BACKGROUND Gastric cancer(GC)is characterized by a low 5-year survival rate.The prognosis is still not satisfactory although it has significantly improved due to developments in medicine.Thus,the identification of mor...BACKGROUND Gastric cancer(GC)is characterized by a low 5-year survival rate.The prognosis is still not satisfactory although it has significantly improved due to developments in medicine.Thus,the identification of more efficient indices for the evaluation of GC prognosis is required.We propose,for the first time,that the alkaline phosphatase(ALP)to prealbumin(PA)ratio(APR)can be used as an independent prognostic factor in GC.AIM To evaluate the prognostic value the APR in GC.METHODS According to the exclusion strategy,we collected the preoperative serologic examination results and clinical information of 409 GC patients treated in Shandong Provincial Hospital from January to December,2016.By calculating the APR,the neutrophil and lymphocyte ratio(NLR),C-reactive protein(CRP)and albumin(ALB)ratio,platelet and lymphocyte ratio,lymphocyte and monocyte ratio,and the relationship with clinical information,we verified the role of preoperative APR ratio in the prognosis of GC.In addition,we used a Cox model combined with the APR and tumor stage to demonstrate its efficacy in assessing the prognosis of GC patients.RESULTS Preoperative APR was an independent prognostic factor for GC.The median age of patients in the APR-high group was greater compared with that in the APR-low group.Patients with a higher APR had a more advanced clinical stage,higher neutrophil to lymphocyte,CRP to ALB,and platelet to lymphocyte ratios,but a lower lymphocyte to monocyte ratio(P<0.05).The APR-high group also had higher glycoprotein antigen 199 and carbohydrate antigen 125 levels than the APR-low group(P<0.05).Median overall survival and disease-free survival were significantly longer in the APR-low group than in the APR-high group.In addition,a Cox model based on the APR and tumor stage was more effective in evaluating the prognosis of patients than models based on stage alone or stage plus the NLR.CONCLUSION A higher APR is an independent and negative prognostic factor for GC.The prognosis of GC can be better evaluated using a Cox model based on the APR and stage.展开更多
Alkaline phosphatase(ALP) plays an integral role in the metabolism of liver and development of the skeleton in humans. To date, the interactions between different-duration terahertz(THz) radiation and ALP activities, ...Alkaline phosphatase(ALP) plays an integral role in the metabolism of liver and development of the skeleton in humans. To date, the interactions between different-duration terahertz(THz) radiation and ALP activities, as well as the influence mechanism are still unclear. In this study, using the para-nitro-phenyl-phosphate(p NPP) method, we detect changes in ALP activities during 40-minute THz radiation(0.1 THz, 13 m W/cm^2). It is found that the activity of ALP decreases in the first 25 min, and subsequently increases in the later 15 min. Compared with the activity of ALP being heated, the results suggest that short-term terahertz radiation induces a decrease in enzyme activity through the non-thermal mechanism. In order to explore the non-thermal effects of THz radiation on ALP, we focus on the impacts of 0.1 THz radiation for 20 min on the activity of ALP in different concentrations. The results reveal that the activity of ALP decreases significantly after exposure to THz radiation. In addition, it could be deduced from fluorescence, ultraviolet-visible(UV-vis), and THz spectra results that THz radiation has induced changes in ALP structures. Our study unlocks non-thermal interactions between THz radiation and ALP, as well as suggests that THz spectroscopy is a promising technique to distinguish ALP structures.展开更多
AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study w...AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genotype 1 were treated for 48 wk with 180 μg PegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 1000 mg/d for those under 75 kg or 1200 mg/d for those over 75 kg. Patients with geno- types 2 and 3 were treated for 24 wk with 180 μgPegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 800 mg/d. RESULTS: In all ETR patients, binary logistic regression analysis identif ied absence of complete early virological response (cEVR) (OR 27.07, 95% CI: 3.09-237.26, P < 0.005), serum alkaline phosphatase (ALP) levels prior to therapy < 75 U/L (OR: 6.16, 95% CI: 2.1-18.03, P < 0.001) and body mass index > 26 kg/m2 (OR: 8.27, 95% CI: 2.22-30.84, P < 0.005) as independent predictors of relapse. When cEVR patients were analyzed exclusively, ALP prior to therapy < 75 U/L remained the only predictor of relapse. CONCLUSION: Lower levels of ALP prior to, during and after therapy seem to be associated with a higher risk of relapse in CHC patients with ETR.展开更多
Background: In temperate-breeding birds, individuals must adjust their physiological states from one life-history stage to another in response to changing conditions to maximize ecological fitness. Previous evidences ...Background: In temperate-breeding birds, individuals must adjust their physiological states from one life-history stage to another in response to changing conditions to maximize ecological fitness. Previous evidences have shown that body mass, size-corrected mass(SCM), and hematocrit(Hct) could be used as estimates of the energetic state of individuals to illustrate life-history trade-offs and individual quality in field physiology. Plasma alkaline phosphatase(ALP) plays critical roles in regulating the metabolism of energy but very limited information is known on its link with body mass or Hct.Methods: We determined the changes of plasma ALP levels in both early breeding and wintering stages of male Eurasian Tree Sparrows(Passer montanus), and examined the relationships between ALP and body mass, SCM, and Hct of the birds.Results: Our study showed that(1) in male Eurasian Tree Sparrows, body mass did not vary with life-history stage but plasma ALP activity significantly increased in the wintering stage compared to the breeding stage;(2) ALP activity was not correlated with individual body mass but was positively correlated with individual SCM and Hct. Such positive correlations, however, only occurred in the wintering but not in the breeding stages.Conclusions: Our results suggest that plasma ALP activities in free-living birds can be used as one of the indicators o body condition or nutritional status for analyzing individual variation in the wintering but not in the breeding stages. The life-history dependent relationships between plasma ALP activity and body condition may contribute to our better understanding of the trade-off between individual survival and reproduction in free-living animals.展开更多
