Histopathologic analysis of stage pT1b kidney neoplasms for optimal surgical margins of nephron-sparing surgery.

OBJECTIVE: To evaluate the pathological features and define the optimal surgical margins (SM) of nephron-sparing surgery (NSS) for kidney neoplasms 4-7 cm (stage pT1b) on preoperative imaging. MATERIALS AND METHODS: The retrospective study included 748 patients who were diagnosed stage pT1b renal tumors and underwent either radical nephrectomy (RN, n = 475) or NSS (n = 273) from January 2004 to March 2017. The tumor size, pathological subtype, Fuhrman grade, status of peritumoral pseudocapsule (PC) and tumor multifocality were recorded. The relationship between peritumoral PC and positive SM was calculated statistically by Fisher's exact probability test. RESULTS: The mean tumor diameter was 5.4 cm (range: 4.1-7.0 cm), 65 (8.7%) cases were discovered with multifocal lesions and 686 (91.7%) were surrounded with peritumoral PC in all 748 specimens. 57 (8.3%) of 686 cases were proved with tumor infiltrated beyond PC [infiltration (+)], and the presence of PC infiltration (+) was significantly correlated with positive SM (p = 0.016). The infiltrative depth of tumor cells into renal parenchyma beyond PC was all limited in 3 mm and the proportion of ≤ 1, 1-2 and 2-3 mm was 21.1% (12/57), 59.6% (34/57) and 19.3% (11/57), respectively. CONCLUSIONS: Our report indicates a 3 mm excisional margin is acceptable to ensure negative SM when operating NSS on stage pT1b kidney neoplasms.

A rapid and high-quality method for total RNA isolation from Haematococcus pluvialis.

Haematococcus pluvialis, as the most potential natural source of astaxanthin, which is a powerful antioxidant with high economic value, has attracted more and more scientific attention in recent years. An in-depth understanding of the mechanism for how H. pluvialis produces astaxanthin requires the intensive investigations on its genetic information. In particular, many reported studies were based on a variety of RNA analyses. However, it is difficult to extract RNA with high quality and quantity from H. pluvialis, because of the blockage from its thick cell wall and contamination by a large quantity of pigments, polysaccharides, and lipids. Therefore, we proposed an optimized Trizol-based RNA extraction method for H. pluvialis by investigating the effect of cell wall broken ways, algal strains, and cell growth status on total RNA isolation. Using this rapid, convenient, and cost-saving method, isolated H. pluvialis RNA had high quantity and quality (with an RNA integrity number of 7.0 and a concentration of 1604.1 ng/µL) equivalent to that isolated by commercial kit, enabling its applications into downstream RNA analyses.

Genetic effect of an A/G polymorphism in the HSP70 gene on thermotolerance in chicken.

Studying thermotolerance is important for the prevention of thermostress in chickens. This study aimed to analyze the effect of mutations in the heat shock protein 70 (HSP70) gene on chicken thermotolerance. The C.-69A>G SNP in the 5'-flanking region of the HSP70 gene was genotyped in Lingshan and White Recessive Rock (WRR) chickens. Association of this SNP with thermotolerance traits revealed it to be significantly associated with CD4+/CD8+, and potentially associated with heterophil-to-lymphocyte ratio in WRR chickens exposed to thermoneutral temperature (15°C). Online prediction detected a putative myeloid zinc finger protein 1 binding factor in the C.-69A>G mutation. Under acute thermostress, mRNA levels of HSP70 in individuals with different C.-69A>G genotypes varied in the heart, leg muscle, and liver tissues. The HSP70 protein was expressed at higher levels in individuals with the GG genotype than in those with the AA genotype. In heart and liver, protein expression of HSP70 in individuals with the GG genotype was significantly higher than in those with the AA genotype. In leg muscle, protein expression was higher in birds with the GG genotype than in those with the AA and AG genotypes. Luciferase activity of the GG genotype was significantly higher than that of the AA genotype, suggesting that the C.-69A>G SNP regulates HSP70 gene expression. These results indicate that the C.-69A>G SNP in the 5'-flanking region of the HSP70 gene might affect chicken thermotolerance and that the GG genotype might be advantageous for the prevention of thermostress.

