ABSTRACT
【Objective】 To explore the effect of B lymphocytes on cardiac structure and function and myocardial immune cells during heart development. 【Methods】 Echocardiography, immunofluorescence staining and flow cytometry were used to evaluate the composition of immune cells of the heart and the cardiac structure and function in wild-type (WT) mice and B-lymphocyte-deficient (μMT) mice, respectively. 【Results】 Compared with those of μMT mice, the ratio of heart weight to mouse weight (P<0.05), left ventricular mass (P<0.05) and the cross-sectional area of myocardial cells WT mice were significantly increased, while the ventricular ejection fraction was significantly decreased (P<0.05). The results of mRNA sequencing showed that WT mice and μMT mice differentially expressed genes were mainly enriched in the signal pathway of heart development and hypertrophic cardiomyopathy. The results of flow cytometry showed that WT mice had more Ly6g+ neutrophils, CD4+ positive T cells (P<0.001) and CD8+T cells (P<0.05) compared with μMT mice. 【Conclusion】 B-lymphocyte depletion alters the composition of cardiomyocyte immune cells, reduces left ventricular mass, and increases myocardial contractility.
ABSTRACT
OBJECTIVES@#To identify, examine and summarize the available evidence on the effectiveness and safety of acupuncture for in vitro fertilisation (IVF) outcomes.@*METHODS@#Eight electronic databases, including PubMed, EMBASE, Cochrane Database of Systematic Review, Cochrane Central Register of Controlled Trials, China National Knowledge Infrastructure, Wanfang Database, Chinese Biomedical Database and VIP Database, were searched, supplemented by manual searches. Two researchers independently conducted the literature screening, data extraction, and methodological quality assessments. A narrative description was provided to show the general information and specific characteristics of the included studies. A bubble plot was used to visually display the overall effects of acupuncture on IVF outcomes.@*RESULTS@#Eighty-two studies were identified, including 64 primary studies and 18 systematic reviews. Transcutaneous electrical acupoint stimulation, electric acupuncture and manual acupuncture were applied in most studies and compared with no acupuncture, sham acupuncture and placebo acupuncture control groups. Sixty-three (98.4%) primary studies reported clinical pregnancy rate, and positive effects of acupuncture were found in 34 studies (54.0%). Live birth rate was reported in only 18 (28.1%) primary studies, of which 10 (55.6%) showed positive results. In addition, only 8 and 2 systematic reviews showed that acupuncture could increase clinical pregnancy events and live birth events, respectively. However, none of these reviews was of high methodological quality.@*CONCLUSIONS@#Available evidence suggests that acupuncture therapy could improve clinical pregnancy rates. However, whether acupuncture could increase live birth events was difficult to determine based on the few studies that have reported this outcome indicator. Furthermore, the methodological quality of most systematic reviews was assessed as critically low or low. Studies with a rigorous design and standardized implementation should be performed to refine the available evidence.
