Measurement of very low-molecular weight metabolites by traveling wave ion mobility and its use in human urine samples

The collision cross-sections(CCS)measurement using ion mobility spectrometry(IMS)in combination with mass spectrometry(MS)offers a great opportunity to increase confidence in metabolite identification.However,owing to the lack of sensitivity and resolution,IMS has an analytical challenge in studying the CCS values of very low-molecular-weight metabolites(VLMs250 Da).Here,we describe an analytical method using ultrahigh-performance liquid chromatography(UPLC)coupled to a traveling wave ion mobility-quadrupole-time-of-flight mass spectrometer optimized for the measurement of VLMs in human urine samples.The experimental CCS values,along with mass spectral properties,were reported for the 174 metabolites.The experimental data included the mass-to-charge ratio(m/z),retention time(RT),tandem MS(MS/MS)spectra,and CCS values.Among the studied metabolites,263 traveling wave ion mobility spectrometry(TWIMS)-derived CCS values(TWCCSN2)were reported for the first time,and more than 70%of these were CCS values of VLMs.The TWCCSN2 values were highly repeatable,with inter-day variations of<1%relative standard deviation(RSD).The developed method revealed excellent TWCCSN2 accuracy with a CCS difference(DCCS)within±2%of the reported drift tube IMS(DTIMS)and TWIMS CCS values.The complexity of the urine matrix did not affect the precision of the method,as evidenced by DCCS within±1.92%.According to the Metabolomics Standards Initiative,55 urinary metabolites were identified with a confidence level of 1.Among these 55 metabolites,53(96%)were VLMs.The larger number of confirmed compounds found in this study was a result of the addition of TWCCSN2 values,which clearly increased metabolite identification confidence.

Altered Levels of Blood Glucose and Serum Lipids in Sudanese Patients with Ovarian Cancer

Background: The etiology of ovarian cancer is not well-understood;numerous metabolomics profiling, epidemiological, and hospital-based case control studies have associated abnormal levels of blood glucose and serum lipids with the risk and the prognosis of various types of cancers including ovarian cancer. The association between the risk of the incidence of ovarian cancer and the alterations in the levels of blood glucose and serum lipids is not well defined. Objective: In this study we aimed to compare the levels of blood glucose, triacylglycerols, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol in patients with different stages of ovarian cancer and healthy controls to determine how they relate to the risk and prognosis of ovarian cancer. Methodology: In a case-control cross sectional study, we enrolled ninety-nine Sudanese women, diagnosed with ovarian cancer but had not received any kind of treatment as the study group, and a control group of forty-one age-matched, apparently healthy women. The patients were classified according to the International Federation of Obstetricians and Gynecologists staging system into two groups: early stages (stage I & II) and late stages (stages III & IV). Blood glucose and serum lipids;triacylglycerols, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol were determined by enzymatic colorimetric methods using commercially available analytical kits. The IBM SPSS version 20 software was used for statistical analysis. A Mann-Whitney U test was used for comparison of the median concentrations of blood glucose, triacylglycerols, total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol in the study groups. Logistic regression model was used to estimate the relative risk of ovarian cancer in relation to levels of blood glucose and serum lipids. P value of 0.05 was considered significant. Results: Our data indicated significantly higher levels of blood glucose (p < 0.001), triacylglycerols (p = 0.002), and low-density lipoprotein cholesterol (p < 0.001), and lower levels of high-density lipoprotein cholesterol (p = 0.023), in ovarian cancer patients compared to the control subjects. No significant difference was found in the levels of blood glucose or any of the serum lipids between patients in the early stages (stage I & II) and those in late stages (stage III & IV) of ovarian cancer. The logistic regression analysis indicated significant association between the elevated levels of the blood glucose, triacylglycerols and low-density lipoprotein cholesterol and the risk of the ovarian cancer. Conclusion: We conclude that the levels of blood glucose, triacylglycerols, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol differ significantly between ovarian cancer patients and the healthy control subjects. The risk of ovarian cancer was positively associated with the levels of blood glucose, triacylglycerols and low-density lipoprotein cholesterol, and negatively associated with levels of high-density lipoprotein cholesterol. Therefore, determination of blood glucose and serum lipids, particularly, triacylglycerols, low-density lipoprotein cholesterol may be helpful as diagnostic indicators of ovarian cancer (OC).

