[Research progress on animal and cell models of pneumoconiosis].

Pneumoconiosis is an occupational lung disease with the highest incidence in China. There is no effective treatment drug at present. Animal and cell models are the basis for exploring its pathogenesis and developing effective drugs. In this paper, we sort out the methods of animal models of pneumoconiosis and the different cell models induced by dust in recent years, by analyzing and summarizing the advantages and disadvantages, modeling time, pathology and changes in important indicators of different preparation methods of animal models, as well as different cell models induced by the dust to simulate different pathological models and pathological stages, to provide basis for the application and improvement of pneumoconiosis model.

[Research progress in pathogenesis of pneumoconiosis].

Pneumoconiosis is an occupational lung disease that is mainly caused by diffuse fibrosis of lung tissue due to long-term inhalation of productive dust during occupational activities and retention in the lungs. Macrophages, epithelial cells and other cells can release a large number of cytokines, such as transforming growth factor-ß, monocyte chemotactic protein-1, etc. These cytokines can participate in pathologies such as local injury, inflammatory response, and pulmonary fibrosis. This article reviews the role of cytokines in the pathogenesis of pneumoconiosis in order to provide a basis for further research.

MicroRNA-135b inhibits odontoblast-like differentiation of human dental pulp cells by regulating Smad5 and Smad4.

AIM: To investigate the function of miRNAs in odontoblast-like differentiation of human dental pulp cells (hDPCs). METHODOLOGY: Integrated comparative miRNA microarray profiling was used to determine the differential miRNAs expression in odontoblast-like differentiation of hDPCs. The abundance of microRNA-135b (miR-135b) was measured by quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR) and in situ hybridization (ISH). Bioinformatic analyses combined with luciferase assays were utilized to identify the targets interacting with miR-135b. Overexpression of miR-135b was performed to investigate the role and mechanism in odontoblast-like differentiation of hDPCs. Statistical analysis was performed by one-way analysis of variance (anova) or Student's t-test. RESULTS: Thirty-six differentially expressed microRNAs in odontoblast-like differentiation of hDPCs were identified. MiR-135b expression was significantly downregulated during hDPCs differentiation (P < 0.05). In addition, miR-135b was able to bind to the 3'-UTR of the Smad5 and Smad4 and repressed these two genes expression (P < 0.05). Furthermore, overexpression of miR-135b suppressed odontoblast-like differentiation of hDPCs and attenuated the expression of Smad5 and Smad4 (P < 0.05). CONCLUSIONS: These observations indicated a potential role of miR-135b in mediating odontoblast-like differentiation of hDPCs and inhibition of miR-135b might be a promising therapeutic way to facilitate dentine tissue engineering.

Effects of glial cell line-derived neurotrophic factor and leukemia-inhibitory factor on the behavior of two calf testis germline stem cell colony types.

Germline stem cells are the only such capable of transmitting genetic information in vivo. The isolation and culture of these cells in vitro provide a unique model to understand sperm differentiation and hence, spermatogenesis and male fertility. In this study, we isolated, purified, and cultured germline stem cells from the testes of newborn calves. Moreover, we investigated the effects of glial cell line-derived neurotrophic factor (GDNF) and leukemia-inhibitory factor (LIF) on their proliferation. Male calf germline stem cells were found to be pluripotent, and able to form grape-like and embryonic stem cell (ES)-like colonies when cultured. GDNF promoted proliferation of the former, whereas LIF induced growth of the latter. The grape-like colonies retained their germline stem cell characteristics, whereas the ES-like colonies demonstrated more primitive attributes. This investigation established a male calf germline stem cell culture model that may serve as a foundation for further studies aiming to understand the properties of such cells.

THIN-LAYER CHROMATOGRAPHY IDENTIFICATION FOR RHUBARB AND PHELLODENDRI AMURENSIS CORTEX IN SHUANG-BAI CATAPLASM AND STUDY OF SKIN IRRITATION ASSAY.

