Relationship between the Expression of Thymidylate Synthase,Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase and Survival in Epithelial Ovarian Cancer

The mRNA and protein expression of thymidylate synthase (TS), thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) and their relationship with prognosis were investigated. Real-time quantitative RT-PCR (Taqman) was used to detect the mRNA expression of TS, TP and DPD in formalin-fixed and paraffin-embedded 106 samples of epithelial ovarian cancer and 29 normal ovaries. A TATA box-binding protein (TBP) was used as an endogenous reference gene. A relationship between TS, TP, DPD expression and clinicopathologic features was investigated. The protein location and expression of TS, TP and DPD was examined in the same patients by an avidin-biotin-peroxidase immunohistochemistry. TS and TP mRNA expression levels were significantly higher in tumor group than in normal controls, with the average value of TS and TP mRNA being 6.14±0.62 and 0.59±0.06 in tumor tissue, and 0.71±0.14 and 0.16±0.04 in normal tissue, respectively. DPD mRNA expression levels were significantly lower in tumor group (0.11±0.02) than in normal controls (0.38±0.05). There was statistically significant difference in TS and TP mRNA expression levels among different pathological grades and clinical stages (P<0.05), but histological subtype was not significantly associated with TS and TP mRNA expression. DPD gene expression was not significantly associated with any clinicopathological parameters. Immunohistochemistry revealed that TP protein was mainly distributed in nucleus, and TS and DPD mainly in cytoplasm. The protein expression intensity of TS, TP and DPD was coincided with the mRNA expression levels. It was concluded that TS, TP mRNA and protein expression levels were significantly higher in epithelial ovarian cancer, and DPD mRNA and protein expression levels were significantly lower. The expression levels of TS and DPD were related to the patients’ prognosis and survival. Combined gene expression levels of TS, TP and DPD represent a new variable to predict the clinical outcome in ovarian cancer. The association of TS, TP and DPD expression levels with survival suggests an importance of these genes for tumor occurrence and progression.

Thymidylate synthase and thymidine phosphorylase gene expression as predictive parameters for the efficacy of 5-fluorouraci-based adjuvant chemotherapy for gastric cancer

AIM： To investigate the prognostic role of thymidylate synthase （TS） and thymidine phosphorylase （TP） mRNA levels in T3 or T4 gastric cancer treated with 5-fluorouraci-based adjuvant chemotherapy. METHODS： Fifty-one patients with T3 or T4 gastric cancer received systemic 5-fluorouraci-based adjuvant chemotherapy, and intratumoral expression of TS and TP in 51 gastric cancer tissue samples was tested by realtime quantitative PCR.RESULTS： The median disease-free survival （DFS） time was 10.2 mo in the patients. There were no significant differences in DFS between the groups with high and low levels of TP. However, the group with low level of TS had a longer DFS （14.4 mo vs 8.3 mo, P = 0.017）. The median overall survival （OS） time was 18.5 mo, and there were significant differences in OS between the groups with high and low levels of TS or TP （for TS, 17.0 mo vs 21.3 mo, P = 0.010; for TP, 16.6 mo vs 22.5 too, P = 0.009）. Moreover, the coupled low expression of these two genes was strongly associated with a longer survival time of patients as compared with that of a single gene.CONCLUSION： Expression of TS and TP mRNA is a useful predictive parameter for the survival of postoperative gastric cancer patients after 5-fluorouracilbased adjuvant chemotherapy.

