Association of Toll-like receptor 7 and 8 gene polymorphisms with Graves' disease in Chinese Cantonese population.

Graves' disease (GD) is a common polygenic multifactorial autoimmune disease. Toll-like receptors (TLRs) play critical roles in the activation of innate and adaptive immune responses. This study investigated the association of TLR7 and TLR8 gene polymorphisms with susceptibility of GD. Five single nucleotide polymorphisms (SNPs), namely, rs179019, rs179010 and rs3853839 in TLR7 and rs3764880 and rs5744088 in TLR8, were evaluated in 332 GD patients and 351 controls using High-Resolution Melting analysis. After adjusting for age, SNP rs179010 was found to decrease the risk of GD in females (OR(T vs C) = 0.64, P = 0.004). In the additive model, the risk of GD decreased significantly as the number of T alleles increased in females [odds ratio (OR) = 0.67 (0.50-0.90), P = 0.007]. The multivariate logistic regression analysis confirmed the independent contribution of rs179010 to the protective effect against GD. This study indicates that rs179010 in TLR7 may be associated with the decreased susceptibility to GD in Chinese Cantonese.

Study of the Effects on the Strengthening Mechanism and Wear Behavior of Wear-Resistant Steel of Temperature Controlling in Heat Treatment.

To improve the wear resistance of the materials used for blades in engineering machinery, this study focused on the microstructural characteristics, mechanical properties, and wear behavior of HB500 grade wear-resistant steel developed using an optimized heat treatment system. To improve the temperature uniformity of the heat treatment furnace, the method of cyclic heating was used to heat the components. Carefully designing the quenching equipment, such as using a cross-shaped press, was employed to enhance the quenching effect and reduce the deformation of the steel plates. The crystal orientation analysis revealed a uniform and fine-grained microstructure, primarily characterized by plate-type tempered martensite, which indicated a good hardenability. The microstructure observations showed that the width of martensite is approximately 200 nm, with a significant presence of dislocations and carbides. Tensile tests and multi-temperature gradient impact tests indicated superior mechanical properties compared to similar grade wear-resistant steels, including a Rockwell hardness of 53, tensile strength of 1610 MPa, yield strength of 1404 MPa, and total elongation around 12.7%. The results of friction and wear experiments indicate that the wear rate decreases as the load increases from 100 N to 300 N, demonstrating an excellent wear resistance under a large load. Observations of the worn surfaces indicated that the wear mainly involved adhesive wear, fatigue wear, and oxidative wear. The properties' improvements were attributed to microstructure refinement and precipitation strengthening. This study indicates that designing a heat treatment system to control temperature uniformity and stability is feasible.

[Association of P53 gene polymorphisms with susceptibility to endometriosis].

OBJECTIVE: To assess the association of a single nucleotide polymorphism(SNP) in tumor suppressor gene P53 with the risk of endometriosis (EM) in Han Chinese women. METHODS: For 460 EM patients, 113 patients with endometrial carcinoma and 530 matched unrelated controls, a rs1042522(C/G) SNP of the P53 gene was genotyped by polymerase chain reaction-single strand polymorphism (PCR-SSP) and DNA sequencing. RESULTS: A significant difference has been detected in the distribution of rs1042522 alleles and genotypes between the EM patients and controls (P< 0.01). Allele G has increased the risk of EM by 1.209 times, while allele C has reduced this risk by 0.837 times. Compared with GG genotype, GC and CC genotypes have both increased the risk for EM (OR=2.073, 95%CI: 1.521-2.820, and OR=1.930, 95%CI: 1.363-2.733, respectively). Significant differences were also detected in the distribution of rs1042522 alleles and genotypes between endometrial carcinoma patients and controls (P< 0.01). Allele G has increased the risk to endometrial carcinoma by 1.311 times, while allele C has reduced this risk by 0.757 times. Compared with GG genotypes, individuals with GC and CC genotypes are more likely to be affected with endometrial carcinoma (OR=2.778, 95%CI: 1.585-4.870, and OR=2.864, 95%CI: 1.557-5.263, respectively). CONCLUSION: Our study has suggested a significant association between the rs1042522(G/C) polymorphism and susceptibility to EM in Han Chinese women. The mechanism of EM is similar to carcinoma from genetics point of view.

Improved glucose yield and concentration of sugarcane bagasse by the pretreatment with ternary deep eutectic solvents and recovery of the pretreated liquid.