Previous clinical study showed that aged garlic extract (AGE) has a preventive effect of vascular calcification in patients with coronary artery diseases. It has been reported that vascular smooth muscle cells (VSMC) ...Previous clinical study showed that aged garlic extract (AGE) has a preventive effect of vascular calcification in patients with coronary artery diseases. It has been reported that vascular smooth muscle cells (VSMC) and inflammatory cells including macrophages and lymphocytes migrate to sub-intimal region where atherosclerotic plaques are formed, and VSMC can differentiate into osteoblast-like cells, which are represented by the induction of alkaline phosphatase (ALP). We found that primary human coronary artery smooth muscle cells (HCASMC) showed the increased ALP activity, when cultured in the medium containing ascorbic acid, β-glycerophosphate, dexamethasone (IM), and supplemented with conditioned medium from macrophages (MCM). Then we tested the effect of AGE subdivided fractions and several compounds found in AGE, and then found that ((+)-(2S,3R)-Dehydrodiconiferyl alcohol, a dilignol compound existed in hydrophobic fraction of AGE, inhibited ALP activity in HCASMC.展开更多
Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease dia...Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods.Terminal deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 30-OH end of single-stranded DNA in the absence of a template.In this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP activity.After ALP hydrolyzes the 30-PO_(4) end of the substrate and generates 30-OH,TdT can effectively elongate the 30-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T probes.Endo Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 30-OH end,leading to the generation of exponential fluorescence signal amplification.The substrate for TdT elongation was optimized,and a limit of detection of 4.3×10^(-3) U/L was achieved for ALP by the optimized substrate structure.This method can also detect ALP in the cell lysate of a single cell.This work has potential applications in disease diagnosis and biomedical detection.展开更多
Dissolved inorganic phosphorus is an important form of directly bioavailable phosphorus for cyanobacteria in natural water.Dissolved organic phosphorus could be used by cyanobacteria via alkaline phosphatase,which is ...Dissolved inorganic phosphorus is an important form of directly bioavailable phosphorus for cyanobacteria in natural water.Dissolved organic phosphorus could be used by cyanobacteria via alkaline phosphatase,which is produced mainly by bacteria and also cyanobacteria itself.Herein,we review the current knowledge of extracellular phosphatase excreted by cyanobacteria,highlighting the development of detection method and its ecological roles in regulating pho sphorus cycling in freshwater systems,which is based on reports for around 100 species of cyanobacteria.Recommendations are suggested concerning the extracellular phosphatase produced by bloom-forming cyanobacteria in terms of the ecological role,followed by a discussion of the future prospects for the study.展开更多
The purpose of this study was to explore the effects of recombinant human intestinal alkaline phosphatase(recIAP) on human neutrophils in vitro, and the migration, phagocytosis, apoptosis in presence and absence of LP...The purpose of this study was to explore the effects of recombinant human intestinal alkaline phosphatase(recIAP) on human neutrophils in vitro, and the migration, phagocytosis, apoptosis in presence and absence of LPS. In this study, freshly extracted human neutrophils were used to establish an inflammatory cell model, and the control group, recIAP group, LPS group and recIAP +LPS group were set up to stimulate the model. The migration of neutrophils was detected by agarose gel drop method. Fluorescent particles and fluorescent probes were added to different treatment groups, and the phagocytic rate of neutrophils and the release of reactive oxygen species(ROS) from neutrophils were detected by flow cytometry. The apoptosis rate of neutrophils was detected by flow cytometry according to Annexin V-FITC apoptosis detection kit. The results showed that regardless of the presence or absence of LPS, recIAP could inhibit the migration of neutrophils, phagocytosis and the release of ROS. In addition, recIAP could weaken the inhibitory effect of LPS on neutrophils apoptosis.展开更多
Objective To investigate the influence of sodium fluoride(NaF)on alkaline phosphatase(ALP)activity and bone gla protein(BGP)synthesis in yellow ligament cells from different surgical simples in vitro.Methods The human...Objective To investigate the influence of sodium fluoride(NaF)on alkaline phosphatase(ALP)activity and bone gla protein(BGP)synthesis in yellow ligament cells from different surgical simples in vitro.Methods The human ligament cells展开更多
BACKGROUND Previous studies have suggested that a low albumin-to-alkaline phosphatase ratio(AAPR)is associated with a lower survival rate in patients with various malignancies.However,the relationship between pretreat...BACKGROUND Previous studies have suggested that a low albumin-to-alkaline phosphatase ratio(AAPR)is associated with a lower survival rate in patients with various malignancies.However,the relationship between pretreatment AAPR and the prognosis of patients with gastric cancer(GC)remains unclear.AIM To investigate the prognostic value of AAPR in distant metastatic GC.METHODS A total of 191 patients with distant metastatic cancer from a single institute were enrolled in this study.Pretreatment clinical data,including serum albumin and alkaline phosphatase levels,were collected.A chi-square test or Fisher’s exact test was applied to evaluate the correlations between AAPR and various clinical parameters in GC patients.The Kaplan-Meier method and Cox proportional hazards regression model were used to evaluate the prognostic efficacy of AAPR in metastatic GC patients.A two-sided P value lower than 0.05 was considered statistically significant.RESULTS A receiver operating characteristic curve indicated that 0.48 was the optimal threshold value for AAPR.AAPR≤0.48 was significantly associated with bone(P<0.05)and liver metastasis(P<0.05).Patients with high levels of AAPR had better survival in terms of overall survival(OS)and progression-free survival(PFS),regardless of the presence of liver/bone metastasis.Pretreatment AAPR was found to be a favorable predictor of OS and PFS based on a multivariate cox regression model.AAPR-M system,constructed based on AAPR and number of metastatic sites,showed superior predictive ability relative to the number of metastatic sites for predicting survival.CONCLUSION Pretreatment AAPR may serve as an independent prognostic factor for predicting PFS and OS in patients with metastatic GC.Furthermore,AAPR may assist clinicians with individualizing treatment.展开更多
