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1.
Article in Chinese | WPRIM | ID: wpr-940760

ABSTRACT

ObjectiveTo investigate the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell and the mechanism. MethodMethyl thiazolyl tetrazolium (MTT) assay, hematoxylin-eosin (HE) staining, 4',6-diamidino-2-phenylindote (DAPI) staining, colony formation assay, scratch assay, and flow cytometry were employed for the analysis of apoptosis and cell cycle. Thereby, the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell was investigated. Then the Pharm Mapper, UniProt, Swissdock, STRING, and Metascape were used for target screening, gene annotation, molecular docking, protein-protein interaction (PPI) network construction, Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis to explore the mechanism. ResultHederasaponin B (15, 30, 60, 120 μmol·L-1) can significantly reduce the survival rate of HGC-27 cell (P<0.01) in a time-dependent and dose-dependent manner compared with the blank group. It had no significant toxicity to normal GES-1 cell at concentration below 120 μmol·L-1. Compared with the blank group, hederasaponin B (30, 60, 120 μmol·L-1) induced cytoplasmic vacuolization, and nuclear deformation and karyopyknosis, inhibited the migration of HGC-27 cell (P<0.01), and brought about the apoptosis (P<0.05, P<0.01) and cell cycle arrest of HGC-27 cell (P<0.05, P<0.01). Hederasaponin B (10, 20, 30 μmol·L-1) also suppressed the independent survival ability and proliferation ability of HGC-27 cell (P<0.01). The possible action targets were kinesin-like protein KIF11, cGMP-specific 3,5 cyclic phosphodiesterase, caspase-3, serine/threonine protein kinase Chk1, proto-oncogene tyrosine protein kinase, epidermal growth factor receptor, and mitogen-activated protein kinase (MAPK) 8. The mechanism may be related to MAPK signaling pathway (pathways in cancer), adhesion connection, focal adhesion and proteoglycans in cancer (epithelial cell signaling pathways in Helicobacter pylori infection). ConclusionHederasaponin B exerts significant inhibitory effect on gastric cancer HGC-27 cell through multiple targets and multiple pathways.

2.
Acta Pharmaceutica Sinica B ; (6): 3124-3138, 2022.
Article in English | WPRIM | ID: wpr-939960

ABSTRACT

Tumor-associated macrophages (TAMs), one of the dominating constituents of tumor microenvironment, are important contributors to cancer progression and treatment resistance. Therefore, regulation of TAMs polarization from M2 phenotype towards M1 phenotype has emerged as a new strategy for tumor immunotherapy. Herein, we successfully initiated antitumor immunotherapy by inhibiting TAMs M2 polarization via autophagy intervention with polyethylene glycol-conjugated gold nanoparticles (PEG-AuNPs). PEG-AuNPs suppressed TAMs M2 polarization in both in vitro and in vivo models, elicited antitumor immunotherapy and inhibited subcutaneous tumor growth in mice. As demonstrated by the mRFP-GFP-LC3 assay and analyzing the autophagy-related proteins (LC3, beclin1 and P62), PEG-AuNPs induced autophagic flux inhibition in TAMs, which is attributed to the PEG-AuNPs induced lysosome alkalization and membrane permeabilization. Besides, TAMs were prone to polarize towards M2 phenotype following autophagy activation, whereas inhibition of autophagic flux could reduce the M2 polarization of TAMs. Our results revealed a mechanism underlying PEG-AuNPs induced antitumor immunotherapy, where PEG-AuNPs reduce TAMs M2 polarization via induction of lysosome dysfunction and autophagic flux inhibition. This study elucidated the biological effects of nanomaterials on TAMs polarization and provided insight into harnessing the intrinsic immunomodulation capacity of nanomaterials for effective cancer treatment.

