Establishment and characterization of a HER2-enriched canine mammary cancerous myoepithelial cell line.

BACKGROUND: Canine mammary tumors (CMTs) have a poor prognosis, along with tumor recurrence and metastasis. Cell lines are vital in vitro models for CMT research. Many CMT epithelial cell lines were reported. However, canine mammary myoepithelial cells, the contractile component of the canine mammary tissue were overlooked. This study aimed at establishing such a cell line. CMT-1 cell line was obtained from a canine mammary tumor CMT-1 and characterized molecularly through qPCR, western blotting, immunochemistry and immunofluorescence. Its doubling time, cytogenetic analysis and migration rate were evaluated using growth study, karyotype analysis and wound healing assay respectively. To determine its tumorigenesis, xenograft transplantation was performed. RESULTS: CMT-1 tumor was a complex canine mammary carcinoma that stained negative to estrogen receptors (ER) and progesterone receptors (PR), but positive to human epidermal growth receptor-2 (HER2), defined as HER2-enriched subtype. In this study, a CMT-1 cell line obtained from CMT-1 tumor was immune-positive to vimentin, α-SMA, p63 and negative to E-cadherin (E-cad), indicating CMT-1 cells were myoepithelial cells. It was successfully cultured for more than 50 passages showing the same immunoreactivity to ER, PR, and HER2 as the primary canine tumor. The doubling time of CMT-1 cell line was 26.67 h. The chromosome number of CMT-1 cells ranged from 31 to 64. A potential spontaneous epithelial to mesenchymal transition (EMT) was noticed during cell cultures. Potential EMT-induced CMT-1 cells showed no significance in migration rate compared to the original CMT-1 cells. CMT-1 cells was able to grow on a 3D culture and formed grape-like, solid, and cystic mammospheres at different time period. Inoculation of CMT-1 cells induced a complex HER2-enriched mammary tumor with metastasis in mice. CONCLUSIONS: A canine cancerous HER2-enriched myoepithelial cell line was successfully established and a canine mammosphere developed from myoepithelial cells was documented in this study. We are expecting this novel cell line and its associated mammospheres could be used as a model to elucidate the role of myoepithelial cells in CMT carcinogensis in the future.

Electro-responsive actuators based on graphene.

Electro-responsive actuators (ERAs) hold great promise for cutting-edge applications in e-skins, soft robots, unmanned flight, and in vivo surgery devices due to the advantages of fast response, precise control, programmable deformation, and the ease of integration with control circuits. Recently, considering the excellent physical/chemical/mechanical properties (e.g., high carrier mobility, strong mechanical strength, outstanding thermal conductivity, high specific surface area, flexibility, and transparency), graphene and its derivatives have emerged as an appealing material in developing ERAs. In this review, we have summarized the recent advances in graphene-based ERAs. Typical the working mechanisms of graphene ERAs have been introduced. Design principles and working performance of three typical types of graphene ERAs (e.g., electrostatic actuators, electrothermal actuators, and ionic actuators) have been comprehensively summarized. Besides, emerging applications of graphene ERAs, including artificial muscles, bionic robots, human-soft actuators interaction, and other smart devices, have been reviewed. At last, the current challenges and future perspectives of graphene ERAs are discussed.

[Arthroscopic reinforced reconstruction of anterior cruciate ligament with autologous hamstring tendon and anchor suture band].

OBJECTIVE: To investigate the effectiveness of arthroscopic reinforced reconstruction of anterior cruciate ligament (ACL) with autologous hamstring tendon combined with anchor suture band. METHODS: Between February 2016 and March 2018, 60 patients who were to be treated with arthroscopic ACL reconstruction and met the selection criteria were selected in the study. Among them, 30 cases were reconstructed with autologous hamstring tendon combined with anchor suture band (trial group), and 30 cases were reconstructed with simple autologous hamstring tendon (control group). There was no significant difference in gender, age, disease duration, cause of injury, injury side, and preoperative Lysholm score, Tegner score, and International Knee Documentation Committee (IKDC) score between the two groups ( P>0.05). After reconstruction, the patients in the trial group were allowed to start knee flexion and extension activities early without wearing an adjustable brace, while the patients in the control group were required to wear an adjustable brace for 12 weeks. The knee joint function (Lysholm score, Tegner score, IKDC score) and stability (Lachman test and pivot shift test) were compared between the two groups after operation. RESULTS: There was no significant difference in graft diameter between the two groups ( t=1.061, P=0.115). Compared with the control group, the operation time of the trial group was significantly different ( t=4.924, P=0.000). All incisions healed primarily. In the control group, the intramuscular venous thrombosis occurred in 2 cases after operation. Both groups were followed up 18 months. The Lysholm score, Tegner score, and IKDC score of the two groups at each time point after operation were significantly higher than those before operation ( P<0.05); the above scores in the trial group were significantly higher than those in the control group at 3, 6, and 9 months after operation ( P<0.05); there was no significant difference between the two groups at 18 months after operation ( P>0.05). There was no significant difference in Lachman test results between the two groups at each time point after operation ( P>0.05). There was a significant difference in pivot shift test results at 6 months after operation between the two groups ( P<0.05); but there was no significant difference at other time points ( P>0.05). CONCLUSION: The effectiveness of ACL reinforcedreconstruction with autologous hamstring tendon combined with anchor suture band is satisfactory. Compared with using autologous hamstring tendon alone, it has better initial strength and joint stability, and is more conducive to early postoperative functional exercise and functional recovery of knee joint.