INTRODUCTIONOnly the liver has the great capability ofregeneration in mammal.Few hepatocytes are inthe phase of division in the normal liver of an adultmammal (including human beings),but theremaining hepatocytes can ...INTRODUCTIONOnly the liver has the great capability ofregeneration in mammal.Few hepatocytes are inthe phase of division in the normal liver of an adultmammal (including human beings),but theremaining hepatocytes can be induced to proliferatequickly by partial hepatectomy (PH),and,to somedegree,they stop dividing and re-differentiate intocells functioning as hepatocytes.This shows展开更多
AIM: To determine the prognostic value of alkaline phosphatase(ALP) and γ-glutamyltransferase(GGT) for hepatocellular carcinoma(HCC).METHODS: We analyzed the outcome of 172 HCC patients who underwent liver resection....AIM: To determine the prognostic value of alkaline phosphatase(ALP) and γ-glutamyltransferase(GGT) for hepatocellular carcinoma(HCC).METHODS: We analyzed the outcome of 172 HCC patients who underwent liver resection. Receiver operating characteristic(ROC) curve analysis was performed to determine the cut-off value of ALP and GGT. Then, preoperative risk factors for survival were evaluated by multivariate analysis. Based on the significant factors, aprognostic score model was established.RESULTS: By ROC curve analysis, ALP > 120 U/L and GGT > 115 U/L were considered elevated. Overall survival(OS) and tumor-free survival(TFS) for patients with elevated ALP and GGT were significantly worse than for patients with ALP and GGT within the normal range. Multivariate analysis showed that the elevated levels of ALP, GGT and tumor size were independent prognostic factors. Giving each positive factor as a score of 1, we established a preoperative prognostic score model. The 5-year OS for patients with a score of 0, 1, 2 and 3 were 84.0%, 45.9%, 44.1% and 0%, respectively, while the TFS was 80.6%, 40.0%, 38.8% and 0%, respectively. When combining patients with scores of 1 and 2 into the middle risk group, and patients with scores of 0 and 3 into the low-risk and highrisk groups, respectively, different outcomes would be significantly distinguished by the risk groups.CONCLUSION: Elevated ALP and GGT levels were risk predictors in HCC patients. Our prognostic model might vary the outcomes of patients from different risk groups.展开更多
BACKGROUND Gamma-glutamyltransferase(GGT) is one of the most important laboratory tests for the evaluation of liver damage. Through a long-term clinical observation of patients with secondary asymptomatic choledocholi...BACKGROUND Gamma-glutamyltransferase(GGT) is one of the most important laboratory tests for the evaluation of liver damage. Through a long-term clinical observation of patients with secondary asymptomatic choledocholithiasis, we found that most patients had abnormal GGT serum levels.AIM To investigate the combination of serum GGT and alkaline phosphatase(ALP) in predicting the diagnosis of asymptomatic choledocholithiasis secondary to cholecystolithiasis.METHODS In this retrospective cohort study, the clinical data of 829 patients with cholecystolithiasis admitted to the Third Affiliated Hospital of Zunyi Medical College from August 2014 to August 2017 were collected. Among these patients,151 patients had secondary asymptomatic choledocholithiasis and served as the observation group, and the remaining 678 cholecystolithiasis patients served as the control group. Serum liver function indexes were detected in both groups,and the receiver operating characteristic(commonly known as ROC) curves were constructed for markers showing statistical significances. The cutoff value,sensitivity, and specificity of each marker were calculated according to the ROC curves.RESULTS The overall incidence of asymptomatic choledocholithiasis secondary to cholecystolithiasis was 18.2%. The results of liver function indexes including serum aspartate aminotransferase, alanine aminotransferase, direct bilirubin and total bilirubin levels showed no significant differences between the two groups(P> 0.05). However, the serum GGT and ALP levels were significantly higher in the observation group than in the control group(P < 0.05). The ROC curve analysis showed that the area under the curve was 0.881(95%CI: 0.830-0.932), 0.647(95%CI: 0.583-0.711) and 0.923(0.892-0.953) for GGT, ALP, and GGT + ALP,respectively. The corresponding cut-off values of GGT and ALP were 95.5 U/L and 151.5 U/L, sensitivity were 90.8% and 65.1%, and specificity were 83.6% and59.8%, respectively. The sensitivity and specificity of GGT + ALP were 93.5% and85.1%, respectively.CONCLUSION An abnormally elevated serum GGT level has an important value in the diagnosis of asymptomatic choledocholithiasis secondary to cholecystolithiasis.The combination of serum GGT and ALP has better diagnostic performance. As a convenient, rapid and inexpensive test, it should be applied in secondary asymptomatic choledocholithiasis routine screening.展开更多
Alkaline phosphatases(APs) include the placental AP(PLAP), germ cell AP(GCAP), intestinal AP(IAP) and tissue nonspecific AP(TNAP). Over expression of TNAP in smooth muscle cells of kidney and vessels provokes the prog...Alkaline phosphatases(APs) include the placental AP(PLAP), germ cell AP(GCAP), intestinal AP(IAP) and tissue nonspecific AP(TNAP). Over expression of TNAP in smooth muscle cells of kidney and vessels provokes the progress of such serious diseases as end-stage renal disease, idiopathic infantile arterial calcification, ankylosis, osteoarthritis and diabetes. In order to design and optimize the potent TNAP inhibitors, comparative molecular field analysis(CoMFA) and comparative molecular similarity indices analysis(CoMSIA) were used to analyze 3D structure-activity relationships(3D-QSAR) of TNAP inhibitors. The 3D-QSAR model(CoMFA with q^2 = 0.521, r^2 = 0.930; CoMSIA with q^2 = 0.529, r^2 = 0.933) had a good predictability. Surflex-dock was used to reveal the binding mode between the inhibitors and TNAP protein. CoMFA, CoMSIA and docking results provide guidance for the discovery of TNAP inhibitors. Finally, eight new compounds as potential TNAP inhibitors were designed.展开更多
Intestinal alkaline phosphatase(IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP's role in the intestine is to dephosphoryl...Intestinal alkaline phosphatase(IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP's role in the intestine is to dephosphorylate toxic microbial ligands such as lipopolysaccharides, unmethylated cytosine-guanosine dinucleotides and flagellin as well as extracellular nucleotides such as uridine diphosphate. IAP's ability to detoxify these ligands is essential in protecting the host from sepsis during acute inflammation and chronic inflammatory conditions such as inflammatory bowel disease. Also important in these complications is IAP's ability to regulate the microbial ecosystem by forming a complex relationship between microbiota, diet and the intestinal mucosal surface. Evidence reveals that diet alters IAP expression and activity and this in turn can influence the gut microbiota and homeostasis. IAP's ability to maintain a healthygastrointestinal tract has accelerated research on its potential use as a therapeutic agent against a multitude of diseases.Exogenous IAP has been shown to have beneficial effects when administered during ulcerative colitis,coronary bypass surgery and sepsis.There are currently a handful of human clinical trials underway investigating the effects of exogenous IAP during sepsis,rheumatoid arthritis and heart surgery.In light of these findings IAP has been marked as a novel agent to help treat a variety of other inflammatory and infectious diseases.The purpose of this review is to highlight the essential characteristics of IAP in protection and maintenance of intestinal homeostasis while addressing the intricate interplay between IAP,diet,microbiota and the intestinal epithelium.展开更多
