Interpreting epigenetic causes of recurrent hypokalemia and seizures: Gitelman syndrome co-exist with pseudohypoparathyroidism type 1B.

We describe a unique case of 27-year-old male with Gitelman syndrome (GS) co-exist with pseudohypoparathyroidism type 1B (PHP1B). The patient presented with a 5-year history of seizures, tetany, and numbness of the extremities. Further examinations showed recurrent hypokalemia, inappropriate kaliuresis, hypocalcemia, hyperphosphatemia, and elevated PTH levels. A novel variant of autosomal recessive GS (p.Val287Met SLC12A3) and a novel 492.3Kb deletion containing the whole of STX16, were discovered by a whole-exome sequencing. Following the diagnosis, calcitriol, calcium, and potassium supplements were started. Hematuria calcium and phosphorus levels, as well as blood potassium levels, have recovered and remained within normal ranges after 3 years of follow-up. Our findings have important consequences for supporting the idea that heterozygosity for variants have effects on the patients' clinical performance with autosomal recessive inheritance disorders. Further study is need for the putative effects of the variant. Likewise, further investigation with regards to the gene-gene interaction relations between GS and other electrolyte imbalance disorders is warranted.

Trichosporon fermentans biomass flocculation from soybean oil refinery wastewater using bioflocculant produced from Paecilomyces sp. M2-1.

The soybean oil refinery (SOR) wastewater contains a high concentration of chemical oxygen demand (COD) and lipid, so the direct emissions of SOR wastewater will result in environmental pollution and waste of resources. Oleaginous yeast Trichosporon fermentans can consume organic materials in SOR wastewater to synthesize microbial oil, which achieves the purpose of SOR wastewater resource utilization. The effective harvesting technology of oleaginous yeasts can improve the utilization efficiency. In this study, Paecilomyces sp. M2-1 with high flocculating activity was isolated. The flocculants produced by M2-1 (MBF2-1) include 75% (w/w) polysaccharides, rely on cations, and display the flocculation percentage of above 77% in the range of pH 2-11. Especially under alkaline conditions, the flocculation percentage can be kept above 97%. The results of scanning electron microscope observation and zeta potential measurements suggested that the bridging, net trapping, and sweeping were the main flocculation mechanism of MBF2-1. MBF2-1 could flocculate T. fermentans that was used to reduce the organic matter in SOR wastewater and to produce microbial oil. Under the optimum conditions, the flocculation percentage of MBF2-1 against T. fermentans from SOR wastewater can reach 95%. Fatty acid content percent in microbial oil from T. fermentans was not almost affected by flocculation of MBF2-1. Moreover, MBF2-1 can further remove 55% and 53% of COD and oil content in the fermented SOR wastewater, respectively. The properties and high flocculating percentage displayed by MBF2-1 indicated its potential application prospect in oleaginous yeast harvest and food industry wastewater treatment.

Wnt/ß-catenin signaling in osteoblasts regulates global energy metabolism.

Obesity, diabetes and osteoporosis have become a major public heath burden, and understanding the underlying mechanisms of these pathophysiological process will benefit their treatment. Osteoblast lineage cells in charge of the bone formation have been showed to participate in the whole-body energy metabolism. In this study, we identify that wnt/ß-catenin signaling in osteoblasts could regulate global energy metabolism, including glucose homeostasis, fat accumulation and energy expenditure. Mice lacking ß-catenin specifically in osteoblasts postnatally exhibit decreased bone mass, increased glucose level, decreased insulin production, decreased fat accumulation and increased energy expenditure. Osteocalcin supplement can rescue the impaired glucose balance by improving insulin production but cannot influence the abnormal fat accumulation and energy expenditure. Osteoprotegerin (OPG) overexpression exclusively in osteoblasts in ß-catenin deletion mice can normalize not only the decreased bone mass but also the decreased fat accumulation and increased energy expenditure. The effect of ß-catenin deletion and OPG overexpression in osteoblasts on global energy metabolism had no relation with inguinal fat browning. These results suggest that the regulation of bone on energy metabolism and fat accumulation is not mediated exclusively by osteocalcin. Our findings may provide a new insight into the regulation of bone on fat accumulation and energy metabolism.

Salvia miltiorrhiza: A source for anti-Alzheimer's disease drugs.

