Intratumoral Polymorphism of Peroxisome Proliferator-Activated Receptor Delta-87 T>C in Colorectal Cancer.

Peroxisome proliferator activated receptor delta (PPARD) is a nuclear receptor transcription factor whose single nucleotide polymorphism (SNP), especially PPARD-87 T>C (rs2016520), may play an important role in expression regulation of PPARD. But its expression patterns as well as contribution in colorectal cancer (CRC) are still controversial. In this study, whether the intratumoral heterogeneity of polymorphism of PPARD-87 T>C (rs2016520) existed and its influence in CRC were investigated. Tumor masses from primary CRC patients were collected during the operation of tumorectomy, specimens at the different sites of the same tumor mass were sampled and stored individually. The SNP of PPARD-87 T>C was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and the expression of PPARD in vivo was observed by immunohistochemistry. The correlation of PPARD -87 T>C intratumoral polymorphism and the clinicopathological parameters of patients was analyzed statistically. Tumor samples were collected from 106 CRC patients (70 males and 36 females) with an average age of 61.04±13.67 years. A total number of 808 samples (7.60±1.60 per patient) were mainly harvested at peripheral superficial (n=376), central superficial (n=163), invasive front (n=112) and mesenteric cancer foci (n=42) of tumor tissues as well as cancerous adjacent mucosa (n=104). PCR-RFLP analysis showed that T/T (n=460, 56.9%) and T/C (n=334, 41.3%) were the main genotypes of -87 T>C among these samples. Furthermore, intratumoral genotype of -87 T>C was homogeneous in 90 patients and heterogeneous in other 16 patients. The intratumoral heterogeneity was related to patients' age (P=0.016), tumor location (P=0.011) and the grade of differentiation (P=0.022). For patients with intratumoral heterogeneity, immunochemistry showed the expressions of PPARD were not influenced by T/T or T/C genotypes. Intratumoral heterogeneity of PPARD-87 T>C wildly existed in CRC, and associated with patients' age, tumor location and differentiation. However, the immunochemistry assay revealed that there's no significant link between heterogeneity and expression of PPARD.

[Value analysis of preoperative and intra-operative ultrasound in diagnosis and treatment of parathyroid adenoma].

Objective:To explore the value characteristics of preoperative and intra-operative ultrasound in the diagnosis and treatment of parathyroid adenoma,and to further clarify the value of ultrasound in the diagnosis and treatment of parathyroid adenoma.Method:A total of 62 cases of parathyroid adenoma confirmed by postoperative pathology from March 2016 to November 2017 were collected, and the pre-operative ultrasound parameters were analyzed;and 26 cases were detected by intra-operative ultrasound.Result:In 62 cases of parathyroid adenoma, 58 cases of parathyroid adenoma were diagnosed by ultrasound before operation,and the sensitivity was 93.54%. Among the 26 cases,the lesions of 24 cases were detected by intra-operative ultrasound,and the sensitivity was 92.31%.Conclusion:Preoperative ultrasonography has a high diagnostic value for the qualitative diagnosis and location diagnosis of parathyroid adenoma, and intra-operative ultrasound can help to detect lesions quickly and safely, which is of great significance to shorten the operation time and improve the safety of operation. It is an important development space of ultrasound in the diagnosis and treatment of parathyroid glands.

LncRNA NBAT-1 is down-regulated in lung cancer and influences cell proliferation, apoptosis and cell cycle.

OBJECTIVE: To explore the expression and role of lncRNA NBAT-1 in lung cancer. PATIENTS AND METHODS: LncRNA NBAT-1 expression in lung cancer tissues and adjacent areas was detected via reverse transcriptase-polymerase chain reaction (RT-PCR). RAC1 protein was analyzed via Western blotting assay. Cell counting kit-8 (CCK-8) and flow cytometry were used to evaluate cell proliferation and apoptosis as well as cell cycle. RESULTS: The expression level of lncRNA NBAT-1 in cancer specimen was remarkably lower than that in adjacent areas. Furthermore, the low expression of lncRNA NBAT-1 had a significant correlation with patient's tumor size, differentiation degree of tumor cells and lymph node metastasis. The overexpression of lncRNA NBAT-1 could inhibit the proliferation and cell cycle, promote the apoptosis of A549 cells, and down-regulate the expression level of RAC1. CONCLUSIONS: The low expression of lncRNA NBAT-1 is involved in the progression of lung cancer.

Development of a novel high-entropy alloy with eminent efficiency of degrading azo dye solutions.