Overexpression of NaKR3 enhances salt tolerance in Arabidopsis.

Salinity is a major abiotic stress in agriculture. Here, we report that SODIUM POTASSIUM ROOT DEFECTIVE3 (NaKR3), which encodes a heavy metal-associated domain protein, is involved in salt tolerance in Arabidopsis. The results of quantitative reverse transcription-polymerase chain reaction analysis revealed that NaKR3 was induced by high salinity and osmotic stresses, but not by Cu(2+) stress. Transient expression of NaKR3-GFP in Arabidopsis protoplasts showed that the NaKR3 protein was localized in the cytosol. Transgenic Arabidopsis plants constitutively expressing NaKR3 under the control of the cauliflower mosaic virus 35S promoter exhibited increased tolerance to salt treatment. Furthermore, overexpression of NaKR3 increased the expression of SOS1 and SOS3, but decreased the accumulation of salt-induced proline. Taken together, our results indicate that NaKR3 is involved in the salt stress response in Arabidopsis.

VDAC2 involvement in the stress response pathway in Arabidopsis thaliana.

The voltage-dependent anion channel (VDAC) is the major transport protein in the outer membrane of mitochondria, and is involved in the formation of a permeable transition pore and metabolite transport. In this study, we explored the role of Arabidopsis thaliana VDAC2 (AtVDAC2) in the signal transduction pathway in Arabidopsis under salt stress. We investigated the germination rates of AtVDAC2 transgenic lines under salicylic acid (SA) treatment, and found that AtVDAC2 can affect the sensitivity of Arabidopsis to SA. Furthermore, the stomatal apertures of AtVDAC2 transgenic Arabidopsis were calculated. Results showed that the over-expression lines showed the obvious stomatal closure, while in the antisense lines, no obvious changes in stomatal apertures were found. In addition, we also detected the expression levels of salt stress and SA response-associated genes in transgenic plants and found that AtVDAC2 affected the expression of these genes. Our study investigated the role of AtVDAC2 in SA and salt stress response in Arabidopsis; our observations provide some helpful information for better understanding the direct and downstream functions of AtVDAC.

Quantitative trait locus analysis for kernel width using maize recombinant inbred lines.

Maize (Zea mays L.) kernel width is one of the most important traits that is related to yield and appearance. To understand its genetic mechanisms more clearly, a recombinant inbred line (RIL) segregation population consisting of 239 RILs was used for quantitative trait locus (QTL) mapping for kernel width. We found four QTLs on chromosomes 3 (one), 5 (two), and 10 (one). The QTLs were close to their adjacent markers, with a range of 0-23.8 cM, and explained 6.2-19.7% of the phenotypic variation. The three QTLs on chromosomes 3 and 5 had positive additive effects, and to a certain extent increased kernel width, whereas the one on chromosome 10 exhibited negative additive effects and decreased kernel width. These results can be used for gene cloning and marker-assisted selection in maize-breeding programs.

Characterization of HSP70 and its expression in tissue: correlation with physiological and immune indices in goose (Anser cygnoides) serum.

We cloned the goose heat shock protein 70 gene (HSP70), to determine its sequence variation and elucidate its mRNA expression. We designed primers to amplify the entire goose HSP70 sequence. We used 10 commercial Wuzong goslings in a heat-stress experiment. We collected tissue samples for RNA extraction and quantitative real-time polymerase chain reaction (qRT-PCR). We analyzed the variation in expression of goose HSP70 before and after heat stress. We constructed a DNA pool from six different species, for single nucleotide polymorphism (SNP) screening. We detected 18 SNPs and selected three of these SNPs for correlation analysis with biological and immune traits in 200 Wuzong geese. We showed that T+237C was significantly correlated with the serum corticosterone level, whereas T+1122C was significantly correlated with the heterophil to lymphocyte ratio. Goose HSP70 contained no introns. The results of qRT-PCR analysis revealed significant gender differences in the expression of goose HSP70 at 40°C but not at 25°C; moreover, in general, expression was significantly higher at 40°C than at 25°C. With the exception of the leg muscle and cerebellum, HSP70 expression was significantly higher in male geese than in female geese. Our results indicate that goose HSP70 plays an important role in response to severe heat stress.