Nickel-Catalyzed Amidoalkylation Reaction of γ-Hydroxy Lactams: An Access to 3-Substituted Isoindolinones.

An efficient Ni(ClO4)2·6H2O-promoted amidoalkylation reaction for the synthesis of 3-substituted isoindolinones involving various γ-hydroxy lactams and nucleophiles has been successfully developed. The transformation proceeds with both carbon (ketones and arenes) and heteroatom (alcohols, thiols, and amines) nucleophiles and in both intermolecular and intramolecular manners. The prominent features of the present strategy are wide substrate scope, excellent group tolerability, and moderate to good yields (up to 96% yield). The present strategy is also characterized by remarkable superiority over the current synthetic methods. Furthermore, the reaction could be scaled up to the multigram scale.

Association of urinary metal profiles with serum uric acid: a cross-sectional study of traffic policemen in Wuhan, China.

OBJECTIVES: Serum uric acid (SUA) is both a strong antioxidant and one of the key risk factors of cardiovascular diseases (CVDs). We aimed to investigate the associations of urinary metal profile with SUA in traffic policemen in Wuhan, China. DESIGN: A cross-sectional study was carried out in traffic policemen. SETTING: A seriously polluted Chinese city. PARTICIPANTS: A total of 186 traffic policemen were recruited in this study. About 56 of them worked in the logistics department and the other 130 maintained traffic order or dealt with traffic accidents on the roads. All these subjects had worked as a policeman for at least 1 year. MAIN OUTCOME MEASURES: SUA. RESULTS: The significantly negative association of lead with SUA was consistent between single-metal and multiple-metal models (p=0.004 and p=0.020, respectively). Vanadium, chromium and tin were reversely associated with SUA levels in the single-metal models after false discovery rate (FDR) adjustment (all P_FDR < 0.05). One IQR increase in vanadium, chromium, tin and lead was associated with 26.9 µmol/L (95% CI -44.6 to -9.2; p=0.003), 27.4 µmol/L (95% CI -46.1 to -8.8; p=0.004), 11.2 µmol/L (95% CI -18.9 to -3.4; p=0.005) and 16.4 µmol/L (95% CI -27.6 to -5.2; p=0.004) decrease in SUA, respectively. Significant interaction between smoking and vanadium on decreased SUV was found (pfor interaction = 0.007 and p_FDR = 0.028). CONCLUSIONS: Urinary vanadium, chromium, tin and lead were negatively associated with SUA. Vanadium and cigarette smoking jointly affected SUA levels. Further studies are needed to replicate these findings and to investigate the potential mechanisms.

Distribution and association study in copy number variation of KCNJ12 gene across four Chinese cattle populations.

Copy number variation is a large genome variation which usually happens in the noncoding-region, and it may occur at the locus associated with the functional gene to further influence the phenotype. Potassium inwardly-rectifying channel, subfamily J 12 (KCNJ12) gene expressed widely in cardiomyocytes and neurons, plays an important role in tumor therapy and muscle movement regulation. In this study, we detected the distribution of CNVs for KCNJ12 gene in 404 individuals belonging to four Chinese cattle breeds (NY, JX, JA and GF). We also investigated the KCNJ12 gene expression in different tissues of JX cattle. Additionally, we examined the association of two CNV regions (CNV1: 1,600 bp, intron 1; CNV2: 4,800 bp, intergenic) with growth traits. The statistical analyses indicated that the CNV1 is associated with the body length, rump length and weight in JX cattle population (P < 0.05); and there has a significant association with the body length, chest circumference, and body weight in GF cattle (P < 0.05).The CNV2 had a significant effect on the body length and body weight in JX cattle (P < 0.05); the body length, chest circumference, rump length and body weight in GF cattle (P < 0.01 or P < 0.05). The copy numbers of KCNJ12 gene presented the negative correlations with the transcript level of gene in skeletal muscles (P < 0.05). Our results provide evidence that CNV1 and CNV 2 in KCNJ12 are associated with growth traits in two cattle populations and may be used as candidates for marker-assisted selection and breeding management in cattle.

l-Arginine and allopurinol supplementation attenuates inflammatory mediators in human osteoblasts-osteoarthritis cells.

