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Aims: Lactobacilli should resist stress environments in industry process and gastrointestinal tract before exerting their beneficial effects. To explore the possible stabilizers in probiotic products, prebiotic oligosaccharides were investigated. Methods and Results: We investigated the effect of four selected oligosaccharides on the survival of probiotic Lactobacillus plantarum and L. acidophilus to simulated stress conditions. It was found that the tolerance of lactobacilli to simulated artificial gastrointestinal juice, heat treatment and phenol solution was obviously enhanced in fructo-oligosaccharides (FOS) and xylo-oligosaccharides (XOS) group. In addition, chito-oligosaccharides (COS), manno-oligosaccharides (MOS) and glucose also had positive effect compared with control group (without sugar). Conclusions: Prebiotic oligosaccharides, especially XOS and FOS added in medium have protection function to lactobacilli in stress environments. The protection function of oligosaccharides may correlate with the bacteria growth, which was stimulated by these oligosaccharides. Significance and Impact of the Study: Prebiotic oligosaccharides may be used as stabilizers in probiotic products.

Aims: To characterize the antimicrobial and adhesion ability of candidate probiotic Clostridium butyricum CB2 for farmed fish in vitro. Methods and Results: The potential probiotic Cl. butyricum CB2 had been evaluated for its adhesion capacity and antagonistic effect against two fish pathogens Aeromonas hydrophila and Vibrio anguillarum by the intestinal cell model. In addition, the aggregation ability and antimicrobial property on agar plate were assayed. The results indicated that the candidate probiotic Cl. butyricum CB2 have strong adhesion property and a higher antagonistic activity to Aer. hydrophila and V. anguillarum both on agar plate and cell model. Clostridium butyricum showed a higher aggregation which might be the reasons for bacteria adhesion and antimicrobial activity. Conclusions: The strain Cl. butyricum CB2 could be used as potential probiotic to inhibit pathogens growth and prevent their colonization in fish intestinal tract. Significance and Impact of the Study: This study revealed the antimicrobial and adhesion characteristic of Cl. butyricum CB2 which was selected as the potential probiotic to farmed fish.

Elastomer compounds reinforced with precipitated silica can exhibit elevated wet sliding friction on a rough surface in comparison with corresponding compounds filled with carbon black particles. The underlying mechanism is currently not well understood. To unravel this puzzling observation, the variation of wet sliding friction with filler volume fraction is examined at the sliding speed of the order of 1 m s−1 under different lubrication conditions. Depending on the lubrication liquid—water or ethanol—a compound that shows both higher bulk hysteretic loss and lower modulus does not always exhibit a higher wet sliding friction. A thorough characterization of the bulk rheology of the compounds investigated fails to provide the rationale for such behaviour, thus constituting an apparent violation of the conventional viscoelastic understanding of rubber friction on a rough surface. On the other hand, the detected lowering of friction when the lubrication liquid is changed from water to ethanol resembles the effect of liquid medium on interfacial adhesion reported in the literature. Hence, it is suggested that a stronger interfacial attractive interaction should exist in water between the road surface and silica particles on the compound surface immediately next to the road surface. This should be related to the elevated wet sliding friction detected for silica-filled compounds under water lubrication.

‘Green’ tire bearing tread rubber reinforced with precipitated silica can exhibit improved wet traction performance. The underlying mechanism is currently not well understood. To improve our capability of rational material design for enhanced driving safety, wet sliding friction for various rubber compounds is tested on a Portland cement concrete surface under varied lubrication conditions. The wetting liquid is either ethanol or water, and the initial amount of liquid on the concrete surface is adjusted. Sliding friction is detected to alter with lubrication condition. Under ethanol lubrication, the sliding friction is markedly lower than that under water lubrication. Additionally the benefit in wet traction from silica is significantly diminished or eliminated in ethanol. Such observations cannot be rationalized with simple considerations of rubber bulk viscoelasticity, liquid viscosity, cavitation, or capillary effect. We believe that these observations strongly indicate the significance of interfacial interactions in determining the wet sliding friction of elastomer compounds. The potential relevance of capillary porosity in concrete and Marangoni drying effect to wet traction is also introduced.

Background Apolipoprotein E (apoE) induces the uptake of vitamin K-rich lipoproteins by the liver, which likely affects inter-individual variation of warfarin dosing requirements. Associations between APOE polymorphisms and warfarin dosing were previously reported inconsistently among different ethnic groups, so the present study investigated this association in northern Han Chinese patients with mechanical heart valve prosthesis. Methods A total of 186 patients who underwent mechanical heart valve replacement and attained a stable warfarin dose were included. APOE single nucleotide polymorphisms (SNPs) rs7412 and rs429358 were genotyped using Illumina SNP GoldenGate Assay. Genotyping results were confirmed by direct sequencing. PHASE v2.1 software was used to construct rs7412 and rs429358 haplotypes. The effects of different APOE genotypes on warfarin dose were analyzed statistically. Results The mean warfarin maintenance dose was 3.10 ± 0.96 mg/day, and the mean international normalized ratio (INR) was 2.09 ± 0.24. APOE E2, E3, and E4 allele frequencies were 11.6 %, 82.5 %, and 5.9 %, respectively. No E2/E2 or E4/E4 genotypes were detected in this population. E2/E3, E3/E3, E2/E4, and E3/E4 genotype frequencies were 21.0 %, 67.2 %, 2.2 %, and 9.7 %, respectively. Significant differences in warfarin dose requirements were observed among patients with E2/E3, E3/E3, and E3/E4 genotypes (p 0.05). Patients were divided into low-intensity anticoagulant treatment group (1.6 ≤ INR

The present study analyzed 5785 vegetables for concentrations of As, Cd, Cr, Pb, Ni and Hg and estimated the health risk to local consumers by deterministic (point estimates) approaches. Levels of elements varied in different vegetables. Average levels of As, Cd, Cr, Ni, Hg and Pb were 0.013, 0.017, 0.057, 0.002, 0.094 and 0.034 mg/kg (fresh weight), respectively. The samples with 0.25% for Cd and 1.56% for Pb were exceeding the maximum allowable concentrations (MACs) set by the Chinese Health Ministry. No obvious regular geographical distribution for these metals in vegetables was found in areas of Zhejiang, China. The mean and 97.5 percentile levels of heavy metal and metalloid were used to present the mean and high exposure assessment. The health indices (HIs) were less than the threshold of 1 both in mean and high exposure assessment. It indicates that for the general people there is very low health risk to As, Cd, Cr, Pb, Ni and Hg by vegetable intake.

