Analysis of the cytochrome c oxidase subunit II (COX2) gene in giant panda, Ailuropoda melanoleuca.

The giant panda, Ailuropoda melanoleuca (Ursidae), has a unique bamboo-based diet; however, this low-energy intake has been sufficient to maintain the metabolic processes of this species since the fourth ice age. As mitochondria are the main sites for energy metabolism in animals, the protein-coding genes involved in mitochondrial respiratory chains, particularly cytochrome c oxidase subunit II (COX2), which is the rate-limiting enzyme in electron transfer, could play an important role in giant panda metabolism. Therefore, the present study aimed to isolate, sequence, and analyze the COX2 DNA from individuals kept at the Giant Panda Protection and Research Center, China, and compare these sequences with those of the other Ursidae family members. Multiple sequence alignment showed that the COX2 gene had three point mutations that defined three haplotypes, with 60% of the sequences corresponding to haplotype I. The neutrality tests revealed that the COX2 gene was conserved throughout evolution, and the maximum likelihood phylogenetic analysis, using homologous sequences from other Ursidae species, showed clustering of the COX2 sequences of giant pandas, suggesting that this gene evolved differently in them.

Expression and clinical implications of enhancer of Zeste homolog 2 and p53 protein in squamous cell carcinoma and precancerous lesions in the cervix.

We investigated the expression and clinical implications of enhancer of Zeste homolog 2 (EZH2) and p53 protein in cervical squamous cell carcinoma (SCC) and precancerous lesions. EZH2 and p53 expressions in SCC (168), cervical intraepithelial neoplasia (CIN)-I (19), CIN-II (35), and normal tissues (30) were detected by streptavidin-peroxidase-conjugation. The correlation between co-expression of EZH2 and p53 protein and the clinic pathological features and prognosis of SCC were discussed. The positive expression rates of EZH2 and p53 were 6.7, 37.0, and 75.6%, and 3.3, 21.1, and 39.3% in normal cervical tissues, CIN, and SCC, respectively, which were significantly different (P < 0.05). The positive expression rate of EZH2 and p53 protein in SCC patients with and without lymph node metastasis was 82.9 and 70.4% (EZH2) and 45.7 and 34.7% (p53), respectively, which was also a significant difference (P < 0.05). The progression-free survival (PFS) rates in followed-up patients (N = 143) who were EZH2- and p53-negative, EZH2- or p53-positive, and EZH2- and p53-positive were 71.3 ± 1.9, 66.1 ± 2.0, and 51.3 ± 3.8 months, respectively, which was a significant difference (P < 0.001); the overall survival among these groups was 72.9 ± 1.1, 68.6 ± 1.8, and 57.4 ± 3.4 months, respectively (P < 0.001). Multivariate analyses revealed that EZH2 expression, lymph node metastasis, and tumor staging were independent prognostic factors of SCC. EZH2 and p53, which affect lymph node metastasis and prognosis of SCC, may play a key role in the occurrence and development of SCC.

Association of the interleukin-10 gene -1082A/G genetic polymorphism with risk of ischemic stroke in a Chinese population.

We investigated the possible association between two single nucleotide polymorphisms of IL10 (-1082A/G and -592C/A) and susceptibility to ischemic stroke. In total, 335 patients with proven ischemic stroke and 335 control subjects were recruited from Xinxiang Central Hospital between March 2013 and May 2015. The IL10 -1082A/G and -529C/A polymorphisms were investigated by polymerase chain reaction-restriction fragment length polymorphism. When compared with the control subjects, patients with ischemic stroke were more likely to be male, have a habit of tobacco smoking, have higher BMI, have hypertension or diabetes mellitus, and have higher levels of TC, LDL-C, HDL-C, and TG. The multivariate logistic regression analyses revealed that the AA genotype of IL10 -1082A/G was significantly associated with development of ischemic stroke in a Chinese population compared with the GG genotype (OR = 1.93, 95%CI = 1.15-3.25). In the dominant model, the association between GA+AA genotype of IL10 -1082A/G and risk of ischemic stroke was also significant compared with the GG genotype, and the adjusted OR (95%CI) for the GA+AA genotype was 1.41 (1.02-1.94). Thus, our study suggests that IL10 gene polymorphisms contribute to the development of ischemic stroke.