This paper aimed to raise a thin-layer chromatography (TLC) identification method for rhubarb and Phellodendri Amurensis Cortex and inspected skin irritation induced by them. It applied the TLC identification for Rhubarb and Phellodendri Amurensis Cortex in Shuang-bai cataplasm prescription. In this study six rabbits were divided into two groups to observe the skin irritation from Shuang-bai cataplasm on intact and defected skin. Another 36 were randomly divided into 6 groups to observe the acute toxicity from Shuang-bai cataplasm on intact and defected skin. Also 30 guinea pigs were divided into 3 groups to observe skin allergy to Shuang-bai cataplasm. The results showed that the average weight of the group of intact-skin rabbits was 2.026±0.10 kg and 2.427±0.023 kg after medication; the average weight of the group of defected-skin rabbits was 2.170±0.05 kg and 2.540±0.15 kg after medication; Shuang-bai cataplasm produced no irritation on intact or defected rabbit skin, no acute toxicity in rabbits and no allergy on the skin of guinea pigs. The skin allergy rate on guinea pigs of the medication group was 0 at each time quantum. Therefore, it can be concluded that this preparation produces no extreme skin irritation for rabbits, guinea pigs or human beings, and it can be safely put into practice.

Associated analysis of single nucleotide polymorphisms found on exon 3 of the IGF-1 gene with Tibetan miniature pig growth traits.

The IGF-1 gene is an important regulating factor that has a growth-promoting effect on growth hormone. The IGF-1 gene promotes muscle cell differentiation in the muscle cell formation process. The IGF-1 gene also regulates the growth of skeletal muscle during skeletal muscle growth. In addition, the IGF-1 gene plays an important role in the formation of mammals and poultry embryos, and the process of postnatal growth. The IGF-1 gene has been implicated as a candidate gene for the regulation of pig growth traits. We analyzed exon 3 of the IGF-1 gene polymorphism in Tibetan miniature pigs (N = 128) by polymerase chain reaction-single-strand conformation polymorphism and DNA sequencing. One single nucleotide polymorphism (T40C) was found on exon 3 of the IGF-1 gene. Statistical analysis of genotype frequencies revealed that the T allele was dominant in Tibetan miniature pigs at the T40C locus. The association analysis showed that the IGF-1 mutation had an effect on the body weight, body length, and chest circumference of pigs aged 6-8 months. In addition, the IGF-1 mutation had an effect on body weight in pigs aged 9-11 months (P < 0.05). We speculated that the pigs with the TT genotype grow more rapidly compared to those with the TC genotype. The TC genotype of the Tibetan miniature pig has a smaller body type. This information provides a theoretical basis for the genetic background of Tibetan miniature pigs.

p38a MAPK is involved in BMP-2-induced odontoblastic differentiation of human dental pulp cells.

AIM: To investigate whether the p38α mitogen-activated protein kinases (MAPK) is involved in bone morphogenetic protein (BMP)-2-induced odontoblastic differentiation of human dental pulp cells (HDPCs). METHODOLOGY: Recombinant retrovirus encoding shRNA against p38α MAPK was constructed to investigate the role of p38α MAPK on BMP-2-induced odontoblastic differentiation of HDPCs. HDPCs were transfected with retrovirus expressing sh-p38α. Activation of p38α MAPK was detected by Western blot. The effects of p38α MAPK on BMP-2-induced odontoblastic differentiation of HDPCs were measured by alkaline phosphatase (ALP) activity, and the expression of odontoblastic markers was identified by quantitative real-time polymerase chain reaction analysis. The effect of SD-282, a p38a-specific inhibitor, on BMP-2-induced odontoblastic differentiation was also investigated. RESULTS: BMP-2 dose- and time-dependently upregulated phosphorylation of p38α of HDPCs. Compared with BMP-2-treatment group, gene knock-down of p38α MAPK significantly inhibited ALP activity and the formation of mineralized nodules in HDPCs. Moreover, suppression of p38α MAPK repressed the odontoblastic differentiation in HDPCs. Consistently, inhibition of p38α by SD-282 also decreased odontoblastic differentiation. CONCLUSIONS: p38α MAPK is involved in BMP-2-induced odontoblastic differentiation of HDPCs.

Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction.

The article "Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction" by G.-X. Hu, J. Zhang, Y.-G. Tian, Y.-H. Li, L. Mou, L.-J. Qiao, published in Eur Rev Med Pharmacol Sci 2016; 20 (19): 4124-4128 has been withdrawn.

Diagnostic value of joint detection of homocysteine and RDW CV on acute myocardial infarction. Retracted.