Effects of thymidine phosphorylase on tumor aggressiveness and 5-fluorouracil sensitivity in cholangiocarcinoma

AIM: To evaluate the role of thymidine phosphorylase (TP) in cholangiocarcinoma using small interfering RNA (siRNA). METHODS: A human cholangiocarcinoma-derived cell line KKU-M139, which has a naturally high level of endogenous TP, had TP expression transiently knocked down using siRNA. Cell growth, migration, in vitro angiogenesis, apoptosis, and cytotoxicity were assayed in TP knockdown and wild-type cell lines. RESULTS: TP mRNA and protein expression were decreased by 87.1% ± 0.49% and 72.5% ± 3.2%, respectively, compared with control cells. Inhibition of TP significantly decreased migration of KKU-M139, and suppressed migration and tube formation of human umbilical vein endothelial cells. siRNA also reduced the ability of TP to resist hypoxia-induced apoptosis, while suppression of TP reduced the sensitivity of KKU-M139 to 5-fluorouracil. CONCLUSION: Inhibition of TP may be beneficial in decreasing angiogenesis-dependent growth and migration of cholangiocarcinoma but may diminish the response to 5-fluorouracil chemotherapy.

Induction of nucleoside phosphorylase in Enterobacter aerogenes and enzymatic synthesis of adenine arabinoside

Nucleoside phosphorylases (NPases) were found to be induced in Enterobacter aerogenes DGO-04, and cytidine and cytidine 5′-monophosphate (CMP) were the best inducers. Five mmol/L to fifteen mmol/L cytidine or CMP could distinctly increase the activities of purine nucleoside phosphorylase (PNPase), uridine phosphorylase (UPase) and thymidine phosphorylase (TPase) when they were added into medium from 0 to 8 h. In the process of enzymatic synthesis of adenine arabinoside from adenine and uracil arabinoside with wet cells of Enterobacter aerogenes DGO-04 induced by cytidine or CMP, the reaction time could be shortened from 36 to 6 h. After enzymatic reaction the activity of NPase in the cells induced remained higher than that in the cells uninduced.

Research development of the relationship between thymidine phosphorylase expression and colorectal carcinoma

Thymidine phosphorylase (TP) is a key enzyme that contributes to the composition and decomposition of pyrimidine nucleotides. TP seems homologous to platelet-derived endothelial cell growth factor, and its effects on inducing vascularization and anti-apoptosis are closely related to growth and metastasis of colorectal carcinoma. In addition, TP is a key enzyme that catalyzes the transformation from 5-fluorouracil (FU) prodrugs of 5′-deoxy-5-fluorouridine (5′- DFUR) to 5-FU. The activity of TP is closely related to the sensitivity of colorectal carcinoma cells to fluorouracil drugs and targeted therapy. Given the important functions of TP in growth, metastasis, tumor treatment, and prognosis, determining its expression mechanism is significant. This article summarizes the research development of TP expression in colorectal carcinoma, tumor neovascularization, cytotoxicity activation of 5′-DFUR, and colorectal carcinoma therapy.

Induction of Recombinant Uridine Phosphorylase and Its Application in Biosynthesis of Pyrimidine Nucleosides

Recombinant Escherichia coli pUDP,which overexpressed uridine phosphorylase（UPase）,was constructed.0.5 mmol·L 1lactose had a similar induction effect as the commonly used inducer IPTG during 2.5-5.5 h of cell growth.The lactose-induced UPase was stable at 50°C.Wet cells of pUDP was used as catalyst to biosynthesize 5-fluorouridine from 30 mmol·L 1uridine and 5-fluorouracil in phosphate buffer（pH 7.0）catalyzed at 50°C for 1.5 h and the yield of 5-fluorouridine was higher than 68%.Under the optimum reaction conditions for production of 5-fluorouridine,5-methyluridine and azauridine were synthesized from uridine by pUDP,the yield was 61.7%and 47.2%respectively.Deoxynucleosides were also synthesized by pUDP,but the yield was only about 20%.

Effects of Trinitrotoluene on Serum Phosphorylase A Activities and on Calcium Contents in Men and Rats

Wistar rats were exposed to trinitrotoluene (TNT) for 6 weeks. After initiation of TNT exposure, serum phosphorylase A activities and calcium contents were assayed for every 2 weeks. Both of these 2 parameters increased in rats treated with 50 and 100 mg TNT/kg b.w. at 3 intervals. Serum phosphorylase A activities and calcium contents of TNT exposure worker increased too.