Delayed versus immediate intervention of ruptured brain arteriovenous malformations:A case report

BACKGROUND Brain arteriovenous malformations(bAVMs)remains one of the most prevalent causes of intracranial hemorrhage and stroke-like syndromes in the young adult population.Although it has been agreed upon that definitive treatment using either single or multi-modal approach is warranted for successful bAVM management,much debate still revolves regarding the optimal timing of definitive treatment.CASE SUMMARY In this report,we present a case of delayed,definitive endovascular treatment for ruptured bAVM in a 21-year-old female,3 mo post-ictus.The bAVM,with a left pericallosal feeding artery and cortical draining veins,was successfully obliterated through embolization using the Onyx 18.On follow-up the patient has recommenced her daily activities and experiences only mild occasional headaches with mild motor deficits.The report leads to our review on an important issue regarding the optimal timing of ruptured bAVM definitive management and bring forward the current evidence available on delayed vs immediate definitive bAVM intervention.We also highlight current issues that need to be addressed for clearer guidelines on definitive therapy initiation.CONCLUSION Current treatment paradigms of ruptured bAVM remains elusive,with substantial heterogeneity in the current literature.A consensus on the definition of“acute”vs“delayed”,management goal,follow-up length and outcome parameters are required to support formation of a clear paradigm.

Diabetes mellitus: The epidemic of the century

The epidemic nature of diabetes mellitus in different regions is reviewed. The Middle East and North Africa region has the highest prevalence of diabetes in adults(10.9%) whereas, the Western Pacific region has the highest number of adults diagnosed with diabetes and has countries with the highest prevalence of diabetes(37.5%). Different classes of diabetes mellitus, type 1, type 2, gestational diabetes and other types of diabetes mellitus are compared in terms of diagnostic criteria, etiology and genetics. The molecular genetics of diabetes received extensive attention in recent years by many prominent investigators and research groups in the biomedical field. A large array of mutations and single nucleotide polymorphisms in genes that play a role in the various steps and pathways involved in glucose metabolism and the development, control and function of pancreatic cells at various levels are reviewed. The major advances in the molecular understanding of diabetes in relation to the different types of diabetes in comparison to the previous understanding in this field are briefly reviewed here. Despite the accumulation of extensive data at the molecular and cellular levels, the mechanism of diabetes development and complications are still not fully understood. Definitely, more extensive research is needed in this field that will eventually reflect on the ultimate objective to improve diagnoses, therapy and minimize the chance of chronic complications development.

Malaria parasite carbonic anhydrase:inhibition of aromatic/heterocyclic sulfonamides and its therapeutic potential

Plasmodium falciparum(P.falciparum) is responsible for the majority of life-threatening cases of human malaria,causing 1.5-2.7 million annual deaths.The global emergence of drug-resistant malaria parasites necessitates identification and characterisation of novel drug targets and their potential inhibitors.We identified the carbonic anhydrase(CA) genes in P.falciparum.The pfGA gene encodes an α-carbonic anhydrase,a Zn^(2+)-metalloenzme,possessing catalytic properties distinct from that of the human host CA enzyme.The amino acid sequence of the pfCA enzyme is different from the analogous protozoan and human enzymes.A library of aromatic/heterocyclic sulfonamides possessing a large diversity of scaffolds were found to be very good inhibitors for the malarial enzyme at moderate-low micromolar and submicromolar inhibitions.The structure of the groups substituting the aromatic-ureido-or aromatic-azomethine fragment of the molecule and the length of the parent sulfonamide were critical parameters for the inhibitory properties of the sulfonamides.One derivative,that is,4-(3,4-dichlorophenylureido)thioureidobcnzcnesulfonamide(compound 10) was the most effective in vitro Plasmodium falciparum CA inhibitor,and was also the most effective antimalarial compound on the in vitro P.falciparum growth inhibition.The compound 10 was also effective in vivo antimalarial agent in mice infected with Plasmodium berghei,an animal model of drug testing for human malaria infection. It is therefore concluded that the sulphonamide inhibitors targeting the parasite CA may have potential for the development of novel therapies against human malaria.