CONTEXT: Alzheimer's disease (AD) is a devastating neurodegenerative disorder that affects millions of elderly people worldwide. However, no efficient therapeutic method for AD has yet been developed. Recently, Salvia miltiorrhiza Bunge (Lamiaceae), a well-known traditional Chinese medicine which is widely used for treating cardio-cerebrovascular, exerts multiple neuroprotective effects and is attracting increased attention for the treatment of AD. OBJECTIVE: The objective of this study is to discuss the neuroprotective effects and neurogenesis-inducing activities of S. miltiorrhiza components. METHODS: A detailed search using major electronic search engines (such as Pubmed, ScienceDirect, and Google Scholar) was undertaken with the search terms: Salvia miltiorrhiza, the components of S. miltiorrhiza such as salvianolic acid B, salvianolic acid A, danshensu, tanshinone I, tanshinone IIA, cryptotanshinone, dihydrotanshinone, and neuroprotection. RESULTS: Salvia miltiorrhiza components exert multiple neuroprotective potentials relevant to AD, such as anti-amyloid-ß, antioxidant, anti-apoptosis, acetylcholinesterase inhibition, and anti-inflammation. Moreover, S. miltiorrhiza promotes neurogenesis of neural progenitor cells/stem cells in vitro and in vivo. CONCLUSIONS: The properties of S. miltiorrhiza indicate their therapeutic potential in AD via multiple mechanisms. In addition, S. miltiorrhiza provides lead compounds for developing new drugs against AD.

Differential therapeutic effects of nateglinide and acarbose on fasting and postprandial lipid profiles: a randomized trial.

BACKGROUND: Dyslipidemia is commonly seen in patients with type 2 diabetes mellitus (T2DM). The current study sought to compare the effects of nateglinide and acarbose, two antihyperglycemic agents, on both fasting and postprandial lipid profiles in Chinese subjects with T2DM. SUBJECTS AND METHODS: For this multicenter, open-label, randomized, active-controlled, parallel-group study, 103 antihyperglycemic agent-naive patients with T2DM were recruited from four hospitals in China. In total, 85 subjects (44 in the nateglinide group, 41 in the acarbose group) with a known complete lipid profile underwent the entire clinical trial and were included in the final analysis. Serum was collected in the fasting state and 30 and 120 min after a standardized meal (postprandial states) to measure the baseline lipid profiles; the same testing was performed upon completion of a 2-week course of nateglinide (120 mg three times a day) or acarbose (50 mg three times a day). RESULTS: Fasting triglyceride (TG) levels were significantly reduced by both nateglinide and acarbose (P<0.001), with acarbose providing a significantly more robust improvement (vs. nateglinide, P=0.005). Additionally, the TG levels at both postprandial times were significantly reduced by acarbose (P<0.001 at 30 min and P=0.002 at 120 min), whereas nateglinide treatment only significantly reduced the 30-min postprandial TG (P=0.029). Neither nateglinide nor acarbose treatment had significant impact on total cholesterol, high-density lipoprotein, low-density lipoprotein, or non-high-density lipoprotein cholesterol. CONCLUSIONS: Compared with nateglinide, acarbose has superior therapeutic efficacy for reducing fasting and postprandial TG levels in patients with T2DM.

Synergistic inhibitory effect of berberine and d-limonene on human gastric carcinoma cell line MGC803.

This study aims at evaluating the anticancer effects of berberine hydrochloride (berberine) and d-limonene, alone and in combination, on human gastric carcinoma cell line MGC803 to determine whether berberine and d-limonene work synergistically and elucidate their mechanisms. MGC803 cells were treated with berberine and d-limonene, alone and in combination, for 24-48 h. The inhibitory effects of these drugs on growth were determined by MTT assay. The combination index and drug reduction index were calculated with the Chou-Talalay method based on the median-effect principle. Flow cytometry and laser scanning confocal microscopy were employed to evaluate the effects of both drugs on cell-cycle perturbation and apoptosis, generation of reactive oxygen species (ROS), mitochondrial membrane potential, and expression of Bcl-2 and caspase-3 in MGC803 cells. Berberine or d-limonene alone can inhibit the growth of MGC803 cells in a dose- and time-dependent manner. Berberine and d-limonene at a combination ratio of 1:4 exhibited a synergistic effect on anti-MGC803 cells. The two drugs distinctly induced intracellular ROS generation, reduced the mitochondrial transmembrane potential (ΔΨm), enhanced the expression of caspase-3, and decreased the expression of Bcl-2. The combination of berberine and d-limonene showed more remarkable effects compared with drugs used singly in MGC803 cells. The combination of berberine and d-limonene exerted synergistic anticancer effects on MGC803 cells by cell-cycle arrest, ROS production, and apoptosis induction through the mitochondria-mediated intrinsic pathway.

