Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina / Propiedades inhibidoras Enzimáticas y Anticancerígenas de la Moringa peregrina

Background: Moringa peregrina is widely used in the traditional medicine of the Arabian Peninsula to treat various ailments, because it has many pharmacologically active components with several therapeutic effects. Objective: This study aimed to investigate the inhibitory effect of Moringaperegrina seed ethanolic extract (MPSE) against key enzymes involved in human pathologies, such as angiogenesis (thymidine phosphorylase), diabetes (α-glucosidase), and idiopathic intracranial hypertension (carbonic anhydrase). In addition, the anticancer properties were tested against the SH-SY5Y (human neuroblastoma). Results: MPSE extract significantly inhibited α-glucosidase, thymidine phosphorylase, and carbonic anhydrase with half-maximal inhibitory concentrations (IC50) values of 303.1 ± 1.3, 471.30 ± 0.3, and 271.30 ± 5.1 µg/mL, respectively. Furthermore, the antiproliferative effect of the MPSE was observed on the SH-SY5Y cancer cell line with IC50 values of 55.1 µg/mL. Conclusions: MPSE has interesting inhibitory capacities against key enzymes and human neuroblastoma cancer cell line.

Antecedentes: La Moringa peregrina se utiliza ampliamente en la medicina tradicional de la Península Arábiga para tratar diversas dolencias, ya que posee numerosos componentes farmacológicamente activos con varios efectos terapéuticos. Objetivo: Este estudio tenía como objetivo investigar el efecto inhibidor del extracto etanólico de semillas de Moringaperegrina (MPSE) frente a enzimas clave implicadas en patologías humanas, como la angiogénesis (timidina fosforilasa), la diabetes (α-glucosidasa) y la hipertensión intracraneal idiopática (anhidrasa carbónica). Además, se comprobaron las propiedades anticancerígenas frente al SH-SY5Y (neuroblastoma humano). Resultados: El extracto de MPSE inhibió significativamente la α-glucosidasa, la timidina fosforilasa y la anhidrasa carbónica con concentraciones inhibitorias semimáximas (IC50) de 303,1 ± 1,3, 471,30 ± 0,3 y 271,30 ± 5,1 µg/mL, respectivamente. Además, se observó el efecto antiproliferativo del MPSE en la línea celular del cáncer SH-SY5Y con valores de IC50 de 55,1 µg/mL. Conclusiones: MPSE posee interesantes capacidades inhibitorias frente a enzimas clave y línea celular de neuroblastoma canceroso humano.

Synthesis of cello-oligosaccharides which promotes the growth of intestinal probiotics by multi-enzyme cascade reaction / 生物工程学报

Soluble cello-oligosaccharide with 2-6 oligosaccharide units is a kind of oligosaccharide with various biological functions, which can promote the proliferation of intestinal probiotics such as Bifidobacteria and Lactobacillus paracei. Therefore, it has a regulatory effect on human intestinal microbiota. In this study, a Cc 01 strain was constructed by expressing cellodextrin phosphorylase (CDP) in Escherichia coli. By combining with a previously constructed COS 01 strain, a three-enzyme cascade reaction system based on strains COS 01 and Cc 01 was developed, which can convert glucose and sucrose into cello-oligosaccharide. After optimization, the final titer of soluble cello-oligosaccharides with 2-6 oligosaccharide units reached 97 g/L, with a purity of about 97%. It contained cellobiose (16.8 wt%), cellotriose (49.8 wt%), cellotetrose (16.4 wt%), cellopentaose (11.5 wt%) and cellohexose (5.5 wt%). When using inulin, xylo-oligosaccharide and fructooligosaccharide as the control substrate, the biomass (OD600) of Lactobacillus casei (WSH 004), Lactobacillus paracei (WSH 005) and Lactobacillus acidophilus (WSH 006) on cello-oligosaccharides was about 2 folds higher than that of the control. This study demonstrated the efficient synthesis of cello-oligosaccharides by a three-enzyme cascade reaction and demonstrated that the synthesized cello-oligosaccharides was capable of promoting intestinal microbial proliferation.

