Effects of different specimen pretreatment methods on the measurement of thyrotropin stimulating hormone in serum.

OBJECTIVE: The pre-analysis processing method of serum samples plays a very important role in the assurance of the quality of the entire test and the accuracy of the results. This study illustrates the importance of pretreatment methods of serum samples for the test results by comparing the effects of different pretreatment methods on the measurement of thyroid stimulating hormone (TSH) concentration in serum. SUBJECTS AND METHODS: In this study, the concentrations of TSH of 37 patients' serum, which were treated in six different ways including the reverse mixing times after blood collection, clotting time and conditions, centrifugal speed and time, were detected on Automatic Chemiluminescence Immunoassay Analyzer, and a comparative analysis of the different results was performed. RESULTS: For serum samples containing coagulants, the test results were significantly affected if the samples were not reversed mixing after collection. The abnormal results would be obtained with insufficient coagulation time, low reaction temperature, low centrifuge speed and insufficient centrifugation time. CONCLUSIONS: The pre-analysis processing of serum samples is the beginning of the entire inspection process. The quality of the entire inspection will not be guaranteed if the pre-analysis processing method is irregular. Therefore, clinical laboratories should pay more attention to the pretreatment process of samples to ensure the quality of the entire inspection process.

s-wave superconductivity in the noncentrosymmetric W3Al2C superconductor: an NMR study.

We report on a microscopic study of the noncentrosymmetric superconductor W3Al2C (withTc= 7.6 K), mostly by means of27Al- and13C nuclear magnetic resonance (NMR). Since in this material the density of states at the Fermi level is dominated by the tungsten's 5dorbitals, we expect a sizeable spin-orbit coupling (SOC) effect. The normal-state electronic properties of W3Al2C resemble those of a standard metal, but with a Korringa product 1/(T1T) significantly smaller than that of metallic Al, reflecting the marginal role played bys-electrons. In the superconducting state, we observe a reduction of the Knight shift and an exponential decrease of the NMR relaxation rate 1/T1, typical ofs-wave superconductivity (SC). This is further supported by the observation of a small but distinct coherence peak just belowTcin the13C NMR relaxation-rate, in agreement with the fully-gapped superconducting state inferred from the electronic specific-heat data well belowTc. The above features are compared to those of members of the same family, in particular, Mo3Al2C, often claimed to exhibit unconventional SC. We discuss why, despite the enhanced SOC, W3Al2C does not show spin-triplet features in its superconducting state and consider the broader consequences of our results for noncentrosymmetric superconductors in general.

Study on effects of miR-141-3p in proliferation, migration, invasion and apoptosis of colon cancer cells by inhibiting Bcl2.

PURPOSE: This study aimed to investigate the relationship between miR-141-3p and B lymphocyte-2 gene (Bcl2) gene and its biological behavior on colon cancer cell line SW480. METHODS: qRT-PCR was used to detect the expression level of miR-141-3p in colon cancer tissues and adjacent tissues, as well as in colon cancer cell line and normal human colonic epithelial cell line FHC. MTT assay, wound assay, and Transwell demonstrated the effects of miR-141-3p on colon cancer proliferation, migration and invasion. Targetscan7.1 predictive software and dual luciferase reporter assays were used to detect the targeted regulation of miR-141-3p on the apoptosis-related gene Bcl2. MTT assay, wound assay, Transwell and flow cytometry were used to detect the effect of Bcl2 on miR-141-3p on colon cancer proliferation, migration, invasion and apoptosis. RESULTS: Compared with adjacent tissues, the expression of miR-141-3p in colon cancer tissues was significantly down-regulated. Colon cancer patients with low expression of miR-141-3p had poorer prognosis. Compared with normal colonic epithelial cells, miR-141-3p expression was significantly down-regulated in colon cancer cell lines, and overexpression of miR-141-3p significantly attenuated the proliferation, migration and invasion of colon cancer cells. Knockdown of miR-141-3p significantly promoted the proliferation, migration and invasion of colon cancer cells. miR-141-3p targets the negative regulation of Bcl2. Knockdown of Bcl2 significantly attenuated the promotion of miR-141-3p inhibitor on proliferation, migration and invasion of colon cancer cells and inhibition of apoptosis. Knockdown of Bcl2 significantly enhanced the inhibition effect of miR-141-3p inhibitor on proliferation, migration and invasion of colon cancer cells. CONCLUSIONS: In conclusion, miR-141-3p can inhibit the cancer by regulating Bcl2, and miR-141-3p has the potential to become a potential therapeutic target for colon cancer.

Signature of type-II Weyl semimetal phase in MoTe2.

Topological Weyl semimetal (TWS), a new state of quantum matter, has sparked enormous research interest recently. Possessing unique Weyl fermions in the bulk and Fermi arcs on the surface, TWSs offer a rare platform for realizing many exotic physical phenomena. TWSs can be classified into type-I that respect Lorentz symmetry and type-II that do not. Here, we directly visualize the electronic structure of MoTe2, a recently proposed type-II TWS. Using angle-resolved photoemission spectroscopy (ARPES), we unravel the unique surface Fermi arcs, in good agreement with our ab initio calculations that have nontrivial topological nature. Our work not only leads to new understandings of the unusual properties discovered in this family of compounds, but also allows for the further exploration of exotic properties and practical applications of type-II TWSs, as well as the interplay between superconductivity (MoTe2 was discovered to be superconducting recently) and their topological order.