3.
Chinese Journal of Biotechnology ; (12): 1039-1049, 2022.
Article in Chinese | WPRIM | ID: wpr-927761

ABSTRACT

Hepatitis B virus core protein (HBc) has become a hot spot in drug carrier protein research due to its natural particle self-assembly ability and ease of modification. The truncation of the C-terminal polyarginine domain (CTD, aa 151-183) of HBc does not affect the self-assembly of the particles. However, it does affect the internal and external charges of the particles, which may subsequently affect drug encapsulation. Thus, the truncated C-terminal polyarginine domain (CTD) of HBc and the inserted RGD peptide were selected to construct and express three HBc variants (RH) encapsulated with ICG (RH/ICG) with different C-terminal lengths to compare the stability and drug activity of their nanoformulations. RH160/ICG was found to have a great advantages in encapsulation efficiency and biological imaging. Compared with other HBc variants, RH160/ICG significantly improved encapsulation efficiency, up to 32.77%±1.23%. Cytotoxicity and hemolysis assays further demonstrated the good biocompatibility of RH160/ICG. Cell uptake and in vivo imaging experiments in mice showed that RH160/ICG could efficiently deliver ICG in tumor cells and tumor sites with good imaging effect. This research provides a new direction for further expanding the diagnosis and treatment application of ICG and development of HBc-based nanoparticle drug carrier platform.


Subject(s)
Animals , Hepatitis B/drug therapy , Hepatitis B Core Antigens , Indocyanine Green/chemistry , Mice , Nanoparticles/chemistry , Viral Core Proteins
4.
Chinese Journal of School Health ; (12): 1007-1010, 2022.
Article in Chinese | WPRIM | ID: wpr-936520

ABSTRACT

Objective@#To analyze the current situation and associated factors of fitness APP usage among college students.@*Methods@#By using three stage compound sampling method, 2 171 college students were selected, and a binary Logistic regression model was established to analyze the influencing factors fitness APP usage.@*Results@#The rate of fitness APP usage among college students was 61.9%, with significant differences by grade, physical health perception, fitness awareness, fitness purpose and exercise frequency( χ 2=28.01, 37.15, 214.12, 32.23, 316.21, P <0.01). The Logistic regression analysis showed that, grade( β = -0.31 ), fitness awareness( β =0.46), fitness purpose (improving health, β =0.52, weight loss, β =0.65), APP user friendliness ( β =0.34) and specific type of sports (running, β =1.24, racket movement, β =0.80) were associated with fitness APP usage( P < 0.05 ). Gender( β =0.30), major ( β =0.01) and exercise frequency ( β =-0.29) have less effect on fitness APP usage among college students( P >0.05).@*Conclusion@#College students have a high level of acceptance and usage of fitness APP. Grade, fitness awareness and purpose, sports type and fitness APP design are prominent factors affecting fitness APP usage among college students. The personalized custom design of fitness APP could be improved for college students.

5.
Chinese Journal of Biotechnology ; (12): 4066-4074, 2021.
Article in Chinese | WPRIM | ID: wpr-921487

ABSTRACT

Different fragments of SARS-CoV-2 nucleocapsid (N) protein were expressed and purified, and a fluorescence immunochromatography method for detection of SARS-CoV-2 total antibody was established. The effect of different protein fragments on the performance of the method was evaluated. The N protein sequence was analyzed by bioinformatics technology, expressed in prokaryotic cell and purified by metal ion affinity chromatography column. Different N protein fragments were prepared for comparison. EDC reaction was used to label fluorescence microsphere on the synthesized antigen to construct sandwich fluorescence chromatography antibody detection assay, and the performance was systemically evaluated. Among the 4 prepared N protein fragments, the full-length N protein (N419) was selected as the optimized coating antigen, N412 with 0.5 mol/L NaCl was used as the optimal combination; deleting 91-120 amino acids from the N-terminal of N412 reduced non-specific signal by 87.5%. the linear range of detection was 0.312-80 U/L, the limit of detection was 0.165 U/L, and the accuracy was more than 95%. A fluorescence immunochromatographic detection method for analysis of SARS-CoV-2 total antibody was established by pairing N protein fragments. The detection result achieved 98% concordance with the commercially available Guangzhou Wanfu test strip, which is expected to be used as a supplementary approach for detection of SARS-CoV-2. The assay could also provide experimental reference for improving the performance of COVID-19 antibody detection reagents.


