The etiology of inflammatory bowel disease (IBD) is not clear and cytomegalovirus (CMV) infection is often associated with IBD patients. The etiologic link between IBD and CMV infection needs to be studied. The objective of the present study is to investigate the prevalence and risk factors of CMV in a cohort of IBD patients from Central China.Two hundred and twenty six IBD patients (189 ulcerative colitis (UC) and 37 patients with Crohn’s disease (CD)), and 290 age and sex matched healthy controls were recruited. CMV DNA was detected by nested PCR, while serum anti-CMV IgG and anti-CMV IgM was determined by ELISAs. Colonoscopy/enteroscopy with biopsy of diseased tissues and subsequent H&E stain were then conducted in IBD patients with positive anti-CMV IgM. Finally, we analyzed the prevalence and clinical risk factors of CMV infection in IBD patients.P < 0.05), However, anti-CMV IgM positive rate was no different with healthy controls . In univariate analysis of risk factors, the recent use of corticosteroid was associated with increase of CMV DNA and IgM positive rate in UC , aminosalicylic acid drug therapy was correlated with positivity of CMV DNA and IgG in UC and CMV DNA in CD , the treatment of immunosuppresent was correlated with CMV IgM (P < 0.001). Furthermore, patients with severe UC were significantly associated with CMV DNA and IgM . Malnutrition  was also found to be related with CMV recent infection (P = 0.031). In multivariate analysis of risk factors in UC, pancolitis was significantly associated with CMV DNA positivity (P = 0.001). Severe UC and pancolitis seemed to be related with IgG positivity. For CD, there was just single factor associated with CMV positive in each group, multivariate analysis was unnecessary.The prevalence of CMV DNA and anti-CMV IgG positive rate in IBD patients were 84.07% and 76.11%, respectively, higher than those in healthy controls (59.66% and 50.69%, respectively, CMV positive rate in IBD patients was significantly higher, than in healthy controls. The use of aminosalicylic acid, corticosteroid, immunosuppressants, pancolitis and severe IBD patients seemed to be more susceptible to CMV infection in univariate analysis of risk factors. However, no risk factor was found to be significantly correlated with CMV infection in multivariate analysis of risk factors. Inflammatory bowel disease (IBD) including ulcerative colitis (UC) and Crohn’s disease (CD), consists of chronic, non-specific inflammatory diseases of the gut with unknown etiology. According to a retrospective survey, the incidence and prevalence of IBD in China are on the rise, and IBDCytomegalovirus (CMV) is a β herpes virus with double-stranded DNA. Worldwide the current infective rate ranges between 40% and 100%. Two stuCMV infection in IBD patients often makes clinical diagnosis and treatment complex. Maher et al. have shoNested PCR of CMV-UL93 is considered to be a highly sensitive method for the detection of CMV. Serum aIn this study, PCR detection of CMV DNA, and serological determinations of IgG and IgM were used to investigate the prevalence of CMV infection in IBD patients from central China. Moreover, risk factors for CMV infection in IBD patients were analyzed to find relationship between IBD and CMV infection.From 2006 to 2011, 226 IBD patients (189 UC and 37 patients with CD) were recruited from Zhongnan Hospital of Wuhan University School of Medicine. The diagnosis of UC and CD was based on clinical, laboratory, imaging, endoscopic and histological examinations, and in accordance to the Chinese Medical Association diagnostic criteria for IBD. ClinicaSimultaneously, 290 sex and age matched healthy volunteers (controls) were recruited in Zhongnan Hospital Medical Center of Wuhan University, who attended this center for constitutional examination of health. All volunteers were unrelated to each other from Hubei province, and had no history of IBD, chronic infectious diseases, immune-mediated, ischemic and radiation-induced diseases. Subjects with history of use of corticosteroids and immunosuppressive agents, of drug abuse and unhealthy living were excluded from the study.The patients and controls were classified as current smokers if they had smoked more than 1 cigarette per day within 6 months before recruited, and nonsmokers if they never or rarely smoked. As for definition of alcohol drinking before the onset of symptoms, 3 categories were included: frequent drinking, light drinking and former drinking. Frequent drinking was defined as alcohol drinking 3 days or more per week for continuous 6 months before recruited, excluded drink everyday to alcoholic intoxication. Light drinking was defined as alcohol beverages less than 3 days per week. Former drinking was defined as patients who had quit drinking for more than 6 months before recruited. They were all defined as drinking. Non-drinking was defined as never or rarely drinking. Mixed food was defined as the composition of diet include vegetable and meat for at least 6 months before recruited. The severity of UC was assessed by Truelove and Witts criteria: Severe The ethic committee of Zhongnan Hospital of Wuhan University approved the study. Consent was written by all subjects. Consent informed in current ethics statement was obtained from all participants involved in this study.5 ml venous blood was taken in anticoagulated tubes with ethylenediamine tetraacetic acid (EDTA) from all subjects, followed by centrifugation. 2 ml of sera was taken and stored at −80 degrees for further anti-CMV IgG and IgM assay. Genomic DNA was extracted using proteinase K and phenol/chloroform method, which was then subsequently stored at −80 degrees.′-GGCAGCTATCGTGACTGGGA-3′, and a downstream primer 5′-GATCCGACC CATTGTCTAAA-3′. PCR conditions included 40 cycles of 95°C for 10 min, 95°C for 30 s, 57°C for 30 s, 72°C for 60 s, and then followed by 72°C for 10 min. Inner primers were an upstream primer 5′-TTAGCGCGT GACCTGTTACG-3′, and a downstream primer 5′-TCTAAATTGTTACGCAGTCCG-3′. PCR conditions included 40 cycles of 95°C for 10 min, 95°C for 30 s, 58°C for 30 s, 72°C for 60 s, and finally followed by 72°C for 10 min. Then electrophoresis using 2.5% agarose gel, of the medial segment was done to identify the products of PCR. Direct sequence for PCR products was done for detection of positive and negative PCR products. DNA-PCR+ was according to the result of electrophoresis of nested PCR, and ensured the result by DNA sequencing.The CMV-UL93 fragment was retrieved from NCBI and BLAST, and was imported into Primer 5.0, according to the primer design principle. Lateral primers consisted of an upstream primer 5ELISA kit  was used to detect serum anti-CMV IgG and IgM in IBD patients and healthy controls. CMV IgG+ and CMV IgM+ were according to the kit: Positive defined as optical density (OD) ratio of sample to standard threshold value greater than 1.1, negative below 0.9.Colonoscopy and/or enteroscopy were conducted in anti-CMV IgM-positive IBD patients, and biopsies from pathological sites were taken. H&E staining for the detection of CMV was done.2(Chi-square) test with Yates continuity correction or Fisher’s exact test was performed to compare frequencies of risk factors between the IBD patients and healthy controls. Multiple logistic regression analysis was performed to evaluate multiple risk factors for IBD. Odds ratio (OR) and 95% confidence intervals (CI) were calculated. All calculated P-values were 2-sided and P < 0.05 was considered significant.SPSS 13.0 software  was used to conduct the statistical analysis. Measurement data are presented as mean ± standard deviation (SD), and numeration data are expressed as percentage and the number of cases. Independent samples were analyzed by Levene’s test. χAs shown in Table P < 0.001; 76.11% vs 50.69%, P < 0.001), However, prevalence of anti-CMV IgM was no different with healthy controls . For UC patients, CMV-UL93 and anti-CMV IgG were all significantly higher than in healthy controls , While there was no difference between UC and healthy controls for anti-CMV IgM (P = 0.344). For CD patients, CMV-UL93 and anti-CMV IgG were higher than in controls ,while anti-CMV IgM was not increased as compared to healthy controls (P = 0.540).As shown in Table However, in biopsies taken from the pathological sites of intestinal mucosa of anti-CMV IgM positive IBD patients, no inclusion bodies were detected in H&E stain.P = 0.048), use of ASA/SASP  and corticosteroids therapy , while other factors, such as age, sex, smoking, alcohol consumption, type of diet, fever, anemia, albumin level, disease location, treatment with immunosuppressive agents had not shown significant association (P > 0.05). As for CD patients, CMV DNA positive was positively associated with use of 5-ASA/SASP (P = 0.014), as seen in Table In UC patients, elevated CMV-DNA was mainly associated with the severity of disease activity (P < 0.001) as shown in Table P = 0.010). Other factors had no statistically significant impact on anti-CMV IgG positive rate (P > 0.05).In UC patients, anti-CMV IgG level was associated with the use of 5-aminosalicylic acid (P = 0.031), severe UC (P = 0.031), use of corticosteroids (P = 0.015), and the use of immunosuppressive agents (P < 0.001), while other factors did not cause any statistically significant changes in anti-CMV IgM positive rate (P > 0.05). In CD patients, no statistically significant association with anti-CMV IgM was found, as shown in Table The positive rate of anti-CMV IgM in UC patients, was associated with low albumin level . Severe UC and pancolitis seemed to be related with IgG positive . For CD, there was just single factor associated with CMV positive in each group, multivariate analysis was unnecessary.As shown in Table In vivo it can proliferate in epitheliums, white blood cells and sperm cells, and it is prone to cause latent infection of salivary gland, mammary gland, kidney and white blood cells. In IBD patients, immunosuppressive therapy, impaired absorption of nutrients, dysfunction of the immune system, render them susceptible to CMV infection[CMV is an opportunistic pathogenic microorganism. nfection, which iThe detection methods of CMV infection in IBD patients included DNA detection, serological tests , histopathology (inclusion bodies detection) in this study. Body fluids or tissue sample was feasible to CMV culture, but it was time-consuming and also had low sensitivity, which limited clinical application. The detThe positive rate of CMV UL-93 or anti-CMV IgG in the healthy controls was about 50–60%, whereas in IBD patients it was around 70–80%. A research conducted in India showed the CMV DNA positive rate was just 12.70% in IBD patients, which wP = 0.235), which was related with small amount of people included.Anti-CMV IgM positive rate is only 1.77% in our research, which was lower as compared to an Indian study, including IBD patients with both active disease and in remission, where tPresence of anti-CMV IgM indicated recent infection of CMV. However in biopsies taken from the pathological sites of intestinal mucosa of anti-CMV IgM positive IBD patients, no inclusion bodies was detected. This was probably due to the low sensitivity of histological examination. The site from where the biopsy was taken, and the amount of tissue retrieved, may also influence the sensitivity of finding inclusion bodies. Next stage we should recruit enough biopsies and screen CMV by immunohistochemistry.In a model of multivariate analysis adjusting for multiple factors for UC. Disease location seemed to be significantly associate with CMV infection or re-activation, this may be associated with the theory that CMV was prone to proliferate in granulation tissue. PancoliCurrently there is no absolute indication for antiviral therapy in CMV-positive IBD patients. However Eddleston recommends antiviral therapy in immunocompetent. Pfau2 found ouIn summary, as compared to healthy individuals, IBD patients have a predisposition to CMV infection. No risk factor was found to be significantly correlated with CMV infection in risk factors analysis.CMV: Cytomegalovirus; IBD: Inflammatory bowel disease; UC: Ulcerative colitis; CD: Crohn’s disease; PCR: Polymerase chain reactio; OR: Odds ratio; CI: Confidence interval; Alb: Albumin; H&E: Hematoxylin and eosin; CDAI: Crohn’s disease activity index; SD: Standard deviation; ASA: Aminosalicylic acid; SASP: Salicylazosulfapyridine.The authors declare that they have no conflict of interests.FY, JZ, YL and BX conceived the study, participated in its design and coordination, and managed the preparation of the manuscript. CW and SH carried out the nested PCR, FY and LS carried out ELISAs, All authors participated in the recruitment of specimens and clinical data of patients and healthy controls. RL and FY participated in the manuscript writing. BX and FY performed the statistical analysis. WW participated in the manuscript revision. All authors read and approved the final manuscript.