Related factors and prevalence of Parkinson's disease among Uygur residents in Hetian, Xinjiang Uygur Autonomous Region.

The aim of this study was to identify the factors related to and determine the prevalence of Parkinson's disease (PD) among Uygur residents in Hetian Prefecture, Xinjiang Uygur Autonomous Region. This population-based prospective cohort study used structured questionnaires to screen for factors related to PD. The prevalence in different age groups was analyzed, and PD risk was assessed using univariate and multivariate conditional logistic regression analysis. In total, 5932 subjects participated in the survey; of these, 88 individuals, all of whom were Uygurs, had PD. The overall prevalence of PD was 1.48% (1.32%) in people over the age of 45; the prevalence was 1.68% (1.59%) in men and 1.28% (1.36%) in women. The Fifth Population Census in China found the prevalence of PD in Hetian, Xinjiang, to be 1.32%. Single-factor results showed that exercise, social activities, and frequent consumption of nuts are protective factors for PD, and long-term pesticide exposure, family history of PD, and consumption of barbecued food were risk factors for this condition. Multi-factor conditional logistic regression analysis showed that participation in sports and social activities can reduce the risk of PD, whereas long-term pesticide exposure and family history of PD increases this risk. These results show that PD is caused by the interaction of several factors.

Association analysis between SNPs in the 5'-flanking region of the chicken GRP78 gene, thermotolerance parameters, and tissue mRNA expression.

Glucose-regulated protein 78 (GRP78) is a molecular chaperone in the endoplasmic reticulum and can be induced by different kinds of environmental and physiological stress. Thus far, the role of the GRP78 gene in thermotolerance in chickens has not been investigated. In the present study, we detected sequence variations in the 5ꞌ-flanking region of the GRP78 gene and evaluated several thermotolerance parameters, such as T3, corticosterone, H/L ratio, and levels of CD3(+), CD4(+), and CD8(+) T cells, to further determine its associations at 35° and 15°C. The sequencing results revealed 10 SNPs in the 5'-flanking region of the GRP78 gene, and seven mutations were chosen for further genotyping in a White Recessive Rock (WRR) chicken population. The SNP C.-744C>G in WRR chickens was significantly correlated with heat tolerance parameters under both conditions; it may therefore exert a potential hereditary effect on heat tolerance, and the genotype GG may be advantageous for thermotolerance. The heart, liver, brain, and leg muscle tissues of 8-day-old WRR chickens were sampled from heat stress groups, which were defined by exposure to 1, 2, 3, and 6 h of persistent thermal stress, and a control group, which was not exposed to thermal stress. Quantitative real-time polymerase chain reaction assay indicated that the mRNA expression level of the GRP78 gene increased gradually under heat stress, peaked at 3 h, and then decreased. We conclude that the mRNA expression of the GRP78 gene is time- and tissue-dependent.

Expression profile of tumor suppressor gene RASSF1 in lacrimal gland carcinoma.

We examined the expression pattern of the tumor sup-pressor gene RAS association domain family gene 1 (RASSF1) in lacri-mal gland carcinoma and analyzed its relationship with the oncogenesis and progression of tumors. Sixty-two patients (30 males, 32 females, average age = 47 ± 3.5 years) admitted with lacrimal gland carcinoma to the Department of Ophthalmology of our hospital between January 2012 and January 2014 were enrolled in this study. Based on tumor ma-lignancy, patients were classified into a malignant group (N = 25) and benign group (N = 37). Healthy lacrimal gland resections from trauma surgery (N = 35) were recruited as a healthy control group. Expres-sion profiles of RASSF1 in all groups were quantified using reverse transcription-polymerase chain reaction and western blotting. Recur-rence of lacrimal gland carcinoma was surveyed through postopera-tive follow-up. Expression levels of RASSF1 in samples from the ma-lignant and benign groups were significantly lower than those in the healthy group (P < 0.05). Furthermore, the malignant group showed lower RASSF1 expression than the benign group (P < 0.05). Postopera-tive follow-up identified 22 cases of recurrence in the malignant group, with a recurrence rate of 88%, while 15 cases in the benign group had a recurrence rate of 40.5%. A direct relationship exists between RASSF1 expression levels and the malignancy grade of lacrimal gland carci-noma. Patients with lower RASSF1 expression showed a higher recur-rence probability, indicating unfavorable prognosis. Therefore, measur-ing RASSF1 expression can be used as a diagnostic method for lacrimal gland carcinoma.

