD in patients with AD. Results:There was no significant statistics differences in the gender, age, body max index, years of education and the medical history of hypertension, diabetes mellitus, hyperlipidemia between the two groups (all P>0.05). Compared with control group, patients with BPSD had lower scores of MMSE and MNA scales [(16.24±7.52) vs (20.81±5.09) points, (21.62±3.75) vs (24.87±2.89) points] (both P<0.001) and higher scores of ADL and CBI scales [29.00 (22.00, 38.50) vs 22.00 (20.00, 25.00) points, 25.00 (12.50, 41.00) vs 3.00 (0.00, 11.00) points](both P<0.001). The multiple logistic regression analysis demonstrated that the decreased MNA scores ( OR=0.762, 95% CI: 0.631-0.922; P=0.005) and elevated CBI scores ( OR=1.077, 95% CI: 1.029-1.128; P=0.002) were associated with BPSD in patients with AD. Conclusion:The malnutrition or the risk of malnutrition and greater caregiver burden can independently contribute to the onset of BPSD in patients with AD.
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Nanodosimetry, a new discipline developed on the basis of microdosimetry, is based on the structural characteristics of particle tracks and studies on the probability distribution of ionization pairs in a specific volume to characterize the damage degree of DNA strand. Now nanodosimetric quantities, which may be measured, gradually form the concept such as ionization cluster size, ionizing cluster size distribution and complementary cumulative probability distribution etc. This paper aims to establish and explain the relationship between nanodosimetry and DNA damage and repair. On the basis of reviewing the current measurement and calculation method of nanodosimetry, this paper summarized the development trend and application prospect of nanodosimetry, and put forward the future research direction of nanodosimetry.
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Objective To observe changes of cell cycle after stable silencing of SNCG gene in human endometrial carcinoma HEC-1A cells.Methods Flow cytometry was used to observe the changes of SNCG stable silencing HEC-1A cell growth cycle.The percentage of G2/M phase of HCE-1A cells after acridine orange staining was observed by laser scanning confocal microscopy.Results Flow cytometry results showed that the group of SNCG stable silencing in G1 and G2/M phase of the cells increased,but the percentage of cells in the S phase were decreased,the difference were both statistically significant (P ＜ 0.05).The results of cells after confocal microscopy observation of acridine orange staining,the group of SNCG stable silencing were observed in G2/M phase high percentage were statistically significant (P ＜ 0.05).Conclusion Two experimental methods commonly validated that the cell cycle of SNCG stable silencing on HEC-1A cells was arrested in G2/M phase,suggesting that SNCG gene is closely related to the cell growth cycle of endometrial carcinoma.
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Objective To investigate the expression of Rap1 GTPase-activating protein 1 (Rap1GAP),matrix metalloproteinase 2 (MMP-2) and matrix metalloproteinase 9 (MMP-9),and their relation with clinical patterns in colorectal carcinoma.Methods Immunohistochemistry was used to detect the expression of Rap1GAP,MMP-2 and MMP-9 in colorectal carcinoma,villous adenoma,tubular adenoma and normal colorectal tissue,and their relationship with clinicopathological parameters was analyzed.Results The positive rate of Rap1GAP expression was 30.4 ％ (14/46),77.8 ％ (14/18),69.6 ％ (16/23) and 95.2 ％ (20/21) in colorectal carcinoma,villous adenoma,tubular adenoma,and normal colorectal tissue,respectively (x2 =30.659,P=0.000).The figures were 71.7 ％ (33/46),55.6 ％ (10/18),52.2 ％ (12/16) and 9.5 ％ (2/21) for the positive rate of MMP-2 expression (x2 =22.459,P =0.000),as well as for 76.1％ (35/46),61.1％ (11/18),56.5 ％ (13/23) and 14.3 ％ (3/21) for the positive rate of MMP-9 expression,respectively (x2 =22.643,P =0.000).In patients with colorectal carcinoma,the expression of Rap1GAP was correlated with tumor differentiation (x2 =5.275,P =0.022),but not sex,age,or lymphatic metastasis (all P ＞ 0.05).The expression of MMP-2 and MMP-9 were correlated with lymphatic metastasis (x2 =6.661,P =0.010;x2 =8.475,P =0.040),but not sex,age or tumor differentiation(all P ＞ 0.05).There was a negative correlation between expression of Rap1GAP and MMP-2,MMP-9 in colorectal carcinoma,respectively (r =-0.424,P =0.003;r =-0.294,P =0.048),but no correlation between the expression of MMP-2 and MMP-9 (r =0.101,P =0.505).Conclusions Rap1GAP,MMP-2 and MMP-9 play important roles in the malignant biological behavior of colorectal carcinoma,and the expression of Rap1GAP is negatively correlated with MMP-2 and MMP-9.The interactions among the three affect the occurrence and development of colorectal carcinoma.