Prevalence of diabetes mellitus (DM) has reached epidemic proportions in Sri Lanka. Presently there are studies on the community prevalence of distal peripheral neuropathy (DPN) in Sri Lanka. We describe prevalence, patterns and predictors of DPN in patients with DM in Sri Lanka. Data were collected as part of a national study on DM. In new cases DPN was assessed using the Diabetic-Neuropathy-Symptom (DNS) score, while in those with established diabetes both DNS and Toronto-Clinical-Scoring-System (TCSS) were used. A binary logistic-regression analysis was performed with ‘presence of DPN’ as the dichomatous dependent variable and other independent co-variants. The study included 528 diabetic patients (191-new cases), with a mean age of 55.0 ± 12.4 years and 37.3% were males, while 18% were from urban areas. Prevalence of DPN according to DNS score among all patients, patients with already established diabetes and newly diagnosed patients were 48.1%, 59.1% and 28.8% respectively. Prevalence of DPN in those with established DM as assessed by TCSS was 24% and the majority had mild DPN (16.6%). The remainder of the abstract is based on subjects with established DM. The prevalence of DPN in males and female was 20.0% and 26.4% respectively. The mean age of those with and without DPN was 62.1 ± 10.8 and 55.1 ± 10.8 years respectively (p < 0.001). The majority of those with DPN were from rural-areas (75.3%) and earned a monthly income < Sri Lankan Rupees 12,000 (87.6%). In the binary logistic-regression presence of foot ulcers , female gender  and smoking  were the strongest predictors followed by insulin treatment , diabetic retinopathy , treatment with sulphonylureas , increasing height , rural residence , higher levels of triglycerides  and longer duration of DM . There is a high prevalence of DPN among Sri Lankan adults with diabetes. The study defines the impact of previously known risk factors for development of DPN and identifies several new potential risk factors in an ethnically different large subpopulation with DM. Diabetes mellitus (DM) has reached epidemic proportions worldwide. Historically, diabetes was considered a disease confined to developed countries and affluent people. However, recent estimates suggest that the prevelence of diabetes is rising globally, particularly in developing countries . DiabeteIncreasing age, longer duration of diabetes and poor glycaemic control are well recognized risk factors for DPN, while cigarette smoking, retinopathy, hypertension, obesity, hyperlipidaemia and microalbuminuria have also been implicated as potential risk markers . Most prSri Lanka is a developing country in the South Asian region with a population of 20.9 million . The preData were collected as part of a wider cross-sectional national study on diabetes conducted in seven of the nine provinces in Sri Lanka between August 2005 and September 2006 (Sri Lanka Diabetes and Cardiovascular Study – SLDCS). In the SLDCS, the research team randomly selected 100 clusters (of 50 adults each) to represent seven out of nine provinces in the country. The sample sizes for each province were determined using a probability-proportional to-size (PPS) technique based on total population of each province. Each cluster was selected by a computer-generated random number list from the ‘Village Office Units’ in each province. Voter registration lists were used to randomly select the first household in each cluster and a uniform criterion was used to select the remaining 49 households. Detailed sampling has been previously reported . Relevan2, based on WHO criteria for Asians ). Details of subject recruitment are presented in Figure 1.9% withThe remainder of this article is based exclusively on subjects with established DM only (n = 337), in whom both symptoms and signs of neuropathy were evaluated. DPN is considered to be present in all subjects with TCSS > 5. In subjects with established DM the prevalence of DPN according to the TCSS score was 24.0% and a majority was suffering from mild neuropathy n = 56, 16.6%)  declare that they have no competing interests.PK, GRC, DRM and MHRS made substantial contribution to conception and study design. PK and GRC were involved in data collection. RJ and PR were involved in refining the study design, statistical analysis and drafting the manuscript. RJ, PR, MHRS, GRC and PK critically revised the manuscript. All authors read and approved the final manuscript.


Helicobacter pylori (H. pylori) infection leads to different clinical and pathological outcomes in humans, including chronic gastritis, peptic ulcer disease and gastric neoplasia and even gastric cancer and its eradiation dependst upon multi-drug therapy. The most effective therapy is still unknown and prompts people to make great efforts to find better and more modern natural or synthetic anti-H. pylori agents. In this report 21 randomly selected herbal methanolic extracts were evaluated for their effect on inhibition of Jack-bean urease using the indophenol method as described by Weatherburn. The inhibition potency was measured by UV spectroscopy technique at 630 nm which attributes to released ammonium. Among these extracts, five showed potent inhibitory activities with IC50 ranges of 18-35 μg/mL. These plants are Matricaria disciforme (IC50:35 μg/mL), Nasturtium officinale (IC50:18 μg/mL), Punica granatum (IC50:30 μg/mL), Camelia sinensis (IC50:35 μg/mL), Citrus aurantifolia (IC50:28 μg/mL). H. pylori, a Gram-negative pathogenic bacterium which specifically colonizes the human gastric mucosa, has been regarded as a primary causative agent of chronic gastritis and peptic ulcer diseases including mucosa-associated lymphoid tissue lymphoma  was shown to contain nickel ions (Ureases (E.C 3.5.1.5), the first enzyme crystallized from Jack been  water. Jack-bean urease was obtained from Merck (5 units/mg).Urease inhibition activity assaybc where c is the concentration of solution (mol/L), b is the length of the UV cell and ε represents molar absorptivity in the specific wavelength, we can calculate the concentration of initially urease solution. After proper dilution, the concentration of enzyme solution adjusts at 2 mg/mL.For urease inhibition assays after addition of 10 mL of phosphate buffer to accurately weight of enzyme, sonication was performed for 60s, followed by centrifugation and evaluating absorbance of upper solution in λ = 280 nm which is attributed to enzyme. By using the following equation A = ε2SO4 acid. Enzyme inhibition activity performed by Berthelot alkaline phenol–hypochlorite method to examine the efficiency of adsorptive immobilization. This method is based on the released ammonia (NH3) which reacts with hypochlorite (OCl−) to form a monochloramine  of Jack-bean urease and 100 μL of the test compound with 0.2 mL of 100 mM phosphate buffer pH 6.8 containing 25 mM urea was pre-incubated for 30 min in water bath at 37°C. The urease reaction was stopped after 30 min incubation with 600 μL of 4% Hloramine . This prThe liberated ammonia was estimated using 500 μL of solution A (contained 5.0 g phenol and 25 mg of sodium nitroprusside) and 500 μL of solution B (contained of 2.5 g sodium hydroxide and 4.2 mL of sodium hypochlorite in 500 mL of distilled water) at 37°C for 30 min and the absorbance was measured at 625 nm against the control. All reactions were performed in triplicate in a final volume of 1 mL. Percentage of inhibitions were calculated using the formula (100 - (OD sample / OD control) × 100).50) was determined through monitoring the inhibition effect of various concentrations of extracts in the assay. The IC50 values were then calculated using above mentioned formula in the previous section.The concentration of compounds that inhibited the hydrolysis of substrate by 50% , Citrus aurantifolia (IC50 = 28 μg/ml), Nasturtium officinale (IC50 = 18 μg/mL), Punica granatum (IC50 = 30 μg/mL) and Nasturtium officinale (IC50 = 35 μg/mL).As shown in H. pylori agents that can help eradicate the invasion and presence of survived H. pylori strains to avoid relapse of gastric ulcer. In this regard, the literature has reported extracts of certain plants such as cashew apple  its H. pylori’s urease inhibition has been proven , is probably due to the diversity of growth, harvesting and extraction methods. Regarding the excess amount of flavon, flavol, catechin epigallocatechin gallate, gallocatechin gallate, gallocatechin, and many other flavonoids in C. sinensis  is attributed to the similar substances found in green tea extract in different amounts. The chemical constituents and functional groups of these compounds play important role in inhibition of urease. The functional groups such as hydroxyl and ketones which are linked with aromatic rings can interact with Ni ions in active site of enzyme, resulting in its inhibition.According to reported investigations on n proven . Achievisinensis , we can Citrus aurantifolia; Omani lime) is a citrus species with a globose fruit, 2.5-5 cm in diameter (1–2 in) that is yellow when ripe but usually picked green commercially. It is smaller, seedier, has a higher acidity, a stronger aroma, and a thinner rind than that of the Persian lime (Citrus x latifolia). It is valued for its unique flavor compared to other limes, with the key lime usually having a more tart and bitter flavor. In Malaya, the juice is taken as a tonic and to relieve stomach ailments and it is given as a vermifuge in combination with oil. In India, the pickled fruit is eaten to relieve indigestion. It have been used as an antiseptic, tonic, an antiscorbutic, an astringent, and as a diuretic in liver ailments, a digestive stimulant, a remedy for intestinal hemorrhage and hemorrhoids, and as a disinfectant for all kinds of ulcers when applied in a poultice . The methanolic extract of dried fruits may contain new substance which is not reported in literature and need more investigation on its isolation and identification.The chemical composition of  amounts . Antibac amounts  but noneMatricaria recutita (Chamomile). Previously, it was shown that the Chamomilla recutita (M. recutita) oil extract is rich in fatty acids, coumarins, terpenes, spiroethers and flavonoids contributing to its medicinal properties   , 37.H. pylori’s growth in very low concentrations of 0.0075% (v/v). The reported MIC50 and MIC90 of Chamomilla recutita (M. recutita) oil extract for H. pylori were 62.5 mg/mL and 125.0 mg/mL, respectively. In addition, it was found that urease production of H. pylori was inhibited by the M. recutita oil extract  could be responsible for the morphological and fermentative changes and subsequent anti-H. pylori activity of the M. recutita ita oil extract. However, a 50 % aqueous methanol extract from M. recutita flowers was also found to be inhibitory against urease, which corroborates previously reported results  are fast-growing, aquatic or semi-aquatic, perennial plants native from Europe to central Asia, and one of the oldest known leaf vegetables consumed by human beings. These plants are members of the family Brassicaceae or cabbage family, botanically related to garden cress and mustard with a peppery, tangy flavor due to high contents of phenethyl isothiocyanate (50 =18 μg/mL) reported in this study among all other extracts, which reported for the first time, is concurrent with its daily usage as digestive aid in people whom suffering gastric upset. Phenethyl isothiocyanate may interact with urease Ni ions and inhibit its catalytic activity. More investigationa are needed to confirm this hypothesis.Watercresses (ocyanate . Many beocyanate -40. The ocyanate . The strPunica granatum (Pomegranate). Previous studies on pomegranate demonstrated its medicinal usage against salmonella  reported here revealed that antibacterial properties of P. granatum against different strands of H. pylori could be at least partially due to inhibition of urease by interaction of ellagitannin and punicalagin with active site of enzyme or modulating its activity by aggregation properties of tannin.The medicinal plant possesses a high amount of tannin (25%). The antimicrobial properties of this substance were well established. Polar fraction of icalagin . The strMedicinal plants, traditional medicinal and other natural sources are still good source for lead discovery. The results of this study revealed that random screening of medicinal plants could lead to introducing new candidate for further studies which, in the end, can help and enhance human health.