Objective This study aimed to investigate the relationship between alkaline phosphatase(ALP) and common carotid intima media thickness(IMT), carotid plaque, and extracranial carotid artery stenosis(ECAS). Methods A to...Objective This study aimed to investigate the relationship between alkaline phosphatase(ALP) and common carotid intima media thickness(IMT), carotid plaque, and extracranial carotid artery stenosis(ECAS). Methods A total of 3,237 participants aged ≥ 40 years were recruited from Jidong community in 2013-2014. Participants were divided into five quintile groups based on their serum ALP levels. Carotid atherosclerosis was assessed using ultrasound. Abnormal IMT, carotid plaque, and ECAS were defined as IMT > 0.9 mm, IMT > 1.5 mm, and ≥ 50% stenosis in at least one extracranial carotid artery, respectively. Results Common carotid IMT values and the prevalence of carotid plaque increased across serum ALP quintiles. Higher ALP quintiles were correlated with an increased risk of abnormal IMT [fourth quintile: odds ratio(OR) 1.78, 95% confidence interval(CI) 1.13-2.82, P = 0.0135;fifth quintile: OR = 1.82, 95% CI: 1.15-2.87, P = 0.0110] and ECAS compared to the lowest quintile(fifth quintile: OR = 1.47, 95% CI: 1.09-1.97, P = 0.0106). The association between ALP and prevalence of carotid plaque became insignificant after adjustment for confounders. Conclusion Serum ALP levels were independently associated with abnormal common carotid IMT and ECAS. These conclusions need to be further corroborated in future prospective cohort studies.展开更多
AIM To clarify the prognostic significance of preoperative albumin-to-alkaline phosphatase ratio(AAPR) in cholangiocarcinoma(CCA) subjects receiving surgery.METHODS In this retrospective study, we included 303 CCA pat...AIM To clarify the prognostic significance of preoperative albumin-to-alkaline phosphatase ratio(AAPR) in cholangiocarcinoma(CCA) subjects receiving surgery.METHODS In this retrospective study, we included 303 CCA patients receiving surgery without preoperative therapy between 2002 and 2014. Clinicopathological characteristics(including AAPR) were analyzed to determine predictors of postoperative overall survival and recurrence-free survival(RFS). In addition,univariate and multivariate Cox proportional hazards models were conducted,followed by application of time-dependent receiver operating curves to identify the optimal cut-off.RESULTS Univariate and multivariate analyses revealed both decreased overall survival[hazard ratio(HR): 2.88, 95%CI: 1.19-5.78] and recurrence-free survival(HR: 2.31,95%CI: 1.40–3.29) in patients with AAPR < 0.41 compared to those with AAPR ≥0.41. The optimal cut-off of AAPR was 0.41. Of the 303 subjects, 253(83.5%) had an AAPR over 0.41. The overall 1-, 3- and 5-year survival rates were 70.2%, 38.0% and 16.5%, respectively in the low(< 0.41) AAPR group, which were significantly lower than those in the high(≥ 0.41) AAPR group(81.7%, 53.9%, and 33.4%,respectively)(P < 0.0001). Large tumor size, multiple tumors, and advanced clinical stage were also identified as significant predictors of poor prognosis.CONCLUSION Our outcomes showed that AAPR was a potential valuable prognostic indicator in CCA patients undergoing surgery, which should be further confirmed by prospective studies. Moreover, it is necessary to investigate the mechanisms concerning the correlation of low AAPR with poor post-operative survival in CCA patients.展开更多
Alkaline phosphatase activity (APA) and dissolved phosphorus were monitored during the batch cultures of two bone microalgae. Results indicate that variation of APA was in the shape of "S" curve. Different s...Alkaline phosphatase activity (APA) and dissolved phosphorus were monitored during the batch cultures of two bone microalgae. Results indicate that variation of APA was in the shape of "S" curve. Different specs of dissolved phaphorus had different effects on APA. The concentrations of dis solved inorganic phosphorus (DIP) and and molecular dissolved organic phosphorus (SDOP) had a sig nificant effect on APA, while the concentration of large molecular dissolved organic phosphorus (LDOP) had a little effect on APA., and the increase of APA could accelerate the decomposing of LDOP in the medium. Results also show that algae species and abundance had why a little effect on APA.展开更多
AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL g...AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by SfiⅠ and NotⅠ, it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E. coli TG1. The positive colonies were screened by colony PCR and their expressions were induced by IPTG. ScFv gene was gained by digesting ScFv expression vector pUC19/119 with Sfi I and NotⅠ restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E. coli TG1. The positive colonies were selected by bacterial colony PCR. The expression of fusion protein (scFv-AP) was induced by IPTG. Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku. Immunofluorescent assay (IFA) demonstrated its reactivity with TfR. The molecular weight of scFv-AP was 75 ku. Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity.CONCLUSION: We have successfully prepared the antihuman TfR scFv and constructed the fusion protein of scFv and AP. It is promising for immunological experiments.展开更多
Alkaline phosphatases(APs) are non-specifi c phosphohydrolases, and they are widely used in clinical diagnostics and biological studies. APs are widespread in nature and exhibit dif ferent structural formulations. Bas...Alkaline phosphatases(APs) are non-specifi c phosphohydrolases, and they are widely used in clinical diagnostics and biological studies. APs are widespread in nature and exhibit dif ferent structural formulations. Based on the diversity of biogenetic sources, APs exhibit temperature-propensity traits, and they are classifi ed as psychrophilic, mesophilic, and thermophilic. In this article, the characteristics of psychrophilic APs from marine organisms were described, accompanied by a simple description of APs from other organisms. This review will facilitate better utilization of marine APs in the biotechnology fi eld.展开更多
基金Supported by the Key Technology Research and Development Program of Shandong,No.2017GSF221018 and No.2017G006007.