This study investigated the synergistic effects of l-arginine and allopurinol on antioxidant and inflammatory mediators in human osteoblasts-osteoarthritis (HOb-OA) cells. The cells were treated with allopurinol (50-150 mg/kg bwt) and l-arginine (50-150 mg/kg bwt) for 72 h. Cell viability, catalase, superoxide dismutase (SOD), glutathione peroxidase (Gpx), reduced glutathione (GSH), lipid peroxidation, and the inflammatory markers interleukin 6 (IL-6), interleukin 1ß (IL-1ß), nuclear factor κB (NF-κB) and tumor necrosis factor alpha (TNF-α) were measured. The combined supplementation with allopurinol and l-arginine increased catalase, SOD, GSH, and Gpx, while it decreased lipid peroxidation, IL-6, IL-1ß, and TNF-α. While TNF-α, IL-6, IL-1ß, and NF-κB mRNA and protein expression were higher in control HOb-OA cells, the combined supplementation with allopurinol and l-arginine substantially reduced their expression in HOb-OA cells by >40%. In summary, combined supplementation with allopurinol and l-arginine might be very effective in osteoarthritis. A search for therapeutic agents that inhibit inflammation could help to prevent and manage osteoarthritis. However, further studies need to determine the biochemical and molecular mechanisms of these agents in osteoarthritis.

Re-characterization of some factors influencing aerosol sampling in the workplace: results from field studies.

OBJECTIVE: Workplace aerosol sampling is challenged by its influencing factors and methodological limitations. Some factors, including blank setting, electrostatic effect, sample mass gain, and limit of detection (LOD), were studied to characterize them further and thereby to improve the sampling method. METHODS: Through a field sampling in two shipyards and by an analysis of a large amount of dust data from different industries, the influencing factors were comparatively studied with emphasis on their effect on the final result. RESULTS: After calibration with field blanks, the concentrations of most sample types in the shipyards decreased significantly, varying by as much as -24.3% of the final measurements. After laboratory blank calibration, dust concentrations increased or decreased without a definite change trend. With a variation of -1.8%, only the measurements of Chinese "total dust" sampled with polypropylene filters were significantly influenced by the electrostatic effect. The LOD coincidence rate was only 17.3% for American respirable dust in different industries and 12.2% for respirable particles collected by normal flow rate samplers (FSP2) in the shipyards. The latter increased to 73.9% when high flow samplers (FSP10) were used. CONCLUSIONS: It was suggested that field blank calibration was the predominant influencing factor in comparison with electrostatic effect and laboratory blank adjustment. The LOD coincidence rate was too low for reliable sampling, and this might be improved by use of high flow samplers.

Genetic variants of nucleotide excision repair genes are associated with DNA damage in coke oven workers.

We explored the effects of single nucleotide polymorphisms (SNP) of nucleotide excision repair (NER) genes on DNA damage caused by exposure to carcinogenic polycyclic aromatic hydrocarbons (PAH) in 475 Chinese workers. We quantified urinary 1-hydroxypyrene using high-performance liquid chromatography, and the DNA damage level of lymphocytes was examined by the comet assay and represented as the Olive tail moment (OTM) value. We genotyped 38 tagSNPs in 10 NER genes. The SNP function was further investigated using luciferase reporter assay in three cell lines. Our results showed that two promoter SNPs, XPA rs1800975 and XPC rs3731055, were associated with lower OTM values (P(trend) = 0.01 and 0.02 respectively). However, another missense SNP rs2228001 in the XPC gene was positively associated with OTM value (P(trend) = 0.01). A stratified analysis found that the association between this SNP and DNA damage was only observed among subjects with higher PAH exposure levels but not among those with lower exposure levels (P(interaction) = 0.018). A dose-response association was found between the combined risk alleles of the above three genetic variants and increased DNA damage levels (P(trend) = 0.004). This association was more pronounced in subjects with higher PAH exposure than those with lower exposure levels (P(interaction) = 0.046). Our functional study indicated that XPA rs1800975G and XPC rs3731055A alleles had a higher luciferase expression than their corresponding SNP alleles (P < 0.05). These results suggested that genetic variations in key NER genes, especially in XPA and XPC genes, may modulate DNA damage levels when exposed to PAHs.

[Co-detection of P21, P53 and HSP70 and their possible role in diagnosis of polycyclic aromatic hydrocarbons (PAHs)-related lung cancer].

OBJECTIVE: To explore the biomarkers of early diagnosis in patients with polycyclic aromatic hydrocarbons (PAHs)-related lung cancer for the application to detection of occupational lung cancer or related lung cancer. METHODS: Western dot blotting was used to explore the expression of ras, p53 and heat stress protein 70 (HSP70) in 29 patients with PAHs-related lung cancer (LC), and 28 patients with non-cancerous pulmonary disease, and 30 healthy controls. RESULTS: The positive detection rates of P21, P53, and HSP70 in LC group (58.62%, 34.48%, 41.38% respectively) were higher than those in non-cancerous pulmonary disease group (14.29%, 7.14%, 10.71% respectively, P < 0.01). The sensitivity of P21, P53 and HSP70 were 58.62%, 34.48% and 41.38% respectively, negative predictive value (NPV) were 68.42%, 78.05% and 63.04% respectively. The co-detection of the three proteins mentioned above produced a sensitivity of 82.76% with a NPV of 78.26% (P < 0.05). Of 18 cases of LC with negative cytology, 13 (72.22%) were found HSP21, P53 or HSP70 positive. CONCLUSIONS: Co-detection of the P21, P53, and HSP70 may be used as the screening marker for diagnosis of PAHs-related lung cancer, and may supplement the diagnostic value of conventional cytology.