Squalidus argentatus (Sauvage and Dabry de Thiersant 1874) is a small-sized freshwater fish which is distributed in Mainland China, Hainan Island and Taiwan. The populations of S. argentatus have dropped sharply probably due to overharvesting and water pollution recently. Eleven polymorphic microsatellite markers were developed for the cyprinid fish S. argentatus. These new markers were tested on 43 individuals collected from Yangtze River and Qiantang River. The number of alleles, observed and expected heterozygosity per locus, in two populations ranged from 3 to 14, from 0.333 to 0.954 and from 0.480 to 0.928, respectively. Only two loci are significantly deviated from Hardy–Weinberg expectations due to the heterozygote deficiency. No significant linkage disequilibrium was detected between the pairwise comparisons of these loci. These polymorphic microsatellite loci will enable us to study the genetic variation, population structure, and conservation genetics of this species in the future. Squalidus argentatus (Sauvage and Dabry de Thiersant 1874)  consists of a group of small-sized freshwater fishes widely found in the lower or middle reaches of Eastern Asia, including Mainland China, Hainan Island and Taiwan. There are at least seven valid species of Squalidus found in mainland China and two endemic species reported from Taiwan [S. argentatus is the only species commonly found in above mentioned areas [S. argentatus has experienced a dramatic decrease probably owing to over-fishing and environmental destruction. In order to develop effective strategies for fishery management and conservation, information, such as the population genetic structure and genetic diversity and divergence of S. argentatus, is important. DNA-based genetic analyses can yield valuable information for assessing levels of variation and population genetic structure in freshwater fishes. Because of their high level of polymorphism and co-dominant inheritance in Mendelian fashion, microsatellites have been widely used as DNA markers in the studies of population structure and conservation genetics of freshwater fishes [S. argentatus, have been conducted, there is no report available on molecular markers isolation for this species. The microsatellite markers identified will allow the undertaking of further genetic studies on S. argentatus and provide valuable genetic information to assist the management and culture of this resource.Genus ed areas . Within r fishes . Althougr fishes  of S. arSqualidus argentatus. In brief, total genomic DNA was extracted from muscle tissue or fins preserved in 95% ethanol, by proteinase K digestion at 55 °C. DNA was purified by traditional phenol-chloroform protocol and ethanol precipitation [2, 0.5 U Taq polymerase (Promega), and 5 pmols of one RAPD primer. Several RAPD primers were used to amplify fragments from the target species’ genome in separate reactions. Reactions of PCR amplification were conducted in a thermal cycler (Eppendorf Mastercycler) using the following conditions: initial denaturing at 94 °C for 3 min; 40 cycles of 94 °C denaturing for 50 s, 37 °C annealing for 1 min, 72 °C extension for 1 min; and 72 °C for 10 min. RAPD-PCR products were size-selected to preferentially obtain small fragments (500–1200 bp). Approximately 100 ng of PCR product was ligated into a pMD 19-T vector (Takara) according to the manufacturer’s instructions, and the ligation mixture was transformed into Escherichia coli DH5α competent cells. Clones were screened of the library for microsatellites by PCR with both vector primers plus a microsatellite specific primer (GT and GA repeats) [In the present study, we developed 11 polymorphic microsatellite loci isolated using PCR-based isolation of microsatellite arrays (PIMA) of pitation . The PCRpitation –9. It tapitation . Amplifirepeats) .2, and 0.12 μM of each primer. PCR reactions were as follows: 94 °C for 4 min followed by 40 cycles at 94 °C for 30 s, 30 s–50 s at primer-specific annealing temperature . Both strands of the DNA insert were sequenced. Sequencing of positive insert PCRs was conducted by Sangon Biotec (Shanghai) Co., Ltd. with an ABI PRISM 3730 sequencer using the BigDye Terminator kit (Applied Biosystems) and by using vector primers. Primer pairs were newly designed for eleven microsatellite sequences with the program PRIMER 3 software . Prelimiperature , 72 °C fHO) and expected (HE) heterozygosities were calculated. All loci were tested for fitness to the Hardy–Weinberg equilibrium (HWE), and all pairwise combinations of loci were tested for linkage disequilibrium. All these parameters and tests were computed using ARLEQUIN 3.5 [Allele frequency, observed  and 0.333–0.857 (average of 0.668), respectively, in Yangtze River. The HE and HO ranged from 0.717–0.928 (average of 0.830) and 0.476–0.954 (average of 0.747), respectively, in Qiantang River. One locus in Yangtze River (MISA10) and in Qiantang River (MISA11) is significantly deviated from Hardy–Weinberg equilibrium after Bonferroni’s correction (P < 0.001) due to the heterozygote deficiency, respectively (P < 0.05), but for stuttering and allelic dropout tests, no evidences were found in all loci (P > 0.05). After applying sequential Bonferroni corrections to compensate for multiple statistical tests, no evidence of genotypic linkage disequilibrium at any pair of loci was found. Microsatellite makers described here should be useful to monitor population dynamics and to determine dispersal patterns and genetic diversity within and between populations of this species. These polymorphic microsatellite loci will be used to investigate population structure and gene flow among S. argentatus populations and are expected to provide a better understanding about the ecology of the species, and consequently a basis for a sustainable management framework in the future.A total of 43 ectively . These dSqualidus argentatus has experienced a dramatic decrease in the last decade. Therefore, there is an urgent need for the development and application of microsatellites to provide an effective tool for investigation of genetic variation and population structure in S. argentatus. In conclusion, we report, these 11 polymorphic microsatellite loci presented in this study were the first set of microsatellite markers designed specifically for S. argentatus, and provides the ground work for further studies on the genetic structure, gene flow, conservation management and molecular evolution of this vulnerable species.The wild resource of


Background: Human milk is a potential source of lead exposure. Yet lactational transfer of lead from maternal blood into breast milk and its contribution to infant lead burden remains poorly understood.Objectives: We explored the dose–response relationships between maternal blood, plasma, and breast milk to better understand lactational transfer of lead from blood and plasma into milk and, ultimately, to the breastfeeding infant.Methods: We measured lead in 81 maternal blood, plasma, and breast milk samples at 1 month postpartum and in 60 infant blood samples at 3 months of age. Milk-to-plasma (M/P) lead ratios were calculated. Multivariate linear, piecewise, and generalized additive models were used to examine dose–response relationships between blood, plasma, and milk lead levels.p = 0.0006) with the M/P ratio decreasing by 16.6 and 0.6 per 0.1 μg/L of plasma lead, respectively, below and above 0.1 μg/L plasma lead. Infant blood lead level (3.4 ± 2.2 μg/dL) increased by 1.8 μg/dL per 1 μg/L milk lead .Results: Maternal lead levels (mean ± SD) were as follows: blood: 7.7 ± 4.0 μg/dL; plasma: 0.1 ± 0.1 μg/L; milk: 0.8 ± 0.7 μg/L. The average M/P lead ratio was 7.7  with 97% of the ratios being > 1. The dose–response relationship between plasma lead and M/P ratio was nonlinear (empirical distribution function = 6.5, Conclusions: The M/P ratio for lead in humans is substantially higher than previously reported, and transfer of lead from plasma to milk may be higher at lower levels of plasma lead. Breast milk is an important determinant of lead burden among breastfeeding infants.http://dx.doi.org/10.1289/ehp.1307187Citation: Ettinger AS, Roy A, Amarasiriwardena CJ, Smith DR, Lupoli N, Mercado-García A, Lamadrid-Figueroa H, Tellez-Rojo MM, Hu H, Hernández-Avila M. 2014. Maternal blood, plasma, and breast milk lead: lactational transfer and contribution to infant exposure. Environ Health Perspect 122:87–92;  Human milk has been suggested as a significant potential source of infant lead exposure due to the redistribution of cumulative maternal lead stores associated with bone resorption of pregnancy and lactation . MaternaThere are some data from rodents on the lactational transfer and uptake of lead in the neonate. Because maternal blood is the matrix from which lead is transferred to breast milk and, ultimately, to the breastfeeding infant, the relationship of lead in maternal blood and in breast milk is of key importance. Early studies supported the belief that human milk levels were one-tenth to one-fifth the levels of lead in maternal blood . HoweverTo better understand the distribution and transfer of lead during lactation, we measured maternal plasma, blood, and breast milk lead concentrations at 1 month postpartum in a study of lactating women in Mexico City, Mexico. We also explored the relationship between breast milk lead and infant blood lead levels among their breastfeeding infants.Study subjects. Subjects were participants in a longitudinal study of lead biomarkers and reproduction which has been described in detail elsewhere  using gravity-assisted procedures to limit potential hemolysis. Samples were processed and shipped frozen to the trace metal facility at the University of California, Santa Cruz, for measurement of plasma lead using ultra-clean methods detailed elsewhere  for blood lead analysis. Graphite furnace atomic absorption spectrophotometry  was used to quantify blood lead according to a technique described by Breast milk lead measurement. Breast milk samples were collected from lactating women using specific techniques designed to minimize potential for environmental contamination and any potential variability in breast milk lead over time . Before manual expression of milk, the breast was washed with deionized water that also was collected and analyzed for lead contamination. Ten milliliters of milk was collected in preleached polypropylene containers. Samples were shipped frozen and stored at –30°C  until analysis. Breast milk sample preparation was performed at University of Massachusetts Research Institute for Analytical Chemistry , and instrumental analysis was performed at the Trace Metals Laboratory of Harvard School of Public Health. Digestion was performed using nitric acid in high temperature high pressure asher (HPA) , and lead content in the samples was analyzed by isotope dilution–inductively coupled plasma mass spectrometry (ID-ICPMS)  by methods previously described in detail  and plasma lead levels > 1 μg/L (n = 2) were excluded. Milk and plasma lead concentrations are expressed in parts per billion (ppb), whereas blood lead concentrations, typically expressed in micrograms per deciliter, were converted to micrograms per liter as necessary to calculate transfer indices . The M/P ratio was calculated by dividing breast milk lead concentration (micrograms per liter) by plasma lead concentration (micrograms per liter).p-values reported. Nonparametric smoothing  was used to explore the shape of the associations between the different lead biomarkers.Bivariate analyses were carried out to explore the relationships between the different biomarkers of lead exposure. Spearman tests of correlation