Application of sucrose phosphorylase in glycosylation / 生物工程学报

Water solubility, stability, and bioavailability, can be substantially improved after glycosylation. Glycosylation of bioactive compounds catalyzed by glycoside hydrolases (GHs) and glycosyltransferases (GTs) has become a research hotspot. Thanks to their rich sources and use of cheap glycosyl donors, GHs are advantageous in terms of scaled catalysis compared to GTs. Among GHs, sucrose phosphorylase has attracted extensive attentions in chemical engineering due to its prominent glycosylation activity as well as its acceptor promiscuity. This paper reviews the structure, catalytic characteristics, and directional redesign of sucrose phosphorylase. Meanwhile, glycosylation of diverse chemicals with sucrose phosphorylase and its coupling applications with other biocatalysts are summarized. Future research directions were also discussed based on the current research progress combined with our working experience.

Characterization of a sucrose phosphorylase from Leuconostoc mesenterides for the synthesis of α-arbutin / 生物工程学报

Sucrose phosphorylase (SPase) gene from Leuconostoc mesenteroides ATCC 12291 was synthesised after codon optimization, and inserted into pET-28a plasmid to generate pET-28a-spase. The recombinant strain Escherichia coli BL21 (DE3)/pET-28a-spase was induced for Spase expression. The recombinant protein Spase was purified and characterized. The specific enzyme activity of SPase was 213.98 U/mg, the purification ratio was 1.47-fold, and the enzyme activity recovery rate was 87.80%. The optimal temperature and the optimal pH of the SPase were identified to be 45 °C and 6.5 respectively, and Km, Vmax and kcat of the SPase for sucrose was 128.8 mmol/L, 2.167 μmol/(mL·min), and 39 237.86 min-1. The recombinant SPase was used for α-arbutin production from hydroquinone and the reaction process was evaluated. The optimal conditions for synthesis of α-arbutin by SPase were 40 g/L hydroquinone, 5:1 molar ratio of sucrose and hydroquinone, and 250 U/mL recombinant SPase at pH 7.0 and 30 °C for 24 h in the dark, and then 500 U/mL glucoamylase was added at 40°C for 2.5 h. Under the optimized process, the yield of α-arbutin reached 98 g/L, and the hydroquinone conversion rate was close to 99%. In summary, the recombinant SPase was cloned and characterized, and its application for α-arbutin production was feasible.

Choice of Capecitabine or S1 in Combination with Oxaliplatin based on Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase Expression Status in Patients with Advanced Gastric Cancer

PURPOSE: To study the efficacy of capecitabine or S-1 plus oxaliplatin (CAPOX or SOX) for treating thymidine phosphorylase (TP)- or dihydropyrimidine dehydrogenase (DPD)-positive advanced gastric cancer.MATERIALS AND METHODS: Eighty-six patients with stage IIIC to IV gastric cancer were assessed for TP and DPD expression by immunohistochemistry. The association between CAPOX or SOX efficacy and TP/DPD expression was retrospectively analyzed.RESULTS: There were no significant differences in the objective remission rate (ORR, 52.27% vs. 47.62%; P>0.05), disease control rate (72.73% vs. 73.81%, P>0.05), progression-free survival (hazard ratio [HR], 1.119; 95% confidence interval [CI], 0.739–1.741; P=0.586), and overall survival (OS; HR, 0.855; 95% CI, 0.481–1.511; P=0.588) between CAPOX and SOX. A higher number of stage IV patients showed TP positivity, while DPD-positive patients predominantly showed intestinal type of gastric cancer. In TP-positive patients, the ORRs associated with CAPOX and SOX treatments were 57.14% and 38.10%, respectively; OS was better with CAPOX than with SOX (HR, 0.447; 95% CI, 0.179–0.978; P=0.046). Among DPD-positive patients, the SOX treatment-associated ORR (60.87%) was significantly higher than the CAPOX treatment-associated ORR (43.48%). Furthermore, SOX treatment resulted in better OS than did CAPOX treatment (HR, 2.020; 95% CI, 1.019–4.837; P=0.049).CONCLUSIONS: No significant difference in clinical efficacy was found between CAPOX and SOX. TP-positive patients might respond better to CAPOX while DPD-positive patients may respond better to SOX. Our findings might serve as a guide for personalized chemotherapy for gastric cancer.

