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Objective:To analyze the distribution characteristics of plasma renin concentration (PRC) in patients with aldosterone-producing adenoma (APA) and its impact on diagnosis.Methods:In this retrospective case series, clinical data from 200 patients with APA (80 men and 120 women; mean age 45.6 years) in the First Affiliated Hospital of Chongqing Medical University from November 2013 to January 2022 were evaluated. PRC was determined by automated chemiluminescence immunoassay. The distribution characteristics of PRC were analyzed, and 8.2 mU/L was used as the low renin cutoff to evaluate whether renin was suppressed.Results:The median PRC was 1.6 mU/L (range, 0.4-41.5 mU/L). There were 116 patients with APA with PRC of ≤2 mU/L, 41 patients with 2<PRC≤4 mU/L. PRC was not suppressed (PRC>8.2 mU/L) in 8.0% (16/200) of the patients with APA. And PRC was not suppressed in 2.5% (5/200) of the patients with APA, resulting in a primary aldosteronism negative screening outcome.Conclusions:Although most patients with APA have low PRC, there are a small number (8%) of patients whose PRC has not been fully suppressed, which can lead to missed diagnoses during primary aldosteronism screening. While primary aldosteronism is highly suspected, further investigations are required to determine the diagnosis, even if PRC is not fully suppressed at screening.
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Age-dependent loss of skeletal muscle mass and function is a feature of sarcopenia, and increases the risk of many aging-related metabolic diseases. Here, we report phenotypic and single-nucleus transcriptomic analyses of non-human primate skeletal muscle aging. A higher transcriptional fluctuation was observed in myonuclei relative to other interstitial cell types, indicating a higher susceptibility of skeletal muscle fiber to aging. We found a downregulation of FOXO3 in aged primate skeletal muscle, and identified FOXO3 as a hub transcription factor maintaining skeletal muscle homeostasis. Through the establishment of a complementary experimental pipeline based on a human pluripotent stem cell-derived myotube model, we revealed that silence of FOXO3 accelerates human myotube senescence, whereas genetic activation of endogenous FOXO3 alleviates human myotube aging. Altogether, based on a combination of monkey skeletal muscle and human myotube aging research models, we unraveled the pivotal role of the FOXO3 in safeguarding primate skeletal muscle from aging, providing a comprehensive resource for the development of clinical diagnosis and targeted therapeutic interventions against human skeletal muscle aging and the onset of sarcopenia along with aging-related disorders.
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Animals , Humans , Sarcopenia/metabolism , Forkhead Box Protein O3/metabolism , Muscle, Skeletal/metabolism , Aging/metabolism , Primates/metabolismABSTRACT
Objective To understand the prevalence of Salmonella and the characteristics of drug resistance genes in General Hospital of People's Liberation Army, and to provide evidence for the prevention and treatment of Salmonella infection. Methods A retrospective study was conducted to collect 78 clinical isolates of Salmonella from 2015 to 2017. The age of the patients was 49 ± 21 years old. The infected patients were mainly young and middle-aged. The clinical samples mainly came from feces and venous blood, accounting for 44.87％(35/78) and 33.33％(26/78), respectively. After serotype identification, drug sensitivity test and whole genome sequencing, multilocus sequence typing and drug resistance genotyping were performed. Cluster of Cefotaxime or Ciprofloxacin resistant Salmonella was analyzed. Results Salmonella group D (53.85％) and Salmonella group C (21.79％) were dominant Salmonella serotype. ST11 was mainly ST type. Drug sensitivity test showed that the multidrug resistance rate of Salmonella was 64.11％ (50/78). The sensitivity to all antimicrobial agents' rate was 25.64 (20/78). The resistance rate of Salmonella to nalidixic acid was 65.38％(51/78). The most common drug resistance gene of Salmonella was extended-spectrum β-lactam drug resistance gene, accounting for 78.21％ (61/78). Conclusions The ST-type and carrying resistance genes of Salmonella in this hospital were diverse. Most pathogens were multi-drug resistant to antimicrobial agents. Molecular typing and drug resistance gene analysis of Salmonella and construction of resistant strains to determine the inheritance of Salmonella relationships have a certain clinical significance.