A sensitive and rapid analytical method based on alkaline diatomaceous earth extraction followed by GC/MS was developed for the quantitative determination of the toxic contaminant ethyl carbamate (EC) in yellow rice wines. The optimal extraction conditions were investigated. With the application of diatomaceous earth extraction, the damage of organic acids to the capillary column was greatly reduced. By using d5-EC as an internal standard for quantitative analysis of EC, the linearity of the calibration curves was good between 10 and 1000 ng/mL. The LOD and LOQ were 1.7 and 5.0 μg/kg, respectively. The spiked level of EC was 5.0–300 μg/kg, and the average recovery of the spikes was between 78.4 and 98.2%, with an RSD between 4.3 and 8.3%. Upon validation by five laboratories when spiked with 50, 100, and 300 μg/kg, the average respective recoveries were 102.9, 102.2, and 98.7% with a RSD between 0.7 and 8.1%. The validation results demonstrated that the method is fast, simple, selective, and suitable for the determination of EC in yellow rice wines.

Familial hypercholesterolaemia (FH) is a serious genetic metabolic disease. We identified a specific family in which the proband had typical homozygous phenotype of FH, but couldn’t detect any mutations in usual pathogenic genes using traditional sequencing. This study is the first attempt to use whole exome sequencing (WES) to identify the pathogenic genes in Chinese FH. The routine examinations were performed on all parentage members and WES on 5 members. We used bioinformatics methods to splice and filter out the pathogenic gene. Finally, Sanger sequencing and cDNA sequencing were used to verify the candidate genes. Half of parentage members had got hypercholesterolaemia. WES identified LDLR IVS8[−10] as a candidate mutation from 222,267 variations. The Sanger sequencing showed proband had a homozygous mutation inherited from his parents and this loci were cosegregated with FH phenotype. The cDNA sequencing revealed that this mutations caused abnormal shearing. This mutation was first identified in Chinese patients and this homozygous mutation is a new genetic type of FH. This is the first time that WES was used in Chinese FH patients. We detected a novel genetic type of LDLR homozygous mutation. WES is powerful tools to identify specific FH families with potentially pathogenic gene mutations.

The receptor for advanced glycation end products (RAGE) is involved in a variety of biological processes, including tumorigenisis. Previous studies have demonstrated that RAGE regulates the neo-angiogenesis related downstream molecule - vascular endothelial growth factor receptor 2 (VEGFR-2). Here, we investigated the potential relationship between RAGE, VEGFR-2 and angiogenesis in 80 renal cell carcinoma (RCC) patients. Real-time quantitative PCR and ELISA analysis were used to explore the RAGE and VEGFR-2 gene expression levels and the protein of VEGFR-2 expression. Meanwhile, angiogenesis was detected by the semi-quantification of endothelial cell marker CD34 combined with caldesmon, which was detected by microvessel density (MVD) technique and immunohistochemistry. Tumors were classified as low or high RAGE-expressing using the median as the cut-off. Immunofluorescence staining for RAGE protein was performed as well. Additionally, the median gene expression levels of VEGFR-2 in the tumors were significantly lower expressing low levels of RAGE expression, 0.34 (95% CI, 0.28-0.39) compared to the expressing high levels of RAGE expression, 0.45 (95% CI, 0.29-0.61), (P=0.03). The median MVD was significantly lower in the tumors expressing low levels of RAGE, 6.5 (95% CI, 6.21-7.43), compared to the expressing high levels, 7.9 (95% CI, 6.25-8.93), (P0.01). Further, a positive association was certified with VEGFR-2 protein levels, P=0.07. Besides, RCC with high levels of RAGE expression are associated with high VEGFR-2 mRNA/protein levels and a higher density of microvessels; conversely, Kaplan-Meier survival analysis suggests that a significant correlation of elevated RAGE expression with decreased overall survival and metastasis-free survival. Our results establish that RAGE was identified as a potential prognostic biomarker for disease prognosis of RCC.

For properly chosen elastomer compounds, thermorheological characterization is combined with an examination of the variation of the wet sliding friction with temperature. A conceptual argument leads to the assumption that the wet sliding friction should maximize at the energy dissipation peak associated with the dynamic softening transition at a characteristic frequency determined by the sliding speed and the effective smallest surface asperity scale. The dynamic softening transition is characterized with the peak in tan δ/G' n , where tan δ is the loss tangent, G' is the elastic modulus, and n is a constant between 0 and 1. The William-Landel-Ferry transform is uncritically applied for extrapolating the position of the peak in tan δ/G' n at high frequencies. Even based on the criterion of tan 8, the results obtained on a concrete surface indicate that the effective smallest asperity scale is of order of 100 μm.

For a vulcanized elastomer compound containing no filler, variation of dynamic moduli with strain was examined in the transition zone approaching glass transition at 0.1, 1, 10, and 15 Hz. In the complete absence of filler-filler/filler-polymer interactions, the nonlinearity in dynamic moduli becomes enhanced upon approaching glass transition regardless of testing frequency. Under the test protocol employed, the extraordinary nonlinearity at 10 Hz and 15 Hz is shown to be associated with a spectacular shear-heating event. At 10 Hz and -19°C, a pronounced yielding-like peak in the in-phase stress component emerges at small strain (< 2%) during testing of increasing strain. However, the corresponding temperature rise here is only about 0.4°C. Interestingly, at or above 0°C, no such peak in the in-phase stress component emerges for compounds filled with reinforcing filler above the percolation threshold. Discussion is also made relating the observations to the sliding friction of rubber materials.

A survey of ethyl carbamate (EC) in fermented foods and beverages was conducted in Zhejiang province, China. The samples comprised 184 alcoholic beverages (rice wine, white spirit, wine, beer, and rice cooking wine), 33 flavorings (soy sauces and vinegars) and 20 fermented sufus. Rice wine contained up to 515 μg/kg with an average of 160 μg/kg, and rice cooking wine up to 206 μg/kg with an average of 87 μg/kg. White spirits, wine and beer contained EC at an average level of 72 μg/kg, 16 μg/kg and 2 μg/kg, respectively. EC was also detected in soy sauces and vinegars at an average level of 47 μg/kg and 27 μg/kg. In fermented sufus, EC was found at an average level of 63 μg/kg in normal sufu and 182 μg/kg in red sufu.