Anti-diabetic activity of quercetin extracted from Phyllanthus emblica L. fruit:In silico and in vivo approaches / 药物分析学报

In this study, molecular interactions of the ligands, quercetin, gallic acid, and metformin with various diabetes mellitus-related protein targets, such as glycogen phosphorylase and peroxisome proliferator-activated receptor gamma, were assessed. It was revealed that quercetin possesses good binding affinity to both targets. Quercetin is a major constituent of methanolic extracts of Phyllanthus emblica fruit. The antihyperglycemic effect of quercetin in streptozotocin (STZ)-induced diabetic rats was examined. The isolated quercetin administered at a dose of 75 mg/kg body weight produced a maximum decrease of 14.78％in blood glucose levels in the diabetic rats after 7 days of treatment. Furthermore, quercetin doses of 50 and 75 mg/kg were shown to significantly improve the profiles of triglycerides, high-density li-poprotein, very-low-density lipoprotein, low-density lipoprotein, and total cholesterol at the end of the study in STZ-induced diabetic rats. The administration of quercetin (25, 50, and 75 mg/kg body weight) daily for 28 days in STZ-induced diabetic rats resulted in a significant decrease in blood glucose and urine sugar levels, with a considerable rise in plasma insulin and hemoglobin levels. Therefore, quercetin is a potential drug with antidiabetic and antihyperglycemic action mediated by changes in the levels of glucose, cholesterol, and triglycerides as indicated by in silico and in vivo studies.

Influence of thymidine phosphorylase polymorphisms on the efficacy of capecitabine-based adjuvant chemotherapy in colorectal cancer patients / 中国肿瘤临床

Objective: To inrestigate the association between thymidine phosphorylase (TYMP) polymorphisms and efficacy of postop-erative capecitabine-based adjuvant chemotherapy in colorectal cancer (CRC) patients. Methods: Two hundred and thirty-five patients with colorectal cancer who received surgical treatment and adjuvant chemotherapy between January 2010 and December 2016 from People's Hospital of Zhengzhou, were included in this study. Peripheral blood and postoperative tissue specimens of the CRC patients were collected for genotyping polymorphisms and measuring TYMP mRNA expression, respectively. The correlation between the poly-morphisms and efficacy of postoperative chemotherapy in CRC patients was analyzed. Results: The prevalence of 5633C>T in TYMP gene among the CRC patients was as follows: CC genotype, 149 cases (63.40%); CT genotype, 73 cases (31.06%); and TT genotype, 13 cases (5.54%); the minor allele frequency of 5633C>T was 0.21. Survival analysis of the patients revealed that the median overall sur-vival (OS) of patients with the CT/TT genotype and those with the CC genotype was 5.9 and 4.5 years, respectively; the result was sta-tistically significant (P=0.009). Following adjustment in multivariate Cox regression analysis, the CT/TT genotype was found to be an in-dependent favorable factor for OS (HR=0.67, P=0.015). Additionally, of the 87 postoperative tissue specimens, results show that the levels of TYMP mRNA in cancer tissues of patients with the CT/TT genotype were significantly higher than those with the CC genotype (P=0.019). Conclusions: TYMP mRNA expression may be influenced by the 5633C>T polymorphism, making CRC patients benefit from capecitabine treatment.