Subject(s)
Antibodies, Viral , COVID-19 , Chromatography, Affinity , Fluorescent Antibody Technique , Humans , Microspheres , SARS-CoV-2 , Sensitivity and Specificity
6.
Article in Chinese | WPRIM | ID: wpr-883311

ABSTRACT

As a non-invasive examination method, anterior segment optical coherence tomography (AS-OCT) has huge potential in detecting neoplastic epithelial lesion.With AS-OCT, detection and differentiation of ocular surface squamous neoplasia (OSSN) within ocular surface pathologies can be achieved and the diagnosis rate is comparable to that of biopsy.With the development of technology, AS-OCT imaging mode has developed from time-domain OCT to spectral-domain OCT with higher penetrance and resolution, the high-quality images of which are helpful to distinguish the benign and malignant OSSN, enhance the detection rate of OSSN, improve the differentiation of ocular surface tumors, and thus improve the diagnostic accuracy of OSSN.In addition, AS-OCT can help optimize the treatment regimen and effectively monitor the outcome of the disease in the clinical management stage, such as monitoring the recurrence of OSSN and evaluating chemotherapy.In this article, the morphological features of OSSN, the advantages and disadvantages of OSSN auxiliary examinations, the development of AS-OCT, the specific manifestation of OSSN on AS-OCT, the application of OCT in atypical OSSN, as well as the value and limitation of AS-OCT in the diagnosis and management of OSSN were reviewed.

7.
Journal of Medical Biomechanics ; (6): E247-E252, 2020.
Article in Chinese | WPRIM | ID: wpr-862320

ABSTRACT

The controllability of hardness and its control method, the corresponding direction of differentiation of mesenchymal stem cells (MSCs) regulated by different hardness, and the role of integrin in the signaling pathway through which hardness regulates MSCs differentiation were briefly described in this paper. Among them, the role of integrin in the signaling pathway of hardness regulation of MSCs differentiation was highlighted. Signal pathways in which hardness regulates MSCs differentiation include Rho/ROCK signal pathway, integrin/FAK signal pathway, ERK signal pathway, JNK signal pathway, Wnt-catenin signal pathway and PI3K/Akt signal pathway, etc. Integrin, as a transmembrane heterodimer glycoprotein, participates in part of the signal pathway to transmit mechanical signals to MSCs. Different integrin families participate in different signaling pathways to regulate the differentiation of MSCs in different directions, and there are certain mutual influences among these signaling pathways. The research findings provide a theoretical basis for the application of tissue repair, organ reconstruction and regenerative medicine.

8.
Chinese Journal of Biotechnology ; (12): 1216-1222, 2020.
Article in Chinese | WPRIM | ID: wpr-826856

ABSTRACT

A rapid and simple method to detect tumor markers in liver cancer was established by combining immunochromatography technique with fluorescent microsphere labeling. According to the principle of double antibody sandwich, the cytoskeleton-associated protein 4 (CKAP4) paired antibody was used as the labeled and coated antibody, and the goat anti-rabbit polyclonal antibody was used as the quality control line coated antibody in the preparation of the CKAP4 fluorescent immunochromatographic test strips. After the preparation, the test strips were evaluated on various performance indicators, such as linearity, precision and stability. The CKAP4 immunochromatographic strip prepared by time-resolved fluorescent microspheres had high sensitivity, and good specificity. Its precision was within 15%, recovery between 85% and 115%, and linear range between 25 and 1 000 pg/mL. The test strip could be kept stable at 37 °C for 20 days, and it correlated well with commercial ELISA kits. The CKAP4 fluorescence immunochromatography method can quantitatively detect the content of CKAP4 in serum. Furthermore, it is rapid, sensitive, simple, economical and single-person operation. This method has the potential of becoming a new method for the diagnosis and treatment of liver cancer.


