[Effect of recombinant human thrombin for hemostasis in liver resection: a randomized controlled phase â ¢ clinical trial].

Objective: To evaluate the hemostatic efficacy, safety and immunogenicity of recombinant human thrombin in the treatment of liver wounds that still ooze after conventional surgical hemostasis. Methods: A multicenter, stratified randomized, double-blind, placebo-controlled phase Ⅲ trial with a planned enrollment of 510 subjects at 33 centers, with a 2∶1 randomization to the thrombin group versus the placebo group. An interim analysis will be conducted after approximately 70% of the subjects have completed the observation period. The primary efficacy endpoint was the rate of hemostasis within 6 minutes at the point of bleeding that could be evaluated. Safety analysis was performed one month after surgery, and the positive rates of anti-drug antibody (ADA) and neutralizing antibody were evaluated. Results: At the interim analysis, a total of 348 subjects had been randomized and received the study drug (215 were male and 133 were female). They were aged 19-69 (52.9±10.9)years. Among them, 232 were in the thrombin group and 116 were in the placebo group, with balanced and comparable demographics and baseline characteristics between the two groups. The hemostasis rate at 6 minutes was 71.6% (95%CI:65.75%-77.36%) in the thrombin group and 44.0% (95%CI: 34.93%-53.00%) in the placebo group, respectively (P<0.001). No grade≥3 drug-related adverse events and no drug-related deaths were reported from the study.No recombinant human thrombin-induced immunologically-enhanced ADA or immunologically-induced ADA was detected after topical use in subjects. Conclusion: Recombinant human thrombin has shown significant hemostatic efficacy and good safety in controlling bleeding during liver resection surgery, while also demonstrating low immunogenicity characteristics.

[Current status of blood lipids in people with hypertension and diabetes in Henan province].

Objective: To investigate the blood lipid levels and prevalence of dyslipidemia in people with hypertension and diabetes in Henan province. Methods: From April 2016 to April 2017, multi-stage cluster sampling was adopted to investigate 71 285 local residents aged between 35 and 75 from 6 districts and counties in Henan province including Zhongmu county of Zhengzhou city, Huojia county of Xinxiang city, Hualong district of Puyang city, Qi county of Hebi city, Xigong district of Luoyang city, and Wugang city of Pingdingshan city. Blood samples were collected. According to the diagnostic criteria of hypertension and diabetes, the study population was divided into control group (n=29 427), hypertension group (n=21 965), diabetes group (n=8 009) and hypertension-diabetes group (n=11 884). Comparisons on blood lipid levels and dyslipidemia between 4 groups were performed. Results: The total cholesterol (TC) level of all subjects was 4.37 (3.78, 5.05) mmol/L. The triglyceride (TG) level was 1.27 (0.97, 1.80) mmol/L, the low-density lipoprotein cholesterol (LDL-C) level was 2.34 (1.88, 2.88) mmol/L and the high-density lipoprotein cholesterol (HDL-C) level was 1.31 (1.08, 1.59) mmol/L. Except for the TC level in women aged 65-75 years and LDL-C levels in women aged 55-64 and 65-75 years, there were significant differences in TC, TG, LDL-C and HDL-C levels between subjects of control group, hypertension group, diabetes group, and hypertension-diabetes group in different age ranges (including 35-44, 45-54, 55-64,and 65-75 years) and genders(all P<0.01).Except for the LDL-C and HDL-C in men aged 35-44 years and LDL-C in women aged 65-75 years, there were significant differences in the dyslipidemia rates of TC, TG, LDL-C and HDL-C between subjects of control group, hypertension group, diabetes group and hypertension-diabetes group in different age ranges and genders(P<0.01 or <0.05). After adjusting for age, gender, smoking, drinking, snoring, region, and body mass index, multivariate logistic regression analysis showed that hypertension (OR=1.221, 95%CI 1.113-1.339, P<0.01), diabetes (OR=1.636, 95%CI 1.461-1.833, P<0.01) and hypertension-diabetes (OR=1.832, 95%CI 1.658-2.023, P<0.01) were independent risk factors for TC abnormality. Hypertension (OR=1.566, 95%CI 1.478-1.659, P<0.01), diabetes (OR=2.182, 95%CI 2.031-2.342, P<0.01) and hypertension-diabetes (OR=2.655, 95%CI 2.492-2.829, P<0.01) were also independent risk factors for TG abnormality. Diabetes (OR=1.510, 95%CI 1.309-1.742, P<0.01) and hypertension-diabetes (OR=1.461, 95%CI 1.285-1.661, P<0.01) were independent risk factors for LDL-C abnormality. Diabetes (OR=1.261, 95%CI 1.180-1.346, P<0.01) and hypertension-diabetes (OR=1.195, 95%CI 1.126-1.268, P<0.01) were independent risk factors for HDL-C abnormality. Conclusion: The prevalence of dyslipidemia in patients with hypertension and diabetes is high in Henan province, so adequate blood lipid education and control should be applied to people with risk factors as soon as possible.