Association of serum lipid levels with diabetic retinopathy

AIM: To assess the association between serum lipids and diabetic retinopathy (DR). · METHODS: Sixty -one diabetic patients without retinopathy (NDR), 55 diabetic patients with non - proliferative retinopathy (NPDR) and 75 diabetic patients with proliferative retinopathy (PDR) according to ETDRS grading scale were enrolled in this study. Total cholesterol, high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL) and triglyceride values were compared between the groups. ·RESULTS: The groups were well-balanced in terms of age and gender (P =0.071, P =0.265 respectively). The mean HbA1c values were significantly lower in NDR group than the NPDR and PDR groups(P =0.004, P =0.009 respectively). Mean total cholesterol, triglyceride, LDL, HDL and VLDL levels were not significantly different between the groups (P =0.693, P =0.774, P =0.644, P = 0.910 and P =0.967 respectively, one way ANOVA). Mean total cholesterol, triglyceride, LDL, HDL and VLDL levels were not significantly different between the patients with ME and patients without ME(P =0.622, P =0.113, P =0.955, P =0.735 and P =0.490 respectively, t -test). The mean blood glucose significantly correlated with total cholesterol (r =0.173, P =0.017) and LDL (r =0.190, P = 0.008). The mean HbA1c significantly correlated with total cholesterol (r =0.158, P =0.030) and triglyceride (r =0.148, P =0.042). ·CONCLUSION: Serum lipid levels were not significantly associated with the severity of DR or existence of ME despite the significant correlation between the mean blood glucose, HbA1c and total cholesterol.

Role of environmental pollution in irritable bowel syndrome

Irritable bowel syndrome(IBS),with the prevalence of 10%-20 % of the population has become an emerging problem worldwide. IBS is a functional gastrointestinal(GI) disorder characterized by abdominal pain or discomfort and altered bowel habits. The etiology of IBS contains genetic,psychological,and immunological factors,and has not been fully elucidated; of note,recent studies also point at environmental pollution and its role in the development of functional GI diseases. In this review we focus on several environmental factors,such as bacterial contamination,air pollution,radiation and even stress as potential triggers of IBS. We discuss associated disturbances in homeostasis,such as changes in intestinal microbiome and related pathophysiological mechanisms. Based on the effect of environmental factors on the GI tract,we also propose novel targets in IBS treatment.

Synergistic growth inhibitory effects of Phyllanthus emblica and Terminalia bellerica extracts with conventional cytotoxic agents:Doxorubicin and cisplatin against human hepatocellular carcinoma and lung cancer cells

AIM: To examine the growth inhibitory effects of Phyllanthus emblica (P. emblica) and Terminalia bellerica (T. bellerica) extracts on human hepatocellular carcinoma (HepG2), and lung carcinoma (A549) cells and their synergistic effect with doxorubicin or cisplatin. METHODS: HepG2 and A549 cells were treated with P. emblica and T. bellerica extracts either alone or in combination with doxorubicin or cisplatin and effects on cell growth were determined using the sulforhodamine B (SRB) assay. The isobologram and combination index (CI) method of Chou-Talalay were used to evaluate interactions between plant extracts and drugs. RESULTS: P. emblica and T. bellerica extracts demonstrated growth inhibitory activity, with a certain degree of selectivity against the two cancer cell lines tested. Synergistic effects (CI < 1) for P. emblica /doxorubicin or cisplatin at different dose levels were demonstrated in A549 and HepG2 cells. The T. bellerica/ cisplatin or doxorubicin also showed synergistic effects in A549 and HepG2 cells. In some instances, the combinations resulted in antagonistic effects. The dose reduction level was different and specific to each combination and cell line. CONCLUSION: The growth inhibitory activity of doxorubicin or cisplatin, as a single agent, may be modified by combinations of P. emblica or T. bellerica extracts and be synergistically enhanced in some cases. Depending on the combination ratio, the doses for each drug for a given degree of effect in the combination may be reduced. The mechanisms involved in this interaction between chemotherapeutic drugs and plant extracts remain unclear and should be further evaluated.