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Objective To investigate the risk factors of restless leg syndrome (RLS) in maintenance hemodialysis patients. Methods The clinical data of 74 maintenance hemodialysis patients in Eastern Hepatobiliary Surgery Hospital of Navy Medical University (Second Military Medical University) from July 2016 to September 2017 were retrospectively analyzed. The gender, age, diabetes mellitus and other biochemical indicators of the patients with or without RLS were compared. The univariate logistic regression analysis was performed with the gender, age, diabetes mellitus and other biochemical indicators as covariates, and the RLS as dependent variable. According to the 2003 International Restless Leg Syndrome Study Group (IRLSSG) criteria, the patients with RLS were divided into mild, moderate, severe and very severe groups, with the patients without RLS as controls; the variables that were significantly different between the groups as analyzed by univariate logistic regression analysis were compared by t-test and multivariate logistic regression analysis. The receiver operating characteristic (ROC) curve for the main relevant factors was plotted, and the area under the curve was calculated to determine the best critical value and the corresponding sensitivity and specificity. Results A total of 74 maintenance hemodialysis patients were enrolled in this study, including 46 males and 28 females; the patients aged from 21 to 87 years, with an average age of (56.70±14.52) years; 55 patients without RLS, and 19 with RLS. The serum calcium in patients with RLS was significantly higher than that without RLS ([2.56±0.38] mmol/L vs [2.26±0.29] mmol/L, t＝2.61, P＝0.02). Multivariate logistic regression analysis showed that serum parathyroid hormone (PTH)515.39 pg/mL was an independent risk factor for RLS in maintenance hemodialysis patients (OR＝1.00, 95% CI 1.00-1.01, P＝0.03). With the cutoff value being 515.39 pg/mL, area under ROC curve performed by PTH for RLS was 0.759, with the 95% CI being 0.610- 0.907, and the sensitivity and specificity being 0.47 and 0.98, respectively. Conclusion RLS is a common complication in maintenance hemodialysis patients. Chronic kidney disease-mineral and bone metabolism disorder may be related to the occurrence of RLS in maintenance hemodialysis patients.
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Age-related macular degeneration (AMD) is the main cause of blindness in the people older than 50 years old,and atrophic age-related macular degeneration accounts for 85％-90％ patients of AMD.Following the application of multi-spectral imaging,fundus autofluorescence,optical coherence tomography,microperimetry,multifocal electroretinogram and other new methods in ophthalmic clinical,the morphological and functional changes of atrophic AMD lesions have been more in-depth and comprehen sive,so this article will give a review on the relevant progress of examination techniques in recent years.
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Age-related macular degeneration (AMD) is the main cause of blindness in the people older than 50 years old,and atrophic age-related macular degeneration accounts for 85％-90％ patients of AMD.Following the application of multi-spectral imaging,fundus autofluorescence,optical coherence tomography,microperimetry,multifocal electroretinogram and other new methods in ophthalmic clinical,the morphological and functional changes of atrophic AMD lesions have been more in-depth and comprehen sive,so this article will give a review on the relevant progress of examination techniques in recent years.
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Objective To introduce JCI standards into medical consumables management in order to improve the overall management of consumables.Methods Root cause analysis (RCA) method in JCI management standards was used to execute root cause analysis of each key point in medical consumables management.PDCA Cycle was involved in to promote medical consumables management.Results JCI management standardized the path,solved some problems and promoted the level during medical consumables management.Conclusion JCI standards contribute to the standardization and humanization of medical consumables management,and also enhances the patient safety at the same time.
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Objective To investigate the expression and variation of MIP 1β,MIP-2,and IL-12p70 in mice with bloodstream infection caused by 4 kinds of bacteria.Methods CD-1 (ICR) mouse models of bloodstream infection with Staphylococcus aureus (S.aureus),Enterococcus f aecalis (E.f aecalis),Escherichia coli (E.coli),and K lebsiella pneumoniae (K.pneumoniae) were established.After mice in each trial group and PBS control group were infected by bacteria for 0.5h,1h,3h,6h,12h,24h,and 48h,concentrations of MIP-1β,MIP-2,and IL-12p70 were detected by Luminex liquid suspension chip system.Results Concentrations of MIP-1β increased significantly 1h after bacteria was in blood,S.aureus,E.faecalis,E.coli,K.pneumoniae,and control groups were (134.5 ± 18.3),(61.5 ± 15.4),(3 354.0 ±809.0),(6 888.4 ± 1 100.2),and (28.9 ± 4.6) pg/mL respectively;the peak values of IL-12p70 were (389.3 ± 118.1),(127.6 ± 10.0),(42.2 ± 3.5),(62.8 ± 8.4),and (4.8 ± 0.3) pg/mL respectively.Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were significantly higher than other trial groups and control group (all P＜0.01),while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than E.coli,K.pneumoniae,and control groups (all P＜0.01).Conclusion Concentrations of MIP-1β and MIP-2 in E.coli and K.pneumoniae groups were both significantly higher than those in S.aureus and E.faecalis groups,while concentrations of IL-12p70 in S.aureus and E.faecalis groups were both significantly higher than those in E.coli and K.pneumoniae groups.The combination detection of multiple cytokines or chemokines are valuable in predicting gram-positive or gram-negative bacterial infection,and can provide basis for treatment of early infection.