In addition to its scientific importance, the degradation of azo dyes is of practical significance from the perspective of environmental protection. Although encouraging progress has been made on developing degradation approaches and materials, it is still challenging to fully resolve this long-standing problem. Herein, we report that high entropy alloys, which have been emerging as a new class of metallic materials in the last decade, have excellent performance in degradation of azo dyes. In particular, the newly developed AlCoCrTiZn high-entropy alloy synthesized by mechanical alloying exhibits a prominent efficiency in degradation of the azo dye (Direct Blue 6: DB6), as high as that of the best metallic glass reported so far. The newly developed AlCoCrTiZn HEA powder has low activation energy barrier, i.e., 30 kJ/mol, for the degrading reaction and thus make the occurrence of reaction easier as compared with other materials such as the glassy Fe-based powders. The excellent capability of our high-entropy alloys in degrading azo dye is attributed to their unique atomic structure with severe lattice distortion, chemical composition effect, residual stress and high specific surface area. Our findings have important implications in developing novel high-entropy alloys for functional applications as catalyst materials.

Observation of clinical efficacy of rt-PA intravenous thrombolytic treatment for patients combined with grade 0-1 diabetic foot by Wagner classification and acute ischemic stroke.

OBJECTIVE: To evaluate the safety and efficacy of rt-PA intravenous thrombolytic treatment for patients with diabetic foot (DF) and acute ischemic stroke. PATIENTS AND METHODS: A retrospective analysis was performed on 76 patients admitted between June 2012 to December 2015 presenting with acute ischemic stroke and Grade 0-1 DF a (Wagner classification). The treatment group consisted of 44 patients who received rt-PA intravenous thrombolytic treatment, while 32 cases in the control group did not. Both groups received monitored dietary therapy and hypoglycemic drugs to control their blood glucose levels. In the treatment group, patients received rt-PA intravenous thrombolytic treatment 4.5h after the onset of ischemic stroke. Physical parameters like color change of sick-foot skin, conditions of ulcer concrescence, changes of skin temperature, and promotion of pain scores were observed in both the groups. RESULTS: The improvement in the rt-PA intravenous thrombolytic treatment group was higher than that in general treatment group, and the difference between them had statistical significance (p<0.01). Though the improvement of foot symptoms of patients who received rt-PA intravenous thrombolytic was better than that of the control group; there was no obvious statistical difference in the incidence of adverse events between the treatment group and the control group. CONCLUSIONS: rt-PA can reduce the level serum fibrinogen, promotes local microcirculation and nutrition metabolism of diabetic foot, and improve the clinical prognosis of patients with diabetic foot, but will not increase the incidence of adverse events at the same time.

Linker length affects expression and bioactivity of the onconase fusion protein in Pichia pastoris.

The aim of this study was to analyze the effect of linker length on the expression and biological activity of recombinant protein onconase (ONC) in fusion with human serum albumin (HSA) in Pichia pastoris. Four flexible linkers with different lengths namely Linker L0, L1: (GGGGS)1, L2: (GGGGS)2, and L3:(GGGGS)3 were inserted into the fusion gene and referred to as HSA-n-ONC, where N = 0, 5, 10, or 15. The sequence of the fusion gene HSA-ONC was designed based on the GC content and codon bias in P. pastoris; the signal peptide of albumin was used as the secretion signal. Gene sequences coding for the fusion protein with different linkers were inserted into pPICZα-A to form recombinant plasmids pPICZα-A/HSA-n-ONC, which were then transformed into P. pastoris X-33 for protein expression. Ideal conditions for expression of the fusion proteins were optimized at a small scale, using shake flasks before proceeding to mass production in 10-L fermenters. The recombinant fusion proteins were purified by aqueous two-phase extraction coupled with DEAE anion exchange chromatography, and their cytotoxic effect on the tumor cell was evaluated by the sulforhodamine B assay. The results showed that the expressed amount of fusion proteins had no significant relationship with the length of different linkers and rHSA-0-ONC had no cytotoxic effect on the tumor cells. While rHSA-5-ONC and rHSA-10-ONC had a weak cytotoxic effect, rHSA-15-ONC could kill various tumor cells in vitro. In summary, the biological activity of the fusion protein gradually improved with increasing length of the linker.

Efficacy of albendazole combined with a marine fungal extract (m2-9) against Angiostrongylus cantonensis-induced meningitis in mice.