Structural Insight into the Binding of Hypoxanthine with Purine Nucleoside Phosphorylase from Streptococcus mutants

Purine nucleoside phosphorylase is a key enzyme in the purine-salvage pathway and an attractive target for drug design. The crystal structure of Streptococcus mutants purine nucleoside phosphorylase（Smu PNP） has been solved by molecular replacement at 1.80 resolution and refined to R factors of 19.9%/23.7%（Rcryst/Rfree） . Sequence alignment and structural comparison show that Smu PNP has more similarity with PNPs isolated from human and malarial sources than the bacterial PNPs. The structure complexed with hypoxanthine（HPA） and sulfate ion was solved at 2.24A resolution and refined to R factors of 21.6%/24.1%（Rcryst/Rfree） . It is interesting to note that the resulting electron density indicated the product,HPA,presents in the active site although inosine was included in the crystallization mixture with Smu PNP. Asn233 and Glu191 are the important residues for ligand binding and recognition. Comparison with PNPs from different species gives detailed information about binding of small molecules on the active site,which is important for the studies of enzymatic mechanism and rational design of specific inhibitors for PNPs.

Case of purine nucleoside phosphorylase deficiency presented with hematuria

Purine Nucleoside Phosphorylase (PNP) deficiency is a rare autosomal recessive metabolic disorder. In PNP-deficiency disorder, the deficient enzyme leads to accumulation of toxic metabolites, especially in lymphocytes and the metabolites exert toxic effect on T-cell generation. Purine nucleoside phosphorylase deficiency causes decreased numbers of T cells and lymphopenia. The patients suffering from PNP-deficiency may be admitted with recurrent infections, as well as neurological and autoimmune findings. We hereby presented a case admitted with the symptom of hematuria in which we established the diagnosis of PNP-deficiency early on the basis of detection of lymphopenia and low level of uric acid.

Methylthioadenosine (MTA) Rescues Methylthioadenosine Phosphorylase (MTAP)-Deficient Tumors from Purine Synthesis Inhibition <i>in Vivo</i>via Non-Autonomous Adenine Supply

Methylthioadenosine phosphorylase, (MTAP) is a key enzyme in the adenine and methionine salvage pathways. MTAP is encoded on human chromosome 9p21 in close proximity to the p16INK4a and p14ARF tumor suppressor genes and is frequently co-deleted with p16INK4a in many cancers. Deletion of MTAP has been reported to create a reliance of MTAP–/– tumors on de novo purine synthesis to maintain adequate pools of AMP, leading to increased sensitivity to purine synthesis inhibitors, such as L-alanosine. The ‘Achilles heel’ created by the loss of MTAP in cancer cells provides a unique therapeutic opportunity whereby MTAP–/– tumors could be selectively targeted with purine synthesis inhibitors and normal tissues could be preferentially rescued with MTAP substrates, such as MTA. We demonstrate that, in contrast to published literature, MTAP–/– cells are not more sensitive to inhibition of de novo purine synthesis than MTAP+/+ cells. Although MTA can preferentially rescue MTAP+/+ cells from purine-synthesis inhibitor toxicity in vitro, MTA protects cells of both genotypes from L-alanosine equivalently in vivo. Our data demonstrate that in vivo, adenine salvaged from plasma and adjacent tissues is sufficient to protect MTAP–/– tumors from the effects of purine synthesis inhibitors. These results suggest targeting MTAP–/– tumors with de novo purine synthesis inhibitors is unlikely to provide significant benefit over other therapeutic strategies and may explain, at least in part, the lack of efficacy of L-alanosine in clinical trials.