were used and correlation coefficients (ρ) with a priori knowledge of biological significance and review of the scientific literature. Covariates were retained in the models if the main effect estimate of plasma or blood lead on breast milk lead changed by > 10% or if it increased the model coefficient of determination (R2) by > 10%. To explore potential nonlinear associations, we also examined the relationships between the variables using nonparametric regression with generalized additive models (GAMs). The GAM models allowed us to examine possible nonlinear effects using a smooth functional term corresponding to plasma lead in the multivariate model. The smoothing term was fitted by using penalized regression splines with degree of smoothness selected by cross-validation technique, allowing us to determine the functional relationship between the different matrices in a flexible, data-adaptive way instead of being restricted to a linear relationship. Where the GAMs indicated a nonlinear relationship, we explored the effects using piecewise linear models to describe the trend.Multivariable linear regression models were fitted using ordinary least-squares methods to estimate associations between the blood or plasma and breast milk lead levels. Given the limited sample size available for analysis, extensive covariate adjustment was not feasible. We chose to investigate covariates  based on We explored the contribution of breast milk lead to infant blood lead levels among a subset of mother–infant pairs using multivariate regression accounting for breastfeeding duration (months of exclusive breastfeeding) over the preceding 3 months of the infant’s life. Stratified and interaction models were also fitted to determine whether breastfeeding status  altered this relationship. Sensitivity analyses of the breast milk–infant blood lead association were carried out to assess the additional contribution of lead from umbilical cord blood (where available) among 36 mother–infant pairs, accounting for infant birth weight (as a proxy for the amount of milk consumed) and breastfeeding status. All statistical analyses were performed using SAS® for Windows, version 9.3  and R Programming Language .Maternal lead levels at 1 month postpartum were as follows: blood: 7.7 ± 4.0  μg/dL; plasma: 0.1 ± 0.1  μg/L; and breast milk: 0.8 ± 0.7  μg/L. The plasma-to-blood ratio ranged from 0.06 to 0.42, and the average M/P lead ratio was 7.7  with 97% of the ratios being > 1. Infant blood lead at 3 months of age was 3.4 ± 2.2  μg/dL . Signifip < 0.0001); however, the multivariate adjusted GAM model indicated that the association was linear [empirical distribution function (EDF) = 1] (data not shown). As p = 0.05). The multivariate linear relationships, adjusted for maternal age, dietary calcium intake and systolic blood pressure, between maternal blood lead, plasma lead, and breast milk lead are reported in p = 0.002).The adjusted plasma–breast milk lead relationship was observed to be nonlinear with considerable variability over the range of plasma lead levels . Below ap-value for interaction = 0.06), indicating effect modification by breastfeeding status  (data not shown). Among infants exclusively breastfed, breast milk and umbilical cord blood explained 37% and 33% of variation in infant blood lead, respectively, compared with 10% and 50% of the variation among infants not exclusively breastfed in the preceding month.Sensitivity analyses indicate that this relationship remained even after accounting for umbilical cord blood lead and birth weight; the increase in infant blood lead per unit change in breast milk lead was higher by 2.3 μg/dL among children exclusively breastfed compared with those not exclusively breastfed in the previous month , and breast milk (the exposure medium for breastfeeding infant) are complex. The relationship between maternal blood and plasma in our population had a curvilinear association best described by a log-linear function, suggesting variability in partitioning of lead between the erythrocyte component of blood and plasma over the range of lead exposure. This is very similar to that observed among 63 nonpregnant  and 237 Our results suggest that the transfer of lead from plasma to breast milk was also variable across the range of plasma lead. The M/P ratios reported here for lead are higher than previously estimated in humans as ≤ 1.0, and may actually be more similar to results reported by earlier animal studies where lead concentrated in breast milk. In fact, the M/P ratio for lead was found to be nonlinear, with the highest M/P ratios at the lowest levels of plasma lead, which dropped off steeply until around 0.1 μg/L plasma lead and then tapered off. This could be explained by passive transfer of lead into breast milk driven by high affinity compartment in milk, thus allowing binding at low concentrations and drawing lead into milk. However, this high affinity compartment would have a low overall capacity, leading to rapid saturation at around 0.1 μg/L plasma lead and the consequent leveling off of the M/P ratio as passive transfer resumes (driven only by the increase in plasma lead levels).A possible candidate for this high-affinity, low-capacity compartment in milk could be the protein casein, which is involved in calcium transport into milk. Several studies indicate that milk casein binds lead with high affinity in rats, mice, cows, and humans [reviewed by Current recommendations for cessation of breastfeeding at maternal blood lead levels > 40 μg/dL are based on an M/P ratio assumed to be < 1 for lead , 2005. WAt high blood lead concentrations (> 40 μg/dL), the relationship between the concentration of lead in serum and its concentration in blood becomes nonlinear . The bloThe proposed nonlinearity in the milk-to-blood lead relationship has been observed in lactating mice and rats with blood lead concentrations > 3 μg/dL  as well The potential magnitude and range of increases in the milk-to-blood lead ratio at maternal blood lead concentrations > 40 μg/dL are not well quantified in humans and are outside the range of blood lead levels in our current study. However, although the milk-to-blood lead ratio may increase as maternal blood lead concentrations exceed 40 μg/dL, the milk-to-plasma lead ratio should not change significantly at higher maternal blood lead concentrations. Nonetheless, the possibility that milk lead increases disproportionately to blood lead in women with high blood lead levels should be considered when discussing breastfeeding with women whose blood lead levels exceed 40 μg/dL.Our findings here do not suggest that measurement of maternal plasma or breast milk lead are likely to become clinically useful environmental health measures because these methods require technically challenging ultra-clean specimen collection and analysis protocols. However, when properly collected and analyzed, these biological markers of exposure are useful tools in research efforts to better understand lead kinetics in the lactating woman and breastfeeding infant. We used different laboratories and analytic procedures for quantification of lead in the various biological matrices to employ the best available methods for the key matrices of interest (plasma and breast milk). Blood lead is a surrogate, measured with error, for the toxicologically available lead fraction, and larger effect size estimates observed in studies using plasma lead suggest many previous studies using blood lead may have had effect estimates downwardly biased by measurement error.Given the unique nutritional characteristics of human milk, breastfeeding is understood to be the optimal mode of nutrient delivery to full-term infants. The benefits of breastfeeding are so compelling that very few situations definitively contraindicate breastfeeding . Howeverin utero exposures. Although a limitation of the present study is that we did not have concurrent infant blood lead levels available at 1 month postpartum, we made the assumption that the ranking of lead levels within the population remained stable over the first 3 months of infant life, and we also accounted for breastfeeding practices in our models.Our results suggest that breast milk lead is a significant and important source of infant lead exposure accounting for approximately 30% of variation in infant blood lead levels. This was higher among infants who were exclusively breastfed (37%) than among those not exclusively breastfed (10%), suggesting that breast milk is an important contributor to infant lead burden on top of any concurrent environmental and r = 0.32, p < 0.0001) with infant blood lead levels at 1 month of age . This was higher among infants exclusively breastfed in the previous month (2.2 μg/dL) compared with breastfeeding infants who were not exclusively breastfed in the preceding month (1.1 μg/dL) .Previously we have reported that even among a population of women with relatively high cumulative lifetime exposures to lead, levels of lead in breast milk were low, influenced both by current lead exposure and by redistribution of bone lead accumulated from past environmental exposures . Although of age . In thath of age . In the r = 0.42, p = 0.0003)  . Despite 0.0003) , the hig 0.0003) . To the  0.0003) . Rabinow 0.0003) . Nonethe 0.0003)  and othein utero exposure. This has implications for policy decisions regarding counseling the lead-exposed woman on breastfeeding, because current recommendations appear to be based on limited empirical evidence.In this study we measured lead in several maternal and infant biomarkers simultaneously, including blood, plasma, and breast milk, using state-of-the-art, ultra-clean methods for specimen collection and laboratory analysis. We found that the M/P ratio for lead in humans is substantially higher than previously reported as being ≤ 1.0, and the transfer of lead from plasma to breast milk may be higher at lower levels of plasma lead. Breast milk represents an additional important source of lead exposure to breastfeeding infants over and above the contribution from