Subject(s)
Female , Humans , Pregnancy , Acupuncture Therapy/methods , China , Fertilization in Vitro , Pregnancy RateABSTRACT
OBJECTIVE@#To investigate the mechanism of Tojapride, a Chinese herbal formula extract, on strengthening the barrier function of esophageal epithelium in rats with reflux esophagitis (RE).@*METHODS@#Ten out of 85 SD rats were randomly selected as the sham group (n10), and 75 rats were developed a reflux esophagitis model (RE) by the esophageal and duodenal side-to-side anastomosis. Fifty successful modeling rats were divided into different medicated groups through a random number table including the model, low-, medium-, and high-dose of Tojapride as well as omeprazole groups (n10). Three doses of Tojapride [5.73, 11.46, 22.92 g/(kg•d)] and omeprazole [4.17 mg/(kg•d)] were administrated intragastrically twice daily for 3 weeks. And the rats in the sham and model groups were administered 10 mL/kg distilled water. Gastric fluid was collected and the supernatant was kept to measure for volume, pH value and acidity. Esophageal tissues were isolated to monitor the morphological changes through hematoxylin-eosin (HE) staining, and esophageal epithelial ultrastructure was observed by transmission electron microscopy. The expressions of nuclear factor kappa-light-chain-enhancer of activated B cells p65 (NF-KBp65), κB kinase beta (IKKß), occludin, and zonula occludens-1 (ZO-1) in the esophageal tissues were measured by immunohistochemistry and Western blot, respectively.@*RESULTS@#The gastric pH value in the model group was significantly lower than the sham group (P<0.05). Compared with the model group, gastric pH value in the omeprazole and medium-dose of Tojapride groups were significantly higher (P<0.05). A large area of ulceration was found on the esophageal mucosa from the model rats, while varying degrees of congestion and partially visible erosion was observed in the remaining groups. Remarkable increase in cell gap width and decrease in desmosome count was seen in RE rats and the effect was reversed by Tojapride treatment. Compared with the sham group, the IKKß levels were significantly higher in the model group (P<0.05). However, the IKKß levels were down-regulated after treatment by all doses of Tojapride (P<0.01 or P<0.05). The occluding and ZO-1 levels decreased in the model group compared with the sham group (Ps0.01 or Ps0.05), while both indices were significantly up-regulated in the Tojapride-treated groups (P<0.01 or P<0.05).@*CONCLUSIONS@#Tojapride could improve the pathological conditions of esophageal epithelium in RE rats. The underlying mechanisms may involve in down-regulating the IKKß expression and elevating ZO-1 and occludin expression, thereby alleviating the inflammation of the esophagus and strengthening the barrier function of the esophageal epithelium.
ABSTRACT
The effect of acupuncture-moxibustion on respiratory system and systemic immune inflammatory response were reviewed to explore the possible role of neuroimmunomodulation in the control of inflammatory response and the effect mechanism of cholinergic anti-inflammatory pathway on coronavirus disease 2019 (COVID-19). Acupuncture-moxibustion could produce the local and systemic anti-inflammatory effect on COVID-19 through the activation of cholinergic anti-inflammatory pathway. Compared with humoral anti-inflammatory pathway, the neuronal anti-inflammatory pathway has earlier initiation, rapider action, and more localization, which play a more important role in the initial stage of inflammatory response. This may be an important basis for acupuncture-moxibustion intervention in the early stage of COVID-19. In addition to cholinergic anti-inflammatory pathway, acupuncture-moxibustion may also play an anti-inflammatory role in activating sympathetic nerve, hypothalamic-pituitary-adrenal axis and other neural anti-inflammatory pathways. How acupuncture-moxibustion play its role in stimulating the vagus nerve and sympathetic nerve in different periods of inflammatory response, and whether the effect is based on the selection of acupoints and the methods of stimulation, will be the research direction of the transformation from basic research to clinical research for acupuncture-moxibustion.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Betacoronavirus , Coronavirus Infections , Therapeutics , Hypothalamo-Hypophyseal System , Moxibustion , Pandemics , Pituitary-Adrenal System , Pneumonia, Viral , TherapeuticsABSTRACT
ObjectiveTo investigate the expression of transforming growth factor β1 (TGF-β1) and collagen I, III (Col I, III) in vulvar lichen sclerosis (VLS) and their role in VLS.MethodsThe specimens of30 VLS tissues (15 in early stage and 15 in progressive stage), and 15 vulvar normal skin tissues were selected by biopsy or surgical excision from 2016 to 2018 in our hospital. The expression of Vimentin was detected by immunohistochemistry. The expression of TGF-β1, TGF-β2, Col I and Col III mRNA was detected through RT-PCR. Simultaneously, the fibroblasts were visualized in diseased tissues by labeling Vimentin.ResultsThe expression of Vimentin in VLS was increased significantly (P0.05). Col I mRNA was up-regulated in VLS, obviously in early stage. Meanwhile, Col III mRNA down-regulated gradually from the early to the progressive stage of VLS. Therefore, the Col I/III ratio increased gradually.ConclusionThe increase of fibroblasts and TGF-β1 in dermis of VLS promotes the synthesis of Col I and reduces the content of Col III, which may be one of the factors leading to the decrease of skin elasticity in VLS.