The critical barrier to progress in dentine bonding with the etch-and-rinse technique.

OBJECTIVES: The lack of durability in resin-dentine bonds led to the use of chlorhexidine as MMP-inhibitor to prevent the degradation of hybrid layers. Biomimetic remineralisation is a concept-proven approach in preventing the degradation of resin-dentine bonds. The purpose of this study is to examine the integrity of aged resin-dentine interfaces created with a nanofiller-containing etch-and-rinse adhesive after the application of these two approaches. METHODS: The more established MMP-inhibition approach was examined using a parallel in vivo and in vitro ageing design to facilitate comparison with the biomimetic remineralisation approach using an in vitro ageing design. Specimens bonded without chlorhexidine exhibited extensive degradation of the hybrid layer after 12 months of in vivo ageing. RESULTS: Dissolution of nanofillers could be seen within a water-rich zone within the adhesive layer. Although specimens bonded with chlorhexidine exhibited intact hybrid layers, water-rich regions remained in those hybrid layers and degradation of nanofillers occurred within the adhesive layer. Specimens subjected to in vitro biomimetic remineralisation followed by in vitro ageing demonstrated intrafibrillar collagen remineralisation within hybrid layers and deposition of mineral nanocrystals in nanovoids within the adhesive. CONCLUSIONS: The impact was realized by understanding the lack of an inherent mechanism to remove water from resin-dentine interfaces as the critical barrier to progress in bonding with the etch-and-rinse technique. The experimental biomimetic remineralisation strategy offers a creative solution for incorporating a progressive hydration mechanism to achieve this goal, which warrants its translation into a clinically applicable technique.

Recombinant human midkine stimulates proliferation of articular chondrocytes.

OBJECTIVES: Midkine, a heparin-binding growth factor, promotes population growth, survival and migration of several cell types, but its effect on articular chondrocytes remains unknown. The aim of this study was to investigate its role on proliferation of articular chondrocytes in vitro and in vivo. MATERIALS AND METHODS: Bromodeoxyuridine incorporation and MTT assays were performed to examine the proliferative effect of recombinant human midkine (rhMK) on primary articular chondrocytes. Activation of extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase (PI3K) was analysed using western blot analysis. Systemic and local delivery of rhMK into mice and rats was preformed to investigate the proliferative effect of rhMK in vivo, respectively. Histological evaluation, including measurement of articular cartilage thickness, cell density, matrix staining and immunostaining of proliferating cell nuclear antigen was carried out. RESULTS: rhMK promoted proliferation of articular chondrocytes cultured in a monolayer, which was mediated by activation of ERK and PI3K. The proliferative role of rhMK was not coupled to dedifferentiation of culture-expanded cells. Consistent with its action in vitro, rhMK stimulated proliferation of articular chondrocytes in vivo when it was administered subcutaneously and intra-articularly in mice and rats, respectively. CONCLUSION: Our results demonstrate that rhMK stimulates proliferation of primary articular chondrocytes in vitro and in vivo. The results of this study warrant further examination of rhMK for treatment of animal models of articular cartilage defects.

P13 of Leucania separata multiple nuclear polyhedrosis virus affected the polyhedra and budded virions yields of AcMNPV.

p13 gene was first described by our laboratory in Leucania separata multiple nuclear polyhedrovirus (Ls-p13, ORF114) back to 1995. However, the functions of Ls-P13 and its reported homologues remained unknown. In order to probe the function of Ls-P13, recombinant Autographa californica nucleopolyhedroviruses (rAcMNPVs) were constructed to express Ls-P13 in the Sf9 cells at early, late or early/late phase. Observations of microscope showed that the expression of Ls-P13 could decrease the yield of AcMNPV polyhedra in Sf9 cells, and early expressed Ls-P13 had stronger inhibition efficiency than that of the late expressed. Results of flow cytometry also indicated that Ls-P13 decreased the yield of AcMNPV polyhedra while increased those of budded virions (BVs) in Sf9 cells, but the efficacy was lost when its leucine zipper-like domain was mutated. Ls-P13 is a transmembrane protein, which was early located in the nucleus and late mainly in the cytoplasm membrane at 48 h. When its transmembrane domains were deleted, Ls-P13 distribution was dramatically diverted from cytoplasm membrane to nucleus, its corresponding efficacy on polyhedra yield was further increased while that on BVs was slightly weakened. Bioassay results indicated that Ls-P13 accelerated the larvae-killing rate. The mechanism might be that Ls-P13 increased BV yield.

[Estimate the safety of baculovirus insecticides: the history and the status Quo].