To identify the predictors of the positive results of transrectal ultrasound (TRUS)-guided biopsy for prostate cancer.We performed univariate and multivariate logistic regression analyses on the relevant data on 385 male patients that underwent TRUS-guided biopsy for prostate cancer, including such potential predictors as age, body mass index (BMI), symptoms, results of digital rectal examination (DRE), tPSA, fPSA, free/total PSA ratio (f/tPSA), prostate volume (PV), and PSA density (PSAD) for identification of the risk factors related to the positive rate of biopsy. Then we constructed a scoring system as a tool for predicting prostate cancer in repeat biopsies and determined the sensitivity of the system by calculating the false positive rate using the receiver operating characteristic curve.Among the 385 patients, 139 (36.1%) were diagnosed with prostate cancer. On multivariate analysis, age (P0.01), DRE (P0.01), tPSA (P0.01), fPSA (P0.01), f/tPSA (P0.01), PV (P0.01), and PSAD (P0.01) were all significant predictors of prostate cancer. Multivariate logistic regression analysis showed age, tPSA, f/tPSA, PV, and PSAD to be independent predictors, with ORs and 95% CIs of 1.07 (1.05-1.16), 1.05 (1.02-1.15), 0.97 (0.86-0.99), 0.98 (0.87-0.96), and 1.79 (1.48-2.06), respectively. Moreover, patients with the risk score of 3-5 had a significantly higher rate of prostate cancer than those with 0-2 (64% vs 11%, P0.001).The scoring system on the key predictors of prostate cancer can help urologists to identify the men in need of prostatic biopsy.

Matrix-dependent signal suppression often occurs in quantitative analysis by ultra-high pressure liquid chromatography tandem mass spectrometry (UPLC-MS/MS). In this study, we investigated three calibration methods for compensation of signal suppression on chloramphenicol (CAP) quantification in chicken muscle. The data showed that the spiking recoveries by solvent standard calibration with a stable isotope labelled internal standard (SIL-IS) and matrix-matched standard calibration with a SIL-IS were significantly higher than by external matrix-matched standard calibration (P0.05). When the SIL-IS was used, standards prepared in the mobile phase solvent showed no significant difference as those prepared in the matrix (P0.05). The limit of detection (LOD) for external matrix matched standard calibration was 0.1 μg kg(-1), and that for SIL-IS calibration (including matrix matched and solvent dissolved standard) was 0.03 μg kg(-1).

Eplets mismatch based on HLAMatchmaker software evaluates the clinical application of kidney transplantation.In 239 cases of renal transplant,merits of methods of the traditional HLA six antigen matcheing criteria, cross reaction groups standard and Eplets mismatch based on HLAMatchmaker standard were compared respectively.The number of mismatchs with three methods in 239 cases, were grouped according to low-high mismatchs. The results revealed that HLAMatchmaker algorithm could significantly increase the number of low mismatchs group 54 (22.6%), compared with the HIA group 19(7.9%) and CREGs group 32 (13.4%). The comparison was discovered statistical significance among the three groups (P0.001), so the comparison between each group was.HLAMachmaker of donor-recipients matching, is a more efficient, time-saving and high sensitivity matching solution to allograft renal transplantation.

Oxymatrine is an alkaloid, which is derived from the traditional Chinese herb, Sophora flavescens Aiton. Oxymatrine has been shown to exhibit anti-inflammatory, antiviral, and anticancer properties. The present study aimed to investigate the anticancer effects of oxymatrine in human pros- tate cancer cells, and the underlying molecular mechanisms of these effects. An MTT assay demonstrated that oxymatrine significantly inhibited the proliferation of prostate cancer cells in a time‑ and dose‑dependent manner. In addition, flow cytometry and a terminal deoxynucleotidyl transferase-medi- ated dUTP-biotin nick end-labeling assay suggested that oxymatrine treatment may induce prostate cancer cell apop- tosis in a dose-dependent manner. Furthermore, western blot analysis demonstrated a significant increase in the expression of p53 and bax, and a significant decrease in that of Bcl-2, in prostrate cancer cells in a dose-dependent manner. In vivo analysis demonstrated that oxymatrine inhibited tumor growth following subcutaneous inoculation of prostate cancer cells into nude mice. The results of the present study suggested that the antitumor properties of oxymatrine, may be associ- ated with the inhibition of cell proliferation, and induction of apoptosis, via the regulation of apoptosis-associated gene expression. Therefore, the results may provide a novel approach for the development of prostate cancer therapy using oxymatrine, which is derived from the traditional Chinese herb, Sophora flavescens

Background Familial hypercholesterolemia (FH) is an autosomal dominant disease that primarily results from mutations in the low-density lipoprotein receptor (LDLR) gene. We investigated two unrelated Chinese FH patients using gene screening and functional analysis to reveal the pathogenicity and the mechanism by which these mutations cause FH. Methods First, the LDLR gene was sequenced in these patients. Then, mutant receptors were transfected into human embryo kidney 293(HEK-293) cells, and a confocal laser-scanning microscope was used to observe the localization of mutant proteins. Further, the expression and the internalization activity were analyzed by flow cytometry. Finally, LDLR protein expression and stability was detected by western blot. Results Two different LDLR class 2B mutations were detected in two patients. The C201F mutation is a known mutation. However, the G615V mutation is novel. Flow cytometry showed that the expression and internalization activity of the mutant LDLRs were reduced to 73.6% and 82.6% for G615V and 33.2% and 33.5% for C201F, respectively. Conclusions This study identified two LDLR mutations in Chinese patients with FH and analyzed the relationship between the genotype and phenotype of these patients. We found that these mutant LDLRs were defective in transport, which led to a reduction in cholesterol clearance. These results increase our understanding of the mutational spectrum of FH in the Chinese population.

Simultaneous measurements of viscoelasticity and electrical conductivity are performed to study the dynamic characteristics of structures formed in a highly concentrated electrorheological suspension under a dc electric field of 1.8 kV/mm. Small amplitude oscillatory shear measurements reveal that there is no direct correlation between the elastic modulus and the electrical conductivity of the suspension. A significant increase in conductivity is observed following large amplitude oscillatory shear and, in this case, the linear elastic modulus is also substantially enhanced. These observations are interpreted according to the concept of contact disorder arising from the anisotropic shape of the particulate phase.