ABSTRACT
Objective To explore the effect of preventive management path of breast cancer related lymphedema based on risk reduction program on breast cancer patients underwent axillary lymph nodes dissection (ALND). Methods This study used convenient sampling method to recruit 60 patients underwent ALND. Patients were randomly divided into the control group (n=31) and the intervention group (n=29). All patients received general care while patients of the intervention group also received the Risk Reduction Program. Lymphedema symptom experience was investigated by Breast Cancer and Lymphedema Symptom Experience Index 1 month, 3 months, 6 months, 12 months after surgery. Results Twelve months after surgery, the intervention group had 4.00(8.00)lymphedema related symptoms while the controll group had 9.00(7.00), the difference was significant (Z=-2.023, P=0.043). The score of lymphedema related symptom distress in the intervention group was 3.00(7.00)while it was 7.00(17.00)in the control group, the difference was significant (Z=-2.159, P=0.031). The scores of functional dimension and sexual dimension in the intervention group were 0(2.00)and 0(0), and in the control group were 3.00(5.00)and 0(1.00), the differences were significant (Z=-2.315,-2.334, P=0.021, 0.020). No significant differences of lymphedema rate existed between groups (P>0.05). Conclusion The preventive management path of breast cancer related lymphedema based on risk reduction program can decrease the number of lymphedema symptoms and release lymphedema symptom distress.
ABSTRACT
Objective To explore the expression and significance of insulin grow th factor 1(IGF-1) and gli-al fibriuary acidic protein(GFAP) in patients with glioma .Methods The serum levels of IGF-1 and GFAP in 40 glioma patients ,30 healthy subjects and 35 patients with other benign intracranial tumors were measured by double antibody sandwich method .Results There was no significant difference in serum IGF-1 and GFAP lev-els between healthy subjects and other benign intracranial tumor patients (P>0 .05) ,and the levels of IGF-1 and GFAP in the serum of glioma patients were significantly higher than those of healthy and other benign in-tracranial tumor patients (P<0 .05) .The serum levels of IGF-1 and GFAP in glioma patients after operation were significantly lower than those before operation (P< 0 .05) .Conclusion The expression of IGF-1 and GFAP in the serum of glioma patients is significantly higher than those of healthy subjects and other benign intracranial tumor patients .It has good sensitivity and specificity .It can be used as a serum marker of glioma patients and has certain clinical value .
ABSTRACT
Objective To investigate the changes in prevalence of nutritional risk and undemutrition in patients with head and neck cancer during radiotherapy.Methods In this longitudinal observational study,a convenience sampling method was used to recruit patients with head and neck cancer who were receiving radiotherapy in Beijing Cancer Hospital.Nutritional Risk Screening 2002 (NRS 2002) was applied to assess the prevalence of nutritional risk in the patients,and Patient-Generated Subjective Global Assessment (PG-SGA) and body composition test to determine the prevalence of malnutrition (undernutrition) before,during and after radiotherapy.Results 56 patients finished the three follow-up exams.Along with the progress of radiotherapy,the scores of NRS 2002 increased gradually (1.64±1.09 vs.2.30 ±1.06 vs.3.14 ±1.07,x2 =46.639,P＜0.001),and the prevalence of nutritional risk also increased gradually (21.43％ vs.37.50％ vs.71.43％,x2 =29.700,P ＜0.001);the total scores of PG-SGA [1 (1-13) vs.6 (1-15) vs.12 (1-18),x2 =63.206,P＜0.001] and dimensions of weight [0 (0-4) vs.1 (0-4) vs.3 (0-6),x2 =40.798,P＜0.001],intake [0 (0-2) vs.1 (0-2) vs.2 (0-4),x2=64.707,P＜0.001] and symptoms [0 (0-7) vs.2 (0-10) vs.6 (0-11),x2 =61.562,P ＜ 0.001] all increased gradually with statistical significance.The prevalence of malnutrition in different stage of radiotherapy were significantly different (x2 =64.999,P ＜ 0.001).The body composition analysis in 40 patients showed that all the indicators of body composition decreased significantly along with the progress of radiotherapy.There was a great loss in patients' body weight during radiotherapy,especially the fat-free mass.Conclusions The prevalence of nutritional risk and undernutrition may increase in patients with head and neck cancer during radiotherapy.Lean body mass accounted for most of the weight loss.We should pay more attention to those patients' nutritional status during radiotherapy.