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Objective To analyze the MRI features of Ewing sarcoma/peripheral primitive neuroectodermal tumors(pPNETs) arising from the meninges.Methods The MRI imaging of 9 patients with Ewing sarcoma/pPNET were reviewed retrospectively,and imaging features and pathological characteristics were analyzed.Results The age of most patients ranged from 10 to 20 years.Magnetic resonance revealed a spindle-like lesion with a wide base in 8 cases.The lesions showed heterogenous iso-or hypo-intense signal on T1 WI in 7 cases,heterogenous hypo-iso-intense signal on T1 WI in 2 case,and iso-or mildly hypderisointense on T2WI in all cases.The solid part of the tumor was heterogeneously enhanced after injection of gadolinium with cyst degeneration or necrosis.The dural tail sign could be seen in 3 cases.The adjacent skull erosion could be seen in 6 cases.The breakthrough of the plate of cranium and soft-tissue invasion was present in 2 cases.The right eye proptosis was present in 1 case.The distant metastasis was found in 3 cases.Pathology showed that the lesions had high cell density.Hemorrhage and necrosis could be observed.The cells were like lymphocytes and spindle cells with transparent cytoplasm.CD99 and Vimentin were expressed in all tumor cells.Conclusion The imaging findings of the meningeal pPNET are different from meningiomas,which could be useful for the diagnosis and differential diagnosis.
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Rap1 is expressed in human umbilical vein endothelial cells (HUVECs). Rap1-GTPase activating protein (Rap1GAP), with its specific target, Rap1, has been shown to be important in the regulation of many physiological and certain pathological processes. In this study, we investigated the effect of Rap1GAP expression on endothelial cell function, or, more specifically, proliferation and migration of endothelial cells. HUVECs were transfected with pcDNA3.1 (empty vector), pcDNA3.1 containing Flag-tagged-Rap1GAP or Myc-tagged-Rap1N17. The proliferation, migration and tube formation were examined and compared among the 3 groups. Expression of Rap1, Rap1GAP, extracellular signal-regulated kinase (ERK), phospho-ERK, Akt, phosphor-Akt was detected by Western blotting. The results showed that the proliferation, migration and tube formation were significantly reduced in Rap1GAP- and Rap1N17-transfected HUVECs as compared with empty vector-transfected control. These changes were coincident with increased expression of Rap1GAP and decreased expression of activated Rap1, phospho-ERK and -Akt. After treatment of Rap1GAP-transfected HUVECs with a stimulator of Rap1 guanine-nucleotide-exchange factor (Rap1GEF) 8CPT-2'OMe-cAMP, it was found that Rap1 activity was decreased as compared with empty vector-transfected control. Pretreatment of HUVECs with an ERK inhibitor PD98059 or a PI3K inhibitor LY294002 prior to stimulation not only blocked 8CPT-2'OMe-cAMP-induced phosphorylation of ERK and Akt, but also significantly reduced cell proliferation and migration. Finally, we examined the effect of vascular endothelial growth factor (VEGF) on HUVECs overexpressing Rap1GAP. VEGF-stimulated Rap1 activity, phosphorylation of ERK and Akt, cyclin D1 expression and cell proliferation were repressed in HUVECs overexpressing Rap1GAP as compared to empty vector-transfected control. Taken together, our findings demonstrate that Rap1GAP/Rap1 and their downstream effectors regulate proliferation and migration of HUVECs via ERK and Akt pathways.