To explore the effects of aging on the levels of reproduction-related mRNA genes including Gnrh, KISS1/KISS1r, estrogen receptor-alpha (ERα), estrogen receptor-beta (ERβ) and progesterone receptor (PR) in hypothalamus.Proestrus and metestrus in young (3-4 months) and middle-aged (10-11 months) female mice and diestrus in senile (18-19 months) female mice were observed. And the levels of related mRNA genes in preoptic area anterior hypothalamus (POA-AH) and medial basal hypothalamus (MBH) were determined by real-time polymerase chain reaction (RT-PCR).In middle-aged mice on proestrus, the level of Gnrh mRNA in POA-AH (0.896 ± 0.049) was significantly lower than that in young mice (1.228 ± 0.147, P = 0.049). The level of ERα mRNA in POA-AH decreased in young mice on proestrus whereas increased in middle-aged mice (0.432 ± 0.063 vs 0.603 ± 0.018, P = 0.016). The level of ERα mRNA of POA-AH, both in middle-aged mice (0.432 ± 0.063, P = 0.014) and senile mice (0.403 ± 0.145, P = 0.020) on diestrus, were significantly lower than that in young mice. The PR mRNA expression in middle-aged mice on proestrus (1.037 ± 0.037) was markedly lower than that in young mice (1.251 ± 0.081, P = 0.031) . In senile mice, the levels of Gnrh mRNA (1.520 ± 0.146, P = 0.004) and ERβ mRNA (1.572 ± 0.184, P = 0.011) increased in POA-AH compared with that in young mice on metestrus. Aging had no effect upon KISS1 and KISS1r mRNA levels in POA-AH. In contrast, KISS1 mRNA level of MBH in middle-aged (1.663 ± 0.398, P = 0.037) and senile (2.622 ± 0.454, P = 0.014) mice obviously increased compared with the young mice group.Higher levels of ERα mRNA and decreases of PR and Gnrh mRNA in POA-AH in middle-aged mice on proestrus may play an important role in declining reproductive function.

Familial hypercholesterolemia is an autosomal dominant inherited disease characterized by elevated plasma low-density lipoprotein cholesterol (LDL-C). It is mainly caused by mutations of the low-density lipoprotein receptor (LDLR) gene. Currently, the methods of whole genome sequencing or whole exome sequencing for screening mutations in familial hypercholesterolemia are not applicable in China due to high cost. We performed targeted exome sequencing of 167 genes implicated in the homozygous phenotype of a proband pedigree to identify candidate mutations, validated them in the family of the proband, studied the functions of the mutant protein, and followed up serum lipid levels after treatment. We discovered that exon 9 c.1268 T>C and exon 8 c.1129 T>G compound heterozygous mutations in the LDLR gene in the proband derived from the mother and father, respectively, in which the mutation of c.1129 T>G has not been reported previously. The mutant LDL-R protein had 57% and 52% binding and internalization functions, respectively, compared with that of the wild type. After 6 months of therapy, the LDL-C level of the proband decreased by more than 50% and the LDL-C of the other family members with heterozygous mutation also reduced to normal. Targeted exome sequencing is an effective method for screening mutation genes in familial hypercholesterolemia. The exon 8 and 9 mutations of the LDLR gene were pedigree mutations. The functions of the mutant LDL-R protein were decreased significantly compared with that of the wild type. Simvastatin plus ezetimibe was proven safe and effective in this preschool-age child.

To explore the impact and related mechanisms of stromal cell-derived factor-1α (SDF-1α) on serum deprivation-induced apoptosis of cardiac stem cells (CSCs).CSCs were isolated from adult mouse heart tissue and cultured in vitro. Obtained cells were purified using magnetic-activated cell sorting (MACS) with c-kit magnetic beads. C-kit(+)CSCs were divided into five groups: normal control group, serum deprivation group, serum deprivation+SDF-1α group, serum deprivation+SDF-1α+AMD3100 group, serum deprivation+SDF-1α+LY294002 group. Cell apoptosis was assessed using the DeadEnd Colorimetric TUNEL System and flow cytometry analyses with an Annexin V-FITC Apoptosis Detection Kit. The viability of CSCs was assessed by CCK-8. The protein expression of Bcl-2 and phosphorylated Akt were detected by Western blot. The caspase-3 activity was determined using caspase-3 Colorimetric Assay Kit.After magnetic separation, more than 85% of cardiosphere derived cells were positive for c-kit expression. Compared with the normal control group, the apoptosis rate of serum deprivation group was significantly increased[(27.03 ± 0.80)% vs. (1.51 ± 0.54)%, P0.01], which could be significantly reduced by SDF-1α in a concentration dependent manner and peak effect was seen with 100 ng/ml SDF-1α[(10.67 ± 1.06)% vs. (27.03 ± 0.80)%, P0.01]. The expressions of p-Akt and Bcl-2 were significantly increased and the activity of caspase-3 was significantly decreased in serum deprivation+SDF-1α group compared to serum deprivation group (P0.01). Further more, the expression of p-Akt and Bcl-2 were significantly decreased and the activity of caspase-3 was increased in both serum deprivation+SDF-1α+AMD3100 group and serum deprivation+SDF-1α+LY294002 group compared to serum deprivation+SDF-1α group (P0.01).SDF-1α reduces serum deprivation induced CSCs apoptosis via modulating PI3K/Akt signaling pathway.

Hyperpolarization-activated currents (Ih) mediated by hyperpolarization-activated cyclic nucleotide-gated (HCN) channels modulate excitability of myelinated A- and Ah-type visceral ganglion neurons (VGN). Whether alterations in Ih underlie the previously reported reduction of excitability of myelinated Ah-type VGNs following ovariectomy (OVX) has remained unclear. Here we used the intact nodose ganglion preparation in conjunction with electrophysiological approaches to examine the role of Ih remodeling in altering Ah-type neuron excitability following ovariectomy in adult rats. Ah-type neurons were identified based on their afferent conduction velocity. Ah-type neurons in nodose ganglia from non-OVX rats exhibited a voltage 'sag' as well as 'rebound' action potentials immediately following hyperpolarizing current injections, which both were suppressed by the Ih blocker ZD7288. Repetitive spike activity induced afterhyperpolarizations lasting several hundreds of milliseconds (termed post-excitatory membrane hyperpolarizations, PEMHs), which were significantly reduced by ZD7288, suggesting that they resulted from transient deactivation of Ih during the preceding spike trains. Ovariectomy reduced whole-cell Ih density, caused a hyperpolarizing shift of the voltage-dependence of Ih activation, and slowed Ih activation. OVX-induced Ih remodeling was accompanied by a flattening of the stimulus frequency/response curve and loss of PEMHs. Also, HCN1 mRNA levels were reduced by ∼30% in nodose ganglia from OVX rats compared with their non-OVX counterparts. Acute exposure of nodose ganglia to 17beta-estradiol partly restored Ih density and accelerated Ih activation in Ah-type cells. In conclusion, Ih plays a significant role in modulating the excitability of myelinated Ah-type VGNs in adult female rats.

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.

Ethyl carbamate (EC) was analyzed during yellow rice wine production and storage. EC increased slowly during fermentation and rapidly after frying and sterilization. Less amount of EC was formed when cooled rapidly to 30 °C than when cooled naturally. High temperature and long storage time increased EC formation. After 400 days storage, EC increased from 74.0 to 84.2, 131.8 and 509.4 μg/kg at 4 °C, room temperature and 37 °C, respectively, and there was significantly difference between the fried wine and the wine on sale from 2011 (p0.01). Urea increased during yellow rice wine fermentation and was above 20 mg/kg after the wine was fried; urea contributed to EC formation when the fried wine was cooled slowly. These results indicate that it is necessary for industry to optimize the wine frying conditions, such as temperature, time and cooling process in order to decrease EC formation.