文摘BACKGROUND Gastric cancer(GC)is characterized by a low 5-year survival rate.The prognosis is still not satisfactory although it has significantly improved due to developments in medicine.Thus,the identification of more efficient indices for the evaluation of GC prognosis is required.We propose,for the first time,that the alkaline phosphatase(ALP)to prealbumin(PA)ratio(APR)can be used as an independent prognostic factor in GC.AIM To evaluate the prognostic value the APR in GC.METHODS According to the exclusion strategy,we collected the preoperative serologic examination results and clinical information of 409 GC patients treated in Shandong Provincial Hospital from January to December,2016.By calculating the APR,the neutrophil and lymphocyte ratio(NLR),C-reactive protein(CRP)and albumin(ALB)ratio,platelet and lymphocyte ratio,lymphocyte and monocyte ratio,and the relationship with clinical information,we verified the role of preoperative APR ratio in the prognosis of GC.In addition,we used a Cox model combined with the APR and tumor stage to demonstrate its efficacy in assessing the prognosis of GC patients.RESULTS Preoperative APR was an independent prognostic factor for GC.The median age of patients in the APR-high group was greater compared with that in the APR-low group.Patients with a higher APR had a more advanced clinical stage,higher neutrophil to lymphocyte,CRP to ALB,and platelet to lymphocyte ratios,but a lower lymphocyte to monocyte ratio(P<0.05).The APR-high group also had higher glycoprotein antigen 199 and carbohydrate antigen 125 levels than the APR-low group(P<0.05).Median overall survival and disease-free survival were significantly longer in the APR-low group than in the APR-high group.In addition,a Cox model based on the APR and tumor stage was more effective in evaluating the prognosis of patients than models based on stage alone or stage plus the NLR.CONCLUSION A higher APR is an independent and negative prognostic factor for GC.The prognosis of GC can be better evaluated using a Cox model based on the APR and stage.
基金Project supported by the National Natural Science Foundation of China(Grant No.61675151)
文摘Alkaline phosphatase(ALP) plays an integral role in the metabolism of liver and development of the skeleton in humans. To date, the interactions between different-duration terahertz(THz) radiation and ALP activities, as well as the influence mechanism are still unclear. In this study, using the para-nitro-phenyl-phosphate(p NPP) method, we detect changes in ALP activities during 40-minute THz radiation(0.1 THz, 13 m W/cm^2). It is found that the activity of ALP decreases in the first 25 min, and subsequently increases in the later 15 min. Compared with the activity of ALP being heated, the results suggest that short-term terahertz radiation induces a decrease in enzyme activity through the non-thermal mechanism. In order to explore the non-thermal effects of THz radiation on ALP, we focus on the impacts of 0.1 THz radiation for 20 min on the activity of ALP in different concentrations. The results reveal that the activity of ALP decreases significantly after exposure to THz radiation. In addition, it could be deduced from fluorescence, ultraviolet-visible(UV-vis), and THz spectra results that THz radiation has induced changes in ALP structures. Our study unlocks non-thermal interactions between THz radiation and ALP, as well as suggests that THz spectroscopy is a promising technique to distinguish ALP structures.
文摘AIM: To investigate relapse predictors in chronic hepatitis C (CHC) patients with end-of-treatment response (ETR), after pegylated interferon-α (PegIFN-α) and ribavirin treatment. METHODS: In a retrospective study we evaluated a spectrum of predictors of relapse after PegIFN-α and ribavirin treatment in 86 CHC patients with ETR. Viral loads were determined with real-time reverse transcrip-tion polymerase chain reaction. Hepatitis C virus geno-typing was performed by sequencing analysis. Patients with genotype 1 were treated for 48 wk with 180 μg PegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 1000 mg/d for those under 75 kg or 1200 mg/d for those over 75 kg. Patients with geno- types 2 and 3 were treated for 24 wk with 180 μgPegIFN-α2a or 1.5 μg/kg PegIFN-α2b once weekly plus ribavirin at a dosage of 800 mg/d. RESULTS: In all ETR patients, binary logistic regression analysis identif ied absence of complete early virological response (cEVR) (OR 27.07, 95% CI: 3.09-237.26, P < 0.005), serum alkaline phosphatase (ALP) levels prior to therapy < 75 U/L (OR: 6.16, 95% CI: 2.1-18.03, P < 0.001) and body mass index > 26 kg/m2 (OR: 8.27, 95% CI: 2.22-30.84, P < 0.005) as independent predictors of relapse. When cEVR patients were analyzed exclusively, ALP prior to therapy < 75 U/L remained the only predictor of relapse. CONCLUSION: Lower levels of ALP prior to, during and after therapy seem to be associated with a higher risk of relapse in CHC patients with ETR.
基金supported by the National Natural Science Foundation of China (Grant Nos. 31672292, 31372201)the Natural Science Foundation of Hebei Province (C2017205059)
文摘Background: In temperate-breeding birds, individuals must adjust their physiological states from one life-history stage to another in response to changing conditions to maximize ecological fitness. Previous evidences have shown that body mass, size-corrected mass(SCM), and hematocrit(Hct) could be used as estimates of the energetic state of individuals to illustrate life-history trade-offs and individual quality in field physiology. Plasma alkaline phosphatase(ALP) plays critical roles in regulating the metabolism of energy but very limited information is known on its link with body mass or Hct.Methods: We determined the changes of plasma ALP levels in both early breeding and wintering stages of male Eurasian Tree Sparrows(Passer montanus), and examined the relationships between ALP and body mass, SCM, and Hct of the birds.Results: Our study showed that(1) in male Eurasian Tree Sparrows, body mass did not vary with life-history stage but plasma ALP activity significantly increased in the wintering stage compared to the breeding stage;(2) ALP activity was not correlated with individual body mass but was positively correlated with individual SCM and Hct. Such positive correlations, however, only occurred in the wintering but not in the breeding stages.Conclusions: Our results suggest that plasma ALP activities in free-living birds can be used as one of the indicators o body condition or nutritional status for analyzing individual variation in the wintering but not in the breeding stages. The life-history dependent relationships between plasma ALP activity and body condition may contribute to our better understanding of the trade-off between individual survival and reproduction in free-living animals.
文摘Previous clinical study showed that aged garlic extract (AGE) has a preventive effect of vascular calcification in patients with coronary artery diseases. It has been reported that vascular smooth muscle cells (VSMC) and inflammatory cells including macrophages and lymphocytes migrate to sub-intimal region where atherosclerotic plaques are formed, and VSMC can differentiate into osteoblast-like cells, which are represented by the induction of alkaline phosphatase (ALP). We found that primary human coronary artery smooth muscle cells (HCASMC) showed the increased ALP activity, when cultured in the medium containing ascorbic acid, β-glycerophosphate, dexamethasone (IM), and supplemented with conditioned medium from macrophages (MCM). Then we tested the effect of AGE subdivided fractions and several compounds found in AGE, and then found that ((+)-(2S,3R)-Dehydrodiconiferyl alcohol, a dilignol compound existed in hydrophobic fraction of AGE, inhibited ALP activity in HCASMC.