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BACKGROUND: Our previous studies demonstrated that cardiac stem cells (CSCs) transplantation could improve cardiac function in rats with myocardial infarction (MI). However, the overall survival and cardiac differentiation of CSCs were low. OBJECTIVE: To investigate the effect of hypoxia preconditioning on CSCs proliferation and cardiogenic differentiation and the role of hypoxia induced factor-1alpha (HIF-1α)/apelin/putative receptor protein related to the angiotensin receptor AT1 (APJ) pathway in the procedure. METHODS: Cells cultured in vitro experienced exposure to hypoxia (1% O2) for 24 hours. Cardiogenic differentiation was induced by using 5-azacytidine for another 24 hours. Then, cells were cultured in normal condition for 2 weeks. Normoxia (20% O2) was used as a negative control during the whole process. Cell proliferation was detected using MTS method and expressions of HIF-1α, apelin, cTnT and APJ were detected using western blot assay after 24 hours of preconditioning and 2 weeks after the induction of differentiation; the percentage of cTnT-positive cardiomyocyte-like cells was observed by immunofluorescence staining. RESULTS AND CONCLUSION: Compared with the normoxia group, the hypoxia group presented a higher proliferation rate and a higher absorbance value at 490 nm (P < 0.01); the protein expressions of HIF-1α, apelin and APJ were all enhanced after hypoxia exposure for 24 hours and 2 weeks after the induction of differentiation (P < 0.01); the percentage of cTnT-positive cells was greatly increased in the hypoxia group (P < 0.01), and the expression of cTnT was also significantly intensified (P < 0.01). To conclude, hypoxia preconditioning could promote the proliferation and cardiogenic differentiation of CSCs, and the activation of HIF-1α/Apelin/APJ pathway might be involved in this process.
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AIM: To synthesis and characterize a multi-functional siRNA delivery agent with effective thera-peutic effects and MR-tracing ability for programmed death ligand-1 ( PD-L1 ) positive gastric cancer SGC-7901 cell line . METHODS:The characterization , binding ability , cytotoxicity , transfection efficiency and cellular internalization of the polyplex were determined .The PD-L1 knockdown effect was analyzed , and cytokines secreted by cocultured T cells were measured.RESULTS:We developed folic acid (FA)-PEG-SS-PEI-SPION as siRNA delivery agent for PD-L1 knockdown. At N/P ratio of 10, the FA-PEG-SS-PEI-SPION bound PD-L1 siRNA to form polyplex in a diameter of (116.7 ±2.5) nm with zeta potential of (9.14 ±0.80) mV.Transfection efficiency of the targeted polyplex was (95.06 ±0.44)%, com-pared with ( 93.87 ±1.05 )% of the untargeted polyplex .Mean fluorescence intensity of the targeted polyplex was 1892.67 ±81.51, significantly higher than 1324.33 ±186.58 of the untargeted.The cellular magnetic resonance (MR) imaging showed the polyplex also acted as T 2 weighted contrast agent for cancer MR imaging .The relative mRNA level of PD-L1 in polymer/siRNA-2 treatment group was (9.07 ±0.79)%.Decreased protein expression of PD-L1 was showed by Western blot .The secretion levels of IFN-γand TNF-αin cocultured T cells increased , while that of IL-10 decreased . CONCLUSION:Our findings highlighted the potential of the multifunctional theranostic nanoparticles for effective targe -ting PD-L1 knockdown therapy and MR imaging diagnosis in gastric cancers .
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Objective To introduce JCI standards into medical consumables management in order to improve the overall management of consumables.Methods Root cause analysis (RCA) method in JCI management standards was used to execute root cause analysis of each key point in medical consumables management.PDCA Cycle was involved in to promote medical consumables management.Results JCI management standardized the path,solved some problems and promoted the level during medical consumables management.Conclusion JCI standards contribute to the standardization and humanization of medical consumables management,and also enhances the patient safety at the same time.