OBJECTIVE: We discussed the diagnostic value of joint detection of homocysteine (HCY) and red blood cell volume distribution width variable coefficient on acute myocardial infarction (AMI). PATIENTS AND METHODS: We collected 300 coronary heart disease cases, among which there were 121 cases of stenocardia, 65 cases of ischemic heart failure, and 114 cases of AMI at the Department of Cardiology of our hospital during the period from January 2012 to June 2013. At the same time, we took 100 normal physical examinees as the control group, used the full-automatic cell-analyzer and the immunization to measure HCY and red blood cell distribution width (RDW) CV respectively and analyze their value in diagnosing AMI. RESULTS: The differences among the four groups of HCY and RDW CV were statistically significant (p < 0.05). The HCY and RDW CV level in the AMI group were significantly higher than those of the other three groups (p < 0.05); the differences between the positive diagnosis rate of HCY, the RDW CV and their joint diagnosis in the AMI group were statistically significant (p < 0.05) while the differences between the positive diagnosis rate of HCY, the RDW CV and their joint diagnosis in the control group were not statistically significant (p > 0.05). The detection sensitivity and specificity of HCY alone were respectively 68.42% and 86.00% with those of the RDW CV alone being 64.91% and 84.00%. The joint detection sensitivity and specificity were 83.33% and 93.00%, statistically different (p < 0.05). The concordance rate, the positive predictive value and the negative predictive value were 87.85%, 93.14% and 83.04%, respectively. CONCLUSIONS: The HCY and RDW CV joint diagnosis of AMI had relatively high sensitivity, specificity, concordance rate, positive predictive value and negative predictive value.

Single-nucleotide polymorphism analysis of GH, GHR, and IGF-1 genes in minipigs.

Tibetan (TB) and Bama (BM) miniature pigs are two popular pig breeds that are used as experimental animals in China due to their small body size. Here, we analyzed single-nucleotide polymorphisms (SNPs) in gene fragments that are closely related to growth traits [growth hormone (GH), growth hormone receptor (GHR), and insulin-like growth factor (IGF)-1)] in these pig breeds and a large white (LW) control pig breed. On the basis of the analysis of 100 BMs, 108 TBs, and 50 LWs, the polymorphic distribution levels of GH, GHR, and IGF-1 were significantly different among these three pig breeds. According to correlation analyses between SNPs and five growth traits--body weight (BW), body length (BL), withers height (WH), chest circumference (CC), and abdomen circumference (AC)--three SNP loci in BMs and four SNP loci in TBs significantly affected growth traits. Three SNP sites in BMs and four SNP sites in TBs significantly affected growth traits. SNPs located in the GH gene fragment significantly affected BL and CC at locus 12 and BL at locus 45 in BMs, and also BW, WH, CC, and AC at locus 45 and WH and CC at locus 93 in TBs. One SNP at locus 85 in the BM GHR gene fragment significantly affected all growth traits. All indices were significantly reduced with a mixture of alleles at locus 85. These results provide more information regarding the genetic background of these minipig species and indicate useful selection markers for pig breeding programs.

Single-nucleotide polymorphism analysis of GH, GHR, and IGF-1 genes in minipigs

Tibetan (TB) and Bama (BM) miniature pigs are two popular pig breeds that are used as experimental animals in China due to their small body size. Here, we analyzed single-nucleotide polymorphisms (SNPs) in gene fragments that are closely related to growth traits [growth hormone (GH), growth hormone receptor (GHR), and insulin-like growth factor (IGF)-1)] in these pig breeds and a large white (LW) control pig breed. On the basis of the analysis of 100 BMs, 108 TBs, and 50 LWs, the polymorphic distribution levels of GH, GHR, and IGF-1 were significantly different among these three pig breeds. According to correlation analyses between SNPs and five growth traits - body weight (BW), body length (BL), withers height (WH), chest circumference (CC), and abdomen circumference (AC) - three SNP loci in BMs and four SNP loci in TBs significantly affected growth traits. Three SNP sites in BMs and four SNP sites in TBs significantly affected growth traits. SNPs located in the GH gene fragment significantly affected BL and CC at locus 12 and BL at locus 45 in BMs, and also BW, WH, CC, and AC at locus 45 and WH and CC at locus 93 in TBs. One SNP at locus 85 in the BM GHR gene fragment significantly affected all growth traits. All indices were significantly reduced with a mixture of alleles at locus 85. These results provide more information regarding the genetic background of these minipig species and indicate useful selection markers for pig breeding programs.