基金financially supported by the National Natural Science Foundation of China(Grant No.:21904045)the Fundamental Research Funds for the Central Universities(HUST:Grant No.:2019kfyXJJS169)Training Program of Innovation and Entrepreneurship for Undergraduates of Hubei Province(Grant No.:S202010487225).
文摘Alkaline phosphatase(ALP)is widely expressed in human tissues.ALP plays an important role in the dephosphorylation of proteins and nucleic acids.Therefore,quantitative analysis of ALP plays a vital role in disease diagnosis and the development of biological detection methods.Terminal deoxynucleotidyl transferase(TdT)catalyzes continuous polymerization of deoxynucleotide triphosphates at the 30-OH end of single-stranded DNA in the absence of a template.In this study,we developed a highly sensitive and selective method based on TdT and endonuclease Ⅳ(Endo Ⅳ)to quantify ALP activity.After ALP hydrolyzes the 30-PO_(4) end of the substrate and generates 30-OH,TdT can effectively elongate the 30-OH end with deoxynucleotide adenine triphosphate(dATP)and produce a poly A tail,which can be detected by the poly T probes.Endo Ⅳ digests the AP site in poly T probes to generate a fluorescent signal and a new 30-OH end,leading to the generation of exponential fluorescence signal amplification.The substrate for TdT elongation was optimized,and a limit of detection of 4.3×10^(-3) U/L was achieved for ALP by the optimized substrate structure.This method can also detect ALP in the cell lysate of a single cell.This work has potential applications in disease diagnosis and biomedical detection.
基金Supported by the National Natural Science Foundation of China(Nos.91951119,42177246,42107279)。
文摘Dissolved inorganic phosphorus is an important form of directly bioavailable phosphorus for cyanobacteria in natural water.Dissolved organic phosphorus could be used by cyanobacteria via alkaline phosphatase,which is produced mainly by bacteria and also cyanobacteria itself.Herein,we review the current knowledge of extracellular phosphatase excreted by cyanobacteria,highlighting the development of detection method and its ecological roles in regulating pho sphorus cycling in freshwater systems,which is based on reports for around 100 species of cyanobacteria.Recommendations are suggested concerning the extracellular phosphatase produced by bloom-forming cyanobacteria in terms of the ecological role,followed by a discussion of the future prospects for the study.
基金the Heilongjiang Natural Science Fund Project (C2017035)。
文摘The purpose of this study was to explore the effects of recombinant human intestinal alkaline phosphatase(recIAP) on human neutrophils in vitro, and the migration, phagocytosis, apoptosis in presence and absence of LPS. In this study, freshly extracted human neutrophils were used to establish an inflammatory cell model, and the control group, recIAP group, LPS group and recIAP +LPS group were set up to stimulate the model. The migration of neutrophils was detected by agarose gel drop method. Fluorescent particles and fluorescent probes were added to different treatment groups, and the phagocytic rate of neutrophils and the release of reactive oxygen species(ROS) from neutrophils were detected by flow cytometry. The apoptosis rate of neutrophils was detected by flow cytometry according to Annexin V-FITC apoptosis detection kit. The results showed that regardless of the presence or absence of LPS, recIAP could inhibit the migration of neutrophils, phagocytosis and the release of ROS. In addition, recIAP could weaken the inhibitory effect of LPS on neutrophils apoptosis.
文摘Objective To investigate the influence of sodium fluoride(NaF)on alkaline phosphatase(ALP)activity and bone gla protein(BGP)synthesis in yellow ligament cells from different surgical simples in vitro.Methods The human ligament cells
文摘BACKGROUND Previous studies have suggested that a low albumin-to-alkaline phosphatase ratio(AAPR)is associated with a lower survival rate in patients with various malignancies.However,the relationship between pretreatment AAPR and the prognosis of patients with gastric cancer(GC)remains unclear.AIM To investigate the prognostic value of AAPR in distant metastatic GC.METHODS A total of 191 patients with distant metastatic cancer from a single institute were enrolled in this study.Pretreatment clinical data,including serum albumin and alkaline phosphatase levels,were collected.A chi-square test or Fisher’s exact test was applied to evaluate the correlations between AAPR and various clinical parameters in GC patients.The Kaplan-Meier method and Cox proportional hazards regression model were used to evaluate the prognostic efficacy of AAPR in metastatic GC patients.A two-sided P value lower than 0.05 was considered statistically significant.RESULTS A receiver operating characteristic curve indicated that 0.48 was the optimal threshold value for AAPR.AAPR≤0.48 was significantly associated with bone(P<0.05)and liver metastasis(P<0.05).Patients with high levels of AAPR had better survival in terms of overall survival(OS)and progression-free survival(PFS),regardless of the presence of liver/bone metastasis.Pretreatment AAPR was found to be a favorable predictor of OS and PFS based on a multivariate cox regression model.AAPR-M system,constructed based on AAPR and number of metastatic sites,showed superior predictive ability relative to the number of metastatic sites for predicting survival.CONCLUSION Pretreatment AAPR may serve as an independent prognostic factor for predicting PFS and OS in patients with metastatic GC.Furthermore,AAPR may assist clinicians with individualizing treatment.
基金China-France Scientific end Technical Cooperation (No.1996-134)Bioengineering Key Laboratory of Henan Province
文摘INTRODUCTIONOnly the liver has the great capability ofregeneration in mammal.Few hepatocytes are inthe phase of division in the normal liver of an adultmammal (including human beings),but theremaining hepatocytes can be induced to proliferatequickly by partial hepatectomy (PH),and,to somedegree,they stop dividing and re-differentiate intocells functioning as hepatocytes.This shows
基金Supported by National natural science foundation of China,No.81272644 and No.81201549
文摘AIM: To determine the prognostic value of alkaline phosphatase(ALP) and γ-glutamyltransferase(GGT) for hepatocellular carcinoma(HCC).METHODS: We analyzed the outcome of 172 HCC patients who underwent liver resection. Receiver operating characteristic(ROC) curve analysis was performed to determine the cut-off value of ALP and GGT. Then, preoperative risk factors for survival were evaluated by multivariate analysis. Based on the significant factors, aprognostic score model was established.RESULTS: By ROC curve analysis, ALP > 120 U/L and GGT > 115 U/L were considered elevated. Overall survival(OS) and tumor-free survival(TFS) for patients with elevated ALP and GGT were significantly worse than for patients with ALP and GGT within the normal range. Multivariate analysis showed that the elevated levels of ALP, GGT and tumor size were independent prognostic factors. Giving each positive factor as a score of 1, we established a preoperative prognostic score model. The 5-year OS for patients with a score of 0, 1, 2 and 3 were 84.0%, 45.9%, 44.1% and 0%, respectively, while the TFS was 80.6%, 40.0%, 38.8% and 0%, respectively. When combining patients with scores of 1 and 2 into the middle risk group, and patients with scores of 0 and 3 into the low-risk and highrisk groups, respectively, different outcomes would be significantly distinguished by the risk groups.CONCLUSION: Elevated ALP and GGT levels were risk predictors in HCC patients. Our prognostic model might vary the outcomes of patients from different risk groups.