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The effect of troglitazone on glucose-stimulated insulin secretion (GSIS) in pancreatic β-cells and its mechanism were investigated.10 μmol/L troglitazone had no effect on basal insulin secretion,but significantly decreased GSIS and stimulated AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) phosphorylations (all P<0.01).These reactions were completely reversed by AMPK inhibitor compound C,suggesting that the troglitazone acutely inhibits insulin secretion via stimulating AMPK activity in beta cells.
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Objective To study the role of pancreatic renin-angiotensin system (RAS) on insulin secretion, proliferation, apoptosis, oxidative stress, and fibrosis of β-cells. Methods The effect of angiotensin Ⅱ on βTC3 cells was studied and the role and mechanism of AT1 R were analyzed with RNAi technology. The expression of AT1 R was measured by Western Blotting. The change of intracellular calcium was detected by microfluorimetry with Furo3-1oaded cells. Peroxide-sensitive fluorescent probe DCFH-DA was used to analyze intracellular ROS by flow cytometry. Real-time PCR was performed to evaluate mRNA levels related to proliferation and fibrosis in βTC3 cells. Apoptosis was detected by flow cytometry and Tunel method. Results Insulin secretion was significantly increased up to four fold and the level of intracellular calcium was sharply increased in response to high glucose in βTC3 cells. Angiotensin Ⅱ has no direct effect on insulin secretion in βTC3 cells and its role in secretion was associated with the role in proliferation. Oxidative stress in βTC3 cells caused by angiotensin Ⅱ may be partially mediated through AT1R, protein kinase C and NAD(P) H. With the decrease of AT1R expression by RNAi technology, apoptosis, and fibrosis of βTC3 cells induced by angiotensin Ⅱ might be ameliorated.Conclusions By means of AT1R, angiotensin Ⅱ plays an important role in insulin secretion, proliferation,apoptosis, oxidative stress and fibrosis in β-cells.
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BACKGROUND: Previous studies have confirmed that embryonic stem cells call be induced to differentiate into insulin-producing cells, but the induction process takes a long time. Most of the processes take about one month.OBJECTIVE: Activin A, all-trans retinoic acid (ATRA), basic fibroblast growth factor (bFGF) and nicotinamide were applied in vitro in combination to observe whether mouse embryonic stem cells could be induct to differentiate into insulin-producing cell in a relatively short time.DESIGN: Cell observation experiment.SETTING: Shanghai Institute of Endocrine and Metabolic Diseases, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.MATERIALS: This study was performed at Shanghai Institute of Endocrine and Metabolic Diseases from October 2004 to February 2006. Two mice of clean grade and of 12.5-14.5 days of gestational age were provided by Shanghai SLAC Laboratory Animal Co., Ltd (Permission No. 2004A034). The protocol was performed in accordance with ethical guidelines for the use and care of animals. Mouse embryonic stem cell lines were supplied by Dr Changxian Zhang (CNRS UMR5641, France). Activin A was the product of the R & D Corporation. ATRA and nicotinamide were supplied by the Sigma Corporation, USA. BFGF was supplied by Gibco Corporaion. METHODS: Head and viscera were removed from embryos of the pregnant mouse. The remaining tissues were cut into pieces and digested with trypsin. Cell suspension was centrifuged and inoculated at 3×108L-1. The cells could be used as mouse feeder layer after 2-3 times of passage. The mouse embryonic stem cells (ESCs) were inoculated onto the feeder layer in knockout Dulbecco's modified Eagle medium (DMEM) supplemented with leukemia inhibitory factors (LIF). ESCs were passaged at 1:3-1:6 after 2-3 days of culture. Culture medium with serum was added into the culture dishes to terminate the digestion. Cell fluid was centrifuged and supernatant was discarded. The sediments were prepared into suspension and inoculated at 2.5×104 with LIF-free culture medium. After 24-48 hours, embryonic bodies (EBs) were collected and replated in 1% Matrigel-coated dishes. When began to adhere to the dishes, EBs were cultured in 10% FBS/DMEM supplemented with 100μg/L activin A for 24 hours. Then EBs were switched to 10% FBS/DMEM for 6-8 hours as an interval. After this interval. EBs differentiated were cultured in 10% FBS/DMEM with 10<-6mol/L RA for another 24 hours followed by culture in 10% FBS/DMEM supplemented with 10μg/L bFGFs for 3-5 