In this study, a novel ternary deep eutectic solvents (DES) consisting of choline chloride/PEG/hydroxyethyl sulfonic acid (HSA) was developed to effectively improve glucose yield and concentration of sugarcane bagasse, and the conditions of the pretreatment were optimized by response surface method (RSM). Under the optimal conditions, the maximum glucose concentration (GC) could reach 12.39 g/L (HSA concentration 1.34 %, PEG400, 2.3 h, 150 °C), and the maximum glucose yield (GY) was 0.2497 g/g (HSA concentration 1.41 %, PEG400, 2.1 h, 150 °C). Hemicellulose was completely removed, and the maximum lignin removal rate was 86.89 %. After pretreatment, 95 % of the pretreated liquid can be recycled. Finally, the structural and morphological changes of bagasse before and after pretreatment were investigated by scanning electron microscopy (SEM), Fourier Transform infrared analyzer (FT-IR) and X-ray diffraction (XRD).

Neuropeptide S receptor 1 is a nonhormonal treatment target in endometriosis.

Endometriosis is a common chronic inflammatory condition causing pelvic pain and infertility in women, with limited treatment options and 50% heritability. We leveraged genetic analyses in two species with spontaneous endometriosis, humans and the rhesus macaque, to uncover treatment targets. We sequenced DNA from 32 human families contributing to a genetic linkage signal on chromosome 7p13-15 and observed significant overrepresentation of predicted deleterious low-frequency coding variants in NPSR1, the gene encoding neuropeptide S receptor 1, in cases (predominantly stage III/IV) versus controls (P = 7.8 × 10-4). Significant linkage to the region orthologous to human 7p13-15 was replicated in a pedigree of 849 rhesus macaques (P = 0.0095). Targeted association analyses in 3194 surgically confirmed, unrelated cases and 7060 controls revealed that a common insertion/deletion variant, rs142885915, was significantly associated with stage III/IV endometriosis (P = 5.2 × 10-5; odds ratio, 1.23; 95% CI, 1.09 to 1.39). Immunohistochemistry, qRT-PCR, and flow cytometry experiments demonstrated that NPSR1 was expressed in glandular epithelium from eutopic and ectopic endometrium, and on monocytes in peritoneal fluid. The NPSR1 inhibitor SHA 68R blocked NPSR1-mediated signaling, proinflammatory TNF-α release, and monocyte chemotaxis in vitro (P < 0.01), and led to a significant reduction of inflammatory cell infiltrate and abdominal pain (P < 0.05) in a mouse model of peritoneal inflammation as well as in a mouse model of endometriosis. We conclude that the NPSR1/NPS system is a genetically validated, nonhormonal target for the treatment of endometriosis with likely increased relevance to stage III/IV disease.

Vacuum-assisted black liquor-recycling enhances the sugar yield of sugarcane bagasse and decreases water and alkali consumption.

Black liquor (BL) remains a critical problem during alkaline pretreatment. To solve this issue, a novel pretreatment strategy termed vacuum-assisted black liquor-recycling pretreatment, was established to pretreat sugarcane bagasse (SCB). Firstly, SCB was pretreated with 2% NaOH at 121 °C for 1 h under vacuum conditions. The produced BL was used for subsequent pretreatments after pH recovery with NaOH. The pretreated SCBs were subject to enzymatic hydrolysis and separate hydrolyzation and fermentation (SHF) without washing to neutral pH. BL was recycled on seven occasions. The results indicated that glucose yields did not significantly differ between pretreatment with NaOH and recovered BL. The enzymatic hydrolysis and the fermentation resulted in maximum 0.35 g/g of glucose yield and 116.5 g/kg of ethanol yield respectively. Compared with conventional pretreatment with NaOH, the VABLR method showed high conversion rates of cellulose into monosaccharaides, whilst preserving ~20% and ~46% of alkali and water usage, respectively.

High-quality genomic DNA extraction from formalin-fixed and paraffin-embedded samples deparaffinized using mineral oil.

Extracting DNA from formalin-fixed and paraffin-embedded (FFPE) tissue remains a challenge, despite numerous attempts to develop a more effective method. Polymerase chain reaction (PCR) success rates with DNA extracted using current methods remain low. We extracted DNA from 140 long-term archived FFPE samples using a simple but effective deparaffinization method, removing the wax with mineral oil, and a commercially available DNA extraction kit. DNA quality was subsequently tested in a genotyping experiment with 14 microsatellite markers. High-quality DNA was obtained with a mean PCR success rate of 97% (range: 88-100%) across markers. The results suggested that DNA extracted using this novel method is likely to be suitable for genetic studies involving DNA fragments <200 bp.