Frontotemporal lobar degeneration (FTLD) represents a group of clinically, neuropathologically and genetically heterogeneous disorders with plenty of overlaps between the neurodegenerative mechanism and the clinical phenotype. FTLD is pathologically characterized by the frontal and temporal lobar atrophy. Frontotemporal dementia (FTD) clinically presents with abnormalities of behavior and personality and language impairments variants. The clinical spectrum of FTD encompasses distinct canonical syndromes: behavioural variant of FTD (bvFTD) and primary progressive aphasia. The later includes nonfluent/agrammatic variant PPA (nfvPPA or PNFA), semantic variant PPA (svPPA or SD) and logopenic variant PPA (lvPPA). In addition, there is also overlap of FTD with motor neuron disease (FTD-MND or FTD-ALS), as well as the parkinsonian syndromes, progressive supranuclear palsy (PSP) and corticobasal syndrome (CBS). The FTLD spectrum disorders are based upon the predominant neuropathological proteins (containing inclusions of hyperphosphorylated tau or ubiquitin protein, e.g transactive response (TAR) DNA-binding protein 43 kDa (TDP-43) and fusedin-sarcoma protein in neurons and glial cells) into three main categories: (1) microtubule-associated protein tau (FTLD-Tau); (2) TAR DNA-binding protein-43 (FTLD-TDP); and (3) fused in sarcoma protein (FTLD-FUS). There are five main genes mutations leading clinical and pathological variants in FTLD that identified by molecular genetic studies, which are chromosome 9 open reading frame 72 (C9ORF72) gene, granulin (GRN) gene, microtubule associated protein tau gene (MAPT), the gene encoding valosin-containing protein (VCP) and the charged multivesicular body protein 2B (CHMP2B). In this review, recent advances on the different clinic variants, neuroimaging, genetics, pathological subtypes and clinicopathological associations of FTD will be discussed.

In many existing elastoplastic plane-strain solutions for a circular tunnel excavated in a homogeneous, isotropic rock mass under a hydrostatic stress field, the longitudinal axial stress (or the out-of-plane stress) {sigma}{sub z} is not correctly evaluated. This paper describes the concepts and a numerical method to calculate this stress component and proposes analytical solutions for practical applications. The computed numerical solutions are compared with theoretical solutions where possible. The relations between rock-mass dilatancy, tunnel displacements, and the in-situ stress field are investigated. Mohr-Coulomb and Hoek-Brown yield criteria with appropriate plastic potential functions and nonassociated flow rules are used in the brittle plastic calculations. The effects of various boundary conditions on the numerical analyses are discussed and several important practical conclusions are drawn from the investigations.

Background Homozygous familial hypercholesterolaemia is a rare life-threatening disease characterised by markedly elevated LDL cholesterol concentrations and accelerated atherosclerosis. Few studies have focused specifically on this group of patients, especially in China. We set out to investigate the phenotypical characteristics and the genotype–phenotype correlations of homozygous familial hypercholesterolaemia in China. Methods In this analysis of cross-sectional data, we collected data from patients with suspected homozygous familial hypercholesterolaemia from ten clinical hospitals in mainland China from 2003 to 2015. Inclusion criteria included total cholesterol greater than 10 mmol/L, presence of xanthomas, and a family history of premature coronary heart disease. Clinical data and DNA sequence of LDLR, APOB , and PCSK9 were obtained in all patients. All participants gave written informed consent using consent forms approved by the research ethics committee of Beijing Anzhen Hospital, China. Findings We collected data from 64 probands with suspected unrelated homozygous familial hypercholesterolaemia (mean age 13 years [range 2–37]). Molecular diagnosis showed that 15 had homozygous familial hypercholesterolaemia, 34 had compound heterozygous disease, nine had heterozygous disease, and six were mutation-negative. 66 LDLR mutations were found, and no APOB or PCSK9 mutations were found. Of the LDLR mutations, 26 were novel, and the most common mutation, occurring in 11 (17%) of 64 patients, was Trp462X (c1448G→A). The mean total cholesterol concentration was 16·6 mmol/L (SD 3·84; range 10·4–25·9), and the mean LDL cholesterol concentration was 13·86 mmol/L (SD 3·50; range 7·1–23·5). Total cholesterol and LDL cholesterol concentrations did not significantly differ between patients with homozygous familial hypercholesterolaemia, compound heterozygous familial hypercholesterolaemia, heterozygous familial hypercholesterolaemia, or those who were mutation-negative. Interpretation This research provides important clinical and genetic information about suspected homozygous familial hypercholesterolaemia in mainland China. Our data suggest that diagnostic criteria need to be developed to screen patients with potential homozygous familial hypercholesterolaemia. Funding National Natural Science Foundation of China (number 81471098).

Background Recent guidelines suggest that more attention should be focused on children with homozygous familial hypercholesterolemia (HoFH). China may have 3.8 million potential FH patients, but there are limited data focused on HoFH children. Objective We systematically analyzed the characteristic phenotype and the relationship between the genotype and the phenotype in HoFH children with the unique Chinese W483X mutation in the low-density lipoprotein (LDL)–receptor gene. Methods A systematic retrospective analysis of the lipid and cardiovascular characteristics of HoFH patients in the atherosclerosis clinic of Beijing Anzhen Hospital was performed. The W483X mutation was confirmed using DNA sequencing of the patients and their parents. Results Two HoFH and 9 compound heterozygous patients (mean age = 14.7 years) with 2 novel mutations, Q254X and c.1363delC, were found. In total, 81.8% of the patients were from southern China. All the patients had xanthoma, and the average TC and LDL-C levels were 16.8 and 14.4 mmol/L, respectively. Echocardiography showed that 63.6% of the patients had aortic calcification, and 54.5% had mild regurgitation of the aortic valve. The coronary flow velocity reserve had a mean value of 2.12, and the cIMT was 0.17 cm. The follow-up period was between 3 months and 8 years. Although all the patients began the lipid-lowering treatment, 2 patients died because of severe cardiovascular disease. The LDL-C levels of 6 patients were slightly decreased by approximately 21% and remained far from the target values, and the other 3 patients' LDL-C levels increased by 13%. Conclusions The results suggest that younger HoFH patients with W483X mutations had a severe phenotype and should receive more aggressive treatment.