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<p><b>OBJECTIVE</b>To observe MET-associated alteration during the trans-differentiation from MSCs to neuron-like cells, and to explore the possible molecular mechanism.</p><p><b>METHODS</b>Bone marrow MSCs were isolated from rat femur and purified in continuous cell culture. After induced differentiation to neuron-like cells by the combination of butylated hydroxyanisole (BHA) and dimethyl sulfoxide (DMSO), cells were tested by comparative polymerase chain reaction (PCR) for the relative expression of MET biomarkers and transcription factors, and for cell cycle by flow cytometry. Meanwhile, target genes of Wnt/β-catenin pathway were also analyzed by comparative PCR to determine the possible involvement.</p><p><b>RESULTS</b>In MSC-induced neuron-like cells, MET-associated transcription factors such as Snail, Slug, ZEB1, ZEB2, and Twist were significantly attenuated in expression level. The Mesenchymal marker Vimentin expression level was increased. Membrane protein E-cad was slightly down-regulated, while N-cad level was marginally elevated. Percentage of proliferating cells (S phase in cell cycle) markedly shrank from 40.42% for MSCs to 6.76% for MSC-derived neuron. Additionally, Wnt/β-catenin target genes β-catenin and c-myc were decreasingly expressed.</p><p><b>CONCLUSION</b>Chemically induced trans-differentiation from MSC to neuron caused similar MET-featured alteration in gene expression and proliferation to known MET, which might be underlied by deactivation of Wnt/β-catenin pathway.</p>
Subject(s)
Animals , Rats , Epithelial-Mesenchymal Transition , Mesenchymal Stem Cells , Cell Biology , Neurons , Cell Biology , Metabolism , Proto-Oncogene Proteins c-myc , Metabolism , Wnt Signaling Pathway , beta Catenin , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the expression of secreted frizzled-related protein 1 (SFRP1), β-catenin and E-cadherin in colorectal carcinoma and its clinicopathological significance.</p><p><b>METHODS</b>The expression of SFRP1, β-catenin and E-cadherin mRNA and protein in tumor and pericancerous tissue samples from 60 cases of colorectal cancer was assayed by reverse-transcription PCR and immunohistochemistry, respectively. The correlation of their expression with clinicopathological factors of colorectal cancer was analyzed.</p><p><b>RESULTS</b>In 52/60 cases the relative mRNA expression of SFRP1 in cancer tissue and pericancerous tissue was 0.4837±0.1532 and 0.7170 ±0.1830; for β-catenin was 0.9293± 0.3705 and 0.6469±0.3166; and for E-cadherin was 0.5556±0.2535 and 0.9422±0.2372 (P<0.01), respectively. SFRP1 mRNA expression was associated with lymphatic metastasis (P<0.05). The positive rate of SFRP1 in colorectal cancer was 31.67% (19/60), and was significantly lower than that in pericancerous colorectal mucosa (75.00%, 45/60). No relationship between SFRP1 protein expression and clinical pathology was found. Abnormal expression rates of β-catenin and E-cadherin in colorectal cancer were 75.00% (45/60) and 58.33% (35/60), respectively, which were significantly higher than that in pericancerous colorectal mucosa (1.67% and 6.67%), respectively. Abnormal β-catenin and E-cadherin expression was associated with tumor differentiation, lymphatic metastasis and Duke's staging. SFRP1 protein expression was negatively correlated with β-catenin and E-cadherin expression (r=-0.517, -0.442, Ps<0.01).</p><p><b>CONCLUSION</b>Down-regulation of SFRP1 in colorectal cancer may cause abnormal Wnt signaling and induce abnormal β-catenin and E-cadherin expression, indicating that SFRP1 might be involved in the development and progression of colorectal cancer, and could be a novel therapeutic target for colorectal cancer.</p>
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Cadherins , Metabolism , Colorectal Neoplasms , Metabolism , Intercellular Signaling Peptides and Proteins , Metabolism , Lymphatic Metastasis , Membrane Proteins , Metabolism , beta Catenin , MetabolismABSTRACT
Objective Peritoneal dialysis can be done at home , however , its clinical follow-up quality is lower compared with hemodialysis due to the lack of detailed follow-up system and professional management team for post-discharge nursing care .The article aimed to explore the effect of a nurse-led integrated specialized disease management model in the follow-up of peritoneal dialysis patients . Methods Nurse-led integrated specialized disease management model was provided for 270 patients with peritoneal dialysis (PD) from January 2012 to May 2013.Self-designed questionnaire on diasease knowledge and self-care assement questionnaire were used for patients to evaluate their self-care ability and understanding of disease knowledge .Regular follow-up continued . Results The patients′understanding of disease knowledge was on the rise at 1st day before discharge, 1st month and 3rd months after discharge. After the application of nurse-led integrated specialized disease management model , the patients′regular clinical follow-up rate was in-creased from 78.3% to 88.6%.The patients′self-care ability improved gradually at 1st day before discharge , 1st month and 3rd months after discharge , which was of significant differences . Conclusion The nurse-led nurse-led integrated specialized disease management model can improve peritoneal dialysis patients′self-care ability and reduce their medical expense , which is of clinical sig-nificance .