Hydroxysafflor yellow A attenuates left ventricular remodeling after pressure overload-induced cardiac hypertrophy in rats.

CONTEXT: Hydroxysafflor yellow A (HSYA), the main chemical component of the safflower yellow pigments, is used extensively in traditional Chinese medicine for the treatment of cerebrovascular and cardiovascular diseases. OBJECTIVE: The present study determined the effects of HSYA on left ventricular hypertrophy after pressure overload and investigated the underlying mechanisms. MATERIALS AND METHODS: Cardiac hypertrophy was induced by the ligation of abdominal aorta in male Wistar rats. The rats were then divided into five groups and treated with captopril (100 mg/kg) or HSYA at different doses (0, 10, 20 and 40 mg/kg). Six weeks after treatment, the weight of left ventricle, LVMI (left ventricular mass index) and pathological changes were measured. MMP-2 (metalloproteinase 2) and MMP-9 (metalloproteinase 9) levels were determined by ELISA. Protein expressions of Bcl-2 and Bax were evaluated by immunohistochemistry. RESULTS: HSYA (20, 40 mg/kg) significantly attenuated the increase of LVMI (ventricular weight/body weight) by 13.04 and 30.43% respectively, when compared with the model group. This was associated with the amelioration of pathological lesion, such as cardiac muscle fibers were smaller and the nuclei of cardiomyocytes were lightly stained in animals treated with HSYA (20, 40 mg/kg). In addition, the administration of HSYA at doses of 20 and 40 mg/kg increased the Bcl-2/Bax ratio (1.17 ± 0.08 and 1.39 ± 0.07 versus 0.71 ± 0.06). In addition, the serum MMP-2 and MMP-9 levels were blocked by the treatment at doses of 20 and 40 mg/kg HSYA (MMP-2, 76.1 ± 9.2 and 65.6 ± 6.8 versus 82.9 ± 6.2, ng/ml; MMP-9, 66.6 ± 4.8 and 57.5 ± 5.0 versus 83.5 ± 6.0, ng/ml). CONCLUSION: These findings indicated that HSYA has beneficial effects on hypertensive ventricular remodeling, which may involve mechanisms of inhibiting cell apoptosis and suppressing metalloproteinases expression.

Nateglinide and acarbose are comparably effective reducers of postprandial glycemic excursions in chinese antihyperglycemic agent-naive subjects with type 2 diabetes.

BACKGROUND: Recent studies have identified postprandial glycemic excursions as risk factors for diabetes complications. This study aimed to compare the effects of nateglinide and acarbose treatments on postprandial glycemic excursions in Chinese subjects with type 2 diabetes. SUBJECTS AND METHODS: This was a multicenter, open-label, randomized, active-controlled, parallel-group study. One hundred three antihyperglycemic agent-naive subjects with type 2 diabetes (hemoglobin A1c range, 6.5-9.0%) were prospectively recruited from four hospitals in China. The intervention was nateglinide (120 mg three times a day) or acarbose (50 mg three times a day) therapy for 2 weeks. A continuous glucose monitoring system was used to calculate the incremental area under the curve of postprandial blood glucose (AUCpp), the incremental glucose peak (IGP), mean amplitude of glycemic excursions, SD of blood glucose, the mean of daily differences, and 24-h mean blood glucose (MBG). Subjects' serum glycated albumin and the plasma insulin levels were also analyzed. RESULTS: Both agents caused significant reductions on AUCpp and IGP. Similarly, both treatment groups showed significant improvements in the intra- and interday glycemic excursions, as well as the 24-h MBG and serum glycated albumin compared with baseline (P<0.001). However, neither of the agents produced a significantly better effect (P>0.05). Moreover, the nateglinide-treated group had significantly increased insulin levels at 30 min and at 120 min after a standard meal compared with baseline, whereas the acarbose-treated group decreased. No serious adverse events occurred in either group. The rates of hypoglycemic episodes were comparable in the two groups, and no severe hypoglycemic episode occurred in either group. CONCLUSIONS: Nateglinide and acarbose were comparably effective in reducing postprandial glycemic excursions in antihyperglycemic agent-naive Chinese patients with type 2 diabetes, possibly through different pathophysiological mechanisms.