The pathogenesis of angiostrongyliasis, resulting from Angiostrongylus cantonensis invasion of the human central nervous system, remains elusive. Anthelmintics are usually used to kill worms, although dead worms in the brain may cause severe inflammation which will lead to central nervous system damage. Therefore, combination therapy with anthelmintics and anti-inflammatory drugs in the treatment of human angiostrongyliasis needs further study. To evaluate the efficacy of albendazole combined with a marine fungal extract (m2-9) in A. cantonensis infection, BALB/c mice infected by the third-stage larvae of A. cantonensis were divided into three groups: mice treated with albendazole or m2-9 alone or in combination from day 5 post-inoculation (PI). Several efficacy parameters were recorded, including weight change, worm recovery, neurological function, behavioural analysis, eosinophil and leucocyte counts. The results showed that combination therapy increased body weight, reduced worm burden, improved learning ability, memory and action, decreased neurological dysfunction and leucocyte response in these mice. The combination of albendazole and m2-9 treatment significantly decreased leucocyte response and increased the frequency of rearing, compared to infected mice treated with either drug alone. Therefore, m2-9 is a natural product with potentially significant therapeutic value for angiostrongyliasis and is worthy of further study.

PLA2G6 gene mutation in autosomal recessive early-onset parkinsonism in a Chinese cohort.

OBJECTIVE: Mutations in the PLA2G6 gene at the PARK14 locus have been reported in complicated parkinsonism. To assess the prevalence of and phenotypes associated with PLA2G6 gene mutations, we screened PLA2G6 mutations in a cohort of patients with autosomal recessive early-onset parkinsonism (AREP). METHODS: We selected 12 families with AREP in which the Parkin, PINK1, DJ-1, ATP13A2, and FBXO7 gene mutations had been previously excluded. All patients came from the mainland of China. The entire PLA2G6 coding region and exon-intron boundaries were sequenced from genomic DNA templates. We then performed PET studies on individuals in the pedigree with a homozygous PLA2G6 mutation, and investigated the enzyme activity level of the mutation. RESULTS: A homozygous missense mutation, c.G991T (p.D331Y), was identified in an autosomal recessive case. A younger sister of the p.D331Y-carrying patient was also homozygous for the mutation, but with no extrapyramidal symptoms. A PET study showed a substantial reduction in dopamine transporter (DAT) binding in the p.D331Y patient, and a slight reduction in DAT binding in his sister. In vitro, we experimentally demonstrate that the D331Y mutation caused an approximately 70%reduction in enzyme activity. CONCLUSIONS: We have confirmed that the PLA2G6 gene allocated PARK14 locus and is associated with AREP.

rSj16, a recombinant protein of Schistosoma japonicum-derived molecule, reduces severity of the complete Freund's adjuvant-induced adjuvant arthritis in rats' model.

Sj16, a 16-kDa protein produced by Schistosoma japonicum, has been demonstrated to have anti-inflammatory effect. However, the possible mechanism of these phenomena has not been discovered. Here, we tried to touch it with arthritis rats' model induced by injection of complete Freund's adjuvant (CFA). A set of pathogenic characters were observed in CFA-treated rat, including local and systematic read-out, which showed the model successfully set up. After administration of rSj16 (recombinant Sj16) in vivo, paw swelling reduced significantly and in a dose-dependent manner, the level of TNF-α, IL-1ß and NO decreased and IL-10 in the serum increased. In vitro, rSj16 reversed the augmented surface expression of CD80, CD86, CD54 and OX6 induced by lipopolysaccharide (LPS) in bone marrow-derived DCs (BMDCs), whereas endocytotic capacity of rSj16-treated dendritic cell (DC) was profoundly increased. IL-12p70 released from rSj16-treated BMDC was decreased but IL-10 increased. Further, following incubation with rSj16 primed BMDCs, the sensitized T cells exhibited increased production of anti-inflammatory IL-10 and IL-4 and decreased production of IL-12p70 and IFN-γ. Collectively, these results implied that rSj16 alleviated CFA-induced arthritis, and the possible mechanisms may be its interruption of maturation and function of DCs. rSj16 could be a potential therapeutic agent against rheumatoid arthritis.

Comparative analysis of Bombyx mori nucleopolyhedrovirus responsive genes in fat body and haemocyte of B. mori resistant and susceptible strains.

The infection profiles of the Bombyx mori nucleopolyhedrovirus (BmNPV) in B. mori larvae revealed that the virus invaded the fat body and haemocyte of both KN and 306 strains, which are highly resistant and susceptible, respectively, to BmNPV infection. However, viral proliferation was notably slowed in the resistant B. mori strain. Using suppression subtractive hybridization, two fat body cDNA libraries were constructed to compare BmNPV responsive gene expression levels between the two silkworm lines. In total, 96 differentially expressed genes were obtained. Real-time quantitative PCR (qPCR) analysis confirmed that eight genes were significantly up-regulated in the fat body and haemocyte of the KN strain following BmNPV injection. Our results suggest that these genes may have potential roles in B. mori antiviral infection mechanisms.