Significance of heart-type fatty acid binding protein and brain glycogen phosphorylase isoenzyme type in septic myocardial injury / 中国小儿急救医学

Objective@#To demonstrate the relevance of heart-type fatty acid binding protein(H-FABP) and brain glycogen phosphorylase isoenzyme type(GPBB) with myocardial injury in sepsis.To explore the effect of H-FABP and GPBB on the severity of disease and clinical prognosis.@*Methods@#A total of 40 cases of children with sepsis were selected in this study from January 1, 2017 to October 31, 2017.According to the illness severity, they were divided into sepsis group(n=15), severe sepsis group(n=13) and septic shock group(n=12), 19 cases of children with non-infectious diseases were selected as the control group for the same period.The levels of serum WBC, C-reactive protein, procalcitonin, cTnI, CK, CK-MB, CK-MB isoenzyme quality and NT-proBNP were collected within 24 hours of admission.The APECHEⅡ scores were calculated for each child.The peripheral blood of the patients within 24 hours after admission was used to determine the levels of serum H-FABP and GPBB by ELISA method.The differences of clinical indicators among the groups were analyzed.According to the survival of patients with sepsis, they were divided into 37 cases in the improved group and 3 cases in the death group.All patients with sepsis were divided into left ventricular ejection fraction(LVEF) normal group(n=26) and LVEF decreased group(n=14). According to whether the serum cTnI was elevated, all children with sepsis were divided into cTnI normal group (n=29) and cTnI increased group (n=11). The differences of H-FABP and GPBB levels were compared.According to the presence or absence of LVEF decline, the area under the ROC curve was used to evaluate the predictive power of each index for myocardial injury in sepsis.Based on the decrease of LVEF, the area under the ROC curve was used to evaluate the prediction of each index for myocardial injury in sepsis.@*Results@#There were significant differences in H-FABP and GPBB levels among the control group, sepsis group, severe sepsis group and the septic shock group (H=42.241, P<0.05; H=32.486, P<0.05). Although there was no significant difference between the improved group and the death group(P>0.05), there was an elevated trend of H-FABP and GPBB in the death group.The levels of H-FABP (t=-3.770, P=0.001) and GPBB (Z=-2.113, P=0.033) were statistically significant in the LVEF normal group and the LVEF decreased group (LVEF≤60%). There were no significant differences in the levels of H-FABP and GPBB between the cTnI normal group and the cTnI increased group (P>0.05), but the cTnI increased group had an increasing trend.The area under the ROC curve in the diagnosis of myocardial injury in sepsis were H-FABP 0.821, NT-ProBNP 0.738, GPBB 0.661, CK 0.560, cTnI 0.512, in which the sensitivity(0.833) and specificity(0.786)of H-FABP were both higher.@*Conclusion@#The serum H-FABP and the GPBB levels can be used to monitor for myocardial damage, and it has a correlation with the severity of the disease and the prognosis.H-FABP has a significant advantage over traditional myocardial markers in sensitivity and specificity to determine the myocardial injury of sepsis.

Expression of phosphorylated protein kinase B1 and human telomere reverse transcriptase in epithelial ovarian carcinoma and their clinical significances / 肿瘤研究与临床

Objective To investigate the expression of phosphorylated protein kinase B1 (pAkt1) and human telomerase reverse transcriptase (hTERT) in ovarian epithelial carcinoma (EOC) and its relationship with prognosis, and to analyze the correlation between pAkt1 and hTERT expression. Methods 92 patients with EOC in Tangshan Gongren Hospital from January 2012 to December 2016 were selected. The expressions of hTERT and pAkt1 proteins were detected by immunohistochemistry. The relationship between the expressions of pAkt1 and hTERT proteins and their relationship with clinical prognosis were analyzed. Results Of the 92 patients with EOC, 68 cases (73.9 %) had positive expression of pAkt and 52 cases (56.5 %) had positive expression of hTERT. There was no significant correlation between expressions of pAkt1 and hTERT proteins in EOC (r= 0.284, P= 0.633). The expressions of pAkt1 and hTERT proteins were not related with age, tumor pathology type and the International Federation of Gynecology and Obstetrics (FIGO) staging (all P> 0.05), but had significant association with tumor pathology differentiation (χ2= 2.694, P= 0.005; χ2=2.284, P=0.018). The disease-free survival of patients with both pAkt1 and hTERT positive was shorter than that of the other groups (P= 0.013). Conclusion The prognosis of EOC patients with high expression of pAkt1 and hTERT proteins is poor.