A method for ethyl carbamate (EC) determination in alcoholic beverages and soy sauce was developed by GC-MS. We adopted the diatomaceous earth solid-phase extraction (SPE) column and elution solvent of ethyl acetate/diethyl ether (5:95 v/v) for sample cleaning. The in-house validation showed the limit of quantification (LOQ) was 5.0 μg/kg. In the accuracy assay, the total average recovery for was 96.7%. The relative standard deviations (RSDs) were5%. Subsequently, a collaborative trial was organized for the further validation. The RSDs for repeatability and reproducibility were 1.2-7.8% and 2.3-9.6% respectively. It indicated that the present method performed well in different laboratories.

We aimed to establish a quantified method for the 17 phthalate acid esters (PAE) in edible vegetable oil by gas chromatography-mass spectrometry (GC-MS) with the pretreatment of acetonitrile extraction and silica/N-(n-propyl)ethylenediamine (silica/PSA) mixed solid phase extraction column and evaluated the PAE of 25 edible oil samples from supermarkets in Hangzhou city.The internal standard solution (D4-DEHP) was added in edible vegetable oil sample. The analytes were extracted by acetonitrile with 1 min vortex, and centrifuged at 3050×g for 5 min. The supernatant was then cleaned with silica/PSA column, and eluted with acetonitrile. The elution was dried with N2 flow at 50°C and diluted to 1.0 ml with hexane. Then, 17 PAE were tested by GC-MS and quantified with internal standards. The repeatability and sensitivity of the assay were evaluated. PAE were then determined in 25 plastic buckets of edible vegetable oil from supermarkets in Hangzhou city.By the quantification of internal standard of D4-DEHP, a good linearity range of related 17 PAE was observed. The correlation coefficient was 0.994-1.000 and the standard lowest quantified level was 0.05-0.15 µg/ml. The spiking recoveries of 17 PAE were 78.3%-108.9% with the RSD of 4.3%-12.1% (n=6). The method detection limits were 0.1-0.2 mg/kg. In 25 plastic buckets of edible vegetable oil from Hangzhou, DMP, DEP, DIBP, DBP and DEHP were detected at the range of0.1-1.8 mg/kg and the detection rates were 12% (3/25), 24% (6/25), 100% (25/25), 96% (24/25) and 100% (25/25), respectively. Other 12 PAE was not detected. For DBP with the level of0.1 to 1.3 mg/kg, the results of 16% (4/25) samples exceeded the regular migrating limit of 0.3 mg/kg. For DEHP of0.2-1.8 mg/kg, the data of 12% (3/25) samples were beyond the regular migrating limit of 1.5 mg/kg.The pretreatment by silica/PSA mixed solid phase extraction column can satisfy the PAE determination requirements in edible vegetable oils. The DMP, DIBP, DEP, DBP and DEHP were detected from the survey of 25 edible oil samples in Hangzhou city.

To observe the ethyl carbamate concentrations in different commercial fermented foods in Hangzhou in 2010.In 2010, 237 commercial fermented food samples of eight categories, including yellow wine, white spirit, wine, beer, cooking wine, sauce, vinegar and fermented bean curd, were purchased from 3 different size markets respectively in Hangzhou. The ethyl carbamate was measured by gas chromatography-mass spectrometry in selection ion mode, after the samples were coupled with D5-ethyl carbamate, and purified by diatomite solid phase extraction column.The results showed that ethyl carbamate was detected in all samples analyzed (100%) with the range from 2.0 µg/kg to 515.0 µg/kg. The ethyl carbamate average (median) levels in 8 food categories were descending with fermented red bean curd (182.2 µg/kg (161.2 µg/kg)), yellow wine (159.6 µg/kg (121.0 µg/kg)), cooking wine (86.8 µg/kg (95.6 µg/kg)), white spirit (72.0 µg/kg (60.5 µg/kg)), soy sauce (47.2 µg/kg (40.7µg/kg)), vinegar (26.7 µg/kg (31.8 µg/kg)), wine (15.7 µg/kg (16.8 µg/kg)) and beer (2.2 µg/kg (2.3 µg/kg)).The ethyl carbamate was detected in all fermented foods in Hangzhou in 2010, and the levels of ethyl carbamate in red bean curd and yellow wine were higher than others.

To investigate the effect of missense mutation in PCSK9 gene R306S on the metabolism of LDL and to analyze its relationship between structure and function. METHODS The wild type PCSK9 gene and the second or the third grade structures of the mutant R306S coding protein were all forecasted with the aid of Expert Protein Analysis System. The PCSK9 gene eukaryocyte expression vector was constructed and the nucleotide sequence was determined; The pathotype recombinant eukaryotic expression plasmid which carrying PCSK9 gene was constructed by directed-mutation method; Liposorne was used as intermediate to transfect the recombinant plasmid into BEL-7402 cell, the cells receiving no plasmid transfection served as control during this process; The LDL-R level was detected by Western Blot. RESULTS Through simulating the second and the third grade structures of protein, conformations changes were found in the two major coding protein parts of mutated PCSK9 gene: carboxy-terminal domain and catalyzing subunit site domain, the space between these two domains increased as well. And that was not found in wild type of PCSK9 gene. Tansfecting wild type of PCSK9 plasmid into BEL-7402 cell could lead to the decrease of LDL-R mature protein expression compared with controlled group(P

To investigate clinical risky factors and improve the ability to discover critically ill patients suffering from influenza A H1N1 in emergency department.A retrospective study of 65 cases with severe influenza A H1N1 was conducted. Univariate analysis and multivariate Logistic regression analysis were used to identify independent predictors of critically ill cases. Goodness of Fit test was performed on an established clinical predicting scoring (prediction of critically ill with influenza score, PCIS) system.Univariate analysis showed that significant difference between critically ill and severe group consisted of eight factors including age distribution, more than one comorbidity, chronic cardiovascular disease, oncologic diseases, sputum, myalgia/arthralgia, dyspnea and oxygen saturation at admission. A multivariate Logistic regression showed an association between development of critical illness and more than one comorbidity [odds ratio (OR)=7.611, 95% confidence interval (95%CI) 1.635-35.429], myalgia/arthralgia (OR=7.523, 95%CI 1.462-38.716), dyspnea (OR=11.090, 95%CI 1.373-89.565), and oxygen saturation0.95 at admission (OR=8.088, 95%CI 1.019-84.969, all P0.05). The prognostic criteria had a good discriminative ability [area under receiver operating characteristic curve (AUC) was 0.922, 95%CI 0.860-0.985, P=0.000]. The PCIS scoring system was established according to the level of four high risk factors. Low risk (PCIS 0-1), intermediate risk (PCIS 2), high risk (PCIS 3), and very high risk (PCIS≥4) were categorized for predicting the occurrence of critical illness, and the Goodness of Fit test was good (R(2)=0.940 6, P=0.030 1).Emergency physician can predict the development of critical condition in patients with influenza A H1N1 by using clinical characteristics including comorbidity, myalgia/arthralgia, dyspnea, and oxygen saturation at admission, and it is helpful in making clinical decision.

