Analysis of alkaline phosphatase and γ-glutamyltransferase after radiofrequency ablation of primary liver cancer: A retrospective study

BACKGROUND Changes in alkaline phosphatase(ALP)andγ-glutamyltransferase(GGT)levels in patients with primary liver cancer(PLC)after radiofrequency ablation(RFA).Hepatocellular carcinoma is a malignant tumor with high incidence worldwide.As a common local treatment,RFA has attracted much attention for its efficacy and influence on liver function.AIM To investigate the effect of serum ALP and GGT levels on the prognosis of patients with PLC treated by RFA.METHODS The preoperative clinical data of 165 patients who were pathologically or clinically diagnosed with PLC and who received RFA in our hospital between October 2018 and June 2023 were collected.The chi-square test was used to compare the data between groups.The Kaplan-Meier method and Cox regression were used to analyze the associ-ations between serum ALP and GGT levels and overall survival,progression-free survival(PFS)and clinical characteristics of patients before treatment.RESULTS The 1-year survival rates of patients with normal(≤135 U/L)and abnormal(>135 U/L)serum ALP before treatment were 91%and 79%,respectively;the 2-year survival rates were 90%and 68%,respectively;and the 5-year survival rates were 35%and 18%,respectively.The difference between the two groups was statistically significant(P=0.01).Before treatment,the 1-year survival rates of patients with normal serum GGT levels(≤45 U/L)and abnormal serum GGT levels(>45 U/L)were 95%and 87%,the 2-year survival rates were 85%and 71%,and the 5-year survival rates were 37%and 21%,respectively.The difference between the two groups was statist-ically significant(P<0.001).Serum ALP[hazard ratio(HR)=1.766,95%confidence interval(95%CI):1.068-2.921,P=0.027]and GGT(HR=2.312,95%CI:1.367-3.912,P=0.002)is closely related to the overall survival of PLC patients after RF ablation and is an independent prognostic factor.The 1-year PFS rates were 72%and 50%,the 2-year PFS rates were 52%and 21%,and the 5-year PFS rates were 14%and 3%,respectively.The difference between the two groups was statistically significant(P<0001).The 1-year PFS rates were 81%and 56%in patients with normal and abnormal serum GGT levels before treatment,respectively;the 2-year PFS rates were 62%and 35%,respectively;and the 5-year PFS rates were 18%and 7%,respectively,with statistical significance between the two groups(P<0.001).The serum ALP concentration(HR=1.653,95%CI:1.001-2.729,P=0.049)and GGT(HR=1.949,95%CI:1.296-2.930,P=0.001)was closely associated with PFS after RFA in patients with PLC.The proportion of male patients with abnormal ALP levels is high,the Child-Pugh grade of liver function is poor,and the incidence of ascites is high.Among GGT-abnormal patients,the Child-Pugh grade of liver function was poor,the tumor stage was late,the proportion of patients with tumors≥5 cm was high,and the incidence of hepatic encephalopathy was high.CONCLUSION Serum ALP and GGT levels before treatment can be used to predict the prognosis of patients with PLC after RFA,and they have certain guiding significance for the long-term survival of patients with PLC after radiofrequency therapy.

Effects of Metal Ions and Organic Solvents on Alkaline Phosphatase from Rice-field Eel

[Objective]The mechanism of alkaline phosphatase（ALP） was studied to promote rice-field eel aquaculture industry. [ Method] The effects of effectors such as multiple metal ions and organic solvents on ALP in viscera of rice-field eel. [ Result] Na^＋ and K ^＋ didn＇t generate big influences on enzyme activity;Mg^2＋ and Ca^2＋ could promote ALP while Li^＋,Cu^2＋ and Zn^2＋ could restrain ALP enzyme activity. Both HPO4^2- and WO4^2- generated by en- zyme catalyzing disodium phenyl phosphate possessed strong inhibitory effects on emzymc, and 9.5 mmol/L HPO4^2 - would make enzyme activity decline by 13% while 9.5 mmol/L WO4^3- would make enzyme decline by 34%. The inhibition types of them were both competitive inhibition on enzyme activity. The organic solvents such as methanol, ethanol,ethylene glycol,isopropannl all generated influences on ALP and the order according to their inhibitory effects was isopropanol 〉 ethanol 〉 methanol 〉 ethylene glycol. [ Conclusion] The inflncnces of various effeetors on ALP aetivity of rice-field eel were studied from dynamics perspective to provide theoretical basis for further clarifying ALP mechanism.