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.

Methylation analysis of CXCR1 in mammary gland tissue of cows with mastitis induced by Staphylococcus aureus.

Mastitis is the most important disease in the global dairy industry, and causes large economic losses. Staphylococcus aureus is one of most common pathogens that cause bovine mastitis. CXCR1 has been implicated as a prospective genetic marker for mastitis resistance in dairy cows; CXCR1 expression significantly increases when cows have mastitis. To investigate the mechanisms involved in its increased expression, bisulfite sequencing polymerase chain reaction (PCR) was used to detect the methylation status of CXCR1 CpG island, and quantitative fluorescence PCR was used to detect CXCR1 expression in bovine mammary tissue induced with S. aureus in three Chinese Holstein cows. No CpG island was found for bovine CXCR1 in the upstream 2-kb region, whereas one CpG island that contained 13 CpG sites was found in exon 1 of CXCR1. All of the CpG sites were under hypermethylation from 90 to 100% in the mammary tissues. When the mammary gland mRNA expression of CXCR1 was 12.10-fold higher in infected cow quarters than in uninfected quarters, the methylation levels of the CpG site at position 519 were significantly lower in the infected quarters than in the uninfected quarters. Pearson correlation analysis showed that the methylation level at position 519 was significantly negatively correlated with the CXCR1 mRNA expression level (P < 0.05). These results indicate that the methylation of the CpG site at position 519 may regulate CXCR1 expression in cows with mastitis induced by S. aureus, but further studies are needed to elucidate the mechanisms involved.

An expression profiling analysis of hybrid millet and its parents at grain filling stage.

Heterosis has been widely used in crop breeding and production. However, a shortage of genes known to function in heterosis significantly limits our understanding of the molecular basis underlying heterosis. Here, we report 740 differentially expressed genes (DEGs) in the leaves of the hybrid millet Zhang No.5 and its parents at the grain filling stage determined using Solexa Illumina digital gene expression. Of the 740 DEGs, 546 were from the hybrid and its parents and most were up-regulated in the hybrid. Particularly, a large number of DEGs related to starch and carbohydrate metabolism and 2 DEGs encoding chlorophyll a/b binding proteins were up-regulated in hybrid millet. Moreover, all DEGs were enriched in the biological process and molecular function, and no DEGs were found to be enriched in the cellular component of GO terms. Pathway enrichment using KEGG showed that several DEGs were enriched in the circadian rhythm pathway. Further analysis revealed that the altered circadian rhythm, which mediates photosynthesis and carbohydrate accumulation, may play an important role in heterosis of the hybrid millet.

Evaluation of a father and son with atypical chronic myeloid leukemia with SETBP1 mutations and a review of the literature

We report the case of a father and son diagnosed with atypical chronic myeloid leukemia (aCML). Both patients harbored SETBP1 mutations, which are present in 24.3% of aCML patients. Moreover, both shared the variant encoding p.Pro737His, but the aCML severity was greater in the son because of the presence of two other missense mutations causing p.Asp868Asn and p.Ser885Arg alterations. SETBP1 mutations may be associated with an adverse prognosis, so their detection would help in the diagnosis of aCML and the determination of a patient's prognosis.

Changes in circulating endothelial progenitor cells predict responses of multiple myeloma patients to treatment with bortezomib and dexamethasone.

Four cycles of chemotherapy are required to assess responses of multiple myeloma (MM) patients. We investigated whether circulating endothelial progenitor cells (cEPCs) could be a biomarker for predicting patient response in the first cycle of chemotherapy with bortezomib and dexamethasone, so patients might avoid ineffective and costly treatments and reduce exposure to unwanted side effects. We measured cEPCs and stromal cell-derived factor-1α (SDF-1α) in 46 MM patients in the first cycle of treatment with bortezomib and dexamethasone, and investigated clinical relevance based on patient response after four 21-day cycles. The mononuclear cell fraction was analyzed for cEPC by FACS analysis, and SDF-1α was analyzed by ELISA. The study population was divided into 3 groups according to the response to chemotherapy: good responders (n=16), common responders (n=12), and non-responders (n=18). There were no significant differences among these groups at baseline day 1 (P>0.05). cEPC levels decreased slightly at day 21 (8.2±3.3 cEPCs/µL) vs day 1 (8.4±2.9 cEPCs/µL) in good responders (P>0.05). In contrast, cEPC levels increased significantly in the other two groups (P<0.05). SDF-1α changes were closely related to changes in cEPCs. These findings indicate that change in cEPCs at day 21 in the first cycle might be considered a noninvasive biomarker for predicting a later response, and extent of change could help decide whether to continue this costly chemotherapy. cEPCs and the SDF-1α/CXCR4 axis are potential therapeutic targets for improved response and outcomes in MM patients.