Intravenous fluid is recommended in international guidelines to improve patient post-operative symptoms, particularly nausea and vomiting. The optimum fluid regimen has not been established. This prospective, randomized, blinded study was designed to determine if administration of equivolumes of a colloid (hydroxyethyl starch 130/0.4) reduced post operative nausea and vomiting in healthy volunteers undergoing ambulatory gynecologic laparoscopy surgery compared to a crystalloid solution (Hartmann’s Solution).-1 per hour of fasting. Patients were interviewed to assess nausea, vomiting, anti-emetic use, dizziness, sore throat, headache and subjective general well being at 30 minutes and 2, 24 and 48 hours post operatively. Pulmonary function testing was performed on a subgroup.120 patients were randomized to receive intravenous colloid (N = 60) or crystalloid (N = 60) intra-operatively. The volume of fluid administered was calculated at 1.5 ml.kgvs 17.9%, P = 0.03) and the mean nausea score were increased in the colloid compared to crystalloid group respectively . The incidence of vomiting and anti-emetic usage was low and did not differ between the groups. Sore throat, dizziness, headache and general well being were not different between the groups. A comparable reduction on post-operative FVC and FEV-1 and PEFR was observed in both groups.At 2 hours the proportion of patients experiencing nausea (38.2 % Intra-operative administration of colloid increased the incidence of early postoperative nausea and has no advantage over crystalloid for symptom control after gynaecological laparoscopic surgery. The incidence of post operative nausea and vomiting (PONV) following ambulatory surgery is 40%–60% and ambulatory gynecological patients are at particularly high risk. PONV continues to occur despite pharmacological prophylaxis in high risk groups. -6 PONV h‘Rehydration’ is a recommended strategy to reduce PONV but have been subject to a relatively small number of studies in ambulatory anesthesia and there is no consensus as to which fluid or volume is optimal. -18 Studi-1.hr-1 of fasting time administered as an intra-operative bolus. Compound Sodium Lactate  contains sodium 131 mmol litre−1, potassium 5 mmol litre−1, calcium 2 mmol litre−1, chloride 111 mmol litre−1 and lactate 29 mmol litre−1. The colloid (CLD) group received 6% Hydroxy Ethyl Starch 130/0.4 in 0.9% saline  at a volume of 1.5 ml.kg-1.hr-1 of fasting time as an intra-operative bolus. To ensure blinding of both the patient and the investigator, all fluid administration was initiated after induction of anesthesia and completed prior to transfer to PACU. No further fluid was administered after this time. Neither the patient nor the investigator was aware of group assignment.Following approval by the Institutional Research Ethics committee , ASA I-II female patients, aged 18–45 years who were scheduled to undergo elective gynecological laparoscopy were invited to participate in the study. The study took place in the operating theatres, recovery room and day ward of the Rotunda Hospital, a tertiary level university teaching hospital. Exclusion criteria included age <18 or >45 years, cardio-respiratory disease, excessive intra-operative blood loss, BMI >30 and relevant drug allergy to medication used in the protocol. Informed written consent was obtained from all patients. Patients were randomized using a table of computer generated numbers each of which was linked to a sealed envelope that contained the allocation to one of two groups. The crystalloid group (CRL) received Compound Sodium Lactate at a volume of 1.5 ml.kg-1, Propofol 2–3 mg.kg-1 and Atracurium 0.35 mg.kg-1 to facilitate airway control. The lungs were ventilated via a supraglottic airway (LMA Supreme™TM size 4) with tidal volume (Vt) 4–6 ml.kg-1, maximum airway pressure (P max) 30 cm H2O and a positive end expiratory pressure (PEEP) of 5 cm H2O. The respiratory rate was adjusted to maintain isocapnia. Anaesthesia was maintained with 1 MAC of Sevoflurane in a mixture of O2 and air to achieve a FiO2 of 0.5. Muscle relaxation was antagonized with neostigmine 2.5 mg and glycopyrrolate 0.5 mg at the end of surgery. Before discontinuation of anesthesia, each patient received rectal Diclofenac 100 mg and Paracetamol 1000 mg i.v. Prophylactic anti-emetics were not administered.All patients received general anesthesia. Intravenous access was obtained after local anesthetic infiltration of the skin. Anesthesia was induced with fentanyl 1.5 mcg.kgi.v. as required to achieve a pain score of 4 or less. On the ward Pethidine 50 mg intramuscularly or simple oral analgesics .  Secondart test or repeated measures analysis of variance as appropriate. Normally distributed data are presented as means and standard deviation and data that was not normally distributed is presented as medians and interquartile range. Categorical data is presented as proportions and percentages.Statistics were performed using Sigma Stat, (Jandel Scientific). Categorical data were analyzed using the χ 2 test or the Fisher's exact test as appropriate. Continuous data were analyzed with the Student-Sample size was calculated as follows: To detect a clinically significant difference of 25% reduction in mean nausea scores based on a baseline score of 40 mm, we calculated a sample size of 60 patients per group .There were no baseline differences between the groups in terms of age, weight, BMI, risk factors for PONV, fasting duration or anaesthetic technique Table .vs 17.9%, P = 0.03)  (Figure At 2 hours the proportion of the patients experiencing nausea (38.2 One patient was withdrawn from the colloid group after developing high airway pressures in the operating room. In PACU a chest x-ray showed pulmonary edema which was treated with diuretics and oxygen. She was well at follow up.Multiple previous prospective randomized clinical trials have demonstrated a reduction in post-operative nausea and vomiting associated with intravenous crystalloid administration which is dose related. ,24,27,28-1 of Ringers lactate or 12 ml.kg-1 4.5% Hydroxyethyl starch prior to induction of anaesthesia. This study was notable for an extremely high incidence of PONV. [-1 of one of three colloidal study solutions of low, medium and high molecular weight respectively. [There have been a small number of related studies in diverse patient populations to date and none in ambulatory care specifically. The results of these studies have been conflicting. Haentjens, in a prospective randomised trial investigated the effect of hydroxyethyl starch 130/0.4 or saline in a mixed surgical population undergoing non-ambulatory surgery and showed no difference between the groups in nausea and vomiting.  The voluof PONV.  The collctively.  The studIn the studies which have been carried out to date on the effects of intravenous fluids on PONV after gynaecological laparoscopy, the specific mechanism of action of fluid has not been determined. Intravenous fluids have not been found to be beneficial in thyroidectomy and therapeutic abortion suggesting that the effect may be specific to laparoscopic surgery. ,36 SpecuFirst, there was no untreated control group and a single dose of colloid was evaluated, therefore beneficial effects of the solution at different doses cannot be excluded. Second, blinding is incomplete as the anaesthetist who administered the fluids was not blinded. Third, the study was conducted in patients without cardiorespiratory disease and use of such volumes of rapidly administered intravenous fluids cannot be recommended to all patients. Fourth, the study was underpowered to demonstrate statistical differences for secondary outcomes, particularly pulmonary function which showed clear trends towards greater impairment in the colloid group. Fifth, following discharge from PACU oral intake was not restricted or measured. Last, prophylactic anti-emetics were not administered, limiting the ability of clinicians to extrapolate of our findings to patients who receive such interventions.Our study has a number of limitations. In conclusion, we were unable to demonstrate a benefit of colloid over crystalloid in PONV after ambulatory gynecologic surgery and cannot recommend it at the investigated dose for this purpose. None of the authors have any financial or non-financial competing interests.IH designed the study, analyzed the data and drafted the manuscript. RR collected data and was involved in drafting the manuscript. KE collected data, was involved in its analysis and manuscript drafting. EM was involved in drafting the manuscript. DK collected data. CMC designed the study, co-ordinated it, analyzed the data and drafted the manuscript. All authors saw and approved the final manuscript.Internal Department only.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2253/12/15/prepub


Type 2 diabetes mellitus is a major global public health problem in the worldwide and is increasing in aging populations. Magnesium intake may be one of the most important factors for diabetes prevention and management. Low magnesium intake may exacerbate metabolic abnormalities. In this study, the relationships of magnesium intake with metabolic parameters, depression and physical activity in elderly patients with type 2 diabetes were investigated.This cross-sectional study involved 210 type 2 diabetes patients aged 65 years and above. Participants were interviewed to obtain information on lifestyle and 24-hour dietary recall. Assessment of depression was based on DSM-IV criteria. Clinical variables measured included anthropometric measurements, blood pressure, and biochemical determinations of blood and urine samples. Linear regression was applied to determine the relationships of magnesium intake with nutritional variables and metabolic parameters.p < 0.05). A positive relationship was found between magnesium intake and HDL-cholesterol (p = 0.005). Magnesium intake was inversely correlated with triglyceride, waist circumference, body fat percent and body mass index (p < 0.005). After controlling confounding factor, HDL-cholesterol was significantly higher with increasing quartile of magnesium intake (p for trend = 0005). Waist circumference, body fat percentage, and body mass index were significantly lower with increase quartile of magnesium intake (p for trend < 0.001). The odds of depression, central obesity, high body fat percentage, and high body mass index were significantly lower with increasing quartile of magnesium intake (p for trend < 0.05). In addition, magnesium intake was related to high physical activity level and demonstrated lower serum magnesium levels. Serum magnesium was not significantly associated with metabolic parameters.Among all patients, 88.6% had magnesium intake which was less than the dietary reference intake, and 37.1% had hypomagnesaemia. Metabolic syndromes and depression were associated with lower magnesium intake (The majority of elderly type 2 diabetes who have low magnesium intake may compound this deficiency with metabolic abnormalities and depression. Future studies should determine the effects of increased magnesium intake or magnesium supplementation on metabolic control and depression in elderly people with type 2 diabetes. Type 2 diabetes mellitus (DM) is a major public health problem in the worldwide and is increasing in aging populations ,2. MoreoMagnesium may be one of the most important factors for diabetes prevention and management ,8. IndeeAssessment of magnesium status is important for determining the association of magnesium with metabolic control and cardiovascular disease. A simple method available in a clinical setting is the measurement of serum magnesium. However, serum magnesium levels correlate poorly with magnesium status . The detNumerous clinical studies on magnesium supplementation have suggested that dietary magnesium deficiency may impair metabolic control ,25. HoweThe present study was conducted to investigate magnesium status in elderly type 2 diabetes patients and to determine its relationship with nutritional variables, metabolic control, depression and physical activity. In this cross-sectional study, information about dietary intake, physical activity, and depression were obtained by questionnaires. In additional, anthropometric values, as well as biochemical determinations of blood and urine samples were also investigated.1) Type 2 diabetes for more than 6 months; 2) no change in any medications for the past 3 months; 3) stable lifestyle for the past 3 months; and; 4) aged 65 years and above. Patients with heart failure, cirrhosis, current malignancy, chronic renal failure, or signs of serious deterioration in comprehension and memory were excluded. Anthropometric measurements, biochemical blood and urine tests, as well as dietary assessment were performed within one month. For elderly patients with a low level of education and who were unable to write their signature, children or family members signed consent forms on their behalf.The cross-sectional study was approved by the Changhua Christian Hospital Institutional Review Board (CCHIRB#090419). Patients were residents in a rural area of central Taiwan, and diabetes at the first the Endocrine and Metabolism clinic was diagnosed according to the American Diabetes Association (ADA) criteria : 1). fasDietary intake was