Hepatocyte growth factor enhances the generation of high-purity oligodendrocytes from human embryonic stem cells.

Generation of homogeneous oligodendrocytes as donor cells is essential for human embryonic stem cell (hESC)-based cell therapy for demylinating diseases. Herein we present a novel method for efficiently obtaining mature oligodendrocytes from hESCs with high purity (79.7+/-6.9%), using hepatocyte growth factor (HGF) and G5 supplement (containing insulin, transferrin, selenite, biotin, hydrocortisone, basic fibroblast growth factor and epidermal growth factor) in a four-step method. We induced hESCs into neural progenitors (NP) with HGF (5ng/ml) and G5 (1x) supplemented medium in an adherent differentiation system. The purified NPs were amplified in suspension as neurospheres for 1 month, and terminal oligodendrocyte differentiation was then induced by G5 supplement withdrawal and HGF treatment (20ng/ml). The cells generated displayed typical morphologies of mature oligodendrocytes and expressed oligodendrocyte markers O4 and myelin basic protein (MBP). Our result revealed that HGF significantly enhanced the proliferation of hESC-derived NPs and promoted the differentiation as well as the maturation of oligodendrocytes from NPs. Further studies suggest that HGF/c-Met signaling pathway might play an important role in oligodendrocyte differentiation in our system. Our studies provide a means for generating the clinically relevant cell type and a platform for deciphering the molecular mechanisms that control oligodendrocyte differentiation.

Predicting disordered regions in proteins using the profiles of amino acid indices.

BACKGROUND: Intrinsically unstructured or disordered proteins are common and functionally important. Prediction of disordered regions in proteins can provide useful information for understanding protein function and for high-throughput determination of protein structures. RESULTS: In this paper, algorithms are presented to predict long and short disordered regions in proteins, namely the long disordered region prediction algorithm DRaai-L and the short disordered region prediction algorithm DRaai-S. These algorithms are developed based on the Random Forest machine learning model and the profiles of amino acid indices representing various physiochemical and biochemical properties of the 20 amino acids. CONCLUSION: Experiments on DisProt3.6 and CASP7 demonstrate that some sets of the amino acid indices have strong association with the ordered and disordered status of residues. Our algorithms based on the profiles of these amino acid indices as input features to predict disordered regions in proteins outperform that based on amino acid composition and reduced amino acid composition, and also outperform many existing algorithms. Our studies suggest that the profiles of amino acid indices combined with the Random Forest learning model is an important complementary method for pinpointing disordered regions in proteins.

[Effect of multi-glycosides of tripterygii on latent autoimmune diabetes in adults in early stage].

OBJECTIVE: To observe the effect of multi-glycosides of tripterygii (MT) on latent autoimmune diabetes of adults (LADA) in early stage. METHODS: The diabetic patients were divided randomly into the control group treated with insulin alone and the treated group treated with insulin and MT. Levels of insulin, C-peptide, glutamic acid decarboxylase antibody (GADAb) and islet cell antibody (ICA) were detected and clinical features of the disease were observed. RESULTS: There was no difference between the control group and the treated group in body mass index (BMI), the occurrence of diabetic keto-acidosis and the function of liver and kidney (P >0.05). After 6 months' treatment, the positive rate of GADAb and ICA decreased, plasma levels of fasting and 2 hrs post-prandial C-peptide and also 2 hrs post-prandial true insulin in the treated group increased (P <0.01), while all the above indexes improved more significantly after 1 year's treatment (P <0.01). CONCLUSION: Compared treatment of insulin with multi-glycosides of tripterygii in early stage of LADA has better effects in relieving autoimmune injury and recovering function of pancreatic island than insulin alone.

Regulation of Cbfa1 expression by total flavonoids of Herba epimedii.

Core binding factor alpha1 (Cbfa1) is a member of the runt family of transcription factors, which appears to play a pivotal role in regulating the differentiation of osteoblastic precursors and the activity of mature osteoblasts. Total flavonoids of Herba epimedii (HEF) is a recognized bone anabolic agent, but there is lack of reports on the modulation of Cbfa1 expression by HEF. Here we investigated the effect of HEF on Cbfa1 expression in the bone of ovariectomized (OVX) rats. HEF could increase the expression of Cbfa1 mRNA in the bone of ovariectomized rats in a dose-dependent manner. Furthermore, the high dose HEF (160 mg/kg) administered for 12 weeks in vivo stimulated osteocalcin expression. These findings suggest that Cbfa1 is required for mediating the anabolic effects of HEF.