Subject(s)
Animals , Antibodies , Metabolism , Chromatography, Affinity , Fluorescence , Humans , Liver Neoplasms , Diagnosis , Membrane Proteins , Molecular Diagnostic Techniques , Methods , Sensitivity and Specificity
9.
Protein & Cell ; (12): 104-119, 2019.
Article in English | WPRIM | ID: wpr-757937

ABSTRACT

Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) is a genetic cardiac muscle disease that accounts for approximately 30% sudden cardiac death in young adults. The Ser358Leu mutation of transmembrane protein 43 (TMEM43) was commonly identified in the patients of highly lethal and fully penetrant ARVD subtype, ARVD5. Here, we generated TMEM43 S358L mouse to explore the underlying mechanism. This mouse strain showed the classic pathologies of ARVD patients, including structural abnormalities and cardiac fibrofatty. TMEM43 S358L mutation led to hyper-activated nuclear factor κB (NF-κB) activation in heart tissues and primary cardiomyocyte cells. Importantly, this hyper activation of NF-κB directly drove the expression of pro-fibrotic gene, transforming growth factor beta (TGFβ1), and enhanced downstream signal, indicating that TMEM43 S358L mutation up-regulates NF-κB-TGFβ signal cascade during ARVD cardiac fibrosis. Our study partially reveals the regulatory mechanism of ARVD development.

10.
Chinese Journal of Biotechnology ; (12): 1012-1018, 2018.
Article in Chinese | WPRIM | ID: wpr-687715

ABSTRACT

To establish a time-resolved fluorescence immunochromatographic assay for quantitative determination of carbohydrate antigen 19-9 (CA19-9) in serum, we prepared CA19-9 test strips by integrating double-antibody sandwich method and fluorescence immunochromatography technique. Carboxy fluorescent microspheres and nitrocellulose membrane were used as carriers for labeling and coating CA19-9 pairing antibodies. We optimized the process by adjusting the amount of labeling and coating antibody. According to the linear range, lowest detection limit and precision, We evaluated the time-resolved fluorescence immunochromatographic assay of CA19-9. When the amount of labeled antibody was 80 μg for 20 μL fluorescent microspheres, and the concentration of coated antibody on the test line was 1.5 mg/mL, the optimal reaction time was 15 minutes. Assay linear range was 12.5 to 800 U/mL and the minimum detection limit was 6.32 U/mL. The Within-run and between-run coefficient of variation were less than 15%. Average recovery rate was 101%. By detecting 50 clinical samples in parallel with Roche electrochemical luminescence detection kit, correlation coefficient was 0.980 6. The experiment, initially established a fluorescence immunochromatographic detection method to quantitative detection of serum CA19-9, which has a good clinical application prospect.