Gastroprotective activity of Nigella sativa L oil and its constituent, thymoquinone against acute alcohol-induced gastric mucosal injury in rats

AIM: To evaluate the role of reactive oxygen species in the pathogenesis of acute ethanol-induced gastric mucosal lesions and the effect of Nigella sativa L oil (NS) and its constituent thymoquinone (TQ) in an experimental model.METHODS: Male Wistar albino rats were assigned into 4groups. Control group was given physiologic saline orally (10 mL/kg body weight) as the vehicle (gavage); ethanol group was administrated 1 mL (per rat) absolute alcohol by gavage; the third and fourth groups were given NS (10 mL/kg body weight) and TQ (10 mg/kg body weight p.o) respectively 1 h prior to alcohol intake. One hour after ethanol administration, stomach tissues were excised for macroscopic examination and biochemical analysis.RESULTS: NS and TQ could protect gastric mucosa against the injurious effect of absolute alcohol and promote ulcer healing as evidenced from the ulcer index (UI) values. NS prevented alcohol-induced increase in thiobarbituric acid-reactive substances (TBARS), an index of lipid peroxidation. NS also increased gastric glutathione content (GSH), enzymatic activities of gastric superoxide dismutase (SOD) and glutathione-S-transferase (GST). Likewise, TQ protected against the ulcerating effect of alcohol and mitigated most of the biochemical adverse effects induced by alcohol in gastric mucosa, but to a lesser extent than NS. Neither NS nor TQ affected catalase activity in gastric tissue.CONCLUSION: Both NS and TQ, particularly NS can partly protect gastric mucosa from acute alcohol-induced mucosal injury, and these gastroprotective effects might be induced, at least partly by their radical scavenging activity.

Drug sensitivity and drug resistance profiles of human intrahepatic cholangiocarcinoma cell lines

AIM: To study the effect of a number of chemotherapeutic drugs on five human intrahepatic cholangiocarcinoma (CCA) cell lines. The expressions of genes that have been proposed to influence the resistance of chemotherapeutic drugs including thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), glutathione-S-transferase P1 (GSTP1), multidrug resistance protein (MDR1) and multidrug resistance-associated proteins (MRPs) were also determined.METHODS: Five human CCA cell lines (KKU-100, KKU M055, KKU-M156, KKU-M214 and KKU-OCA17) weretreated with various chemotherapeutic drugs and growth inhibition was determined by 3-(4,5-dimethylthiazol-2-yl)5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assay. Semi-quantitative levels of gene expression were determined by a reverse transcriptase polymerase chain reaction (RT-PCR). Results of IC50 values and the ratios of gene expression were analyzed by linear regression to predict their relationship. RESULTS: Among five CCA cell lines, KKU-M055 was the most sensitive cell line towards all chemotherapeutic drugs investigated, particularly taxane derivatives with IC50 values of 0.02-3 nmol/L, whereas KKU-100 was apparently the least sensitive cell line. When compared to other chemotherapeutic agents, doxorubicin and pirarubicin showed the lowest IC50 values (＜5 μmol/L) in all five CCA cell lines. Results from RT-PCR showed that TS, MRP1, MRP3 and GSTP1 were highly expressed in these five CCA cell lines while DPD and MRP2 were only moderately expressed. It should be noted that MDR1 expression was detected only in KKU-OCA17 cell lines. A strong correlation was only found between the level of MRP3 expression and the IC50 values of etoposide, doxorubicin and pirarubicin (r = 0.86-0.98, ,P＜0.05). CONCLUSION: Sensitivity to chemotherapeutic agents is not associated with the histological type of CCA. Choosing of the appropriate chemotherapeutic regimen for the treatment of CCA requires knowledge of drug sensitivity. MRP3 was correlated with resistance of CCA cell lines to etoposide, doxorubicin and pirarubicin, whereas other chemotherapeutic drugs showed no association. The roleof this multidrug resistance-associated protein, MRP3, in chemotherapeutic resistance in CCA patients needs to be further investigated.