Association between PARK16 gene polymorphisms and susceptibility of Parkinson's disease in a Chinese population.

Recent genome-wide association studies identified 11 risk loci in different populations of familial and sporadic Parkinson's disease (PD) patients. Few loci have been verified in different European and Asian populations. We also validated 2 new single-nucleotide polymorphisms, rs947211 and rs823144, in PARK16 to explore their association with susceptibility to PD in the Xinjiang Uygur and Han populations. This case-control study included 312 PD patients (130 Uygur and 182 Han) and 359 control subjects (179 Uygur and 180 Han). Polymerase chain reaction-restriction fragment length polymorphism analysis and DNA sequencing were used to detect the rs947211 and rs823144 polymorphism in the PARK16 gene between the Xinjiang Uygur and Han populations. Frequencies of the A allele and AA genotype (42.1 and 15.7%, respectively) of rs947211 in PD patients were significantly lower than those in the control group (54.7 and 28.7%, respectively, P < 0.01). A allele and AA genotype frequencies of rs823144 were 56.8 and 31.8% in the PD patients group and were 54.1 and 29.3% in the control group; no significant difference was found (P > 0.05). In both the Han and Uygur groups, the rs947211 polymorphism was associated with PD. Haplotype analysis also indicated that the A-A and G-A haplotypes were associated with PD. We found that the rs947211 polymorphism may be a susceptibility marker for PD in the Chinese population; the A-A and G-A haplotypes may be a protective factor and a risk factor, respectively, for PD in the Chinese population.

Analysis of genetic diversity of the heat shock protein 70 gene on the basis of abundant sequence polymorphisms in chicken breeds.

This study was designed to detect the sequence variation of the chicken heat shock protein 70 (HSP70) gene. A total of 102 individuals from 8 native Chinese breeds together with Dwarf White Chicken and Red Junglefowl were used to detect sequence variations. The coding regions of the chicken HSP70 gene from 102 individuals were cloned and sequenced. Thirty-six variations were identified, which included 34 single nucleotide polymorphisms and 2 indel mutations. Fifty-seven haplotypes were observed, of which, 43 were breed-specific and 14 were shared. There were 7 Red Junglefowl-specific haplotypes, while Haidong and Silkie only had 2 specific haplotypes. Eleven and 3 haplotypes were shared between and within species, respectively. The variation in nucleotide diversity (Pi) and average number of nucleotide differences (K) among species were consistent. The total Pi of HSP70 was 0.0016, and the total K was 4.1998. The Pi value of Red Junglefowl was the highest (0.0018) and K was 4.8000, while the Pi of Silkie was the lowest (0.0010) and K was 2.5000. These results demonstrated that variation in chicken HSP70 was abundant between and within species.

Functional characterization and analysis of the Arabidopsis UGT71C5 promoter region.

In the present study, we isolated an Arabidopsis promoter, UGT71C5, and analyzed its role in the regulation of the light response mechanism. We constructed a fusion vector pBI121-pU-GUS by integrating the UGT71C5 promoter upstream of the GUS reporter gene in pBI121, and then transferred this vector into Arabidopsis plants. The GUS activity of the transgenic plants was detected using a spectrophotometer under normal growth conditions as well as under light, drought, and ABA stress-treatments. The obtained results indicated that the GUS activity of transgenic plants ranged in between the activities observed in wild-type and 35S transgenic plants, which were used as positive control. Light stress for 8 and 12 h increased the GUS activity in transgenic plants by 3 and 4 times, respectively, compared to the activity in these plants under normal conditions. No such change in the GUS activity was observed under drought and ABA-treated conditions. This suggests that the UGT71C5 promoter is light inducible. Our study provides helpful insights into the elucidation of inducible promoters in Arabidopsis and the molecular mechanisms of light response.