11.
Article in Chinese | WPRIM | ID: wpr-657180

ABSTRACT

Objective:To study the effects of down-regulated Smad4 expression on the proliferation and apoptosis of breast carcinoma MCF-7 cells,and to explore their mechanisms.Methods:The human breast carcinoma MCF-7 cells and MDA-MB-231 cells were cultured in vitro.RT-PCR method was used to detect the expression levels of Smad4 mRNA in MCF-7 cells and MDA-MB-231 cells.The Smad4-shRNA plasmid and Scramble-shRNA plasmid were respectively stably transfected into the MCF-7 cells with high expression of Smad4.The experiment was divided into non-transfected MCF-7 cells (normal control) group,Smad4 gene silencing group,Scramble(negative control) group.The proliferation abilities of the cells in various groups were detected by CCK-8 method.The apoptotic rates of the cells in various groups were detected by flow cytometry.Real-time PCR method wasused to detect the mRNA expression levels of the proliferation-related genes CDKN1A,CDK1 and CDK2 and the apoptosisrelated genes Suvivin,bcl-2,caspase 3 and caspase 9.Results:The proliferation abilities of cells had no statistical significance between various groups (P>0.05).The mRNA expression levels of CDKN1A,CDK1 and CDK2 in the cells had no statistical significance between various groups (P>0.05).Compared with normal control group and negative control group,the apoptotic rate of the cells in Smad4 gene silencing group was significantly decreased (P<0.01),the expression levels of Suvivin and bcl-2 mRNA in Smad4 gene silencing group were significantly increased (P<0.01),and the mRNA expression levels of caspase 3 and caspase 9 in Smad4 gene silencing group were significantly decreased (P<0.05).Conclusion:Smad4 could induce the apoptosis of MCF-7 cells by downregulating the expressions of Suvivin and bcl-2 and up-regulating the expressions of caspase 3 and caspase 9.

12.
Article in Chinese | WPRIM | ID: wpr-658987

ABSTRACT

Objective:To study the effects of down-regulated Smad4 expression on the proliferation and apoptosis of breast carcinoma MCF-7 cells,and to explore their mechanisms.Methods:The human breast carcinoma MCF-7 cells and MDA-MB-231 cells were cultured in vitro.RT-PCR method was used to detect the expression levels of Smad4 mRNA in MCF-7 cells and MDA-MB-231 cells.The Smad4-shRNA plasmid and Scramble-shRNA plasmid were respectively stably transfected into the MCF-7 cells with high expression of Smad4.The experiment was divided into non-transfected MCF-7 cells (normal control) group,Smad4 gene silencing group,Scramble(negative control) group.The proliferation abilities of the cells in various groups were detected by CCK-8 method.The apoptotic rates of the cells in various groups were detected by flow cytometry.Real-time PCR method wasused to detect the mRNA expression levels of the proliferation-related genes CDKN1A,CDK1 and CDK2 and the apoptosisrelated genes Suvivin,bcl-2,caspase 3 and caspase 9.Results:The proliferation abilities of cells had no statistical significance between various groups (P>0.05).The mRNA expression levels of CDKN1A,CDK1 and CDK2 in the cells had no statistical significance between various groups (P>0.05).Compared with normal control group and negative control group,the apoptotic rate of the cells in Smad4 gene silencing group was significantly decreased (P<0.01),the expression levels of Suvivin and bcl-2 mRNA in Smad4 gene silencing group were significantly increased (P<0.01),and the mRNA expression levels of caspase 3 and caspase 9 in Smad4 gene silencing group were significantly decreased (P<0.05).Conclusion:Smad4 could induce the apoptosis of MCF-7 cells by downregulating the expressions of Suvivin and bcl-2 and up-regulating the expressions of caspase 3 and caspase 9.

13.
Article in Chinese | WPRIM | ID: wpr-505840

ABSTRACT

Purpose Glioblastoma and metastasis are the common and aggressive type of brain tumors in adults,and a definitive diagnosis has an important guiding significance to the clinical practice.Our purpose is to investigate the value of post contrast-enhanced T1WI-based histogram analysis in the differential diagnosis of glioblastoma and solitary brain metastasis.Materials and Methods Sixty-eight consecutive patients with glioblastoma or solitary metastasis confirmed pathologically or surgically at Meizhou People's Hospital between January 2012 and November 2015 were included in the study.Glioblastoma was diagnosed in 34 patients and solitary brain metastasis was diagnosed in the rest 34 patients.All patients had undergone routine brain MR examination before surgical resection and the imaging data were retrospectively analyzed.The ROIs were manually placed in enhanced solid parts of the tumors on the maximal cross-sectional post contrasted-enhanced T1WI by Image J.Then the mean,standard deviation (SD),minimum,maximum,skewness and kurtosis were calculated respectively.Results Kurtosis of post contrast-enhanced T1WI of metastasis (1.260 ± 1.271) was statistically higher than that of glioblastoma (0.071 ± 0.667)(P<0.05).When the optimal threshold criterion ofkurtosis was set at 0.736,the areas under ROC curve of kurtosis was 0.792 (95% CI:0.676-0.881),and the sensitivity,specificity,positive predictive value and negative predictive value at the optimum cutoff value were 76.47%,88.24%,80.65%,78.38%,respectively.Conclusion Kurtosis of post contrastenhanced T 1 WI-based histogram is valuable in differential diagnosis of glioblastoma and brain metastasis,and thus provides reliable and objective basis for differentiating the two kinds of tumors.