days. Finally, EBs differentiated were cultured in DMEM/F12 supplemented with N2 supplement, B27 supplement, 1μg/L laminin, 10μg/L bFGFs, and 10mmol/L nicotinamide for 3-5 days. Dithizone (DTZ) staining, inununofluorescent staining and reverse transcription-polymerase chain reaction (RT-PCR) were applied to detect insulin expression in the differentiated cells.MAIN OUTCOME MEASURES: Induction of ESCs, DTZ staining and immunofluorescent staining as wel as RT-PCR detection.RESULTS: Mouse ESCs growing on a feeder layer formed many colonies with clear boundary and dense structure. However, there was no obvious outer limit between these ESCs. EBs began to adhere to the dishes, which were coated with matrigel, on the 2nd day. After activin A and ATRA interval induction, EBs spread, and most of the living cells were epithelial cell-like when cultured in 10% FBS/DMEM supplemented with 10μg/L bFGFs. After culturing in DMEM/F12 supplemented with N2, B27, nicotinamide, bFGFs and laminin, the cells formed small clusters. The insulin-producing cells were stained dark red with DTZ, and the cells stained with primary antibody to insulin were insulin-positive. After 2 weeks of induction of activin A, ATRA, bFGFs and nicotinamide, the insulin-producing cells expressed insulin 2, Pdxl, Nkx6.1, Nkx2.2, PP, IAPP, Glut2, Somastatin, Hnf3β and Neuro D mRNA but did not express insulin 1 mRNA.CONCLUSION: Mouse ESCs call be induced to differentiate into insulin-producing cells by activin A, ATRA, bFGFs and nicotinamide in vitro. Induction time call be shortened to 2 weeks.
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BACKGROUND: Vascular endothelial growth factor receptor 2 (VEGFR2) is primarily involved in vascular endothelial growth factor (VEGF)-mediated signal transduction and plays a critical role in the pathological angiogenesis that occurs in a number of diseases, including leukemia. Besides, VEGF secreted by leukemia cells also induces its own expression which leads to an enhanced production of VEGFR2 which contributes to the survival and proliferation of leukemia cells.OBJECTrVE: To evaluate the inhibitive effect of Lenti6/shVEGFR2 on the VEGFR2 expression and leukemia growth in mouse.DESIGN: A randomized, parallelized, controlled and open trial.SETTING: Department of Pediatrics, the Second Affiliated Hospital of Sun Yat-sen University; Biotechnology Research Center, Sun Yat-sen University.MATERIALS: The experiment had been done in the laboratories for Medical Research Center of the Second Affiliated Hospital, Sun Yat-sen University and Biotechnology Research Center, Sun Yat-sen University from May 2004 to January Lentiviral RNAi Expression System was purchased from Invitrogen, Co.,Ltd.; human VEGFR2 Mcb (PE) was purchased from R&D; CD31 immunohistochemistry kit was purchased from Boster, Co.,Ltd.; CD33-PE fluorescence labeled antibody was purchased from BD, Co.,Ltd.transiently and expression clone (Lenti6/shVEGFR2) was constructed, then cotransfected with ViraPowerTM Packaging Mix pU6/shVEGFR2 entry clone and transducting with Lenti6/shVEGFR2 expression clone, the effect on the development of intravenous xenograft leukemia mouse model, the distribution of microvessels in mouse bone marrow was observed after leukemia model mouse injected with recombinant lentivirus (group B); leukemia model mouse injected with recombinant lentivirus and endothelial cell (group C); leukemia model mouse injected with endothelial cell (group D). Through detecting changes of CD33 positive cells and microvessel density (MVD) in bone marrow, observing peripheral blood cell (PBC)smear and slice of liver, spleen, the effect of Lenti6/shVEGFR2 recombinant lentivirus on mouse leukemia was evaluated.mediated with lentivirus on VEGFNEGFR2 paracrine and autocrine loops in leukemia mouse.effective in inhibiting HL60 cell. pU6/shVEGFR2 entry clone constructed according to it had cell inhibitory rate as high as after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone: 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell growth inhibitive rates were similar. However, the cell growth inhibitive rate of entry clone descended rapidly after 48 hours (P＜0.01); which of expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance mouse: The amount of HL60 cells in bone marrow of groups A, B and C detected with flow cytometry were (25.8%±4.9)%, (14.3%±5.1)%, (8.4±2.6)%, respectively (P＜0.05); MVD in group C was obviously less than that in group D (P＜0.05); The amount of HL60 cells in leukemia model mouse injected with recombinant lentivirus and endothelial cell was the lowest as compared with the other groups.