Establishment of a novel hyperuricemiaanimal model using mice and assessment ofhyporuricemia action of PNP inhibitor Ulodesine / 中国药理学通报

Aim To establish a novel acute hyperuricemia mouse model and apply it to evaluate the hyporucicemia effects of Ulodesine, a purine nucleoside phosphorylase(PNP) inhibitor.Methods The mice were intraperitoneal injected inosine and subcutaneous injected Oteracil potassium to induce accumulation of uric acid, and the animal blood was collected from eyeball or vena angularis in different time points.The levels of serum uric acid were measured and determined to test whether the acute hyperuricemia mouse model were successful or not.In order to verify the hyperuricemia seen in the model was associated with the accumulation of inosine, which was converted to uric acid by action of PNP,hyporucicemia effects of Ulodesine, a PNP inhibitor, was assessed in an enzyme assay and confirmed by using the newly established model.Result Accumulation of uric acid in the blood of mouse models was observed by combined injections of intraperitoneal 200 mg·kg-1 inosine and subcutaneous 200 mg·kg-1 Oteracil potassium respectively after 1.5 h.The enzyme assay indicated that Ulodesine was a potently PNP inhibitor with IC50 of 2.293 nmol·L-1.IV injection of Ulodesine eliminated uric acid accumulations in blood of the mouse model, which was expected as the in vivo action of Ulodesine.Conclusions A novel acute hyperuricemia mouse model is established.This is a relatively easy and more effective protocol to generate the hyperuricemia in mice, which will be a useful platform to assess the anti-hyperuricemia activity of PNP-target drugs in vivo.

Correlation of heart-type fatty acid-binding protein and glycogen phosphorylase enzyme BB in brain with septic myocardial injury / 中国小儿急救医学

The incidence of sepsis is on the rise, and the mortality is also high. Myocardial injury caused by sepsis is one of the important causes of death. However,there is no clear and unified diagnostic criteria to sepsis complicated with myocardial injury. The clinical diagnostic criteria of myocardial injury is mainly based on cardiac ultrasound,myocardial markers and so on. But the measurement of ejection fraction is needed to consider the impact of preload and afterload of left ventricula,also the subjective level of opera-tors. So cardiac markers play an important role in the detection of myocardial injury in sepsis and can reflect the severity of myocardial injury and sepsis,especially in the early diagnosis and prognosis. Heart-type fatty acid binding protein and brain glycogen phosphorylase isoenzyme type exhibit higher sensitivity, specificity and stability compared to other traditional cardiac markers in the clinical diagnosis of myocardial injury, which are worth to be studied and applied in the clinical diagnosis of myocardial injury.

Falta de evidencia en la determinación de la isoenzima BB de la fosforilasa de glucógeno (GPBB) como prueba diagnóstica de preeclampsia / Lack of evidence for offering the qualitative determination of glycogen phosphorylase isoenzyme BB (GPBB) in capillary blood of pregnant women as diagnostic test of pre-eclampsia