Role of serum interleukin-18 as a prognostic factor in patients with hepatocellular carcinoma

AIM:To determine whether serum interleukin-18 (IL-18) levels correlated with clinicopathologic features and prognosis in patients with hepatocellular carcinoma (HCC). METHODS:Serum IL-18,IL-6 and IL-12 levels were measured by enzyme-linked immunosorbent assay (ELISA) from 70 patients with HCC and 10 healthy controls. RESULTS:Serum IL-18,IL-6 and IL-12 levels of patients with HCC were significantly higher that those of the controls. The levels of IL-18 correlated significantly with the presence of venous invasion and advanced tumor stages classified by Okuda's criteria. Patients with high serum IL-18 levels (≥ 105 pg/mL) had a poorer survival than those with low serum IL-18 levels (< 105 pg/mL) (4 and 11 mo,respectively,P = 0.015). Multivariate analyses showed that serum IL-18 level,but not IL-6 and IL-12 levels,was a significant and independent prognostic factor of survival. CONCLUSION:These findings demonstrate that serum IL-8 may a useful biological marker of tumor invasiveness and an independent prognostic factor of survival for patients with HCC. Thus,the detailed mechanisms of IL-18 involving in tumor progression should be further investigated.

Gender difference in clinicopathologic features and survival of patients with hepatocellular carcinoma

AIM: To determine the influence of gender on the clinicopathologic characteristics and survival of patients with hepatocellular carcinoma (HCC). METHODS: A retrospective analysis of medical records was performed in 299 patients with HCC and their clinicopathologic features and survival were compared in relation to gender. RESULTS: There were 260 male (87%) and 39 female patients (13%), with a male-to-female ratio of 6.7:1. Female patients had lower mean serum bilirubin levels (P=O.03), lower proportion of alcohol abuse (P=O.O02), smaller mean tumor size (P=O.02), more frequent nodular type but less frequent massive and diffuse types of HCC (P=O.01), wereless advanced in Okuda's staging (P=O.04), and less frequently associated with venous invasion (P=O.03). The median survivals in females (14 mo) were significantly longer than that of male patients (4 mo) (P=O.O04, log-rank test). Multivariate analysis demonstrated that high serum alphafetoprotein levels, venous invasion, extrahepatic metastasis and lack of therapy were independent factors related to unfavorable prognosis. However, gender did not constitute a predictive variable associated with patient survival. CONCLUSION: Female patients tend to have higher survival rates than males. These differences were probably due to more favorable pathologic features of HCC at initial diagnosis and greater likelihood to undergo curative therapy in female patients.