ABSTRACT
Objective To investigate the expression of syndecan-1 (SDC1) in glioma cells and the effects of synde?can-1 knockdown on the proliferation and invasion of A172 cells. Methods The expression of syndecan-1 in glioma cells was analyzed using quantitative Real-time PCR and Western blotting. A172 cells were transfected with lentiviral vector carrying SDC1 shRNA to establish a stable SDC1-silencing cell line. The cell proliferation was analyzed by MTT assay. Trypan blue exclusion assay and flow cytometry, and Transwell assays were performed to measure the migration and invasion abilities, respectively. The mRNA and protein and expression levels of SDC1, Proliferation Cell Nuclear An?tigen (PCNA) and Matrix Metalloproteinase 9 (MMP-9) were detected by using qRT-PCR and Western blotting. Results The expression levels of SDC1 were significantly different in different glioma cell lines. The stable SDC1-silencing cell line was successfully established, in which the mRNA and protein expression levels of SDC1 were significantly decreased (P<0.05). SDC1 knockdown significantly reduced the cell proliferation, migration(58.40±5.24 vs. 255.8±16.09、226.5± 22.84,F=126.4,P<0.05)and invasion(61.67 ± 16.26 vs. 233.70 ± 17.24、244.30 ± 28.15,F=69.87,P<0.05)compared with either control group or blank group. SDC1 knockdown also significantly decreased the mRNA and protein expression levels of PCNA and MMP-9 (P<0.05). Conclusion:SDC1 knockdown suppresses the capacities of proliferation, invasion and migration of glioma A172 cell, implying that SDC1 may serve as a novel target in the biotherapy of glioma.
ABSTRACT
Objective To investigate the effect of different gene expression levels of Syntenin on invasion and mi?gration of glioma cells and the underlying molecular mechanisms. Methods Lentiviral RNA interference was used to knockdown the expression of syntenin in U-87 cells. Real-time quantitative PCR was used to detect mRNA expression levels of syntenin . Transwell assay and adhesion assay were used to examine the invasion, migration and adhesion, re?spectively. Western-blot was used to detect the protein expression levels of Syntenin, AKT, p-AKT, and MMP-9. Re?sults The mRNA expression level of Syntenin was greatly reduced in interference group compared with empty vector group (P0.05). Conclusion Syntenin may enhance the invasion and migration ability of glioma though up-regulation of p-AKT, which in turn pro?motes MMP-9 expression in a corresponding signal transduction pathway.