RNAi in the striped stem borer, Chilo suppressalis, establishes a functional role for aminopeptidase N in Cry1Ab intoxication.

The striped stem borer, Chilo suppressalis, is a major target pest of transgenic rice expressing the Cry1Ab protein from the bacterium Bacillus thuringiensis (Bt) in China. Evolution of resistance in this pest is a major threat to the durability of Bt rice. Since Bt exerts its activity through binding to specific receptors in the midgut of target insects, identification of functional Cry1Ab receptors in the midgut of C. suppressalis larvae is crucial to evaluate potential resistance mechanisms and develop effective strategies for delaying insect resistance. In this work, we identified the putative Cry1Ab toxin-binding protein, aminopeptidase-N (APN), in the midgut of C. suppressalis by ligand blot and mass spectrometry. After cloning the full-length cDNAs encoding APN isoforms from the C. suppressalis larval midgut, we studied their spatiotemporal expression in different gut tissues and developmental stages. Furthermore, RNA interference (RNAi) against C. suppressalis aminopeptidases (CsAPNs) was employed to illustrate a functional role for CsAPNs in Cry1Ab toxicity to C. suppressalis larvae using injection and oral delivery of Stealth™ siRNA. Down-regulating the expression of CsAPNs by RNAi was closely associated with reduced susceptibility of C. suppressalis to Cry1Ab. These data provide the first direct evidence that CsAPNs participate in the mode of Cry1Ab action and may act as the functional receptor of Cry1A in C. suppressalis larvae.

[The role of hexokinase 2 in the metastasis of hepatocellular carcinoma cells].

Objective: To investigate the regulatory role of HK2 in the metastasis of hepatocellular carcinoma (HCC). Methods: The protein expressions of HK2 in 73 HCC tumor tissues and paired adjacent non-tumor tissues were evaluated by using immunohistochemical analysis. The scratch wound healing assay and Transwell assay had been used to analyze the migration and invasion of HCC cells with HK2 knockdown. Expressions of epithelial-to-mesenchymal transition (EMT)markers, such as E-cadherin, ZO-1, N-cadherin and vimentin, in HCC cells with HK2 knockdown were determined by qRT-PCR and Western blot analysis. Results: The expression levels of HK2 in tumor tissues and adjacent non-tumor tissues were 5.39±3.40 and 2.16±1.55, respectively. The protein expression of HK2 was significantly higher in tumor tissues compared with adjacent non-tumor tissues of HCC (P<0.05). Knockdown of HK2 in HCC cells decreased the cell motility from 1.00±0.54 to 0.56±0.09 (P<0.05), andknockdown of HK2 in HCC cells decreased the number of invaded cells form 345±42 to 215±34 (P<0.05). The expression of epithelial markers ZO-1 and E-cadherin were up-regulated, while mesenchymal markers vimentin and N-cadherin were down-regulated in HCC cells when HK2 was knockeddown. Conclusions: HK2 is up-regulated in HCC and promotes cell motility by stimulating EMT.

Low temperature superplasticity and thermal stability of a nanostructured low-carbon microalloyed steel.