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Acute kidney injury (AKI), associated with significant morbidity and mortality, is widely known to involve epithelial apoptosis, excessive inflammation, and fibrosis in response to ischemia or reperfusion injury, which results in either chronic pathological changes or death. Therefore, it is imperative that investigations are conducted in order to find effective, early diagnoses, and therapeutic targets needed to help prevent and treat AKI. However, the mechanisms modulating the pathogenesis of AKI still remain largely undetermined. MicroRNAs (miRNAs), small non-coding RNA molecules, play an important role in several fundamental biological and pathological processes by a post transcriptional regulatory function of gene expression. MicroRNA-21 (miR-21) is a recently identified, typical miRNA that is functional as a regulator known to be involved in apoptosis as well as inflammatory and fibrotic signaling pathways in AKI. As a result, miR-21 is now considered a novel biomarker when diagnosing and treating AKI. This article reviews the correlative literature and research progress regarding the roles of miR-21 in AKI.
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Animals , Humans , Acute Kidney Injury , Diagnosis , Drug Therapy , Genetics , Pathology , Apoptosis , Biomarkers , Metabolism , MicroRNAs , Genetics , Metabolism , Molecular Targeted TherapyABSTRACT
<p><b>BACKGROUND</b>The regulation of endometrial physiology and morphogenesis by the paracrine effectors has been well established using in vivo studies. A more complete understanding of the endometrial function has been delayed due, in part, to a lack of appropriate culture models. In this study, we aimed to simulate the in vivo three-dimensional (3-D) growth pattern of endometrial cells using a 3-D in vitro culture system.</p><p><b>METHODS</b>Isolated endometrial epithelial cells, stromal cells and RL95-2 cells were seeded into culture chambers coated with the extracellular matrix Matrigel and observed using light microscopy. Fluorescence staining and immunohistochemistry were used to assess the morphology.</p><p><b>RESULTS</b>Depending on the culture conditions, epithelial cells and RL95-2 cells formed multicellular structures on Matrigel; stromal cells remained individually distinguishable or grew together to form 3-D lattice-like structures.</p><p><b>CONCLUSIONS</b>Matrigel provided a good microenvironment for culturing endometrial cells. The cells cultured in the Matrigel-coated chambers closely resembled those seen in vivo.</p>
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Female , Humans , Cell Culture Techniques , Methods , Cell Line , Cells, Cultured , Endometrium , Cell Biology , ImmunohistochemistryABSTRACT
BACKGROUND: Conventional hemodialysis mainly for cleaning uremic micro molecule substance, such as urea nitrogen or creatinine; however, few hemodialyses can clean uremic middle molecule substances (MMS). With prolonged dialysis duration, MMS accumulates in vivo and induces a series of complications. OBJECTIVE: To compare the efficiency of adsorptive dialysis (hemoperfusion unites hemodialysis) and conventional hemodialysis in cleaning uremic MMS. METHODS: Totally 60 maintenance hemodialysis patients were averagely divided into the adsorptive dialysis group and conventional hemodialysis group. First of all, hemoperfusion apparatus and dialyser were connected in series to take the adsorptive dialysis in the adsorptive dialysis group (hemoperfusion apparatus were equipped before dialyser). 120 minutes later, the hemoperfusion apparatus was toke off and continues to hemodialysis for 120 minutes. Duration of conventional hemodialysis was 240 minutes. Changes in clinical symptoms and levels of liver function, kidney function, serum electrolytes, hemocytes and uremic MMS were observed prior to and after treatment. RESULTS AND CONCLUSION: Adsorptive dialysis could remove the MMS notably. Compared with the conventional hemodialysis group, a single 120 minutes treatment could decrease MMS significantly (P < 0.05). The platelet levels were obviously decreased in the adsorptive dialysis group after treatment (P < 0.05), which were significantly different from the conventional hemodialysis group (P < 0.05). There was no significant difference in liver function, kidney function or serum electrolytes concentration. But related symptoms, such as the skin itch, sleep disorders and myalgia, were relieved more or less.