ABSTRACT
The main objective of this study was to prepare sustained release metformin hydrochloride microcapsules by the Wurster fluidized bed and to obtain the optimized coating process and formulation. Fine microcapsules without agglomeration were obtained in a continuous coating process with the atomization air pressure of 0.2Mpa and an appropriate coating speed temperature. With other design variables of coating process fixed, the effects of different fluidizing air volume, coating temperature, coating speed, coating material, coating materials amount, plasticizer type and plasticizer amount on drug release were investigated respectively. Coating solution was achieved by dissolving EC45cps of 21 g, EC100cps of 7 g, DBS of 2.8 g and talcum powder of 8 g in ethanol to get a final volume of 500 ml. Particles of 150g along with 500mL coating solution would be fine. The results showed that with the air volume of 35 m3·h-1, coating temperature of 35o, coating speed of 6 mL·min-1 and proper amount of coating solution, fine microcapsules were obtained. The mean diameter of the microcapsules obtained eventually were 213 micro m and the drug content were 23%, which was suitable for producing a suspension. Particle diameter distribution corresponded to the normal distribution and obviously prolonged drug-release was achieved
ABSTRACT
ObjectiveTo analyze characteristic changes of shoulder muscles by investigating surface electromyographic regularity changes before and after the treatment of Frozen shoulders. Methods7 cases of frozen shoulders were selected, surface electromyography (sEMG) before and after treatment were recorded by AMT-8 EMG recorder. The collected data was processed using MATLAB software integrated EMG (IEMG) value were obtained. Then the IEMG data were statistically analyzed using Stata11.0 software and compared. Results ①The differences of abduction deltoid and infraspinatus muscle IEMG values before and after treatment were statistically significant(P ＜0.05), while IEMG values changes of biceps, triceps, pectoralis major, latissimus dorsi and trapezius before and after treatment were not statistically significant (P＞0.05); changes of adduction IEMG values of all muscles were not statistically significant (P＞0.05). ②Changes of flexion IEMG values of biceps before and after treatment were statistically significant (P＜0.05), while IEMG values changes of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid, triceps before and after treatment were not statistically significant (P＞0.05). Changes of extension IEMG values of triceps before and after treatment were statistically significant (P＜0.05), while those of pectoralis major, infraspinatus muscle, latissimus dorsi, trapezius, deltoid and biceps were not statistically significant (P＞0.05). ③IEMG values of all the muscles during external rotation, internal rotation before and after treatment IEMG were not significantly different. Conclusion①After treatment, the outreach functions of infraspinatus and deltoid muscles were improved, while adduction functions of all muscles were not improved. ②Flexion function of biceps was improved significantly after treatment as well as extension function of triceps.③Extemal rotation, internal rotation functions of all muscles were not improved significantly.
ABSTRACT
Hepatitis C virus (HCV) infection is the leading cause of chronic liver diseases worldwide.There is no vaccine to prevent HCV infection.Current standard of care (SOC) for hepatitis C is pegylated interferon-α (pegIFN-α) in combination with ribavirin (RBV).However,the efficacy of pegIFN-α and RBV combination therapy is less than 50％ for genotype 1 HCV,which is the dominant virus in human.Additionally,IFN and RBV are highly toxic,causing severe side effects.Therefore,it is urgent to develop safer and more efficacious anti-HCV drugs.Over the last decade,a number of HCV-specific inhibitors have been discovered with many of them reached to late stages of clinical trials.Recently,2 HCV NS3 protease inhibitors,telaprevir and boceprevir,have been approved by the Unite States Food and Drug Administration (FDA).This opens up a new era for anti-HCV therapy.Several new classes of antiviral drugs targeting HCV NS3 protease,NS5A and NSSB RNA-dependence RNA polymerase (RdRp) are currently at various stages of preclinical and clinical studies.Upon approval of more NS3 protease,NS5A and NS5B polymerase inhibitors,future clinical studies will lead to optimal combination therapies which will have desirable parameters such as IFN-free,higher efficacy,safe,one daily dose and short duration.
ABSTRACT
There are problems in traditional medical education in China. It is important to cultivate medical talents who are adaptable to social changes and medical advancement. However, the scientific quality and innovative medical students have long been neglected. This paper discussed the essence of scientific quality and innovative talents, and introduced the experience of the program of "long term clinical medicine education" at Peking University Health Science Center. We here delineateded the key points of the scientific quality and innovation education that may provide new ideas for the training of the medical talents.