This article reviews the evaluation of security of baculovirus used as a pesticide. Since 1970s, the scholars have done a lot of experiments with kinds of baculovirus to test the security of a large number kinds of living things even our human beings. Almost all experiments proved that baculovirus is secure, but some experiments came to different conclusions. These gave rise to great debate twice when they were published, but these conclusions have been proved to be wrong with later test by other scientists or the author himself. Since 90s baculovirus have been used a great deal as the vector to express the foreign gene. Some of them reported the expression in mammalian cells, which brought the suspicious of the baculovirus safety. This article made an analysis and a conclusion about them. Also, this article laid emphasis on the security of recombination baculovirus with the results of the security experiment of self-made recombination baculovirus pesticide. In the last analysis, this article draws a conclusion that the baculovirus and recombinant baculovirus insecticides are secure.

[Construction and bioassay of secondary recombinant baculovirus with two foreign gene].

Plasmid pAcLEneo which bears neomycin gene derived by the baculovirus IE1 promotor was digested and the gene was harvested. A transfer vector pAc34DZ2 in which the polyhedrin envelope gene has been inactivated by insertion of expression cassette of PIE1/neo was constructed by inserting the neo cassette into the SacI site of plasmid pAc34DZ1. We have constructed the polyhedrin positive recombinant virus (I) vAcPhBtT which was able to express Bt truncated endotoxin gene. In order to improve the insecticide efficiency of recombinant virus (I), an anti-neomycin recombinant virus (II) vAcPhBtTPE- was obtained by second co-transfection into the Sf9 cells with pAc34DZ2 and recombinant virus (I) DNA. Southern blot and SDS-PAGE analysis indicated that the recombinant virus (II) was still able to express the 80 kD Bt truncated delta-endotoxin, but did not express the 34 kD polyhedrin envelope protein. The recombinant virus (II) without envelope released virion faster than recombinant virus (I) after alkaline lysis. Bioassay was carried out using Spodoera exigua. LC50 of the recombinant virus (II) was about half of wild type virus and LT50 reduced about two days.

A baculovirus vector derived from immediately early gene promoter of Autographa californica nuclear polyhedrosis virus.

A transfer vector was constructed in which the neomycin resistance (neo) gene was under the control of a copy of Autographa californica nuclear polyhedrosis virus (AcMNPV) IE1 gene promoter at the p10 locus. After cotransfection of Spodoptera frugiperda (Sf9) cells with the transfer vector and infectious AcMNPV DNA, the recombinant virus-containing neo gene was selected by serial passage of the mixed progenies from cotransfection. This was done at low MOI in the presence of G418 in growth medium and was followed by limited dilution. RNA dot hybridization showed that the neo gene was transcribed from immediately early phase to very late phase, in infected Sf9 cells. These results demonstrate that a new baculovirus vector system had been constructed in infected cells. Furthermore, a new method for selection of positive recombinant baculovirus had been developed.

Nucleotide sequence of a 1446 base pair SalI fragment and structure of a novel early gene of Leucania separata nuclear polyhedrosis virus.

A 1446 bp SalI fragment of LsNPV was sequenced by the silver staining method, and two large open reading frames (ORFs, ORF1 and ORF2) were found, both contain typical characteristics of the 5' regulatory elements of baculovirus early genes. ORF1 is 345 bp long with the capacity to encode a putative protein of 114 amino acid residues with MW about 13 kDa and was designated p13 gene, ORF2 comprises 248 bp from the 3' end of the fragment. In the untranslated region (UTR) of ORF1, a 33 bp mini cistron (ORF3), a core recognition sequence (CGTCG) for many bHLHzip transcription factors and a late promoter sequence TTAAG are present. In the UTR of ORF2, two host transcription factor binding elements (CACGTG and GATA motif) and two CGT motifs were found. Some regular leucine zipper-like structures, designated leucine trans-conformation structure and LVT repeat, were found near the N-terminus and the middle of p13 protein. The leucine trans-conformation structure that is near the N-terminus consists of 4 leucines and 7 other amino acids between every two leucines, and every leucine is located at a conformation shift point of the predicted secondary structure of the p13 protein. In LVT repeat, L-6aa-V-6aa-T-6aa is repeated once. The functions of those structures remain unclear, and the two ORFs, not found in the genome of Autographa californica nuclear polyhedrosis virus, are possibly two new genes.

[Studies on the chemical constituents of the herb huanghuaren (Sida acutá Burm. f.)].

Seven constituents were isolated from the dry herb Huanghuaren. On the basis of spectroscopic analysis and physico-chemical constants four of them were identified as heraclenol, beta-sitosterol, acanthoside B and daucoglycoside.

Coliphage M13 cloning system and ddXTP chain-termination method for DNA sequencing.

Two DNA fragments of cytochrome b gene in yeast mitochondrial DNA were sequenced by Messing's M13 cloning system and Sanger's ddXTP chain-termination method. M13mp8 and M13mp9 serve as vectors for insert fragment. The recipient strain is E. coli JM103 for transfection. When the ratio of insert/vector was 3:1, high frequencies of recombination and positive recombination were obtained. The two fragments, which have 575 bp and 709 bp, were sequenced. To read more bases, the ratio of ddXTP/dXTP must be reduced. On one X-ray film of 80 x 17 cm 394 bp were read.