Digestive enzyme and alkaline phosphatase activities during the early stages of Silurus soldatovi development

To provide a theoretical basis to improve the survival and growth rate and optimize diet of sheatfish （Silurus soldatovi）, the activities of certain digestive enzymes and alkaline phosphatases were investigated during larval development of one-ten day old individuals. Results indicated that sheatfish larva （～ three days after hatching） had high levels of alkaline protease activity, which peaked at five days old and dipped by eight days old, although the trend was generally upward. Acid protease activity at one-eight days old was low, after which it increased rapidly. Amylase activity reached the highest value at five days old, after which it began to decline. Lipase activity fluctuated markedly and showed two peaks at three-four days old and six-eight days old. Larval digestive enzyme activity and alkaline phosphatase activity were higher when fed live food than when fed an artificial diet. Throughout the early development process, alkaline protease activity was higher than acid protease, alkaline protease and amylase specific activity decreased significantly for eight-day-old transition larvae, while acid protease activity increased rapidly. These results indicate that the changes in digestive enzyme activity were relevant to digestive function conversion during fish larvae development. Alkaline phosphatase activity showed an upward trend over the first ten days of life, which indicated that the gastrointestinal function of sheatfish improved gradually.

Construction of single chain Fv antibody against transferrin receptor and its protein fusion with alkaline phosphatase

AIM: To construct fusion protein of a single-chain antibody (scFv) against transferrin receptor (TfR) with alkaline phosphatase(AP). METHODS: The VH-linker-VL,namely scFv gene,was prepared 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 5/71 and Not I,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 5/71 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 anti-human TfR scFv and constructed the fusion protein of scFv and AP.It is promising for immunological experiments.

Model based on γ-glutamyltransferase and alkaline phosphatase for hepatocellular carcinoma prognosis

AIM: To determine the prognostic value of alkaline phosphatase (ALP) and &#x003b3;-glutamyltransferase (GGT) for hepatocellular carcinoma (HCC) .

Interplay between intestinal alkaline phosphatase, diet, gut microbes and immunity

Intestinal alkaline phosphatase (IAP) plays an essential role in intestinal homeostasis and health through interactions with the resident microbiota, diet and the gut. IAP&#x02019;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&#x02019;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&#x02019;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&#x02019;s ability to maintain a healthy gastrointestinal 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.

Use of the alkaline phosphatase to prealbumin ratio as an independent predictive factor for the prognosis of gastric cancer

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.

Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease

AIM： To investigate intestinal alkaline phosphatase （iAP） in the intestinal mucosa of children with inflammatory bowel disease （IBD）. METHODS： Colonic biopsy samples were taken from 15 newly diagnosed IBD patients and from 10 healthy controls. In IBD patients, specimens were obtainedboth from inflamed and non-inflamed areas. The lAP mRNA and protein expression was determined by reverse transcription-polymerase chain reaction and Western blotting analysis, respectively. Tissue localiza- tion of lAP and Toll-like receptor （TLR） 4 was investi- gated by immunofluorescent staining. RESULTS： The lAP protein level in the inflamed muco- sa of children with Crohn＇s disease （CD） and ulcerative colitis （UC） was significantly decreased when compared with controls （both P 〈 0.05）. Similarly, we found a significantly decreased level of lAP protein in the in- flamed mucosa in CD compared with non-inflamed mucosa in CD （P 〈 0.05）. In addition, the iAP protein level in inflamed colonic mucosa in patients with UC was decreased compared with non-inflamed mucosa in patients with CD （P 〈 0.05）. lAP protein levels in the non-inflamed mucosa of patients with CD were similar to controls, lAP mRNA expression in inflamed colonic mucosa of children with CD and UC was not significant- ly different from that in non-inflamed colonic mucosa with CD. Expression of lAP mRNA in patients with non- inflamed mucosa and in controls were similar. Co-local- ization of lAP with TLR4 showed intense staining with a dotted-like pattern, lAP was present in the inflamed and non-inflamed mucosa of patients with CD, UC, and in control biopsy specimens, irrespective of whether it was present in the terminal ileum or in the colon. However, the fluorescent signal of TLR4 was more pro- nounced in the colon compared with the terminal ileum in all groups studied. CONCLUSION： Lower than normal lAP protein levels in inflamed mucosa of IBD patients may indicate a role for lAP in inflammatory lesions in IBD. Based on our results, administration of exogenous lAP enzyme to pa- tients with the active form of IBD may be a therapeutic option.