Pre-treatment of sugarcane bagasse with aqueous ammonia-glycerol mixtures to enhance enzymatic saccharification and recovery of ammonia.

In this work, an efficient aqueous ammonia with glycerol (AAWG) method to improve the digestibility of sugarcane bagasse (SCB) was developed. Response surface methodology was utilized to optimize the AAWG parameters to achieve the maximum total fermentable sugar concentration (TFSC) and total fermentable sugar yield (TFSY). Under optimal AAWG conditions, 13.59 g/L TFSC (9.25% ammonia, 1.86 h, 180 °C) and 0.4449 g/g TFSY (9.51% ammonia, 1.78 h, 180 °C) were achieved, with delignification of 77.81% and 70.91%, respectively. Compared to pretreatment with glycerol or aqueous ammonia, the AAWG method significantly enhanced the enzymatic efficiency of SCB. The ammonia was recovered from the pretreatment liquid by distillation, and about one-third of the ammonia was retained. The overall results indicate that AAWG is effectively used as a pretreatment method for recovering ammonia, which would largely contribute to the economic benefits of biomass biorefinery.

The distribution of HLA alleles revealed a founder effect in the geographically isolated Chinese population, Drung.

The Drung ethnic minority is one of the smallest ethnic groups of China, geographically isolated by mountains and rivers. Before 1949, Drung society maintained many vestiges of the primitive commune system. The origin and migration of the Drung and their genetic background are still unknown because of limited records about this population. Here, we for the first time demonstrated the unique distribution of HLA alleles in the Drung by high-resolution sequence-based typing (SBT) method. Number of alleles detected is obviously less than expected and only a few alleles with a high homozygosity in each locus are predominant in this minority. The characteristics of HLA allele distribution in the Drung could reflect founder effects, suggesting the Drung probably descended from very few ancestors. The statistical analysis based on allele frequencies indicated that the Drung was an isolated ethnic group, but it also provided the clue that the Drung was genetically related to Chinese southwestern ethnic groups. Significant reduced allelic diversity and genetic isolate in the Drung make it an ideal homogeneous population and very useful model to study the evolution of HLA and the origin and migration of Chinese ethnic groups. The research paved a way to elucidate the genetic background of this mysterious minority and disease predisposition.

Enhanced enzymatic saccharification of sugarcane bagasse pretreated by sodium methoxide with glycerol.

Sodium methoxide (CH3ONa) with glycerol pretreatment (CWGP) was performed to improve the enzymatic digestibility of sugarcane bagasse (SCB). Response surface methodology was utilized to optimize the CWGP parameters for pretreating SCB from the perspective of total fermentable sugar yield (TFSY) and total fermentable sugar concentration (TFSC). Under the optimal CWGP conditions, 0.5666g/g of TFSY (0.82% CH3ONa, 1.11h, 150°C) and 17.75g/L of TFSC (0.87% CH3ONa, 1.38h, 149.27°C) were achieved, corresponding to delignification of 79.05% and 79.34%, respectively. Compared the pretreatment using glycerol or CH3ONa alone, the CWGP has significant synergies to enhance the enzymatic efficiency of SCB. The physical and chemical characteristics of untreated and pretreated SCBs were analyzed using FT-IR, XRD, and SEM, and the results suggest that CWGP significantly increased the susceptibility of the substrates to enzymatic digestibility. Ultimately, CWGP might be a prospective candidate for the pretreatment process of enzyme-based lignocellulosic biorefineries.

HLA class II polymorphisms associated with the physiologic characteristics defined by Traditional Chinese Medicine: linking modern genetics with an ancient medicine.

OBJECTIVES: The aim of this study was to test whether human leukocyte antigen (HLA) polymorphism contributes to the physical constitutions classified in Traditional Chinese Medicine (TCM). DESIGN: Seven hundred six (706) individuals of the Han ethnic group inhabiting South China were classified into 7 TCM constitution groups, according to the criteria described in Theories of Physical Constitutions of Traditional Chinese Medicine, and the distributions of HLA-DRB1, DPB1, and DQB1 were investigated using the polymerase chain reaction-sequencing-based typing method. RESULTS: The allele frequencies of DPB1*0501 in the Yin-deficiency group, DRB1*09012 in the Phlegm-wetness group, and DQB1*03032 in the Qi-deficiency and Phlegm-wetness groups were significantly different from that of the corresponding alleles in the Normality constitution, suggesting those alleles might be group-specific alleles and thus related to a particular constitution. Based on our analysis of serological groups of HLA, the associations of DR*04 with the Blood-stasis group and DQ*09 with the Qi-deficiency and Phlegm-wetness groups were observed. CONCLUSIONS: This was the first study to systematically investigate the relationship between HLA and TCM constitution using a high-resolution typing technique. The results suggested a genetic basis for the classification of physical constitution in TCM. This study laid the foundation, for the first time ever, toward gaining insight into the theory of traditional medicine using modern biological approaches.