Resumen OBJETIVO: estructurar una pregunta clínica para conocer la utilidad de la prueba cualitativa de GPBB en sangre capilar de pacientes embarazadas, con la finalidad de establecer el diagnóstico de preeclampsia. MATERIALES Y MÉTODOS: estudio ambispectivo efectuado con base en las respuestas a la pregunta clínica estructurada: ¿cuál es la validez diagnóstica de la prueba cualitativa de la isoenzima BB de la fosforilasa de glucógeno (GPBB) en sangre capilar para preeclampsia? se buscaron las fuentes primarias en PubMed, mediante las palabras clave (MeSH): "Glycogen phosphorylase isoenzyme BB AND Pregnancy" y "Glycogen phosphorylase isoenzyme BB AND Pre-eclampsia". Se seleccionaron los artículos más relevantes, se les asignó nivel de evidencia, se investigó si la muestra y metodología fueron iguales, comparables o diferentes de la prueba cualitativa en sangre capilar, y se analizaron sus resultados con la finalidad de establecer el diagnóstico clínico de preeclampsia. RESULTADOS: no se encontraron estudios de la determinación cualitativa de GPBB en sangre capilar. En tres ensayos de casos y controles (nivel de evidencia 4) se compararon las concentraciones de GPBB en suero mediante ELISA y se encontraron cifras elevadas al momento de establecer el diagnóstico clínico; sin embargo, se consideró como un rasgo fenotípico de preeclampsia clínica, pero no como una prueba diagnóstica. CONCLUSIONES: no existe suficiente evidencia médica que respalde la utilidad de la determinación cualitativa de GPBB en sangre capilar de pacientes embarazadas para establecer el diagnóstico de preeclampsia; por lo tanto, no se sustenta su aplicación en ningún escenario clínico.

Abstract OBJECTIVE: To establish if the clinical use of the test for the diagnosis of pre-eclampsia is scientifically supported by evidence-based medicine. MATERIALS AND METHODS: Ambispective study based on the responses to the PICO clinical question was constructed, then primary sources were searched in PubMed by the strategies Glycogen phosphorylase isoenzyme BB AND Pregnancy, and Glycogen phosphorylase isoenzyme BB AND pre-eclampsia, the relevant ones were selected, with allocation of level of evidence, determining if samples and methodology were similar, comparable or different to the capillary blood test, and results were analized under diagnosis clinical intention. RESULTS: No studies were found on qualitative GPBB detection in capillary blood. Three case-control studies (evidence level 4) compared GPBB maternal serum levels determined by ELISA, and described an elevation of GPBB at clinical diagnosis, rather as a phenotypical feature of clinical Pre-eclampsia, but not a diagnostic test. CONCLUSIONS: There is no evidence at all to offer the qualitative determination of GPBB in capillary blood of pregnant women; its clinical use is not supported whatsoever.

The level of glycogen phosphorylase isoenzyme BB and ischemia modified albumin in newborn with asphyxia and myocardial injury and its clinical significance / 国际检验医学杂志

Objective To explore the clinical significance of early biomarkers in neonatal asphyxia diagnosis with myocardial damage by detection of ischemia modified albumin in neonatal serum(IMA) and glycogen phosphorylase isoenzyme BB(GPBB) for screening sensitive markers with direct myocardial injury.Methods 166 neonates were selected in our hospital as the research object,in which 136 cases with myocardial injury dividend into the experimental group and 30 cases into the control group.The experimental group were divided into mild group and severe group according to the degree of asphyxia.All the children were tested for GPBB and IMA 1 h after admission.Results The levels of GPBB in neonatal asphyxia myocardial injury group and severe asphyxia group were significantly higher than those in the control group,the difference was statistically significant(P<0.05).The sensitivity of GPBB in diagnosis of asphyxia was higher than that of IMA,the difference was statistically significant(P<0.05).Conclusion The degree of asphyxia is closely related to serum IMA,GPBB level in neonatal asphyxia complicated with myocardial injury.The sensitivity and specificity of GPBB in diagnosis asphyxia is better than IMA in children complicated with myocardial damage.

Synthesis and biological evaluation of novel amide derivatives of pentacyclic triterterpenes as inhibitors of glycogen phosphorylase / 中国药科大学学报

@#Eight novel amide derivatives of corosolic acid were synthesized by acylation, amidation and hydrolysis reactions, etc. Six novel amide derivatives of oleanolic acid were synthesized by acylation, alkylation and reduction reactions, etc. Biological evaluation of the target compounds as inhibitors of glycogen phosphorylase(GP)has been described. Results indicated that eleven derivatives exhibited moderate-to-good inhibitory activities against glycogen phosphorylase. Among them, four target compounds possess better inhibition effects than lead compounds and compound 6 show the best inhibition effect with the IC50 of 11. 2 μmol/L.