assessed using 24-hour recall . In addiAssessment of physical activity is based on a slightly modified version of the method using in a Finnish study . BrieflyAssessment of depression was based on the criteria in Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV), and included the number and severity of symptoms, duration of the current episode, and course of illness. Depression was diagnosed in patients with 5 or more symptoms including at least one key symptom, most days, most of the time for at least 2 weeks, as defined by the American Psychiatric Association .subjects were asked to fast for 8–10 hours on the night prior to measurement. Study measurements were performed by the hospital medical laboratory (certified ISO15189) using standard methods. The glycated hemoglobin (HbA1C) (CV < 3.0%) was measured by ion-exchange chromatography . Serum magnesium, albumin, lipids, urea nitrogen, creatinine, high-sensitivity CRP (hsCRP), microalbumin using a Dimension RxL autoanalyzer . Serum magnesium (CV < 3.0%) was measured using the methylthymol blue method. Serum albumin (CV < 2.0%) was estimated using the bromcresol purple method. Lipids  were assessed using standard enzymatic methods. The high-sensitivity C-reactive protein (hsCRP) (CV < 3.0%) was measured by particle enhanced turbidimetric immunoassay. The levels of creatinine (CV < 2%) in urine and serum were used the alkaline picrate-kinetic method. Blood urea nitrogen (CV < 3.0%) was measured using the methylthymol blue method. Estimated glomerular filtration rate (eGFR) was calculated as eGFR (mL/min/1.73 m2) ) = 186× serum creatinine-1.154 × Age-0.203 in men, and 186× serum creatinine-1.154× Age-0.203× 0.742 in women according to the formula recommended by the Taiwan Society of Nephrology.For biochemical determinations of blood, 2).Anthropometric measurements included height, weight, blood pressure, waist circumference (WC) and body fat percentage. As for blood pressure measurement, prior to measurement, the subject removed any clothing in the arm area and was left to rest for five minutes to avoid stimulation. A mercury sphygmomanometer was used, which was held at the same height as the arm and heart during measurement, with palms facing upward. Percentage of body fat was estimated with bioelectrical impedance analysis using TBF-410 . Body mass index (BMI) was also calculated in weight (kg)/height (m2); stage 2 (eGFR 60–89 mL/min/1.73 m2); stage 3 (eGFR 30–59 mL/min/1.73 m2); stage 4 (eGFR 15–29 mL/min/1.73 m2); and stage 5 (eGFR <15 mL/min/1.73 m2) [Clinical criteria for blood, urine and anthropometric measurements were defined the following ranges: 1) patients with hypomagnesaemia, defined as serum magnesium < 0.75 mmol/L ; 2) goal1.73 m2) .Data were analyzed by 2-tailed t-test and one-way ANOVA for continuous variables and frequency distribution for categorical variables. Correlation of magnesium intake levels with metabolic parameter, body fat percentage and body mass index were examined in multiple linear regression analysis. Tests for linear trend were conducted by using the statement contrast in linear regression models. Logistic regression models were used to examine the relationship between magnesium intake levels with depression, metabolic risk factors, high body fat percentage and high body mass index status. Data are expressed as odds ratios and 95% CIs. Tests for linear trend were conducted by modeling the odds of each quartile -defined category of magnesium intake as a continuous variable in logistic regression models. All statistical procedures were performed using SPSS (version 17.0) statistical software, , and a p-value of 0.05 was considered statistically significant.p < 0.001). The low physical activity group displayed lower magnesium intake than those of the moderate and high physical activity group (p = 0.002). However, the high physical activity group displayed lower serum magnesium levels than those of the moderate physical activity group (p = 0.024). Compared with non-depressed patients, depression patients consumed significantly lower magnesium intake (p = 0.001). Mean magnesium intakes were significantly lower in patients with low HDL (p < 0.001), central obesity (p = 0.051), high body fat percent (p = 0.000) and high BMI (p < 0.001). Compared with non- metabolic syndromes patients, patients with metabolic syndromes displayed significantly lower magnesium intake (p = 0.013). Estimated GFR showed marginal positivity relationships with magnesium intake (p = 0.067). Magnesium intake was not significantly correlated with serum albumin, HbA1C, blood pressure, triglyceride or hsCRP. However, serum magnesium levels were not significantly associated with other variables.The clinical characteristics of the elderly patients with type 2 DM are shown in Table p = 0.000) and protein intake . A borderline positive relationship was observed between magnesium intake and serum albumin . Furthermore, magnesium intake was positively correlated with HDL . Magnesium intake had significant inverse relationships with triglyceride  and WC . Magnesium intake showed marginal inverse associations systolic blood pressure  and diastolic blood pressure . However, there was no significant relationship between magnesium intake and HbA1C. In addition, magnesium intake had an inverse relationship with body fat percentage  and BMI . A marginal positive relationship were observed between magnesium intake and eGFR . No significant relationship was found between magnesium intake and hsCRP.Simple linear regression was applied to determine the relationship of magnesium intake to serum magnesium, nutritional variables, metabolic parameters, body fat percent, body mass index, hsCRP, and eGFR. Serum magnesium levels were not significantly correlated with magnesium intake. Magnesium intake had positive association with energy intake . High physical activity demonstrated lower serum magnesium levels in men and female (p < 0.05). In addition, man and female were similar to unstratified analysis that serum magnesium was not significantly correlated with energy intake, protein intake, and eGFR, metabolic parameter, body fat percentage and body mass index. For the elderly men with Type 2 diabetes, magnesium intake was significantly lower in patient with metabolic syndrome (0.022). Physical activity level was significantly associated with magnesium intake in men (p = 0.010). However, physical activity level were not significantly associated with magnesium intake in female (p = 0.218). For the elderly female with Type 2 diabetes, magnesium intake was significantly lower in patient with depression (p = 0.003). Central obesity showed marginal inverse relationships with magnesium intake (p = 0.056).These data were also analyzed and stratified by sex (data not shown). The information remains about the same. For both men and female, lower magnesium intake was significantly associated with lower intake of energy and protein and high body mass index (p for trend = 0.006). In addition, adjusted mean of waist circumference, body fat percentage, and body mass index were significantly lower with increasing quartile of magnesium intake (p for trend < 0.001). The adjusted mean of systolic blood pressure was marginal linear trend with quartile of magnesium intake (p for trend < 0.089). The adjusted mean of triglyceride and diastolic blood pressure was not significantly linear trend with quartile of magnesium intake, but the adjusted mean of triglyceride and diastolic blood pressure were lower with increasing quartile of magnesium intake (p for trend < 0.120). The odds of depression, central obesity, high body fat percentage, and high body mass index were significantly lower with increasing quartile of magnesium intake (p for trend <0.005). Although the odds of low HDL and metabolic syndrome was not significantly linear trend with quartile of magnesium intake, the odds of low HDL and metabolic syndrome were lower with increasing quartile of magnesium intake (p for trend < 0.165). In addition, adjusted mean of metabolic parameters, body fat percentage and body mass index and adjusted Odd ratio of depression and metabolic risk factors was not significantly correlated with levels of serum magnesium.Adjusted mean of metabolic parameters, body fat percentage and body mass index and adjusted Odd ratio of depression and metabolic risk factors by quartile of magnesium intake after controlling confounding factors are shown in Table Our data suggest that (1) the majority of elderly type 2 diabetes patients may have low magnesium intake; (2) about one-third of elderly diabetes patients had hypomagnesaemia; (3) low magnesium intake was associated with metabolic abnormalities and depression; (4) magnesium intake was related to high physical activity level and demonstrated lower serum magnesium levels. However, serum magnesium was not significantly associated with metabolic parameters.In general, dietary magnesium does not meet the DRI for elderly subjects and may be prone to chronic latent magnesium deficiency ,42. DailIt is generally accepted that magnesium deficiency is closely related to metabolic syndromes. Increased dietary magnesium intake and/or magnesium supplementation may improve metabolic syndromes and type 2 DM ,25. Our Depression is the fourth-leading cause of disability worldwide according to the World Health Organization . PrevaleA review study indicated that 14-48% of diabetes suffered from hypomagnesaemia . Many stThe relationship between magnesium status and physical activity has received much attention in the past decade . Our datThere were several limitations of this study. First, the analyses were highly dependent on self-reported dietary intake and lifestyle data. Overestimation, underestimation, and poor recall might therefore have confounded the results. Fortunately, these elderly DM patients lived in rural areas and mostly of them had a simple lifestyle and eating patterns. The use of traditional quantitative tools, food models, food pictures and photos helped these elderly patients to recall the amount of consumed food and may also increase the effectiveness of the 24-hour recall. Moreover, questionnaires were also used to valid dietary patterns during the previous week. Second, serum magnesium may not be sufficient for indicating magnesium deficiency . TherefoMajority of elderly diabetes patients have low magnesium intake and may exacerbate metabolic abnormalities and depression. Clinical care should therefore focus on increasing dietary magnesium intake or magnesium supplementation to improve metabolic control, depression, and physical performance in elderly diabetes patients.BMI, Body mass index; BP, Blood pressure; CKD, Chronic kidney disease; DRI, Dietary reference intakes; DSM-IV, Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition; GFR, Glomerular filtration rate; FPG, Fasting plasma glucose; HbA1C, Glycated hemoglobin; HDL, High-density lipoprotein; hsCRP, High-sensitivity C-reactive protein; OGTT, Oral glucose tolerance test; WC, Waist circumference.The authors declare that they have no competing interests.J-HH has substantial contributions to conception and design of study, implementation of study, analysis and interpretation of data. Y-FL has substantial contributions to review the design and edit the manuscript. F-CC has substantial contributions to interpret original data and initiate the first draft of manuscript. JN-YL has substantial contributions to review and give comments on this manuscript. L-CT has substantial contributions to the conception and the original design of study, conducting the clinical study, and furnishing the final manuscript. All authors have read and approved the final manuscript.