Alteration of HSF3 and HSP70 mRNA expression in the tissues of two chicken breeds during acute heat stress.

This study aimed to estimate changes in HSF3 and HSP70 mRNA expression in stress-sensitive tissues of 2 chicken breeds during acute heat stress. Lingshan chickens (LSC) and White Recessive Rock (WRR) (24 chickens of each breed) were randomly divided into 4 groups (0, 2, 3, and 6 h of heat treatment). With increasing heat treatment time, both HSF3 and HSP70 expression first declined and then showed a significant increase in both breeds. However, HSP70 expression decreased in the heart following 6 h of heat treatment, whereas HSF3 expression continued to increase. After 2 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.05, P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and leg muscle of WRR compared to LSC (P < 0.01, P < 0.05). After 3 h of heat treatment, HSF3 expression was significantly higher in the brain and leg muscle of LSC compared to WRR (P < 0.01). In comparison, HSP70 expression was significantly higher in the liver and heart of LSC compared to WRR (P < 0.01). These results indicate that the expression of HSF3 and HSP70 mRNA in LSC and WRR exhibit species-specific and tissue-specific differences during heat treatment.

Correlation between DQB1 genetic polymorphism and genetic susceptibility in patients diagnosed with irritable bowel syndrome with diarrhea.

We examined patients of Han nationality diagnosed with irritable bowel syndrome with diarrhea (IBS-D) in Guangdong, China, to analyze the correlation between DQB1 allele polymorphisms and the genetic susceptibility to IBS-D. A total of 120 IBS-D patients of Han nationality in Guangdong, China, and 60 healthy control volunteers were included. DQB1 allele polymorphisms were investigated by polymerase chain reaction. Subjects' serum interleukin (IL)-10 level, colonic permeability, and tight junction marker zonula occludens 1 (ZO1) mRNA level were also investigated. Our data showed that the DQB1*02 allele frequency was significantly higher in IBS-D patients, while the DQB1*0603 frequency was lower than in healthy volunteers. The DQB1*03, DQB1*04, DQB1*05, DQB1*0601, DQB1*0602, and DQB1*0604 alleles did not show significant differences between IBS-D patients and healthy controls. Furthermore, patients with DQB1*03- positive and DQB1*0603-negative alleles showed more severe colonic permeability and lower serum IL-10 level and ZO1 level compared to healthy controls or even IBS-D patients with other genotypes. The present study indicated the DQB1*02 or DQB1*0603 alleles are related to IBS-D occurrence in Guangdong, China, and the mechanism of the disease may be related to reduced serum IL-10 levels.

Effects of destrin pathway mutations on the gene expression profile.

This study aimed to explore the interaction and crosstalk between pathways in response to destrin mutations. All the pathways from the MINT database were downloaded, a protein-protein interaction network was then constructed, and the crosstalk between pathways was investigated, in particular, the overlap of 2 significant pathway analysis results. As expected, the results showed that regulation of the actin cytoskeleton was the significant pathway of destrin mutations in mice. Further analysis indicated that 28 significant pathways cross-talked with the pathway regulating the actin cytoskeleton. Importantly, 3 pathways, including regulation of actin cytoskeleton pathway, pathways in cancer, and the B cell receptor signaling pathway were linked by inositol phosphate metabolism based on crosstalk analysis of Gene Ontology relationships among pathways. All of these pathways have been demonstrated to participate in cytoskeleton dynamics. These findings might provide valuable insights into cytoskeleton dynamic abnormalities in destrin mutations of corneal diseases.

Development and characterization of SSR markers from Pinus massoniana and their transferability to P. elliottii, P. caribaea and P. yunnanensis.