ABSTRACT
<p><b>BACKGROUND</b>The CKLF-like MARVEL transmembrane domain-containing family (CMTM) is a novel family of proteins linking chemokines and TM4SF. Different members exhibit diverse biological functions. In this study, the effect of intracellular CMTM2 on regulating human immunodeficiency virus type-1 (HIV-1) transcription was evaluated.</p><p><b>METHODS</b>The effects of CMTM2 on regulating full-length HIV-1 provirus and the HIV-1 long terminal repeat (LTR)-directed transcription were assessed by luciferase assay. Transcription factor assays, using the luciferase reporter plasmids of AP-1, CRE, and NF-κB were conducted to explore the signaling pathway(s) that may be regulated by CMTM2. The potential relationship between CMTM2 and the transcription factor AP-1 was further analyzed by Western blotting analyses to investigate the effect of CMTM2 on PMA-induced ERK1/2 phosphorylation.</p><p><b>RESULTS</b>The results from the current study revealed that CMTM2 acts as a negative regulator of HIV-1 transcription. CMTM2 exerted a suppressive action on both full-length HIV-1 provirus and HIV-1 LTR-directed transcription. Transcription factor assays showed that CMTM2 selectively inhibited basal AP-1 and CREB activity. Co-expression of HIV-1 Tat, a potent AP-1 and CREB activator, can not reverse CMTM2-mediated AP-1 and CREB inhibition, suggesting a potent and specific effect of CMTM2 on negatively regulating these two signaling pathways.</p><p><b>CONCLUSION</b>Intracellular CMTM2 can negatively regulate HIV-1 transcription, at least in part, by targeting the AP-1 and CREB pathways. Exploring the mechanisms further may lead to new ways to control HIV-1 replication.</p>
Subject(s)
Humans , Chemokines , Physiology , Cyclic AMP Response Element-Binding Protein , HIV Long Terminal Repeat , HIV-1 , Genetics , Intracellular Space , Metabolism , Jurkat Cells , MARVEL Domain-Containing Proteins , Tetradecanoylphorbol Acetate , Pharmacology , Transcription Factor AP-1 , Transcription, Genetic , U937 CellsABSTRACT
Objective To investigate the inactivating effect of heat and ultraviolet(UV) light on HCV JFH-1 strain using the cell culture system. Methods The HCV JFH-1 virus stock, with an initial titer of 2.5 × 104 FFU/ml, was exposed in 56℃ water bath or to UV light for varying durations of time for explo-ring their inactivating effects on the virus. The kinetics of virus titer reduction was determined by an indirect immuno-fluorescence assay (IFA). If the cells infected with the exposed virus stock were IFA negative after three blind passages, the virus stock was considered to be inactivated completely. Results After incubation of the HCV JFH-1 virus stock (2.5 × 104 FFU/ml)in 56℃ water bath for 10 min, 20 min and 30 min, the virus titers were reduced to 1.6 × 103 FFU/ml, 3.1 × 102 FFU/ml and 3.3 × 10 FFU/ml, respectively. The exposure of the virus stock to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2, 30 cm below the UV lamp) for 15 s, 30 s and 45 s resulted in virus fiter reduction to 1.0 × 103 FFU/ml, 1.1 × 102 FFU/ml and 2.7 × 10 FFU/ml, respectively. After 40 min incubation of the virus stock at 56℃, or 1 min exposure to UV light (wavelength 253.7 nm, intensity ≥60 μW/cm2) the virus infectious titer was reduced below the detection limit of IFA, and the IFA was still negative even after three blind passages, indicating that the virus was inactivated completely. Conclusion HCV is sensitive to heat and UV light treatment. For HCV JFH-1 virus stock containing 2.5 × 104 FFU/ml virus, heat treatment at 56℃ for 40 min, or UV light expo-sure at an intensity of ≥60 μW/cm2 for 1 min, resulting in complete virus inactivation.