Evaluation of a father and son with atypical chronic myeloid leukemia with SETBP1 mutations and a review of the literature.

We report the case of a father and son diagnosed with atypical chronic myeloid leukemia (aCML). Both patients harbored SETBP1 mutations, which are present in 24.3% of aCML patients. Moreover, both shared the variant encoding p.Pro737His, but the aCML severity was greater in the son because of the presence of two other missense mutations causing p.Asp868Asn and p.Ser885Arg alterations. SETBP1 mutations may be associated with an adverse prognosis, so their detection would help in the diagnosis of aCML and the determination of a patient's prognosis.

Estimation of the growth curve and heritability of the growth rate for giant panda (Ailuropoda melanoleuca) cubs.

Giant panda cubs have a low survival rate during the newborn and early growth stages. However, the growth and developmental parameters of giant panda cubs during the early lactation stage (from birth to 6 months) are not well known. We examined the growth and development of giant panda cubs by the Chapman growth curve model and estimated the heritability of the maximum growth rate at the early lactation stage. We found that 83 giant panda cubs reached their maximum growth rate at approximately 75-120 days after birth. The body weight of cubs at 75 days was 4285.99 g. Furthermore, we estimated that the heritability of the maximum growth rate was moderate (h(2) = 0.38). Our study describes the growth and development of giant panda cubs at the early lactation stage and provides valuable growth benchmarks. We anticipate that our results will be a starting point for more detailed research on increasing the survival rate of giant panda cubs. Feeding programs for giant panda cubs need further improvement.

Overwhelming postsplenectomy infection.

This report aims to deepen the understanding of the pathogenesis, diagnosis, clinical characteristics, and treatment of overwhelming postsplenectomy infection (OPSI). A patient treated at Taihe Hospital for tuberculous OPSI is described, and relevant literature is reviewed. Broad-spectrum antibiotics, suppression of the systemic inflammatory reaction, and anti-shock measures were the keys to the successful treatment of this condition. OPSI is a life-threatening condition and has a high mortality rate. Early diagnosis, use of anti-inflammatory glucocorticoids, and administration of high-dose gamma globulin and ulinastatin for the treatment of OPSI may improve outcomes.

Post-surgical treatment of a patient with ectopic pheochromocytoma.

We report here the course and treatment of a patient with ectopic pheochromocytoma. The patient was cured after treatment with respiratory and circulatory support, multiple-organ protection, and continuous renal replacement therapy (CRRT) for approximately 2 weeks. After misdiagnosis, a patient with ectopic pheochromocytoma who is being treated should undergo aggressive fluid supplementation and CRRT instead of central venous pressure measurement, which has limited value in guiding fluid supplementation. The main priority is maintaining hypotension. Hypertension may be controlled with rapid-acting agents. A good outcome can be anticipated for patients who undergo comprehensive intensive care unit therapy.

Cloning and characterization of a farnesyl pyrophosphate synthase from Matricaria recutita L. and its upregulation by methyl jasmonate.

Matricaria recutita (L.), commonly known as chamomile, is one of the most valuable medicinal plants because it synthesizes a large number of pharmacologically active secondary metabolites known as α-bisabolol and chamazulene. Although the plant has been well characterized in terms of chemical constituents of essential oil as well as pharmacological properties, little is known about the genes responsible for biosynthesis of these compounds. In this study, we report a new full-length cDNA encoding farnesyl diphosphate synthase (FPS), a key enzyme in the pathway of biosynthesis of isoprenoids, from M. recutita. The cDNA of MrFPS comprises 1032 bp and encodes 343 amino acid residues with a calculated molecular mass of 39.4 kDa. The amino acid sequence homology and phylogenetic analysis indicated that MrFPS belongs to the plant FPS super-family and is closely related to FPS from the Asteraceae family. Expression of the MrFPS gene in Escherichia coli yielded FPS activity. Using real-time quantitative PCR, the expression pattern of the MrFPS gene was analyzed in different tissues of M. recutita as well as in response to methyl jasmonate. The expression analysis demonstrated that MrFPS expression varies in different tissues (with maximal expression in flowers and stems) and was significantly elevated in response to methyl jasmonate. This study will certainly enhance our understanding of the role of MrFPS in the biosynthesis and regulation of valuable secondary metabolites in M. recutita at a molecular level.