Plasmodium falciparum continues to shape the human genome. Severe disturbances within the microcirculation result from the adhesion of infected erythrocytes to host receptors on monocytes, platelets, and endothelium. In this prospective study, we compared expression of all major host cytoadhesion receptors among Ugandan children presenting with uncomplicated malaria (n = 1078) versus children with severe malaria (n = 855), including cerebral malaria (n = 174), severe anaemia (n = 522), and lactic acidosis (n = 154). We report a significant survival advantage attributed to blood group O and increased monocyte expression of CD36 and ICAM1 (CD54). The high case fatality rate syndromes of cerebral malaria and lactic acidosis were associated with high platelet CD36 expression and thrombocytopenia, and severe malaria anaemia was characterized by low ICAM1 expression. In a logistic regression model of disease severity, odds ratios for the mitigating effects of blood group O, CD36, and ICAM1 phenotypes were greater than that of sickle haemoglobin. Host genetic adaptations to Plasmodium falciparum suggest new potential malaria treatment strategies.As a leading cause of childhood mortality worldwide, selection pressure by  Plasmodium falciparum continues to kill large numbers of children born within malaria endemic regions characterized by limited health resources  within the spleen as a result of modified red cell fitness   Study in a cohort of two thousand children in Uganda. We compared expression of the major host cytoadhesion receptors across a spectrum of malaria severity, quantified their relative effect size compared with other recognized factors that influence severity (such as sickle trait), and examined their distributions in three severe malaria syndromes.P falciparum malaria to Mulago Hospital's Acute Care Unit in meso-endemic Kampala, Uganda between October 15, 2007 and October 30, 2009. Children with severe malnutrition were ineligible for enrolment. The malaria diagnosis was established first with clinical symptoms and a positive screening thick smear. Two blinded reviewers of thin and thick smears at a reference parasitology laboratory independently confirmed the diagnosis and parasite density using leucocyte counts  reviewed logistics, tabulated accrual statistics, and monitored study quality control. The accuracy of data transfer to the digital CRF was audited on each 25th CRF quarterly. The digital CRF  mirrored the paper CRF with multiple features to prevent transcription errors. Paper and digital CRFs were kept in a secure, locked environment during enrollment and transferred at the conclusion of enrollment to study headquarters for use in resolution of any discrepancies in the database.Each enrolled patient was evaluated and treated by physicians experienced in malaria care, using all available clinical resources to assess the presence of other conditions. Data were recorded on a hard-copy Case Report Form (CRF), available at The electronic study database, prepared from exports of the digital CRF, underwent extensive testing for data integrity, consistency, and accuracy. Scrutiny included range value testing, missing data testing, logical tests on data consistency across fields, and multiple comparisons with the original paper CRF record. Strict version control was used. All data analyses were done on a single version of a finalized corrections dataset.Patients were observed until discharge or death by two study physicians. Prior to analysis, each patient was categorized at final disposition as a case or control, and if failing to meet pre-specified definitions, was excluded. Controls were those with uncomplicated malaria (UM), defined as the absence of any impairment of consciousness or hypoxia, with peripheral blood lactate <5 mmol/l and haemoglobin (Hb) >70 g/l without transfusion. Cases were those with severe malaria (SM) including severe malaria anaemia (SMA), defined as Hb < 50 g/l (or <60 g/l after transfusion); lactic acidosis (LA), defined as blood lactate >5 mmol/l; hypoxia, defined as oxygen saturation <90% while breathing ambient air; or cerebral malaria (CM), as defined below.et al, et al, WHO guidelines provide general criteria to characterize cerebral malaria, i.e., Blantyre Coma Scale (BCS) ≤2 with depressed levels or loss of consciousness (LOC) persisting >1 h after any possible prior convulsions  if the patient had coma plus >3 of the following 10 WHO malaria severity criteria: >2 seizures in 24 h, respiratory distress, jaundice, haemoglobinuria, spontaneous bleeding, hypoglycaemia (glucose < 2·2 mmol/l), lactic acidosis (lactate >5 mmol/l), normocytic severe anaemia, hyperparasitaemia >5%, or new acute renal failure; or if the patient had a cumulative score of ≥3 points (n = 169) using the following scale:To increase the specificity of our categorization of CM cases, we used objective clinical criteria that included both WHO features of severe disease, and a case report form-derived scoring system. A case was designated as probable to definite CM  in 24 h.2 points: fatality with ≤3 observed seizures (or number not reported) in 24 h.6 points: >3 observed seizures in 24 h.Blood sugar:0 points: blood sugar not tested at time of coma or Blantyre assessment.0 points: blood sugar ≥3 mmol/l at time of coma or Blantyre score assessment.−1 point: blood sugar <3 mmol/l at time of coma or Blantyre score assessment.n = 56):Lumbar puncture (performed in 0 points: not performed.1 point: performed and did not show signs of meningitis.n = 12):Retinoscopy (performed in 0 points: if negative or not performed.2 points: if positive.Patients were excluded if human immunodeficiency virus (HIV) positive. 740 patients were tested for HIV in Kampala using a screening assay . Negative screening tests were re-tested with a second assay , and positive screening tests confirmed with a second enzyme-linked assay . Archived plasma aliquots from the remaining 1331 who were not tested in Kampala  were tested with a high-sensitivity commercial assay for antigen and antibody at Mount Sinai Hospital's Microbiology Laboratory in Toronto, Canada . 45 HIV+ subjects were excluded from analysis. Of the 733 subjects identified as HIV-negative by laboratory testing in Kampala, 0·5% of samples were randomly re-assessed in Toronto to validate the negative result.ABO blood groups were determined locally on peripheral blood collected in EDTA- anticoagulated tubes. ABO Blood Group Reagents  were applied to whole blood for direct haemagglutination typing according to manufacturer's directions.et al, et al, et al, Monocyte CD54, monocyte CD36, and platelet CD36 were determined in a blinded fashion by flow cytometry  according to previously published methods established for malaria patients . Because group A and AB individuals strongly express the A antigen, they were entered together in the model. Because CD36 and CD54 MFI approximate a log-normal distribution . Of the 2011, 78 were excluded from analysis because at final disposition, they did not meet pre-study definitions of uncomplicated malaria (UM) or severe malaria (SM). Controls included UM outpatients  and UM inpatients . Cases comprised SM survivors  and fatalities   A. MedianIllness in the great majority was attributable exclusively to malaria rather than to other infections. None of the 56 lumbar punctures in suspected cases of cerebral malaria showed meningitis. In four chest radiographs of suspected pneumonia, only one showed infiltrates. Parenteral antibiotics were empirically offered in one case of unconfirmed meningitis and 38 cases of unconfirmed pneumonia.P = 0·0031]. As shown in n = 558), lactic acidosis , and cerebral malaria  with some expected overlap (n = 174); SMA with or without LA ; and isolated LA without either SMA or CM , P < 0·0001).SM cases reflected severe malaria anaemia  compared with UM (28·0%), P < 0·0001; while group O prevalence was 9·3% higher in UM (49·9%) compared with SM (40·6%), P < 0·0001. This was further reflected in a consistent gradient of changing ABO proportions across severity subgroups   after controlling for the effect of age, gender, HbS, platelet count, leucocyte count, hyperparasitaemia, monocyte CD54, monocyte CD36 and platelet CD36  A. Group let CD36 . The ABOlet CD36  and the let CD36 B.2 = 10·36, P < 0·0157) (P = 0·033), and correspondingly more group B (B or AB) among those with SMA . When group A or AB cases were further sub-classified according to the presence or absence of the A1 antigen, a larger proportion of the patients with CM or isolated LA were represented among those positive for A1  A. AlthouP < 0·0001 (P < 0·0001) or in cases compared with controls (P < 0·0001)  B.P < 0·0001) (P = 0·0009), (P < 0·0001), and with CM and isolated LA.Monocyte CD36 levels were significantly higher in UM versus SM ( 0·0001) C. Among 0·0009), A. Given 0·0009), B. This rThrombocytopenia is recognized by the WHO as a manifestation of severe malaria WHO, . The preP = 0·005) in SM-f versus UM, while monocyte ICAM1 and CD36 expression were significantly higher in UM compared with SM-f (P < 0·0001).P. falciparum infection demonstrated a relationship between the expression of host receptors for ligands of cytoadhesion and disease severity. The malarial syndromes marked most by hypoperfusion, namely CM and LA, are of particular significance owing to their unrivalled case fatality rates  (Rowe et al, et al, et al, et al, et al, et al, et al (et al (et al (et al, et al, This study is novel for the simultaneous analysis of the major malaria cytoadhesion receptors found on erythrocytes, leucocytes, and platelets together with established clinical factors across a spectrum of disease severity. In a logistic regression model, the odds ratio for the harm of blood group A or AB or the benefit of group O remained significant even after controlling for numerous clinical factors including HbS . Consistl, et al  concludel (et al  proposedl (et al  inferredet al, P. falciparum, especially in rural areas with higher endemicity than present in our study. Thus, malaria selection may be a plausible explanation for the shift in ABO distributions that we observed within a single nation and generation.That malaria is likely to be changing global distributions of ABO types (Cserti & Dzik, et al, et al, et al, et al, et al, ICAM1 is expressed on the endothelium and may serve as a sequestration receptor for iRBC. The level of expression of ICAM1 on monocytes has had an uncertain overall role in malaria. Monocyte ICAM1 may indirectly enhance iRBC immune clearance but may conversely co-localize with iRBC in CM (Stevenson & Riley, et al, et al, et al, et al (in-vitro that binding of iRBCs to brain endothelial cells was mediated by platelets and was CD36-dependent. Because genetic studies (Fry et al, et al, et al, et al, et al, The effects of CD36 in malaria have likewise been particularly controversial. On monocytes, CD36 supports phagocytosis of iRBC (McGilvray l, et al  demonstrPlasmodial infections, with several mechanisms unique in P. falciparum, including platelet binding to iRBCs and endothelial cells (de Mast, et al, Thrombocytopenia is a hallmark of et al, The strengths of our study include its mechanism-driven design of case evaluation, large-volume accrual, completeness of case reporting, laboratory blinding, and use of validated assays for host receptors of adhesion ligands. In this largest prospective study of malaria outcomes according to host adhesion receptors, limitations nevertheless included: not characterizing parasite-encoded adhesins or performing adhesion assays with patient-derived iRBC isolates (Ochola in vitro studies document that soluble blood group substances can block the formation of iRBC rosettes (Barragan et al, In summary, we have demonstrated that host phenotypes for receptors of parasite cytoadhesion ligands associate with the clinical severity of malaria, as well as with high mortality manifestations. Specifically, SMA exhibited low ICAM1 expression, whereas CM or isolated LA demonstrated a high prevalence of group A1 and an elevated ratio of platelet-to-monocyte CD36. Informed by the molecular interactions of parasite and human, the measurable “panel” of ABO type, monocyte ICAM1 and CD36 expression, and platelet CD36 expression translates clinically. The determination of ABO type, an unchanging attribute and a categorical measure of the host, is of particular appeal because of its ease of testing in resource-limited settings, and its significance at the level of malaria mortality. The observation that ABO is an independent risk factor for survival among children with malaria provides the most compelling explanation to date for evolution away from the A1 wild-type and towards the weaker subgroups of A, group B or AB, and most importantly group O. Finally, these results invite approaches to malaria treatment that transcend the challenge of developing new cost-prohibitive drug therapies to which the parasite may also gain resistance. Re-directing our focus on co-evolutionary interactions between host and parasite suggests new lines of research. For example, the use of “universal” group O RBCs rather than type-specific RBCs may prove especially advantageous for group A1 patients. Because