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the related factors of portal vein tumor thrombosis (PVTT) in patients with HCC.</p><p><b>METHODS</b>A total number of 234 patients with hepatocellular carcinoma (HCC) were included in this retrospective study. Uni-variate and multi-variate logistic regression analysis were employed to analyze the association between PVTT and 18 routine clinical parameters.</p><p><b>RESULTS</b>Among the 234 patients with HCC, 15% of patients (35/235) had PVTT. Univariate logistic regression analysis revealed significant association of age (P = 0.016), gamma glutamyl transferase (GGT, P = 0.003), number of segmental invasion (P = 0.007), microvascular invasion (P < 0.01), segment location of S2 (P = 0.001), S3 (P = 0.000), S4 (P = 0.004) and S6 (P = 0.016). Multivariate analysis shows potential significant predictors of PVTT in HCC were age (RR: 0.373; 95% CI: 0.146-0.954; P = 0.040), the tumor location of S3 (RR: 4.625; 95% CI: 1.916-11. 165;P = 0.001), GGT (RR: 4.091; 95% CI: 1.448-11.553; P = 0.008) and microvascular invasion (RR: 20.912; 95% CI: 4.745-92.172; P < 0.01).</p><p><b>CONCLUSIONS</b>PVTT occurred more commonly in the younger (< 50 years old), and those with high level of GGT, segment location of S3 and microvascular invasion.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Carcinoma, Hepatocellular , Pathology , Embolism , Follow-Up Studies , Liver Neoplasms , Pathology , Logistic Models , Portal Vein , Retrospective Studies , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To study differentiation of human bone marrow-derived mesenchymal stem cells (BDMSC) into blood vessel endothelial cells for ideal cell origin of complex organ tissue engineering vascularization and injured tissue repairing by cell transplantation.</p><p><b>METHOD</b>After different days of induction with vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in 3D fibrin-gels and matrigel, BDMSC and angiogenesis were determined by the utilization of morphological observation, tissue section and CD34, CD31, vascular endothelial growth factor receptor-1 (VEGFR-1, Flt-1), VEGFR-2 (Flk-1), and vWF that were special for blood vessel endothelial cells.</p><p><b>RESULT</b>After 3D-cultured and induced with VEGF and bFGF in vitro in fibrin-gels and matrigel for 3-21 days, BDMSC expressed CD34, CD31, Flt-1, Flk-1, and vWF came into vessel-like configuration.</p><p><b>CONCLUSION</b>VEGF, bFGF as well as Flt-1 and Flk-1, expressed by BDMSC, may form a feasible microenviroment after induction and play an important role during processes of blood vessel endothelial cell differentiation and vessel-like configuration forming of BDMSC. Mesenchymal stem cells may be applied to tissue engineering vascularization and injured tissue repairing by cell transplantation.</p>
Subject(s)
Humans , Cell Culture Techniques , Methods , Cell Differentiation , Endothelial Cells , Cell Biology , Fibrin , Fibroblast Growth Factor 2 , Pharmacology , Gels , Mesenchymal Stem Cells , Cell Biology , Vascular Endothelial Growth Factor A , Pharmacology , Vascular Endothelial Growth Factor Receptor-1 , Metabolism , Vascular Endothelial Growth Factor Receptor-2 , Metabolism , von Willebrand Factor , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To prepare the polyclonal anti-peptide antibody against chemokine-like factor1 (CKLF1) and apply it to the expression and functional studies of CKLF1.</p><p><b>METHODS</b>CKLF1 was analyzed with bioinformatics methods. The 16 amino acids sequence peptide was selected from CKLF1 C terminal end. Antibody was raised by immunizing rabbits with the peptide conjugated to keyhole limpet hemocyanin (KLH).</p><p><b>RESULTS</b>A high titer polycolonal antibody was obtained from the rabbit against the peptide. ELISA analysis proved that the titer of rabbit serum against anti-peptide of CKLF1 was up to 10(-4). Western blot analysis revealed that it could react not only with recombinant CKLF1 expressed in a cell-Free Protein Biosynthesis System and Drosophila S2 cells, but also recognize the endogenous CKLFs in the tissue array. Positive staining was detected in the normal bronchial cartilage, gastric mucosa, and gastric smooth muscle tissues. Normal rectum and well-differentiated rectal carcinoma showed strong positive staining, but the poor-differentiated rectal carcinoma samples revealed negative staining.</p><p><b>CONCLUSION</b>The anti-peptide antibody can specifically recognize CKLFs and may be a useful reagent for the detection of CKLF1.</p>