[Study on influence of different dosage of vitamin E on peripheral blood cell activities in rats].

OBJECTIVE: To investigate the influences of vitamin E (VE) at different dosage on peripheral blood lymphocyte (PBL) proliferation and anti-DNA oxidative damage activities and erythrocyte membrane fluidity. METHODS: 48 Wistar rats were randomly divided into four groups including control group, VE1, VE2, and VE3 groups supplemented with 7.5, 50, 200, 750 IU/kg bw x d VE, respectively. The trial lasted 8 weeks and the blood samples were collected at the end of the trial. The level of plasma VE was analyzed by fluorescent spectrometry. Plasma MDA and membrane GSH-Px were analyzed by kits. The blood erythrocyte membrane fluidity was detected by fluorescence polarization method, lymphocyte transformation rate by MTT method and DNA oxidative damage by comet assay. RESULTS: The results showed that plasma VE levels significantly increased in VE1, VE2, and VE3 groups. Plasma MDA and erythocyte membrane GSH-Px activity in the rats in 50 IU/kg bw x d (VE1) group were (2.29 +/- 0.55) nmol/ml and (367.17 +/- 129.86) U/mg prot, respectively. P (fluorescence polarization) and eta(microviscosity), which were inversely related with membrane fluidity, in VE1 group were significantly lower. Lymphocyte transformation rate was significantly increased by 261.86%, 199.23% and 412.97% and H2O2 induced DNA damage significantly decreased compared with the control, VE2, and VE3 groups. CONCLUSION: It is indicated that an effective intake of VE for enhancing erythrocyte membrane fluidity, lymphocyte proliferation and DNA stability was 50 IU/kg bw x d, while too excessive intake of VE could not be found to be beneficial.

[Effect of multiple micronutrients supplementation on anti-oxidative activity and oxidized DNA damage of lymphocytes in children].

OBJECTIVE: To examine the effect of multiple micronutrients supplementation on anti-oxidative activity and decreasing oxidized DNA damage of lymphocytes in Chinese children. METHODS: 82 healthy children in a rural areas, aged 9-11 years, were selected and randomized allocated into group receiving supplements and control group with each of them 41. 24-hour dietary recall was used to collect data on daily nutrient intakes of the research subjects. The subjects in the supplement group were given vitamin A (VA) 600 microg, beta-carotene (beta-C) 1.0 mg, vitamin E (VE) 100 mg, vitamin C (VC) 300 mg and Na2SeO3(Se) 200 microg in a tablet on daily base while those in the control group took a same-sized color placebo tablet. The trial lasted 8 weeks. 5 ml blood samples from each subject were taken during 7 to 9 o'clock in the morning. DNA damage of lymphocytes and levels of plasma VA, VE, VC, beta-C, Se, malondialdehyde (MDA), activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were then analyzed twice before and after the 8-week of trial. RESULTS: The low intakes of VA, VC and Se only accounted for 50.6%, 65.6% and 67.3% of their recommended nutrient intake (RNI) respectively. After the trial, levels of plasma beta-C, VA, VE, VC and Se in the supplemented group increased by 13.4%, 32.8%, 11.5%, 46.9% and 24.6% respectively, compared with the control group, indicating that nutritional status regarding antioxidant nutrients had largely been improved. GSH-Px activity had a significant increase in the supplement group than before the supplement and in the control group (P < 0.01). GSH-Px before the trial (the 100.4 U/ml) also showed significant increase after the trial (161.7 U/ml) (P < 0.01). However, the values of SOD and MDA significantly decreased after the trial. Analysis of DNA damage indicated that there was no significant difference in the intrinsic damage of DNA (P > 0.05). Significant decreases of oxidized DNA damage induced by 5 micromol/L, 10 micromol/L and 25 micromol/L H2O2 were found more in peripheral lymphocytes of the supplemented group, than in pre-supplement and the control group after the trial (P < 0.01). CONCLUSION: Supplementation of multiple micronutrients could effectively increase the levels of beta-C, VA, VE, VC and Se in plasma, and GSH-Px activity. In the meantime, MDA and oxidized DNA damage induced by a low level H2O2 decreased significantly after the trial. The reason accounted for the decrease of SOD activity after the trial needs to be further studied.