14.
Chongqing Medicine ; (36): 2495-2496,2499, 2017.
Article in Chinese | WPRIM | ID: wpr-620332

ABSTRACT

Objective To investigate the clinical preemptive analgesic efficacy of parecoxib sodium(PS) at different administration timing in the patients with total hip arthroplasty(THA).Methods Sixty patients receiving THA were prospectively enrolled and randomized into three groups.The group A started to be intravenously injected by PS 40mg/d on preoperative 3 d until operation day;the group B was intravenously injected by parecoxib sodium 40mg at preoperative 30 min;the group C began to be intravenously injected by the same dosage of normal saline at the same time point as the group A.The rest pain was assessed by using the visual analog scale(VAS) at postoperative 6,24,48,72 h.The duration of patient-controlled intravenous analgesia(PCIA) and total dosage were recorded.The first time unaided ambulation time was observed.Results The VAS scores at various postoperative time points in the group A and B were significantly lower than those in the group C(P<0.05),the VAS scores at postoperative 6,24 h in the group A were remarkably lower than those in the group B.The PCIA duration in the group A,B and C were 25.05±10.32),(36.75± 13.91),(50.40 ± 15.17)h,respectively,the pair-wise comparison of the group A,B and C showed statistical difference(P<0.05).The total dosages of PCIA drug in the group A,B and C were(29.25 ± 4.58),(34.50 ± 5.09),(62.65 ±10.52)tg,respectively,the dosage in the group A and B was significantly lower than that in the group C(P<0.05).The first time unaided ambulation time in the group A,B and C were (2.75 ± 0.81),(3.05 ± 1.08),(4.10 ±-0.92)d respectively,which in the group A and B was earlier than that in the group C (P<0.05).Conclusion Continuously using PS on preoperative 3 d can increase the analgesic effect in THA patients and is conducive to the functional rehabilitation and increase the patient satisfaction.

15.
Chinese Journal of Immunology ; (12): 712-714,720, 2017.
Article in Chinese | WPRIM | ID: wpr-613977

ABSTRACT

Objective:To establish quantitative a fluorescence immunochromatographic assay detection method for the heat shock protein 90α(HSP90α)in human serum.Methods: Indirect the technology of fluorescence microshers immunochromatographic assay was used to research fluorescent microsphere activation,ensure optimal testing time,determine linearity range,precision,recovery experiments,testing clinical samples and that sort of thing in the fluorescence immunochromatographicassay experiment.Results: In all the reaction conditions,it was determined that the optimal teaction time was 5 minutes,and in the best line range (0.39-100 ng/ml) precision quality control materials of high recovery (30 ng/ml) Intra-assay CV was 8.14%;quality control materials of low recovery (10 ng/ml) Intra-assay CV was 9.26%.With high,medium and low recovery of serum recovery was 100.56%,99.76%,94.1%,separately.Foreign kit(ELISA) for detection of 40 cancer patients clinical serum,the result showed that the correlation was 0.968 5.Conclusion: Establishing the method initially quantitative fluorescence immunochromatographic assay for the heat shock protein 90α(HSP90α)in human serum have high performance and linear,clinic accordance rate also can satisfy the demand of clinic quantitative detection,and will improve hopeful to applied to clinic after apprroved.