To explore the possible mechanism of tert-butyl hydroperoxide (t-BHP)-induced apoptosis in murine MIN6 pancreatic beta-cells.MIN6 cells were cultured in vitro. Cell damage was evaluated by epifluorescence microscopy after staining with AO-EB. The percentage of cell apoptosis was determined by flow cytometric assay after Annexin- V-PI staining. Nitric oxide levels were measured by Griess assay. Inducible nitric oxide synthase(iNOS) protein and NF-kappaBp65 fragment were detected by Western blot.Exposure of 25 micromol/L t-BHP to MIN6 cells for 60 min, cell viability was reduced and the percentage of apoptosis was increased significantly. The levels of cytoplasmic iNOS protein and nitrite were elevated. Meanwhile, treatment with t-BHP resulted in nucleus NF-kappaBp65 fragment peaking at 20 min. Both L-NAME and N-Acetyl-l-cysteine (NAC) attenuated the elevated levels of nitrite and percentage of apoptosis due to t-BHP alone.NF-kappa-iNOS-nitric oxide signalling pathway can mediated t-BHP induced apoptosis in MIN6 cells .

This study is to explore whether the Wnt/beta-catenin signaling pathway is involved in the process of tripchlorolide (T4) protecting against oligomeric Abeta(1-42)-induced neuronal apoptosis. Primary cultured cortical neurons were used for the experiments on day 6 or 7. The oligomeric Abeta(1-42) (5 micromol x L(-1) for 24 h) was applied to induce neuronal apoptosis. Prior to treatment with Abeta(1-42) for 24 h, the cultured neurons were pre-incubated with T4 (2.5, 10, and 40 nmol x L(-1)), Wnt3a (Wnt signaling agonists) and Dkk1 (inhibitors) for indicated time. Then the cell viability, neuronal apoptosis, and protein levels of Wnt, glycogen synthase kinase 3beta (GSK3beta), beta-catenin and phospho-beta-catenin were measured by MTT assay, TUNEL staining and Western blotting, respectively. The result demonstrated that oligomeric Abeta(1-42) induced apoptotic neuronal cell death in a time- and dose-dependent manner. Pretreatment with T4 significantly increased the neuronal cell survival and attenuated neuronal apoptosis. Moreover, oligomeric Abeta(1-42)-induced phosphorylation of beta-catenin and GSK3beta was markedly inhibited by T4. Additionally, T4 stabilized cytoplasmic beta-catenin. These results indicate that tripchlorolide protects against the neurotoxicity of Abeta by regulating Wnt/beta-catenin signaling pathway. This may provide insight into the clinical application of tripchlorolide to Alzheimer's disease.

The spatial non-uniformity in the photo-response of the detectors strongly influences the quality of infrared image. There was no one effective means to evaluate the quality of infrared imaging systems and the performance of the NUC algorithms. A real-time testing system was designed and implemented, which could be used to acquire the data from infrared imaging system, measure the system and analyze the data, and provide researching platform both on software and hardware for developing the infrared processing system. The experimental results show that the system implemented acquiring infrared data in real-time with high speed and high precision. The system has been applied and it was helpful for the researcher to exploit and improve the NUC algorithms.

To develop a simple, accurate, rapid, economic, large-scale detection method for the detection of single nucleotide polymorphisms (SNPs) metabolic enzymes, using polymerase chain reaction with confronting two-pair primers (PCR-CTPP).The primers of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were designed for PCR-CTPP, and the PCR conditions were optimized. The results of genotyping were verified by DNA sequencing. The above SNPs were detected by the PCR-CTPP detection method in a randomly selected 183 healthy individuals of Han ethnicity. The genotype frequencies were analyzed and compared with people from other ethnicities.The allele-specific bands of CYP1A1 (A4889G), EPHX1 (A416G) and NQO1 (C609T) were successfully amplified by PCR-CTPP under the optimal conditions and the results of genotyping were consistent with DNA sequencing. Among 183 healthy Han individuals, the genotypic distributions of CYP1A1 (A4889G) , EPHX1 (A416G) and NQO1 (C609T) showed that the wild-type, homozygous variants, and heterozygotes were 103 (56.3%), 8 (4.4%), 72 (39.3%) and 142 (77.6%), 4 (2.2%), 37(20.2%), 60(32.8%), 32 (17.5%), 91 (49.7%) respectively. The distributions of genotypes were all in accordance with the Hardy-Weinberg equilibrium (P0.05), with statistical differences and with other ethnic populations (P0.05).The SNPs of metabolic enzymes can be detected by PCR-CTPP method which is simple, accurate, rapid, economic and with large scale. PCR-CTPP can be used for large scale clinical and epidemiological screening.

Extracts of Chinese herb Tripterygium wilfordii Hook. f. (TWHF) have been found to have potent anti-inflammatory and immunosuppressive functions and widely used in China for treatment of rheumatoid arthritis. Also they have been considered to be the potential drugs in the treatment of tumor and acute graft rejections. With the progress of neuroimmunological research on neurodegenerative diseases, such as Alzheimer's disease (AD), Parkinson disease (PD) and multiple sclerosis (MS), the neuroprotective strategies to rescue neurons from immunological injury are currently being explored. Recently, studies have indicated that extracts of TWHF such as triptolide, tripchlorolide and (5R)-5-hydroxytriptolide are able to attenuate progression of these neuroimmunologic disorders in vitro and in vivo. Accumulating evidence has shown that they can promote neuronal survival and neurite growth and facilitate functional recovery of brain injury by invoking distinct mechanisms that are related to their neuroprotective roles as inhibitor of neuroinflammatory toxicity of activated-microglia, antioxidants, calcium channel blockers, neurotrophic actions, modulating T cell functions, inhibitor of transcriptional activation of NF-kappaB on genes and signaling. Significant pharmaceutical strategies against neuroimmunologic disorders will hopefully be discovered by understanding the valuable components of TWHF.

To explore the effect of glucagon-like peptide-1 receptor agonist--Exendin-4 (Ex-4) on murine MIN6 pancreatic beta-cells apoptosis induced by oxidative stress, the morphological changes of cell damage were evaluated by epifluorescence microscopy after staining with AO-EB. The percentage of cell apoptosis was determined by flow cytometric assay after Annexin-V-FITC-PI staining. Nitric oxide level was measured by Griess reagent assay. Inducible nitric oxide synthase (iNOS) protein and NF-kappaBp65 fragment were detected by Western blotting. Ex-4 inhibited the increase of nitrite level and percentage of apoptosis induced by t-BHP in MIN6 cells. Furthermore, Ex-4 partly reduced the expression of iNOS protein and the ratio of NF-kappaBp65 protein in nucleus:cytosol induced by t-BHP. These results suggest that Ex4 protects MIN6 pancreatic kappa-cells from oxidative stress-induced apoptosis via down-regulation of NF-kappaB-iNOS-nitric oxide pathway.