Clinical, Biochemical, and Genetic Characterization of Glycogen Storage Type IX in a Child with Asymptomatic Hepatomegaly / 대한소아소화기영양학회지

Glycogen storage disease type IX (GSD IX) is caused by a defect in phosphorylase b kinase (PhK) that results from mutations in the PHKA2, PHKB, and PHKG2 genes. Patients usually manifest recurrent ketotic hypoglycemia with growth delay, but some may present simple hepatomegaly. Although GSD IX is one of the most common causes of GSDs, its biochemical and genetic diagnosis has been problematic due to its rarity, phenotypic overlap with other types of GSDs, and genetic heterogeneities. In our report, a 22-month-old boy with GSD IX is described. No other manifestations were evident except for hepatomegaly. His growth and development also have been proceeding normally. Diagnosed was made by histologic examination, an enzyme assay, and genetic testing with known c.3210_3212del (p.Arg1070del) mutation in PHKA2 gene.

Purification and Characterization of α-Glucan Phosphorylase Isoform Pho 2 from Spinach Leaves.

α-Glucan phosphorylase is an important enzyme of carbohydrate metabolism. In spinach leaves, it has been reported in two multiple forms viz. Pho 2 (cytosolic) and Pho 1 (plastidial). Here, we extracted and purified Pho 2 form of α-glucan phosphorylase from spinach using salting out with ammonium sulfate, desalting using Sephadex G-25 chromatography, anion exchange chromatography using DEAE-Sepharose and gel filtration chromatography using Sepharose-4-B. The purified enzyme had a specific activity, 150 units/mg protein. There was 38% recovery and 652 fold purification after final Sepharose-4B chromatography. The purified enzyme showed a single protein band on SDS sodium dodecyl sulfate polyacrylamide gel electrophoresis having molecular weight 94,000±2000 daltons. The native molecular weight is found to be 188,000±3000 daltons as determined using gel filtration chromatography over Sephadex G-200. The Pho2 exhibited optimum pH at pH 6.0 with two half pH optima at pH 5.2 and pH 7.0. The optimum temperature of Pho2 is found to be 37ºC with two half temperature optima at 30ºC and 40ºC. The Km value of the enzyme for starch and glucose-1-phosphate is found to be 116 μg/mL and 0.55 mM, respectively.

Starch Phosphorylase: An Overview of Biochemical Characterization, Immobilization and Peptide Mapping.

Starch phosphorylase (EC 2.4.1.1) is the key enzyme which catalyzes the reversible conversion of starch and inorganic phosphate into glucose-1-phosphate (G-1-P) and has been reported in many higher plants. It can be exploited for industrial purposes for tailoring starch and for the synthesis of biopolymers which may be useful in food industry and are also of clinical importance. Starch in the form of insoluble granules accumulates in chloroplasts as the primary product of photosynthesis. Multiple forms of the enzyme have been reported in different plant tissues which have been predicted to have different roles in starch metabolism. Here, various biochemical properties have been reviewed. Various techniques for enzyme immobilization have been discussed. Besides, reports on immobilization of starch phosphorylase from different sources and on different solid matrices have also been reviewed. Peptide mapping reports of various proteins have also been assessed in this review.