We describe here for the first time the low temperature superplasticity of nanostructured low carbon steel (microalloyed with V, N, Mn, Al, Si, and Ni). Low carbon nanograined/ultrafine-grained (NG/UFG) bulk steel was processed using a combination of cold-rolling and annealing of martensite. The complex microstructure of NG/UFG ferrite and 50-80 nm cementite exhibited high thermal stability at 500 °C with low temperature elongation exceeding 100% (at less than 0.5 of the absolute melting point) as compared to the conventional fine-grained (FG) counterpart. The low temperature superplasticity is adequate to form complex components. Moreover, the low strength during hot processing is favorable for decreasing the spring back and minimize die loss.

Genetic variability and individual assignment of Chinese indigenous sheep populations (Ovis aries) using microsatellites.

The purpose of this study was to assess the genetic characteristics of six breeds of Chinese local sheep using 19 microsatellite loci and to effectively validate statistical methods for individual assignment based on informative microsatellites. All the six breeds deviated from Hardy-Weinberg equilibrium expectations, while the majority of markers complied. The polymorphism information content (PIC) of overall loci for the six populations ranged from 0.283 (SRCRSP5) to 0.852 (OarVH72). Tibetan sheep were the most diverse population with the highest mean allelic richness (6.895), while Ujmuqin (UQ) harboured the lowest allelic richness (6.000). The F-statistics for the six populations were F(IS) = -0.172, F(IT) = -0.082 and F(ST) = 0.077, respectively. Furthermore, the pair-wise F(IS) revealed a moderate genetic differentiation among populations (P < 0.01), indicating that all breeds can be considered genetically independent entities. The lowest genetic differentiation was between Tengchong (TC) and UQ (F(ST) = 0.041), and the highest one was between TC and Fat-tailed Han (F(ST) = 0.111). In comparing the three statistical models, we note that the seven microsatellite loci (MAF65, OarJMP58, SRCRSP9, MCM140, OarAE129, BM8125 and SRCRSP5) commonly used for individual assignment will ensure a powerful detection of individual origin, with accuracy up to 91.87%, when the likelihood-based method is used. Overall, these findings shed light onto the genetic characteristics of Chinese indigenous sheep and offer a set of microsatellite loci that is simple, economic and highly informative for individual assignment of Chinese sheep.

Optimal Design for Marker-assisted Gene Pyramiding in Cross Population.

Marker-assisted gene pyramiding aims to produce individuals with superior economic traits according to the optimal breeding scheme which involves selecting a series of favorite target alleles after cross of base populations and pyramiding them into a single genotype. Inspired by the science of evolutionary computation, we used the metaphor of hill-climbing to model the dynamic behavior of gene pyramiding. In consideration of the traditional cross program of animals along with the features of animal segregating populations, four types of cross programs and two types of selection strategies for gene pyramiding are performed from a practical perspective. Two population cross for pyramiding two genes (denoted II), three population cascading cross for pyramiding three genes(denoted III), four population symmetry (denoted IIII-S) and cascading cross for pyramiding four genes (denoted IIII-C), and various schemes (denoted cross program-A-E) are designed for each cross program given different levels of initial favorite allele frequencies, base population sizes and trait heritabilities. The process of gene pyramiding breeding for various schemes are simulated and compared based on the population hamming distance, average superior genotype frequencies and average phenotypic values. By simulation, the results show that the larger base population size and the higher the initial favorite allele frequency the higher the efficiency of gene pyramiding. Parents cross order is shown to be the most important factor in a cascading cross, but has no significant influence on the symmetric cross. The results also show that genotypic selection strategy is superior to phenotypic selection in accelerating gene pyramiding. Moreover, the method and corresponding software was used to compare different cross schemes and selection strategies.

Expression analyses and antiviral properties of the Beijing-You and White Leghorn myxovirus resistance gene with different amino acids at position 631.

Influenza is a topic of wide public concern, particularly because of the recent emergence of avian flu. The myxovirus resistance (Mx) protein has been shown to have an inhibitory effect on influenza virus and is therefore of great interest. This study examines the Mx protein in 8 local Chinese chicken breeds and 2 exotic chicken breeds. Amino acid 631, found in the Mx GTPase effector domain, was examined in 534 individuals by comparing PCR results, and individuals were separated into the A/A genotype or the G/G genotype, depending on whether amino acid 631 is an Asn or Ser. In the native breed, the frequency of G/G homozygotes is 0.780 (294/377). The Mx expression levels in tissues and chicken embryo fibroblast cells with different genotypes were also studied. The A/A individuals from Beijing-You and White Leghorn breeds had higher Mx expression levels than G/G individuals. The liver, heart, and spleen had higher expression levels than muscle or kidney. The A/A chicken embryo fibroblast cells had higher antiviral activity against vesicular stomatitis virus and Newcastle disease. We provide the first report examining the expression level and antiviral activity of different Mx alleles of nucleotide 2216(S631N) genotypes. This study lays a good foundation for correlative studies examining genotype and antiviral function.