A description of alkaline phosphatases from marine organisms

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.

Effect of dissolved phosphorus on alkaline phosphatase activity in marine microalgae

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.

Alkaline phosphatase activity in ornithogenic soils in polar tundra

Phosphatase plays an important role in the microbial liberation of phosphorus in soil systems. In this study, alkaline phosphatase activity （APA） was investigated from tundra ornithogenic soil profiles in Antarctica and Arctic. The organic carbon （TOC）, total nitrogen （TN）, and phosphorus fractions and pH were also analyzed in these soils. The correlation between APA and soil chemical properties is discussed. In almost all the soil profiles, APA showed the same variation patterns. The maximum APA appeared in the surface layers, and decreased with soil depth. The APA ranged from 1.00 ppm to 1 403.5 ppm with an average of approximately 408.3 ppm. The APA showed a significant positive correlation with TOC （r =0.70, p 〈0.001）, TN （r =0.43, p =0.002）, total phosphorus （r =0.39, p 〈0.05）and inorganic phosphorus （r =0.40, p =0.037）, indicating that they were predominant factors affecting APA in the polar tundra soils. In addition, APA showed a significant negative correlation with Cu and Zn concentrations in the soils, indicating that Cu and Zn may inhibit APA. Our results showed that APA could be used as an important indicator for soil fertility in polar tundra ecosystems.

Non-thermal effects of 0.1 THz radiation on intestinal alkaline phosphatase activity and conformation

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.

Alkaline phosphatase predicts relapse in chronic hepatitis C patients with end-of-treatment response

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.

Cyanobacterial extracellular alkaline phosphatase:detection and ecological function

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.

Electrochemical Determination of Alkaline Phosphatase in Human Serum by Differential Pulse Voltammetry

Differential pulse voltammetry(DPV) was applied to the determination of alkaline phosphatase(ALP) activity in human serum with phenyl phosphate as the substrate. Phenyl phosphate can enzymatically be hydrolyzed to produce phenol which is quantified by DPV at +660 mV(vs.Ag/AgCl) in the concentration range of 2.0_100 μmol/L. The standard curve for ALP is linear over the range from 0.06 to 1000 U/L with a relative standard deviation of 3.0%. The conditions for the enzymatic reaction and voltammetric detection were optimized and the kinetic constants were also examined.The human serum samples were tested by this method and the results were in good agreement with those obtained by the routine p-nitrophenyl phosphate spectrophotometric method.

Electrochemical Detection of Alkaline Phosphatase in BALB/c Mouse Fetal Liver Stromal Cells with Capillary Electrophoresis

A method for determination of alkaline phosphatase (ALP) in BALB/c mouse fetal liver stromal cells has been described based on the catalytic reaction. After the cell extract is incubated with the substrate disodium phenyl phosphate, the reaction product phenol generated by ALP is determined by capillary electrophoresis with electrochemical detection.

Regulation of Migration, Phagocytosis and Apoptosis of Human Neutrophils by Recombinant Human Intestinal Alkaline Phosphatase

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.

Life-history dependent relationships between plasma alkaline phosphatase activity and body condition in male Eurasian Tree Sparrows

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.

Effect of noise exposure(85 dB) on testicular adrenocortical steroidogenic key enzymes, acid and alkaline phosphatase activities of sex organs in mature albino rats

Changes in the activities of Δ 5\|3β\|hydroysteroid dehydrogenase(HSD) in testis and adrenal gland, 17β\|hydroxysteroid dehydrogenase in testis, acid and alkaline phosphatase in testis, prostate and seminal vesicle were observed in noise exposed mature rats at the intensity of 85 dB for 8 h/day for 45 days. The results indicated that noise exposed group showed a significant diminution in the activities of androgenic key enzymes Δ 5\|3β and 17β\|HSD, acid phosphatase in testis, prostate and seminal vesicle. There was a significant elevation in the activities of adrenal Δ 5\|3β\|HSD, alkaline phosphatase in testis and other accessory sex organ in noise exposed group. Gonadosomatic, prostatosomatic and seminal vesiculo\|somatic indexes were decreased significantly in noise exposed group. Therefore, it is evident that noise exposure at 85dB exerts a deleterious effect on testicular and adrenocortical activities.

Aged Garlic Extract and One of the Constituent, (+)-(2S,3R)-Dehydrodiconiferyl Alcohol, Inhibits Alkaline Phosphatase Activity Induced by Inflammation Factors in Human Vascular Smooth Muscle Cells

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.