Diagnostic role of serum interleukin-18 in gastric cancer patients

瞄准：在胃的癌症病人的各种各样的方面决定当前的地位并且在泰国病人与浆液 interleukins 的预示的角色发现临床的关联。方法：68 个病人在 2003 年 4 月期间在楚拉隆克恩国王纪念物医院在这研究被注册到 2005 年 5 月。胃的癌症组织学地在 17 个病人在 51 个病人和胃溃疡被证明。浆液 IL-6， IL-10， IL-12，和 IL-18 层次被连接酶的免疫吸着剂试金(ELISA ) 测量。结果：有 26 男性(55.32%) 和 21 女性(44.68%) ，他们的年龄从 33 ～ 85 年(意味着年龄 64.49 +/- 13.83 年) 。普通演讲是重量损失(41.2%) ，消化不良(39.2%) ，并且上面的胃肠的流血 15.7%) 。35.3% 胃的癌症病人的一个总数并且 6.3% 胃溃疡病人是吸烟者(P = 0.029 ) 。而且， 32.4% 胃的癌症病人和 6.3% 胃溃疡病人是含酒精的喝酒者(P = 0.044 ) 。损害地点在 39.4% 是幽门窦的，在 39.4% 的胃的身体，在 12.2% 的上面的胃和在 6.1% 病人的全部胃。H pylori 感染在 44.4% 被检测。糟糕区分的腺癌是最普通的病理学的发现(60.7%) 。外科疗法在 44.1% 病人被执行(在 5.9% 的全胃切除术，在 32.4% 的大部胃切除术和 palliative 在 5.9% 绕过外科) 。全身的化疗在 8.8% 病人作为辅助治疗被给。癌病 peritoneii 在 18.8% 病人被发现。吝啬的生存时间是 13.03 +/- 9.75 瞬间。在胃的癌症病人组的 IL-18 水平(58.54 +/- 43.96 pg/mL ) 在胃溃疡病人组比那显著地高(30.84 +/- 11.18 pg/mL )(P = 0.0001 )(95% CI 是 42.20， 13.19 ) 。为胃的癌症的诊断的 IL-18 的伤口点是 40 pg/mL，积极预兆的价值是 92.31% 。在有远转移的胃的癌症病人的 IL-6 水平(20.21 +/- 9.37 pg/mL ) 在没有转移的那些比那显著地高(10.13 +/- 7.83 pg/mL )(P = 0.037 )(95% CI 是 19.51， 0.65 ) 。在胃的癌症病人的 IL-10 和 IL-12 层次的角色是没有有效差量提供数据。结论：这些调查结果表明那浆液 IL-6 和 IL-18，然而并非 IL-10 和 IL-12 层次可以在有胃的癌症的病人是临床的关联和预示的因素的有用生物标记。而且， IL-18 能与高积极的预兆的价值为胃的癌症用作一个诊断标记。

Putative metabolic roles of the mitochondria in asexual blood stages and gametocytes of Plasmodium falciparum

Upon infection into human red cell,Plasmodium falciparum differentiates into asexual and sexual(gametocyte) stages.The mitochondrion is a tubular-cristate organelle,functionally and structurally different between the two stages.Genes and proteins involving metabolic and functional roles,protein targeting and import to this organelle, are comprehensively reviewed.The genes and proteins of the electron transport system are identified, partially characterized in human and rodent malaria parasites consisting of a single subunit of NADH dehydrogenase, two subunits of succinate dehydrogenase,cytochrome C reductase and cytochrome Coxidase.One of the primary functional roles of the mitochondrion in the parasite is the coordination of pyrimidine biosynthesis, the electron transport system and oxygen utilization through dihydroorotate dehydrogenase.All enzymes of tricarboxylic acid cycle,pyruvate dehydrogenase complex and some enzymes of ATP synthase,are identified and partially characterized using the completed P.falciparum genome.Some metabolic and functional roles of the organelle include oxidative phosphorylation,ubiquinone and heme biosynthesis,antioxidant defense and redox balance.Recent physiological studies involve membrane potential maintenance,cellular signaling and cation homeostasis.The organelle is a target for antimalarial drug,i.e.atovaquone.Based on the lines of evidence, we hypothesize that the parasite exhibits metabolic adaptation of the underdeveloped mitochondrial organelle to life in the mosquito vector and the human host.

Noninvasive assessment of hepatic fibrosis in Egyptian patients with chronic hepatitis C virus infection