Vacuum-assisted alkaline pretreatment as an innovative approach for enhancing fermentable sugar yield and decreasing inhibitor production of sugarcane bagasse.

Sodium hydroxide pretreatment of sugarcane bagasse under vacuum conditions was established and evaluated in this study. Compared to pretreatment under conventional moderate pressure conditions, only half of the total phenolic compounds and less than half of the formic acid were produced under vacuum conditions, while the yield of total fermentable sugar was significantly increased by 31.38%. The pretreatment parameters: NaOH concentration, pretreatment time, and pretreatment temperature, were optimized using response surface methodology based on the response values of the total fermentable sugar yield (TFSY) and the total fermentable sugar concentration (TFSC), respectively. Under the optimal conditions, the TFSY of 0.5146g/g and the TFSC of 17.37g/L were achieved, respectively. By adjusting the ratio of cellulases to xylanase, the TFSY reached a maximum of 0.5213g/g when the ratio was 1:1, while the maximum TFSC of 17.71g/L was achieved when the ratio was 1:4.

Comparative study of different alcoholate pretreatments for enhanced enzymatic hydrolysis of sugarcane bagasse.

Pretreatment of sugarcane bagasse (SCB) with alcoholates, sodium methoxide (CH3ONa), potassium methoxide (CH3OK) and sodium ethoxide (C2H5ONa), was investigated. Analyses of lignocellulose composition and enzymatic saccharification indicated that C2H5ONa showed the highest enzymatic efficiency of 102.1%. The response surface optimization of C2H5ONa pretreatment showed that under optimal conditions (4% of C2H5ONa, 121°C, 1h), 65.4% of lignin was removed and the enzymatic efficiency reached 105.2%. Hydrolysis of SCB with cellulases and xylanase at a ratio of 4:1 showed the strongest synergism with reducing sugar production of 21g/L and conversion rates of cellulose and xylan reaching 110.4% and 94.5%, respectively. These results indicated that C2H5ONa is a promising alkali to pretreat SCB and the synergism between cellulases and xylanase has a significant effect on enzymatic saccharification of the pretreated SCB.

Polymorphisms in TLR1, TLR6 and TLR10 genes and the risk of Graves' disease.

Graves' disease (GD) is postulated to be caused by the combined effects of susceptibility genes and environmental triggers. Toll-like receptors (TLRs) play a role in the activation of innate and adaptive immune responses in mammalians. The aim of this study was to evaluate the potential association of polymorphisms in TLR1, TLR6 and TLR10 genes with GD in Chinese Cantonese population. Seven single nucleotide polymorphisms (i.e. rs4833095 and rs5743565 in TLR1; rs5743808 in TLR6; and rs4504265, rs11466655, rs11096957 and rs10856839 in TLR10) were evaluated in 332 GD patients and 351 unrelated controls from Chinese Cantonese population. SNP rs5743565 in TLR1 conferred a protective effect against GD. The minor allele G of rs5743565 decreased the risk of GD in all cases (odds ratio; ORG vs. A=0.72 (0.58-0.91); p=0.005; ptrend=0.004) and early onset patients (ORG vs. A=0.72 (0.56-0.91); p=0.007; ptrend=0.006). This study provided evidence that genetic variation rs5743565 in TLR1 might be associated with the decreased susceptibility of GD.

EST analysis of gene expression in the tentacle of Cyanea capillata.

Jellyfish, Cyanea capillata, has an important position in head patterning and ion channel evolution, in addition to containing a rich source of toxins. In the present study, 2153 expressed sequence tags (ESTs) from the tentacle cDNA library of C. capillata were analyzed. The initial ESTs consisted of 198 clusters and 818 singletons, which revealed approximately 1016 unique genes in the data set. Among these sequences, we identified several genes related to head and foot patterning, voltage-dependent anion channel gene and genes related to biological activities of venom. Five kinds of proteinase inhibitor genes were found in jellyfish for the first time, and some of them were highly expressed with unknown functions.

Molecular profile of the unique species of traditional Chinese medicine, Chinese seahorse (Hippocampus kuda Bleeker).