Pinus massoniana (Masson's pine) is a widespread tree species in central and southern China and northern Vietnam; it is valued for rosin and paper production. Despite the significant economic value of Masson's pine, little work has been done on its molecular genetics. We developed 318 SSR primers from genome sequences of P. massoniana, and we identified 10 polymorphic markers. The number of alleles in the population of P. massoniana that we examined ranged from two to four, and the Shannon diversity index ranged from 0.150 to 1.133. Cross-species transferability of the 318 SSRs was also analyzed in the slash pine (Pinus elliottii), the Caribbean pine (Pinus caribaea) and the Yunnan pine (Pinus yunnanensis); 15, 10, and 10 primer pairs generated polymorphic amplification, respectively. These sets of polymorphic SSR markers will be useful for population genetics studies of P. massoniana, for genetic identification of interspecific hybridization, and for phylogeographic studies of Pinus spp.

Descriptive statistics and correlation analysis of agronomic traits in a maize recombinant inbred line population.

Maize (Zea mays L.) is one of the most important crops in the world. In this study, 13 agronomic traits of a recombinant inbred line population that was derived from the cross between Mo17 and Huangzao4 were investigated in maize: ear diameter, ear length, ear axis diameter, ear weight, plant height, ear height, days to pollen shed (DPS), days to silking (DS), the interval between DPS and DS, 100-kernel weight, kernel test weight, ear kernel weight, and kernel rate. Furthermore, the descriptive statistics and correlation analysis of the 13 traits were performed using the SPSS 11.5 software. The results providing the phenotypic data here are needed for the quantitative trait locus mapping of these agronomic traits.

Genetic diversity of Saccharum spontaneum from geographical regions of China assessed by simple sequence repeats.

Saccharum spontaneum is the most variable wild relative of sugarcane with potential for use in sugarcane improvement programs. In order to help preserve and exploit this species, 152 accessions from eight major geographical regions in China, including Hainan, Guangdong, Guangxi, Yunnan, Sichuan, Guizhou, Fujian, and Jiangxi provinces, were investigated by analyzing 20 simple sequence repeats (SSRs), including 11 genomic SSRs (gSSRs) and nine SSRs developed from expressed sequence tags (EST-SSRs). A total of 454 alleles were generated by the 20 SSRs, with 295 and 159 alleles detected by gSSRs and EST-SSRs respectively. The Mantel test showed significant correlation between genetic matrixes among the studied accessions revealed by gSSRs versus EST-SSRs, although the average polymorphism of EST-SSRs (17.7) was much lower than that of gSSRs (26.8). Among the eight provinces, collections from Guizhou were the most diverse and those from Guangdong were the most distinct. Clustering analysis and principal component analysis accordantly classified the accessions into four groups, which were "Southwest group", "Hainan group", "Guangdong group", and "Guangxi group", based on the geographical origin of the major accessions in each group, demonstrating that geographical factors play an important role in the pattern of genetic structure of Chinese S. spontaneum. As two (Guizhou and Yunnan) of the three provinces with highest genetic diversity are located in southwest China, we concluded that southwest China is the region with the highest genetic diversity of S. spontaneum.

Promoter methylation negatively correlated with mRNA expression but not tissue differential expression after heat stress.

DNA methylation plays a central role in gene expression. In this study, we detected the promoter methylation pattern of the chicken heat shock protein 70 (HSP70) gene and its association with messenger RNA (mRNA) expression before and after heat shock. The results showed that mRNA expression increased in response to heat stress and peaked at 3 h before dropping. Hypomethylation of the HSP70 promoter occurred in all of the groups studied, but the difference between groups within tissue type was not significant. The DNA methylation level of the control and the 6-h treatment groups was slightly higher than that of the 3-h treatment group in brain tissue and leg muscle. Correlation analysis between mRNA expression and DNA methylation of HSP70 showed that DNA methylation was negatively associated with mRNA expression in leg muscle (P = 0.0124), indicating that DNA methylation may be negatively associated with the expression of HSP70, although the difference was not significant. We concluded that the expression of HSP70 is heat inducible and tissue dependent and that heat induction may correlate with DNA methylation pattern in the HSP70 promoter, whereas tissue dependence is unrelated to DNA methylation pattern.