Clostridium butyrium CB2 isolated from chickens was tested as a potential fish probiotic in the Chinese drum, Miichthys miiuy. Fish were fed live (CB), dead CB2 (D-CB) cells (10(8) cells g(-1)) or spent culture supernatant (SCS), for 30 days and challenged with Vibrio anguillarum or Aeromonas hydrophila. Survival was higher in both the CB and the CB-D fed groups, but the SCS group was not significantly different from the control. After feeding live or dead CB2 cells, there was increase in phagocytic activity of the head kidney macrophages, the lysozyme activity of serum and gut mucosa and immunoglobulin (Ig) level. The SCS group showed no obvious change in immune parameters. The results suggest that live or dead Clostridium butyrium CB2 has an immunomodulatory effect on fish.

We investigated the effects of fish protein hydrolysate (FPH) on growth performance and humoral immune response of the large yellow croaker (Pseudosciaena crocea R.). One thousand and two hundred large yellow croakers [initial average weight: (162.75+/-23.85) g] were divided into four groups and reared in floating sea cages (3 m x 3 m x 3 m). The animals were fed with 4 diets: basal diet only (control) or diets supplemented with 5%, 10% and 15% (w/w) FPH. The results show that dietary FPH levels significantly influenced the growth and immunity of the large yellow croaker. Compared with the control group, total weight gain (TWG) in all treatment groups, relative weight gain (RWG) and specific growth rate (SGR) in fish fed with diets supplemented with 10% and 15% FPH were significantly increased (P0.05). Similar results were observed in immune parameters [lysozyme activity, serum complements, immunoglobulin M (IgM)]. Lysozyme activity, complement C4 and IgM were also significantly increased (P0.05) in fish fed with diets supplemented with 10% and 15% FPH, while complement C3 level was significantly increased (P0.05) in all treatment groups. In general, with the supplementation of FPH, particularly at dose of 10%, the growth performance and immunity of the large yellow croaker can be improved effectively.

Background Familial hypercholesterolemia (FH), caused by low density lipoprotein (LDL) receptor (LDL-R) gene mutations, is associated with increased risk of premature coronary heart disease. Until now, limited molecular data concerning FH are available in China. The present study described the clinical profiles and cell biological defects of a Chinese FH kindred with novel LDL-R gene mutation. Methods The patient's LDL-R gene coding region was sequenced. The patient's lymphocytes were isolated and the LDL-R expression, binding and up-take functions were observed by immunohistochemistry staining and flow cytometry detection. The patient's heart and the major large vessels were detected by vessel ultrasound examination and myocardial perfusion imaging (MPI). Results The patient's LDL-R expression, LDL binding and up-take functions were significantly lower than normal control (39%, 63% and 76% respectively). A novel homozygous 1439 C-->T mutation of the LDL-R gene was detected in the patient and his family. ECG showed atypical angina pectoris. Echocardiogram showed stenosis of the coronary artery and calcification of the aortic valve and its root. Blood vessel ultrasound examination showed the thickness of large vessel intima, and the vessel lumen was narrowed by 71%. MPI showed ischemic changes. Conclusions The LDL-R synthesis dysfunction of FH patients leads to arterial stenosis and calcification, which are the major phenotype of the clinical disorder. The mutation of the LDL-R gene is determined. These data increase the mutational spectrum of FH in China.

This study is to explore the effect of ginsenoside Rb1 on the process of beta-amyloid peptide(25-35) (Abeta(25-35)) -induced hyperphosphorylation of tau protein, and on the level of cyclin-dependent kinase 5 activator, p25/p35. Western blotting and/or immunocytochemical staining were used to detect the levels of phosphorylation of tau protein at the sites of Thr205, Ser396, Ser404 in hippocampal neurons, cdk5 and p25/p35. After exposure to Abeta(25-35) (20 micromol x L(-1)) for 12 h, the levels of tau protein phosphorylation at the sites of Thr205, Ser396, Ser404 were enhanced, the level of p25 was increased, but the level of protein cdk5 was not changed markedly. Pretreatment with ginsenoside Rb1 reduced Abeta(25-35) -induced hyperphosphorylation of tau protein and decreased the lever of p25, but had no effect on cdk5. Ginsenoside Rb1 can attenuate Abeta(25-35) -induced hyperphosphorylation of tau protein through CDK5 signal pathway.

The variation of the dynamic modulus of a model electrorheological fluid with strain amplitude is shown to closely resemble that of traditional physical gels. Comparison of the in-phase stress component in each system indicates that the material strength of electrorheological fluids in shear is limited by the small strain amplitude to structural failure. An anisotropic network model is proposed for electrorheological fluids, in which the primary structure consists of chains of particles spanning the electrode gap along the field direction, while many-body interactions form a secondary structure of short chains tilted with respect to the field direction and interconnecting the primary chains. A geometrical argument shows that the tensile strain in the secondary structure can be an order of magnitude larger than that in the primary chains. This nonuniform strain distribution poses an inherent structural limitation on the shear material strength of electrorheological fluids.

To identify the mutation of low density lipoprotein receptor(LDLR) gene in a large Chinese family with familial hypercholesterolemia(F H) and make a discussion on the pathogenesis of FH at the molecular level.Investigations were made on a patient with the clinical phenotype of homozygous FH and his parents for mutations of promoter and all 18 exons of LDLR gene. Screening was carried out using Touch down PCR and a g arose gel electrophoresis, combined with DNA sequence analysis. The results were compared with the normal sequences in GenBank and FH database (www.ucl.uk/fh) t o find the mutation. Then the mutation was identified in other members of the family. In addition, the authors screened the apolipoprotein B(100) (apoB(100)) gene f or known mutations (R3500Q) that cause familial defective apoB(100) (FDB) by PCR-RFLP.A novel homozygous IN III 5' GT --AT mutation in the splice donor of LDLR intron 3 was detected in the homozygote propositus with FH. The mutation was also identified in four heterozygous carriers in his family. No mutations R3500Q of apoB(100)were observed.A homozygous G --A splice mutation in LDLR gene was first reported. The change of the splice donor in LDLR intron 3 may cause skipping of exon 3, which is responsible for FH. Perhaps it is a particular pathogenesis for Chinese people.