AIM: To evaluate the accuracy of specific biochemical markers for the assessment of hepatic fibrosis in patients with chronic hepatitis C virus (HCV) infection. METHODS: One hundred and fifty-four patients with chronic HCV infection were included in this study; 124 patients were non-cirrhotic, and 30 were cirrhotic. The following measurements were obtained in all patients: serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), albumin, total bilirubin, prothrombin time and concentration, complete blood count, hepatitis B surface antigen (HBsAg), HCVAb, HCV-RNA by quantitative polymerase chain reaction, abdominal ultrasound and ultrasonic-guided liver biopsy. The following ratios, scores and indices were calculated and compared with the results of the histopathological examination: AST/ALT ratio (AAR), age platelet index (API), AST to platelet ratio index (APRI), cirrhosis discriminating score (CDS), Pohl score, G teborg University Cirrhosis Index (GUCI). RESULTS: AAR, APRI, API and GUCI demonstrated good diagnostic accuracy of liver cirrhosis (80.5%, 79.2%, 76.6% and 80.5%, respectively); P values were: < 0.01, < 0.05, < 0.001 and < 0.001, respectively. Among the studied parameters, AAR and GUCI gave the highest diagnostic accuracy (80.5%) with cutoff values of 1.2 and 1.5, respectively. APRI, API and GUCI were significantly correlated with the stage of fibrosis (P < 0.001) and the grade of activity (P < 0.001, < 0.001 and < 0.005, respectively), while CDS only correlated significantly with the stage of fibrosis (P < 0.001) and not with the degree of activity (P > 0.05). In addition, we found significant correlations for the AAR, APRI, API, GUCI and Pohl score between the non-cirrhotic (F0, F1, F2, F3) and cirrhotic (F4) groups (P values: < 0.001, < 0.05, < 0.001, < 0.001 and < 0.005, respectively; CDS did not demonstrate significant correlation (P > 0.05). CONCLUSION: The use of AAR, APRI, API, GUCI and Pohl score measurements may decrease the need for liver biopsies in diagnosing cirrhosis, especially in Egypt, where resources are limited.

Inhibition of ileal bile acid transporter:An emerging therapeutic strategy for chronic idiopathic constipation

Chronic idiopathic constipation is a common disorder of the gastrointestinal tract that encompasses a wide profile of symptoms. Current treatment options for chronic idiopathic constipation are of limited value; therefore, a novel strategy is necessary with an increased effectiveness and safety. Recently, the inhibition of the ileal bile acid transporter has become a promising target for constipation-associated diseases. Enhanced delivery of bile acids into the colon achieves an accelerated colonic transit, increased stool frequency, and relief of constipationrelated symptoms. This article provides insight into the mechanism of action of ileal bile acid transporter inhibitors and discusses their potential clinical use for pharmacotherapy of constipation in chronic idiopathic constipation.

Effects of α-mangostin on apoptosis induction of human colon cancer

AIM:To investigate the effect of α-mangostin on the growth and apoptosis induction of human colon cancer cells.METHODS:The three colorectal adenocarcinoma cell lines tested (COLO 205,MIP-101 and SW 620) were treated with α-mangostin to determine the effect on cell proliferation by MTT assay,cell morphology,chromatin condensation,cell cycle analysis,DNA fragmentation,phosphatidylserine exposure and changing of mitochondrial membrane potential.The molecular mechanisms of α-mangostin mediated apoptosis were further investigated by Western blotting analysis including activation of caspase cascade,cytochrome c release,Bax,Bid,p53 and Bcl-2 modifying factor.RESULTS:The highest inhibitory effect of α-mangostin on cell proliferation of COLO 205,MIP-101 and SW 620 were 9.74 ± 0.85 μg/mL,11.35 ± 1.12 μg/mL and 19.6 ± 1.53 μg/mL,respectively.Further study showed that α-mangostin induced apoptotic cell death in COLO 205 cells as indicated by membrane blebbing,chromatin condensation,DNA fragmentation,cell cycle analysis,sub-G1 peak (P < 0.05) and phosphatidylserine exposure.The executioner caspase,caspase-3,the initiator caspase,caspase-8,and caspase-9 were expressed upon treatment with α-mangostin.Further studies of apoptotic proteins were determined by Western blotting analysis showing increased mitochondrial cytochrome c release,Bax,p53 and Bmf as well as reduced mitochondrial membrane potential (P < 0.05).In addition,up-regulation of tBid and Fas were evident upon treatment with α-mangostin (P < 0.01).CONCLUSION:α-Mangostin may be effective as an anti-cancer agent that induced apoptotic cell death in COLO 205 via a link between extrinsic and intrinsic pathways.