Molecular cloning and characterization of the full-length Hsp90 gene from Matricaria recutita.

Heat shock protein 90 (Hsp90) is one of the most abundant and conserved chaperone proteins and plays important roles in plant growth and responses to environmental stimuli. However, little is known regarding the sequence and function of Hsp90s in Matricaria recutita. In the present study, we cloned the full-length cDNA sequence of the hsp90 gene from this species. Using rapid amplification of cDNA ends technologies with 2 degenerate primers that were designed based on the hsp90 gene sequence from other members of Asteraceae, we isolated and characterized an Hsp90 homolog gene from M. recutita (Mr-Hsp90). The full-length Mr-hsp90 cDNA sequence, containing 2097 base pairs, encodes a protein of 698 amino acids. Based on amino acid sequence identity, Mr-Hsp90 showed high similarity to other cloned Hsp90 proteins. The Mr-Hsp90 protein was closely clustered with the Lactuca sativa in a phylogenetic tree. These results indicate that the cloned sequence of Mr-Hsp90 is a member of the Hsp90 family, which is reported for the first time in M. recutita. Next, we conducted a salt stress experiment to determine the protein's function under salt stress conditions. Survival of chamomile seedlings subjected to heat-shock pretreatment was significantly increased compared with groups that had not undergone heat-shock pretreatment in a salt stress environment. This indicates that Mr-Hsp90 plays an important role in the salt resistance of chamomile seedlings.

Effect of atorvastatin on vascular endothelial function in moderately nicotine-dependent smokers.

We investigated the effect of atorvastatin on vascular endothelial function in moderately nicotine-dependent smokers. One hundred and sixty moderately nicotine-dependent smokers were randomly divided into the atorvastatin group (N = 80) and the control group (N = 80). Total cholesterol (TC), triglyceride, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol (LDL-C), fasting plasma glucose levels, aspartate aminotransferase, total bilirubin, creatine phosphokinase, and brachial artery flow-mediated vasodilation function (FMD) were measured before and 12 months after atorvastatin treatment. After a 12-month atorvastatin therapy, the TC and LDL-C levels of patients were decreased significantly (P < 0.05) and the FMD of patients were improved significantly (P < 0.05). Compared with the control group, TC and LDL-C of the patients were significantly decreased (P < 0.05) and the FMD of the patients were significantly improved (P < 0.05). Atorvastatin may significantly improve endothelial function in moderately nicotine-dependent smokers.

Quantitative trait locus analysis for ear height in maize based on a recombinant inbred line population.

Maize (Zea mays L.) is among the crops with the greatest worldwide economic importance. Ear height is a very important trait that is considered necessary in maize and is related to morphology, lodging, and yield. To realize its genetic basis, an F9 recombinant inbred line population and a genetic map consisting of 101 simple sequence repeat markers were used to detect the quantitative trait locus (QTL) for ear height, and the result showed that one QTL on chromosome 1 was identified with a mapping interval of 5 cM to its linked marker Umc1358. The QTL from elite inbred line Mo17 could explain 9.55% of the phenotypic variance, and because of the additive effect, it could result in an ear height increase of 4.86 cm. This result was beneficial for understanding the genetic basis of ear height in maize.

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.

Application of genomics and proteomics in drug target discovery.

Gene medicine is making breakthroughs in health questions that have baffled humanity for centuries. To understand and utilize gene medicine, it is necessary to realize its action against targets at the molecular level. Currently, many methods can be used to discover drug targets; among these, genomic and proteomic methods are the two most important. In this study, we introduced how to discover drug targets by genomic and proteomic methods in detail. These contents are beneficial for understanding and utilizing the two methods to discover new drug targets of gene medicine.