Results of Combining Phosphorylase Kinase Inhibition with Removal of Precipitating Factors in Large Cohort of Psoriatic Patients: A Proof of Concept Study

Background: Phosphorylase kinase (PhK) activity is induced by injurious stimuli, which is known to precipitate psoriasis. We had previously reported that elevated PhK activity in psoriatic epidermis correlated with increased psoriatic activity, and that suppression of PhK activity by its inhibitor, curcumin gel, correlated with disease resolution. Objective: We evaluated the efficacy of a strategy of combining PhK inhibition by topical curcumin with elimination of PhK-generating precipitating factors from various injurious stimuli in producing improvement of psoriatic activity, aiming at complete resolution. Patients and Methods: We studied a cohort of 647 consecutive patients with mild to severe psoriasis in a single center. Our therapeutic regimen consisted of curcumin gel, topical steroids, strict avoidance of contact allergens, avoidance of dairy products in lactose-intolerant patients, and treatment of infections to eliminate bacterial superantigens. Results: PASI scores at 0 wk was 24.7 +/– 17.1 (SD), n = 647. PASI scores improved significantly at 4 weeks to 11.5 +/– 8.1 (n = 638;p < 0.0001), at 8 weeks to 4.5 +/– 4.2 (n = 636, p < 0.0001), and at 16 weeks to 0.9 +/– 2.5 (n = 641, p < 0.0001). At 16 weeks, 72.2% of patients were completely clear of psoriatic activity (PASI = 0). Conclusion: Our results indicate that a regimen of PhK inhibition by topical curcumin with elimination of PhK-generating factors is effective in producing significant reduction of psoriatic activity at 16 weeks, with complete clearance of psoriasis in 72.2% of patients.

Immobilization of Starch Phosphorylase from Germinating Wheat Seeds

Starch phosphorylase is an industrially important enzyme used in the production of glucose-l-phosphate. Here, we extracted the enzyme from the germinating wheat seeds and partially purified using ammonium sulfate fractionation. The partially purified enzyme showed maximum enzyme activity at pH 6.2 and pH 7.2 in the polysaccharide and glucose-l-phosphate formation directions, respectively. The enzyme showed maximum enzyme activity at 37 ℃ temperature with 50% of the enzyme activity at 32 ℃ and 42 ℃. The desalted ammonium sulfate fractionated enzyme has been immobilized on brick dust with nearly 60% enzyme activity retention. The specific activity of the immobilized starch phosphorylase increased from 0.410 to 0.925. There was a slight alkaline shift in the optimum pH when assayed in both the directions. The immobilized starch phosphorylase also displayed increased optimum temperature and thermo-stability and could be reused many times. The desalted ammonium sulfate fractionated enzyme exhibited a half life of 4 h at 30 ℃ and 30 rain at 50 ℃ whereas brick dust immobilized enzyme exhibited a half life of 7 h at 30 ℃ and 45 min at 50 ℃. The immobilized enzyme may be exploited for glucose-l-phosphate production.

Endometrial Cancer Research Based on Gut Microbiomics and Metabolomics:An Analysis of Correlation and Differences

Endometrial cancer(EC)is a malignant tumour that occurs in the epithelial cells of the endometrium and represents one of the most common malignancies involving the female reproductive system,with endometrioid adenocarcinoma as the most common type.In recent years,with an increasingly aging society and the growing number of obese people,the incidence of EC is constantly rising,posing a serious threat to women’s health.Some studies have reported that the interruption of digestion and absorption caused by imbalance in intestinal microbiota may lead to conditions such as obesity,hypertension,diabetes,and hormone imbalance,which are all risk factors for EC.Meanwhile,intestinal bacteria produce a series of metabolites during colonization and reproduction,which can rapidly respond to changes in the microenvironment of the body.Changes in their types and quantities can serve as sensitive indicators of physiological and pathological changes in the body.Patients with EC often suffer from metabolic diseases,which can lead to metabolic disorders involving carbohydrates,fats,and amino acid in their bodies.

Changes in Urinary Metabolomics of Female Kashin-Beck Disease Patients in Qinghai-Tibet Plateau,China

Kashin-Beck disease(KBD)is a chronic,endemic,degenerative osteoarthropathic condition that is predominantly found in China's Mainland.KBD is a progressive disease,and the signs and symptoms become more severe in adulthood owing to the continuous development of lesions in the joint cartilage[1].Although childhood KBD has largely been controlled,adult KBD is an important health concern in rural regions.Unfortunately,in the last century,many adult KBD patients still remain during serious epidemics.