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<p><b>AIM</b>To observe the change of potassium current on cultured neurons differentiated from hippocampus neural stem cells of the newborn rat.</p><p><b>METHODS</b>Neural stem cells from newborn rat hippocampus were cultured in vitro and passaged continuously. Differentiation of the cell was induced by serum and removing mitogens. After differentiation cells were plated on plastic dishes and cultured for 1 d, 7 d, 14 d and 21 d. Whole-cell voltage patch clamp recording was used respectively to detect voltage-dependent K+ current.</p><p><b>RESULTS</b>After 1 d culture, no current was detected, and on the 7th d, 14th d, 21st d after differentiation, the amplitude of K+ currents was (18.077 +/- 2.789)pA/pF, (13.099 +/- 2.742)pA/pF, (34.045 +/- 8.067)pA/pF at +50 mV. The recorded K+ current included two components that could be blocked by TEA and 4-AP separately, assumed the slowly inactivating delayed rectifier K+ current (IK) and the fast inactivating transient outward K+ current (IA).</p><p><b>CONCLUSION</b>The function of potassium channels on the hippocampus neural stem cells of the newborn rat approaches mature gradually when the time of differentiation becomes longer in vitro.</p>
Subject(s)
Animals , Rats , Animals, Newborn , Cells, Cultured , Delayed Rectifier Potassium Channels , Physiology , Hippocampus , Cell Biology , Neural Stem Cells , Cell Biology , Metabolism , Physiology , Patch-Clamp Techniques , Potassium Channels , Physiology , Potassium Channels, Inwardly Rectifying , Physiology , Rats, Sprague-DawleyABSTRACT
<p><b>AIM</b>To learn the electrophysiological interference of levofloxacin (LVFX) to heart in guinea pig.</p><p><b>METHODS</b>High, moderate and low doses of LVFX were given to the anesthetic guinea pig via i.p., and QT interval span and corrected QT-interval span in the II leading lines of ECG were recorded and analyzed from 5 min to 360 min after the drug administration. Single ventricular myocytes were obtained and impacted by LVFX solution of different concentrations. Then delayed rectifier potassium currents (I(K)) on single cells were recorded with whole-cell patch clamp technique, and compared with control group(without impact of LVFX).</p><p><b>RESULTS</b>(1) After the administration of LVFX, at the dose of 200 mg/kg. QT-interval span was significantly elongated, and the increasing rate is 19.38% +/- 3.15% (P < 0.05). While at the relatively lower doses of 50 mg/kg and 100 mg/kg, the elongation is of low/no significance (P > 0.05). (2) LVFX inhibited I(K) dose-dependently and time-dependently.</p><p><b>CONCLUSION</b>LVFX might prolong the QT-interval span by the mechanism of inhibiting I(K), which implies a potential risk in clinical application.</p>
Subject(s)
Animals , Guinea Pigs , Levofloxacin , Membrane Potentials , Myocytes, Cardiac , Physiology , Ofloxacin , Pharmacology , Patch-Clamp TechniquesABSTRACT
To construct a scFv library by phage display technique from the spleen cells of mice immunized with B3HM cells. Three mice were immunized with B3HM cells, and their spleen cells were harvested. The genes of VH and Vk were amplified by RT-PCR from the cDNA of the immunized spleen cells and a scFv-phage display antibody library was constructed. The capacity of library was measured,and the variety of the library was analyzed by digesting with restriction endonuclease BstNI.ScFv phage clones were randomly picked and identified phage-scFv clone by binding B3HM cells using immunofluorescein.A scFv library containing 5?106 individual clones which showed different patterns after digested with restriction endonuclease BstNI was produced. Individnal phage-scFv clone showed B3HM cells positive using immunofluorescein. A scFv library of anti-B3HM cell surface molecules has been constructed. It will be useful for finding out some novel genes of causing leukemia, and establishs the infarctate foundation of clarifying the pathogenesis of leukemiagenesis.