Combination therapy using evening primrose oil and electrical stimulation to improve nerve function following a crush injury of sciatic nerve in male rats

Peripheral nerve injuries with a poor prognosis are common.Evening primrose oil（EPO） has beneficial biological effects and immunomodulatory properties.Since electrical activity plays a major role in neural regeneration,the present study investigated the effects of electrical stimulation（ES）,combined with evening primrose oil（EPO）,on sciatic nerve function after a crush injury in rats.In anesthetized rats,the sciatic nerve was crushed using small haemostatic forceps followed by ES and/or EPO treatment for 4 weeks.Functional recovery of the sciatic nerve was assessed using the sciatic functional index.Histopathological changes of gastrocnemius muscle atrophy were investigated by light microscopy.Electrophysiological changes were assessed by the nerve conduction velocity of sciatic nerves.Immunohistochemistry was used to determine the remyelination of the sciatic nerve following the interventions.EPO ＋ ES,EPO,and ES obviously improved sciatic nerve function assessed by the sciatic functional index and nerve conduction velocity of the sciatic nerve at 28 days after operation.Expression of the peripheral nerve remyelination marker,protein zero（P0）,was increased in the treatment groups at 28 days after operation.Muscle atrophy severity was decreased significantly while the nerve conduction velocity was increased significantly in rats with sciatic nerve injury in the injury ＋ EPO ＋ ES group than in the EPO or ES group.Totally speaking,the combined use of EPO and ES may produce an improving effect on the function of sciatic nerves injured by a crush.The increased expression of P0 may have contributed to improving the functional effects of combination therapy with EPO and ES as well as the electrophysiological and histopathological features of the injured peripheral nerve.

Effect of vitamin E on oxidative stress status in small intestine of diabetic rat

AIM:To investigate the effect of vitamin E on oxidative stress status in the small intestine of diabetic rats. METHODS:Twenty-four male Wistar rats were randomly divided into three groups:Control (C),non-treated diabetic (NTD) and vitamin E-treated diabetic (VETD) groups. The increases in lipid peroxidation,protein oxidation and superoxide dismutase (SOD) in these three groups was compared after 6 wk. RESULTS:There was no significant difference in catalase activity between NTD and control rats. Compared to NTD rats,the treatment with vitamin E significantly decreased lipid peroxidation and protein oxidation,and also increased catalase activity and SOD. CONCLUSION:The results revealed the occurrence of oxidative stress in the small intestine of diabetic rats. Vitamin E,as an antioxidant,attenuates lipid peroxidation and protein oxidation,and increases antioxidant defense mechanism.

Effect of mesenchymal stem cells on anti-Thy1,1 induced kidney injury in albino rats

Objective:To evaluate the effect of mesenchymal stem cells(MSCs)in rats with anti-Thy 1,1nephritis.Methods:Female albino rats were divided into three groups,control group,anti-Thy 1,1group and treatment with i.v.MSCs group.MSCs were derived from bone marrow of male albino rats,Y—chromosome gene was detected by polymerase chain reaction in the kidney.Serum urea and creatinine were estimated for all groups.Kidney of all studied groups was examined histologically and histochemically(total carbohydrates and total proteins).DNA fragmentation and expression ofα-SMA were delected.Results:Kidney of animals injected with anti-Thy 1,1showed inflammatory leucocytic infiltration,hypertrophied glomeruli,tubular necrosis and congestion in the renal blood vessels.The kidney tissue also showed reduction of carbohydrates and total proteins together with increase in apoptosis and in expression ofα-SMA.Moreover,the levels of urea and creatinine were elevated.Treating animals with MSCs revealed that kidney tissue displayed an improvement in the histological and histochemical changes.Apoptosis andα-SMA expression were decreased,and the levels of urea and creatinine decreased.Conclusions:The obtained results demonstrated the potential of MSCs to ameliorate the structure and function of the kidney in rats with anti-Thy 1,1 nephritis possibly through the release of paracrine growth factor(s).