A cDNA library of male Chinese seahorse (Hippocampus kuda Bleeker) was constructed to investigate the molecular profile of seahorse as one of the most famous traditional Chinese medicine materials, and to reveal immunological and physiological mechanisms of seahorse as one of the most primitive vertebrates at molecular level. A total of 3372 expressed sequence tags (ESTs) consisting of 1911 unique genes (345 clusters and 1566 singletons) were examined in the present study. Identification of the genes related to immune system, paternal brooding and physiological regulation provides not only valuable insights into the molecular mechanism of immune system in teleost fish but also plausible explanations for pharmacological activities of Chinese seahorse. Furthermore, the occurrence of high prevalent C-type lectins suggested that a lectin-complement pathway might exert a more dominant function in the innate immune system of teleost than mammal. Carbohydrate recognition domain (CRD) without a collagen-like region in the lectins of seahorse was likely an ancient characteristic of lectins similar to invertebrates.

Comparing the base usage frequency between bacteria DNA double strand.

During the Evolution, effected by select pressure or nature mutation, the compositions of bacteria genomes is various. And many experiences prove that the genes between leading strand and lagging strand is distinctly in copy, transcription and repair. Some scholars presume that the bases distribution is difference between the two strand, to verify the guess, we using the technology of bioinformatics, compare the base usage between the DNA double strand in 17 species bacteria. The result show: 1. there is same bases usage frequency in coding sequence between leading strand and lagging strand 2. There also same bases usage frequency in first codon, second codon and third codon. It suggest that there is a equilibrium between the two strand by the effect of select pressure and nature mutation.

[Comparative study of HLA-DQA1 and HLA-DRB1 allele in patients with endometriosis and adenomyosis].

OBJECTIVE: To make a comparative study of HLA-DQA1 and HLA-DRB1 allele frequencies in the cases of endometriosis and adeonmyosis. METHODS: The allelic types of HLA-DQA1 and HLA-DRB1 were detected by polymerase chain reaction-sequence specific primers (PCR-SSP) technique in 51 cases of endometriosis, 45 cases of adenomyosis, and 44 normal individuals as the control. RESULTS: The frequencies of HLA-DQA1*0401(7.8%, 10.0%) were significantly increased in the endometriosis group and the adenomyosis group (Pc=0.03, Pc=0.01), and the frequencies of HLA-DQA1*0301(8.8%, 5.6%) were significantly decreased in these two groups (Pc=0.00, Pc=0.00).There was no significant difference between the frequencies of HLA-DQA1 and HLA-DRB1 of endometriosis and adenomyosis. CONCLUSION: The results indicate that HLA-DQA1*0301 and *0401 alleles are associated with both endometriosis and adenomyosis, and there is perhaps common mechanism involved in both endometriosis and adenomyosis based on HLA-DQA1 and HLA-DRB1 allele frequencies.

[Determination of HLA-DRB1 gene polymorphism by PCR-SBT in Lahu ethnic group of Yunnan, China].

The HLA-DRB1 gene polymorphism in Lahu ethnic of Yunnan, China was the first time investigated using high resolution PCR-SBT method, which is based on sequences of HLA-DRB1 Intron 1 and Intron 2 and with our improvement. From 55 individuals of Lahu ethnic 16 DRB1 alleles were detected. The three most common alleles were HLA-DRB1 * 12021(30.909%), 09012(15.455%), 15011(13.636%), and they covered 60% of the total alleles detected from Lahu ethnic.HLA-DRB1 * 1413, * 11081, * 1312, * 1418, * 1504 were the first time detected in the Chinese, and were very rare in worldwide ethnic groups. With comparison of HLA-DRB1 gene frequencies between various ethnic groups we analyzed the characteristics of HLA-DRB1 gene distribution in worldwide populations,and constructed the phylogenetic tree by Neighbor-joining method and Nei measure of genetic distance. The result showed Lahu ethnic obviously belong to the Chinese South ethnic groups and can't trace its origin from northern groups with the HLA-DRB1 genetic data. The preliminary explanations about the contradiction were given in this paper.

Association of HLA-DPB1 gene with endometriosis in women of Guangdong Province in China.

OBJECTIVE: To analyze the hereditary susceptibility in patients with endometriosis by way of genotyping of HLA-DPB1 alleles. METHODS: The allelic types of HLA-DPB1 were detected by sequence-based typing (SBT) in 38 patients with endometriosis and 36 healthy women as the control. RESULT: Significant differences in the frequency of HLA-DRB1 allele was not observed between endometriotic patients and normal subjects. CONCLUSION: HLA-DPB1 allele may not be related to endometriosis.