There have not been yet enough studies about effects of beta glucan and gliclazide on oxidative stress created by streptozotocin in the brain and sciatic nerve of diabetic rats. The aim of this paper was to investigate the antioxidant effects of gliclazide and beta glucan on oxidative stress and lipid peroxidation created by streptozotosin in brain and sciatic nerve. Total of 42 rats were divided into 6 groups including control, diabetic untreated (DM) , STZ (DM) + beta glucan, STZ (DM) + gliclazide, only beta glucan treated (no diabetic), and only gliclazide treated (no diabetic). The brain and sciatic nerve tissue samples were analyzed for malondialdehyde (MDA), total oxidant status (TOS), total antioxidant status (TAS), oxidative stress index (OSI), and paraoxonase (PON-1) levels. We found a significant increase in MDA, TOS, and OSI along with a reduction in TAS level, catalase, and PON-1 activities in brain and sciatic nerve of streptozotocin-induced diabetic rats. Also, this study shows that in terms of these parameters both gliclazide and beta glucan have a neuroprotective effect on the brain and sciatic nerve of the streptozotocin-induced diabetic rat. Our conclusion was that gliclazide and beta glucan have antioxidant effects on the brain and sciatic nerve of the streptozotocin-induced diabetic rat.  Diabetes mellitus (DM) is one of the most common chronic metabolic disorders leading to complications in a number of organ and systems. DM characterised by disturbed glucose metabolism due to an absolute or relative insulin deficiency affects the central and peripheral nervous systems . It is rβ-glucan is an antioxidant with the scavenging ability lying between that of α-tocopherol, which is known to be incorporated in the lipid bilayer, and the water-soluble antioxidant, mannitol [One of the important goals of DM treatment is to prevent its complications. To this end, there is accumulating evidence that supplementation antioxidant compounds may offer some protection against diabetic complications . Therefomannitol . TherefoGliclazide which is another possible protective material of this study, is a second-generation sulfonylurea hypoglycemic agent. It may show effect as reduction in free radical generation or an increase in free radical scavenging. Also, gliclazide may contribute to the control of the physiopathological mechanisms underlying both the process of aging and type 2 diabetes by reducing oxidant stress and DNA damage, improving antioxidant status . In diabThis study was approved by Dicle University Animal Ethical Committee and was carried out in accordance with the ‘‘Animal Welfare Act and the Guide for the Care and Use of Laboratory animals prepared by the Dicle University, Animal Ethical Committee.” Female Wistar Albino rats (250 ± 50 g) were obtained from the Animal labaratuary of Dicle University. The rats were housed in clean polypropylene cages having six rats per cage and maintained under temperature controlled room (23 ± 2°C) with a photoperiod of 12 h light and 12 h dark cycle. The rats were given standard pellets diet and water ad libitum throughout the experimental period. All efforts were made to minimize animal suffering and to use only the number of animals necessary to produce reliable scientific data.The rats were fasted overnight, and diabetes was induced by a single dose via intraperitoneal injection of streptozotocin solution (STZ). The experiment was designed as a total of 28 days. A week after STZ administration, their blood glucose values were measured and were defined as diabetic rats, that is, their blood glucose values were above 250 mg/dL. STZ caused the death of some rats. At the end, 42 rats were divided into 6 groups including control, diabetic untreated , diabetics treated with beta glucan (STZ + beta glucan), diabetics treated with gliclazide (STZ + gliclazide), only beta glucan treated , and only gliclazide treated . The beta-glucan  used in this study is 1,3-1,6 beta-D-glucan in the microparticulate form, which was prepared from the S. cerevisiae yeast. Beta glucan was administered orally (50 mg/kg body weight) with a gavage for 21 days in the STZ + beta glucan and beta glucan groups. The gliclazide  used in the tablet form was administered orally (10 mg/kg body weight) with a gavage for 21 days in the STZ + gliclazide and only gliclazide groups. STZ solution was prepared as follows: 24 mg of STZ was dissolved in 1 mL of 5 mM citrate buffer (pH 4.5) before the injection, and a volume of 2.5 mL/kg was administrated into each rat. The animals were fasted overnight and diabetes was induced by a single i.p. injection of a freshly prepared solution of STZ  (50 mg/kg body weight) in 0.1 M cold citrate buffer (pH 4.5). One week after STZ administration, blood was taken from the lateral veins of the tail, their blood glucose levels were measured by a glucometer  using a glucose oxidase method and the rats whose blood glucose values were above 250 mg/dL were accepted as diabetic [2O2 equivalent/mg protein [μmole H2O2 Equiv./gram protein and mmole H2O2 Equiv./gram protein, respectively. The cerebrum tissue OSI value was calculated as follows: OSI = / × 100) [The excised cerebrum and sciatic nerve tissue samples for biochemical analyses were weighed, immediately stored at −50°C. The cerebrum tissues cleaned with 1.15% ice-cold KCl, minced, then homogenized in five volumes (w/v) of the same solution. Assays were performed on the supernatant of the homogenate that is prepared at 14.000 rpm for 30 min at +4°C. The protein concentration of the tissue was measured by the method of Lowry . Serum p protein . The TOS) × 100) .U test. A value of P < 0.05 indicates a significant difference. Data are expressed as mean ± S.D.The data was analyzed by using Statistical Package for the Social Sciences version 11.5 . The variables between the groups were tested by Mann Whitney P < 0.05). However, beta glucan-treated diabetic rats significantly reversed the catalase, PON-1 and TAS back to normal levels compared to untreated diabetic rats (for both parameters P < 0.05). PON-1 activity was significantly higher in gliclazide-treated diabetic group than untreated diabetic group in the brain (P < 0.05), but catalase activity and TAS level were not significant (for both parameters P > 0.05). As can be seen from P < 0.05). However, MDA, TOS, and OSI levels were significantly reduced to gliclazide-treated diabetic group compared with the untreated-diabetic group in brain tissue (for both parameters P < 0.05). Likewise, MDA, TOS, and OSI levels were significantly reduced to beta-glucan-treated diabetic group compared with the untreated-diabetic group in brain tissue (for both parameters P < 0.05).In the rat brain, the MDA, TOS, TAS, and OSI level, catalase and PON-1 enzyme activities are presented in P > 0.05). There was a significant depletion in the PON-1, catalase, and TAS levels in the sciatic nerve of the diabetic rat compared to the control groups (for both parameters P < 0.05). However, beta-glucan-treated diabetic rats significantly reversed catalase, PON-1, and TAS back to normal levels compared to untreated diabetic rats (for both parameters P < 0.05). PON-1 activity was significantly higher in gliclazide-treated diabetic group than untreated diabetic group in the brain (P < 0.05), but catalase activity and TAS level were not significant (for both parameters P > 0.05). As can be seen from P < 0.05). However, MDA, TOS, and OSI levels were significantly reduced to gliclazide-treated diabetic group compared with the untreated-diabetic group in sciatic nerve tissue (for both parameters P < 0.05). Likewise, MDA, TOS, and OSI levels were significantly reduced to beta-glucan-treated diabetic group compared with the untreated-diabetic group in sciatic nerve tissue (for both parameters P < 0.05).It was observed that medication similarly affected both tissue in the rat sciatic nerve compared with brain tissue. In the rat sciatic nerve, the MDA, TOS, TAS, and OSI levels as well as catalase and PON-1 enzyme activities are presented in DM is a metabolic disorder with a globally rising prevalence which can affect the peripheral and central nervous system , 22. It Previous experimental studies in STZ-induced diabetic rats have been suggested that the increased oxidative stress is an important factor in the pathogenesis of the complications . For exaOwing to their useful effects on the immune system, antioxidant, and anti-inflammatory properties, and because they lack any toxic effects, beta glucans have been used in previous many studies , 33. It Gliclazide which is a another marker of drug of the present study, second-generation sulfonylurea, might exert antioxidant effect. It has been demonstrated that gliclazide prevents, through its antioxidant properties, oxidized low-density lipoprotein-associated endothelial dysfunction, apoptosis, and plaque rupture . It has β-glucan and gliclazide correlated in a dose-dependent pattern with therapeutic response, and they penetrated to blood-brain barrier [Paraoxonase is one of the important antioxidant enzymes. PON-1 hydrolyses lipid peroxides in oxidized lipoproteins. A close relationship between PON-1 deficiency and accelerated progression of arteriosclerosis has been found in experimental and human studies , 43. Alt barrier , 46. To Hyperglicemia in STZ-induced diabetic rats can cause oxidative damage in brain and sciatic nerve tissues, which may play vital roles in the pathogenesis of diabetic neuropathy and encephalopathy. Also treatment of beta glucan and/or gliclazide inhibits lipid peroxidation and regulates total oxidant/antioxidant status in diabetic rat brain and sciatic nerve tissues. This study results suggested that beta glucan and gliclazide may be considered to reduce oxidative stress in diabetic brain and sciatic nerve and may be used as a protective agent against diabetic damage of brain and sciatic nerve.