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Objective To study the characteristics of serious central nervous system(CNS) involvement in childhood systemic lupus erythematosus.Methods We made a comparison on the level of ANA、dsDNA and positive rate of Sm、C3 between primary and secondary CNS involvement and analysed the clinical manifestations between two groups.Results The level of ANA、dsDNA and ositive rate of Sm、C3 were not related with SLE encephalopathy;EEG was useful to the diagnosis of SLE.Conclusion The differiential diagnosis between primary and secondary CNSD in volvement of SLE must be analysed according to clinical manifestations and other laboratory findings.
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AIM: To observe the effects of valproic acid(VPA) and HA14-1 on Bcl-2 overexpressed leukemia cells in vitro and in vivo.METHODS:(1) Cells were divided into control group,HA14-1 group,VPA group and HA14-1+VPA group.The apoptotic rate,the mean fluorescent index(MFI) of Bcl-2,the levels of caspase 3,8 and 9 were detected with FCM.(2) 24 h after transplantation with BALL-1,the NOD/SCID mice were divided into 4 groups as(1),then the survival time was compared.The expression of CD19 in the peripheral blood cells,bone marrows,livers,spleens and lungs of each group was detected.RESULTS:(1) The apoptotic rate in HA14-1+VPA group was(76.5?6.9)%,this was significantly elevated compared to the data in other groups(P
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AIM: To look for harmfulless anti-leukemia drug with selective high performance, lethal effect of small hairpin RNA (shRNA) on VEGFR2 gene expression of tumor cell line HL60 in vitro.METHODS: The most effective VEGFR2 siRNA was designed and screened. The shRNA oligo was designed and pU6/VEGFR2 entry clone was constructed. HL60 was transfected transiently and vascular endothelial growth factor receptor 2(VEGFR2) expression was tested with MTT assay, RT-PCR and Western blotting. The expression clone was constructed and cotransfected with ViraPowerTM Packaging Mix into 293FTTM cells to produce Lentiviral vectors harboring Lenti6/shVEGFR2. The virion supernatant was added into HL60 cells and VEGFR2 gene inhibitory effect was determined. RESULTS: The inhibitory rates of VEGFR2 siRNA c were high. VEGFR2 expression in HL60 was inhibited by using pU6/VEGFR2 entry clone constructed with shRNA and pENTRTM/U6. For HL60 cells, the inhibitory rate was 84.9%. The expression of VEGFR2 mRNA and protein decreased significantly. 48 hours after transfection of pU6/shVEGFR2 entry clone and transduction of Lenti6/shVEGFR2 expression clone, the cell inhibitory rates were similar. Cell growth inhibitory rate of entry clone descended rapidly after this time point, the expression clone changed slowly, reaching the peak at 96 hours, dropped slightly, having no significance deviation. CONCLUSION: in vitro, VEGFR2 shRNA using lentiviral vector blocks VEGF/VEGFR2 self-secretion in HL60 cells, which inhibits leukemia development.