A novel maternal lineage revealed in sheep (Ovis aries).

It is generally believed that domestic sheep have two maternal lineages (haplotypes A and B), based on mitochondrial DNA analysis. In the present study, we provide evidence that a novel maternal lineage (haplotype C) is exhibited in Chinese native sheep. To verify this finding, 231 samples were collected from six Chinese local breeds, which cover the vast geographical region of sheep inhabitation in China. For comparison, 50 samples were collected from two Western breeds collected in China. Mitochondrial DNA was screened by PCR single-strand conformational polymorphism (SSCP), leading to the identification of novel band patterns in ND2 and ND4 genes in the Chinese breeds. Interestingly, mutations at the two loci were in strong linkage disequilibrium. Direct sequencing of the DNA fragments revealed a non-synonymous substitution in ND2. Furthermore, two synonymous mutations were identified by comparisons of the novel type (haplotype C) and the established types (haplotypes A and B). The entire mitochondrial control region for 55 samples was then sequenced to construct a phylogenetic tree and median joining network. Both the tree and network demonstrated a topology of three groups, which is in consistent with the SSCP analysis. Unlike Western breeds, Chinese breeds are composed mainly of haplotypes A and B, but with a small fraction of haplotype C. According to Fu's test and mismatch distribution, haplotype C has not been subject to a recent population expansion. Based on these results, we propose a novel origin for Chinese sheep.

[Analysis on single nucleotide polymorphisms of porcine myostatin gene in different breeds].

By PCR-RFLPs and PCR-SSCP approach, three single nucleotide polymorphisms (SNPs) of porcine myostatin gene (MSTN) were analyzed in different breeds including "doubled-muscled" Yorkshire, Yorkshire, Landrace, Hamshire, Duroc, Piteran, Erhualian, Min, Hubei White and some hybrids. The three SNPs were located in the 3' encoding region, 5' promoter region and intronl region respectively. For the SNP in the 3' encoding region, which was caused by C-->T transition, the mutation frequency was relatively low: no TT genotype was detected in 274 individuals of different breeds. For the SNP in the 5' promoter region, 560 pigs were investigated. The allele T dominates in the imported lean-type pig breeds such as Yorkshire, Landrace, Duroc, Hampshire, Piteran and hybrid, however, in Erhualian and Hubei White pigs, the allele A was in majority. Polymorphism showed the similar pattern for the SNP in intron 1 region. G was the dominant allele in Yorkshire, Landrace and their hybrids, while in Erhualian and Hubei White pigs the frequency of A was much higher. Obviously they were not in Hardy-Weinberg equilibrium state. For Min and Yorshire x Erhualian pigs, they were in Hardy-Weinberg equilibrium state for the SNPs in the 5' promoter region and (or) intron 1 region. The frequency for the A alleles of SNPs in the 5' promoter region and intron 1 region was higher for "double-muscled" Yorkshire than for Yorkshire and linkage for these two mutation sites was also observed.

[Compound RSA 11 beta-hydroxylation with Curvularia lunata].

Compound RSA 11 beta-Hydroxylation of Curvularia lunata was studied. The composition of hydroxylation products was analyzed. It was found that the conversion rate with protoplasts was increased conspicuously, the construction and quantity of hydroxylation products was not alteration, but the percentage was changed greatly. The 14 alpha-OH percentage of protoplasts was 3.03-fold than that of mycelia, the 11 alpha-OH percentage was decreased 60%, and the conversion ratio of 11 beta-OH was more 1.29-fold than that of the mycelia. It affected the percentage of hydroxylation products when 2 mg/mL polyoxins were added into the mycelia conversion system.