Impeded Carbohydrate Metabolism in Rice Plants under Submergence Stress

The detrimental effects of submergence on physiological performances of some rice varieties with special references to carbohydrate metabolisms and their allied enzymes during post-flowering stages have been documented and clarified in the present investigation. It was found that photosynthetic rate and concomitant translocation of sugars into the panicles were both related to the yield. The detrimental effects of the complete submergence were recorded in generation of sucrose, starch, sucrose phosphate synthase and phosphorylase activity in the developing panicles of the plants as compared to those under normal or control （i.e. non-submerged） condition. The accumulation of starch was significantly lower in plants under submergence and that was correlated with ADP-glucose pyrophosphorylase activity. Photosynthetic rate was most affected under submergence in varying days of post-flowering and was also related to the down regulation of Ribulose bisphosphate carboxylase activity. However, under normal or control condition, there recorded a steady maintenance of photosynthetic rate at the post-flowering stages and significantly higher values of Ribulose bisphosphate carboxylase activity. Still, photosynthetic rate of the plants under both control and submerged conditions had hardly any significant correlation with sugar accumulation and other enzymes of carbohydrate metabolism like invertase with grain yield. Finally, plants under submergence suffered significant loss of yield by poor grain filling which was related to impeded carbohydrate metabolism in the tissues. It is evident that loss of yield under submergence is attributed both by lower sink size or sink capacity （number of panicles, in this case） as well as subdued carbohydrate metabolism in plants and its subsequent partitioning into the grains.

Metabolomic Profiling of Mice Exposed toα-amanitin Using Ultra-performance Liquid Chromatography Quadrupole Time-of-flight Tandem Mass Spectrometry

Numerous instances of poisoning caused by the ingestion of mushrooms are reported annually worldwide,with most mushroom poisoning deaths caused by the ingestion of toxic amanita fungi.Amatoxin is the main toxic component of amanita,andα-amanitin is one of the most potent toxins present in amatoxin[1].

Metabotropic glutamate receptor 1 alpha:a unique receptor variant with variable implications for Alzheimer’s disease pathogenesis

Alzheimer’s disease (AD) is the leading neurodegenerative disorder globally.Despite this,there is minimal effective therapeutics proven to reduce or prevent the progression of this disease.Glutamate is the main excitatory neurotransmitter within the central nervous system and plays a crucial role in neuronal and synaptic functions.

Metabolomic profiling provides new insights into blood-brain barrier regulation

Early blood-brain barrier(BBB) disturbance contributes to many different neurological diseases including Alzheimer 's disease, Parkinson's disease and stroke, particularly in the etiology and early stages(Abbott et al., 2010).

Long-term Di-(2-ethylhexyl) phthalate Exposure Disturbs the Lipid Metabolism Profiles and Hepatic Enzymes in Male Rats:A UPLC-MS-based Serum Metabolomics Analysis

In recent years,many investigations have indicated that environmental endocrine disruptors(EEDs)may accelerate the development of obesity[1].One kind of EEDs,di-(2-ethylhexyl)phthalate(DEHP),is ubiquitous in natural environments due to its role in the manufacture and use of plastic products[2].

Bone Metabolic Disorders due to Antiepileptic Drugs and Therapeutic Effects of Vitamine D_2

Bone Mineral Content(BMC), serum cal-cium, phosphorus and AlP were measured in64 epileptic patients on long term treatmentwith one or two kinds of antiepileptic drugs(AEDs) ,and in 14 epileptic patients not tak-ing AEDs. In the epileptic patients takingAEDs, the BMC value was 0.645±0.020g/cm^2,significantly lower than 0.693±0.022g/cm^2, theBMC value of the control group (P【0.01).