Application of the glycogen phosphorylase BB and heart-type fatty acid binding protein to the early diagnosis of neonatal myocardial injury / 实用医学杂志

Objective To evaluate the efficiency of glycogen phosphorylase BB (GPBB) and heart-type fatty acid binding protein (H-FABP) in the early diagnosis of neonatal myocardial injury (NMI). Methods The GPBB and H-FABP levels in the blood were detected at 3, 6, 12 and 24 h after onset of the NMI. The GPBB and H-FABP levels were compared among NMI, Non-NMI and control groups. The NMI diagnosis sensitivity and specificity for GPBB, H-FABP, GPBB combined with H-FABP were compared. The relationships of GPBB, H-FABP levels with the risk factors were analyzed. Results The GPBB and H-FABP levels of NMI group were higher than those of Non-NMI and control groups (P < 0.01). The diagnosis sensitivity of four time points (3, 6, 12 and 24 h) of combined GPBB and H-FABP were higher than those of the GPBB and H-FABP (P < 0.05). The GPBB and H-FABP were negatively correlated with blood glucose level, positively correlated with asphyxia degree and septicemia degree. Conclusion The combined application of GPBB and H-FABP can improve sensitivity in the early diagnosis of NMI.

Transfected thymidine phosphorylase cDNA with lentiviral vector and INF-α2a enhance the anticancer effects of 5'-deoxy-5-fluorouridine on colorectal cancer cell line LOVO / 中华普通外科杂志

Objective Thymidine phosphorylase (TP) cDNA was transfected into colorectal cancer cell lines LOVO with the lentiviral vector,the anticancer effeciency of 5'-deoxy-5-fluorouridine (5'-DFUR)and 5-fluorouracil (5-FU) on LOVO cells were evaluated.Methods TP cDNA were transfected into LOVO cell line with the lentiviral vector pLenti6.3_MCS_IRES2-EGFP,and the transfection efficiency was analyzed by flow cytometer and immunohistochemistry.Cells were divided into six groups:LOVO,LOVO-TP,LOVO-control,LOVO + INF-α2a,LOVO-TP + INF-α2a,LOVO-control + INF-α2a.TP protein expression and the relative quantitative analysis for TP mRNA in transfections cells were detected by Western blot and RT-PCR respectively.Volumes of converted 5-FU from either in the medium containing different concentration of 5'-DFUR,in which all cells were cultured,or in cells lysate,were detected by high performance liquid chromatography (HPLC).Results The transfection efficiency of TP cDNA in LOVO cells was 95％.The Mean gray value of TP protein expression in LOVO-TP and LOVO-TP + INF-α2a were 198.15 folds and 243.22 folds higher than LOVO cell,respectively (P ＜ 0.01).The RQ values of TP mRNA expression in LOVO-TP and LOVO-TP + INF-α2a were also 18.56 folds and 59.61 folds higher than wild LOVO cell,respectively (P ＜ 0.01).The IC50 values of 5'-DFUR on LOVO-TP and LOVO-TP + INF-α2a were 1 660 μ mol/L and 813 μ mol/L,respectively,significantly lower than 4 462.59 μ mol/L in wild LOVO (P ＜ O.01).The 5-FU volumes detected from media contained series concentration of 5'-DFUR for culturing LOVO-TP and LOVO-TP + INF-α2a were 2.0-5.3 folds,and 2.9-10.4 folds more than wild LOVO,respectively (P ＜ 0.01).Conclusions Transfected TP cDNA into colorectal cancer cell line LOVO with lentiviral vector increases the expression of TP mRNA and TP protein and the amount of 5-FU converted from 5'-DFUR,enhancing its anticancer effect.

The Significance of Clinical and Laboratory Features in the Diagnosis of Glycogen Storage Disease Type V: A Case Report

Glycogen storage disease type V (GSD-V) is the most common disorder of muscle glycogenosis with characteristic clinical and laboratory findings. A 32-yr-old woman complained of exercise intolerance and myoglobulinuria since early adolescence. She reported several episodes of second-wind phenomenon. Physical examination did not show any neurological abnormality, including fixed muscle weakness or atrophy. Serum creatine kinase level was 1,161 IU/L at rest. The result of the non-ischemic forearm exercise test was compatible with GSD-V. Mutation analysis identified the compound heterozygous mutations of the PYGM, p.D510fs and p.F710del, which has not yet been reported in Korea. The present case recognizes that detail clinical and laboratory analysis is the first step in the diagnosis of GSD-V.