16.
Article in Chinese | WPRIM | ID: wpr-494398

ABSTRACT

Objective:To investigate the expressions of Smad2 and Smad4 in breast carcinoma tissue,and to analyze their relationships with oncogenesis and development of breast carcinoma and significances.Methods:Fifty-three samples of breast ductal carcinoma tissue and 50 samples of surrounding normal tissue were selected.The expression levels of Smad2 and Smad4 in cancer tissue and surrounding normal tissue were detected with immunohistochemical S-P method,and the relationships between the expression levels of Smad2 and Smad4 and the clinicopathologic parameters of breast carcinoma were evaluated.Results:The expression level of Smad2 protein in the breast carcinoma tissue was significantly higher than that in the surrounding normal tissue (z = - 2.08,P 0.05;r=-0.077,P >0.05),tumor size (r= 0.128,P >0.05;r=0.133,P >0.05),lymph node invasion (r =0.163,P >0.05;r =0.006 P >0.05),distant metastasis (r =0.113,P >0.05;r = 0.126,P > 0.05),ER expression (r = 0.056,P > 0.05;r = 0.047,P > 0.05) and PR expression (r=0.129,P >0.05;r=0.107,P >0.05).However,the expression levels of Smad2 and Smad4 were negatively correlated with the expression of HER2 (r = - 0.388,P < 0.01;r = - 0.360,P < 0.01 ) and pathological grade (r = - 0.331,P < 0.05;r = - 0.388,P < 0.01 ).The expression of Smad2 was positively correlated to the expression of Smad4 in breast carcinoma (r=-0.83,P <0.01).Conclusion:The expressions of Smad2 and Smad4 may play an important role in the development of breast carcinoma,and they may be used as the potential biological markers for evaluating the degree of malignancy and prognosis of breast carcinoma.

17.
Article in Chinese | WPRIM | ID: wpr-484499

ABSTRACT

Objective:To study the differentiation capacity of the fibroblast-like cells isolated from human skin dermis into mesenchymal stem cells, and to explore the feasibility to use these cells as alternative cell source of autologus bone marrow mesenchymal stem cells (BMSCs ) for regeneration of tissue inj uries and defects. Methods:Full thickness skin samples were obtained from the abdomen of surgical patients, then digested with dispase and collagenase Ⅰ subsequently. Thereafter, the digested cells were collected and cultured, followed by suspension with serum free medium containing N2,B27,basic fibroblast growth factor (bFGF),and epidermal growth factor (EGF).The skin dermis derived spheroids (SDDSs)were collected and monolayer cultured in serum-containing medium.Finally,the cells were characterized by immunofluorescence staining and differentiation assays.Results:The dermis derived cells proliferated and formed SDDSs in the suspension of serum-free medium. After monolayer cultivation in serum-containing medium, the cells from spheroids were successfully expanded to large number. The cells expressed mesenchymal stem cells markers CD90, CD105 and vimentin. Under osteogenic,chondrogenic and adipogenic differentiation conditions,these cells were differentiated into the alizarin red,safranin O, and oil red O staining positive cells, displayed similar differentiation traits with BMSCs. However,safranin O staining was weaker in the dermis derived cells than BMSCs. Conclusion:A kind of fibroblast-like cells exist in human skin dermis, and have osteocytic, chondrogenic and adipogenic differentiation potentials,demonstrating that these cells will be utilized as a novel cell source for repairing the tissue injury and defect in clinic.