基金Supported by the Guizhou Provincial Department of Health Science and Technology Fund,No.GZWJKJ2014-2-151the Science and Technology Fund of Guizhou Province,No.QKH LH [2016]7421the Zunyi Science and Technology Research and Development Fund,No.ZSKHS[2016] 06
文摘BACKGROUND Gamma-glutamyltransferase(GGT) is one of the most important laboratory tests for the evaluation of liver damage. Through a long-term clinical observation of patients with secondary asymptomatic choledocholithiasis, we found that most patients had abnormal GGT serum levels.AIM To investigate the combination of serum GGT and alkaline phosphatase(ALP) in predicting the diagnosis of asymptomatic choledocholithiasis secondary to cholecystolithiasis.METHODS In this retrospective cohort study, the clinical data of 829 patients with cholecystolithiasis admitted to the Third Affiliated Hospital of Zunyi Medical College from August 2014 to August 2017 were collected. Among these patients,151 patients had secondary asymptomatic choledocholithiasis and served as the observation group, and the remaining 678 cholecystolithiasis patients served as the control group. Serum liver function indexes were detected in both groups,and the receiver operating characteristic(commonly known as ROC) curves were constructed for markers showing statistical significances. The cutoff value,sensitivity, and specificity of each marker were calculated according to the ROC curves.RESULTS The overall incidence of asymptomatic choledocholithiasis secondary to cholecystolithiasis was 18.2%. The results of liver function indexes including serum aspartate aminotransferase, alanine aminotransferase, direct bilirubin and total bilirubin levels showed no significant differences between the two groups(P> 0.05). However, the serum GGT and ALP levels were significantly higher in the observation group than in the control group(P < 0.05). The ROC curve analysis showed that the area under the curve was 0.881(95%CI: 0.830-0.932), 0.647(95%CI: 0.583-0.711) and 0.923(0.892-0.953) for GGT, ALP, and GGT + ALP,respectively. The corresponding cut-off values of GGT and ALP were 95.5 U/L and 151.5 U/L, sensitivity were 90.8% and 65.1%, and specificity were 83.6% and59.8%, respectively. The sensitivity and specificity of GGT + ALP were 93.5% and85.1%, respectively.CONCLUSION An abnormally elevated serum GGT level has an important value in the diagnosis of asymptomatic choledocholithiasis secondary to cholecystolithiasis.The combination of serum GGT and ALP has better diagnostic performance. As a convenient, rapid and inexpensive test, it should be applied in secondary asymptomatic choledocholithiasis routine screening.
基金supported by the Key Project of Natural Science Foundation of Chongqing(No.cstc2015jcyjBX0080)Science and Technology project of Chongqing Education Commission(KJ1600907)
文摘Alkaline phosphatases(APs) include the placental AP(PLAP), germ cell AP(GCAP), intestinal AP(IAP) and tissue nonspecific AP(TNAP). Over expression of TNAP in smooth muscle cells of kidney and vessels provokes the progress of such serious diseases as end-stage renal disease, idiopathic infantile arterial calcification, ankylosis, osteoarthritis and diabetes. In order to design and optimize the potent TNAP inhibitors, comparative molecular field analysis(CoMFA) and comparative molecular similarity indices analysis(CoMSIA) were used to analyze 3D structure-activity relationships(3D-QSAR) of TNAP inhibitors. The 3D-QSAR model(CoMFA with q^2 = 0.521, r^2 = 0.930; CoMSIA with q^2 = 0.529, r^2 = 0.933) had a good predictability. Surflex-dock was used to reveal the binding mode between the inhibitors and TNAP protein. CoMFA, CoMSIA and docking results provide guidance for the discovery of TNAP inhibitors. Finally, eight new compounds as potential TNAP inhibitors were designed.
基金Supported by Crohn’s and Colitis Foundation of Canada,No.11883 1486 RR 0001Natural Science and Engineering Re-search Council and the Intestinal Diseases Education Awareness Society,No.86435 5557 RR0001
文摘Intestinal alkaline phosphatase(IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP's role in the intestine is to dephosphorylate toxic microbial ligands such as lipopolysaccharides, unmethylated cytosine-guanosine dinucleotides and flagellin as well as extracellular nucleotides such as uridine diphosphate. IAP's ability to detoxify these ligands is essential in protecting the host from sepsis during acute inflammation and chronic inflammatory conditions such as inflammatory bowel disease. Also important in these complications is IAP's ability to regulate the microbial ecosystem by forming a complex relationship between microbiota, diet and the intestinal mucosal surface. Evidence reveals that diet alters IAP expression and activity and this in turn can influence the gut microbiota and homeostasis. IAP's ability to maintain a healthygastrointestinal tract has accelerated research on its potential use as a therapeutic agent against a multitude of diseases.Exogenous IAP has been shown to have beneficial effects when administered during ulcerative colitis,coronary bypass surgery and sepsis.There are currently a handful of human clinical trials underway investigating the effects of exogenous IAP during sepsis,rheumatoid arthritis and heart surgery.In light of these findings IAP has been marked as a novel agent to help treat a variety of other inflammatory and infectious diseases.The purpose of this review is to highlight the essential characteristics of IAP in protection and maintenance of intestinal homeostasis while addressing the intricate interplay between IAP,diet,microbiota and the intestinal epithelium.