18.
Article in Chinese | WPRIM | ID: wpr-481923

ABSTRACT

Objective To discuss the clinical value of high sensitivity C‐reactive protein(hs‐CRP) ,fibrinogen(Fib) and D‐dimer (D‐D) measurement for patients with acute myocardial infarction(AMI) before and after the treatment with the anticoagulation and thrombolysis therapy .Methods 110 patients with AMI were recruited in the study and the plasma hs‐CRP ,Fib ,D‐D and myocardi‐al damage markers were measured before and after the treatment .Results 66 of the 110 patients′plasma hs‐CRP ,Fib ,D‐D concen‐trations elevated(higher than the threshold) before treatment and after treatment within 24 h ,while 44 patients′plasma hs‐CRP , Fib concentrations increased ,but D‐D didn′t .Conclusion The measurement of hs‐CRP is helpful for the diagnosis and treatment of AMI .Hs‐CRP is another good myocardial injury marker ,and the plasma hs‐CRP concentration after treatment for 24 -48 h could reflect the severity and prognosis of AMI better than after treatment within 12 h .Fib decreases relatively slowly after the treat‐ment ,so it cannot be used for curative effect observation for AMI patients;D‐D concentration dosen′t have the determined negative predictive value for the diagnosis of AMI ,so it cannot be used as screening out indicator for AMI ,but D‐D concentration can be used as therapeutic effect monitoring indicator for AMI patients with D‐D positive .

19.
Chinese Journal of Immunology ; (12): 517-521,526, 2015.
Article in Chinese | WPRIM | ID: wpr-601052

ABSTRACT

Objective:To investigate the best preservation conditions of N-terminal pro-brain natriuretic peptide( NT-proBNP) and provide detection references with stable performance for detection kits.Methods: ELISA was used to quantitatively detect the changes in NT-proBNP contents in various preservation solutions.The effects of basic buffer system, preservative Proclin300 and antibiotics on the preservation of NT-proBNP were analyzed using univariate analysis.The combination of various factors was then optimized using orthogonal experiments, to identify the best preservation system for NT-proBNP.Results: The univariate analysis determined that the basic buffer system for NT-proBNP was 0.02 mol/L phosphate buffered saline(PBS) at pH7.2,the addition of pre-servative Proclin300 could extend the preservation time of NT-proBNP at 37℃ by one day, the combined addition of penicillin and streptomycin prolonged the preservation time of NT-proBNP by one day compared with individually adding penicillin or streptomycin.The orthogonal experiments identified a preservation solution for NT-proBNP as 20%calf serum,1/1 000 Proclin300,120 U/ml penicillin and 80 U/ml streptomycin in a basic buffer system of 0.02 mol/L PBS at pH7.2.This solution was used to preserve an NT-proBNP reference sample at 37℃.Seven days later,the calibrated fixed-value of the sample at 37℃was only 1.3%lower than that at 4℃.Conclusion:Optimized NT-proBNP serum preservation solution could preserve NT-proBNP standard sample at 37℃ for seven days.

20.
Chinese Journal of Immunology ; (12): 1088-1092, 2014.
Article in Chinese | WPRIM | ID: wpr-454852

ABSTRACT

To prepare monoclonal antibody of carbohydrate antigen 19-9(CA19-9).Methods: Based on the titer test results of mouse ascites and its IC 50 values ,the mouse that prepare for fusion was identified.Positive monoclonal cell strains were established by cell fusing and screening.Monoclonal antibody from ascites was produced by peritoneal injection monoclonal cell , and then purified by octoic acid-ammonium sulfate precipitation method.After determine the protein concentrations by UV-spectrophotometry ,the monoclonal antibody against CA 19-9 was labelled with horseradish peroxidase.Based on antibody pairing test , DAS-ELISA method was established .To compared with abroad kit , analyzing performance of this method.Results: Three strains of monoclonal antibody were obtained.And the optimal working concentrations of mAb (ZJY3-1G9) ,as coated antibody,McAb(ZJY2-7F10),as HRP-IgG,were assured.Limit of detection was 26.4 U/ml.Linear range was 30-300 U/ml.By detecting patients with serum 33 , confirmed the correlation coefficient of r=0.950 4 , compared with abroad kit that measure simultaneously.Conclusion:Monoclonal antibody prepared for CA 19-9 can be used to develop a kit.

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