文摘Objective This study aimed to investigate the relationship between alkaline phosphatase(ALP) and common carotid intima media thickness(IMT), carotid plaque, and extracranial carotid artery stenosis(ECAS). Methods A total of 3,237 participants aged ≥ 40 years were recruited from Jidong community in 2013-2014. Participants were divided into five quintile groups based on their serum ALP levels. Carotid atherosclerosis was assessed using ultrasound. Abnormal IMT, carotid plaque, and ECAS were defined as IMT > 0.9 mm, IMT > 1.5 mm, and ≥ 50% stenosis in at least one extracranial carotid artery, respectively. Results Common carotid IMT values and the prevalence of carotid plaque increased across serum ALP quintiles. Higher ALP quintiles were correlated with an increased risk of abnormal IMT [fourth quintile: odds ratio(OR) 1.78, 95% confidence interval(CI) 1.13-2.82, P = 0.0135;fifth quintile: OR = 1.82, 95% CI: 1.15-2.87, P = 0.0110] and ECAS compared to the lowest quintile(fifth quintile: OR = 1.47, 95% CI: 1.09-1.97, P = 0.0106). The association between ALP and prevalence of carotid plaque became insignificant after adjustment for confounders. Conclusion Serum ALP levels were independently associated with abnormal common carotid IMT and ECAS. These conclusions need to be further corroborated in future prospective cohort studies.
基金National Key Project Research and Development Projects,No.S2016G9012International Science and Technology Cooperation Projects,No.2015DFA30650the Capital Special Research Project for Clinical Application,No.Z151100004015170
文摘AIM To clarify the prognostic significance of preoperative albumin-to-alkaline phosphatase ratio(AAPR) in cholangiocarcinoma(CCA) subjects receiving surgery.METHODS In this retrospective study, we included 303 CCA patients receiving surgery without preoperative therapy between 2002 and 2014. Clinicopathological characteristics(including AAPR) were analyzed to determine predictors of postoperative overall survival and recurrence-free survival(RFS). In addition,univariate and multivariate Cox proportional hazards models were conducted,followed by application of time-dependent receiver operating curves to identify the optimal cut-off.RESULTS Univariate and multivariate analyses revealed both decreased overall survival[hazard ratio(HR): 2.88, 95%CI: 1.19-5.78] and recurrence-free survival(HR: 2.31,95%CI: 1.40–3.29) in patients with AAPR < 0.41 compared to those with AAPR ≥0.41. The optimal cut-off of AAPR was 0.41. Of the 303 subjects, 253(83.5%) had an AAPR over 0.41. The overall 1-, 3- and 5-year survival rates were 70.2%, 38.0% and 16.5%, respectively in the low(< 0.41) AAPR group, which were significantly lower than those in the high(≥ 0.41) AAPR group(81.7%, 53.9%, and 33.4%,respectively)(P < 0.0001). Large tumor size, multiple tumors, and advanced clinical stage were also identified as significant predictors of poor prognosis.CONCLUSION Our outcomes showed that AAPR was a potential valuable prognostic indicator in CCA patients undergoing surgery, which should be further confirmed by prospective studies. Moreover, it is necessary to investigate the mechanisms concerning the correlation of low AAPR with poor post-operative survival in CCA patients.
基金National Natural Science Foundation of China ! 49206063.
文摘Alkaline phosphatase activity (APA) and dissolved phosphorus were monitored during the batch cultures of two bone microalgae. Results indicate that variation of APA was in the shape of "S" curve. Different specs of dissolved phaphorus had different effects on APA. The concentrations of dis solved inorganic phosphorus (DIP) and and molecular dissolved organic phosphorus (SDOP) had a sig nificant effect on APA, while the concentration of large molecular dissolved organic phosphorus (LDOP) had a little effect on APA., and the increase of APA could accelerate the decomposing of LDOP in the medium. Results also show that algae species and abundance had why a little effect on APA.
基金Supported by Natural Key and Basic Research Development Program,No.2002CB513109
文摘AIM: To construct fusion protein of a single-chain antibody(scFv) against transferrin receptor (TfR) with alkalinephosphatase (AP).METHODS: The VH-linker-VL, namely scFv gene, wasprepared by amplifying the VH and VL genes from plasmid pGEM-T-VH and pGEM-T-VL with splicing overlap extension polymerase chain reaction (SOE PCR). After the ScFv gene was modified by SfiⅠ and NotⅠ, it was subcloned into the secretory expression vector pUC19/119, and then was transformed into E. coli TG1. The positive colonies were screened by colony PCR and their expressions were induced by IPTG. ScFv gene was gained by digesting ScFv expression vector pUC19/119 with Sfi I and NotⅠ restriction enzymes, then subcloned into expression vector pDAP2, followed by transformation in E. coli TG1. The positive colonies were selected by bacterial colony PCR. The expression of fusion protein (scFv-AP) was induced by IPTG. Its activity was detected by enzyme immunoassay. The molecular weights of scFv and scFv-AP were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).RESULTS: The product of SOE PCR formed a band of 700 bp in agarose gel electrophoresis. SDS-PAGE demonstrated the molecular weight of scFv was 27 ku. Immunofluorescent assay (IFA) demonstrated its reactivity with TfR. The molecular weight of scFv-AP was 75 ku. Enzyme immunoassay showed that scFv-AP could specifically bind to human TfR and play AP activity.CONCLUSION: We have successfully prepared the antihuman TfR scFv and constructed the fusion protein of scFv and AP. It is promising for immunological experiments.
基金Supported by the National Natural Science Foundation for Creative Research Groups(No.41221004)the Natural Science Foundation of Shandong Province(No.ZR2011DQ005)+6 种基金the Key Laboratory of Marine Ecology and Environmental Science and Engineering,SOA(No.MESE-2013-03)the Key Laboratory for Ecological Environment in Coastal Areas,State Oceanic Administration(No.201306)the Major International Joint Research Project of NSFC(No.41320104008)the Changjiang Scholars Program,Ministry of Education of Chinathe Taishan Scholars Program of Shandong Provincethe Key Lab of Marine Bioactive Substance and Modern Analytical Technique,SOA(No.MBSMAT-2013-05)This is MCTL Contribution No.81
文摘Alkaline phosphatases(APs) are non-specifi c phosphohydrolases, and they are widely used in clinical diagnostics and biological studies. APs are widespread in nature and exhibit dif ferent structural formulations. Based on the diversity of biogenetic sources, APs exhibit temperature-propensity traits, and they are classifi ed as psychrophilic, mesophilic, and thermophilic. In this article, the characteristics of psychrophilic APs from marine organisms were described, accompanied by a simple description of APs from other organisms. This review will facilitate better utilization of marine APs in the biotechnology fi eld.
