Objective To investigate the effects of chronic fluorosis on the expressions of matrix metalloproteinase-9 (MMP-9) mRNA and protein and the differentiation and maturation process of bone cell in the osteoclast of bone tissue of rats.Methods According to body weight,thirty-six healthy SD rats(body mass 100-120 g) were divided into three groups by random number table,twelve in each group,half male and half female.The rats of control group were given tap water(NaF ＜ 1 mg/L),and rats of low-fluorine and high-fluorine groups were fed with tap water containing 5 and 50 mg/L NaF to establish chronic fluorosis model.Rats were sacrificed after eight months; the contents of urinary fluoride in 24 hours and bone fluoride were analyzed by fluoride selective electrode.Serum content of tartrate resistant acid phosphatase 5b(TRACP5b)was detected by enzyme-linked immunosorbent assay (ELISA).The paraffin section of bone tissue was stained by hematoxylin-eosin (HE) and pathological morphometry was observed under optical microscope.The protein and mRNA levels of MMP-9 in the osteoclast of bones were detected by immunohistochemistry (IHC) and in situ hybridization (ISH),respectively.Results The differences of fluoride contents of urine and bone in rats were statistically significant between groups(F =400.612,48.229,all P ＜ 0.05).Fluoride contents of urine and bone were increased in lowfluorine and high-fluorine groups[(6.09 + 0.56),(7.69 + 0.64)mg/L,(12.65 ± 3.07),(26.53 + 5.88)mg/kg] compared to the control groups[(1.36 ± 0.51)mg/L,(0.67 ± 0.16)mg/kg,all P ＜ 0.05],and the fluoride contents of urine and bone were gradually increased with increasing fluoride doses(all P ＜ 0.05).The difference of TRACP5b content in serum was statistically significant between groups (F =9.607,P ＜ 0.05),in low-fluorine and high-fluorine groups,the TRACP5b contents[(1.86 ± 0.13),(1.92 ± 0.22)U/L] were higher than that of control group [(1.57 + 0.20)U/L,all P ＜ 0.05].The pathological examination showed osteosclerosis in fluoride exposed groups.The differences of MMP-9 mRNA and protein expressions were statistically significant between groups (F =365.727,331.382,all P ＜ 0.05).Compared to the control groups(97.22 ± 2.24,78.51 ± 1.16),the expressions of MMP-9 protein(108.18 ± 1.97,119.28 ± 1.76) and mRNA(89.44 ± 2.86,102.14 ± 2.39) were increased(all P ＜ 0.05),and the expressions of MMP-9 mRNA and protein were gradually increased with increasing fluoride doses (all P ＜ 0.05).Conclusions Chronic fluorosis might influence osteoclast differentiation and maturation process through regulating the expression levels of MMP-9 protein and mRNA.

Ascites ; Humans ; Language ; Quality of Life ; Reproducibility of Results ; Surveys and Questionnaires ; Translations

Objective To discover the influence of thiomersalate exposure on the behavior and humoral immunity of premature rats.Methods According to common thiomersalate dose of vaccine used in human,the surface conversation method was used to change the dose to 1,2,3,4 times for rats(32.8,65.6,98.4 and 131.2 μg/kg),and the thiomersalate was separately injected to the gluteus maximus of premature rats respectively,which were delivered on the 20th day of gestation by cesarean for the injection on day 1 (premature).Simultaneously,saline was injected to the control group.On day 44 to 48 after injection,the Morris water maze test was used to evaluate the spatial learning and memory abilities of rats and on the post-injection day 49,immunoglobulin content in rat blood and cerebrospinal fluid was measured.Results The spatial learning abilities of rats in 131.2 μg/kg group and the memory abilities in 65.6 μg/kg,98.4 μg/kg and 131.2 μg/kg group were significantly weaker than those in 9 g/L saline group(all P ＜0.001).The immunoglobulin content in blood and cerebrospinal fluid changed in different groups exposed to thiomersalate.The levels of IgG,IgA,IgE,IgM in 98.4 μg/kg group and 131.2 μg/kg group rose significantly higher than those in 9 g/L saline group(all P ＜ 0.01).Analysis of rectilinear correlation between the immunoglobulin and learning abilities of rats revealed that IgG,IgA,IgE levels in the blood were correlated with the spatial learning abilities (r =0.36,0.47,0.50,all P ＜ 0.05) and the levels of IgG,IgA,IgE,IgM in blood were rectilinear correlated with the memory abilities of rats (r =-0.39,-0.43,-0.49,-0.38,all P ＜ 0.05).In rats'cerebrospinal fluid,IgG,IgA,IgE,IgM levels were rectilinear correlative with both the spatial learning and memory abilities (r =0.48,0.59,0.54,0.41,all P＜0.05;r=-0.39,-0.61,-0.57,-0.44,all P＜0.05).Conclusion With the dose ofthiomersalate increased,the learning and memory and humoral immunity are sustained damaged for premature rats.

Retinopathy of prematurity screening is very important for early identification of retinopathy of prematurity in premature infants, but procedural pain caused by screening process seriously affects the growth and development of premature infants. This paper reviews the non-pharmacological intervention methods for relieving procedural pain of retinopathy of prematurity screening at home and abroad, so as to provide theoretical basis for medical staff to carry out procedural pain management of retinopathy of prematurity screening.

Objective:To investigate the risk factors and treatment outcome of recurrent Acanthamoeba keratitis (AK) after corneal transplantation. Methods:A serial case-observational study was carried out.Twenty-eight eyes of 28 patients with AK who underwent corneal transplantation in Shandong Eye Hospital from January 2012 to January 2019 were enrolled.All the eyes received corneal transplantation from failing to respond to topical and systemic anti- Acanthamoeba medical therapy, including 13 eyes that received penetrating keratoplasty (PKP) and 15 eyes that received lamellar keratoplasty (LKP). The corneal lesion was removed by a trephine with a diameter of 0.5 mm over infiltration area during PKP or LKP.The clinical features of recurrent AK were summarized, including recurrence time, site and signs, and the risk factors of AK recurrence were analyzed.Local and systemic anti- Acanthamoeba medical therapy was performed in all relapsed eyes, and secondary surgery was performed for the eyes with poor response to medication.The therapeutic outcome of recurrent AK was evaluated.The study adhered to the Declaration of Helsinki.This study protocol was approved by an Ethics Committee of Shandong Eye Hospital (No.201112). Results:In the 28 eyes, 7 eyes (25%) appeared recurrent AK after keratoplasty, including 2 eyes after PKP and 5 eyes after LKP.There was no significant difference in the recurrence rate between the two methods ( P=0.396). The recurrence rate of eyes that had used glucocorticoids drugs before operation was 57.14% (4/7), which was significantly higher in comparison with 14.29% (3/21) of eyes without glucocorticoids before surgery ( P=0.043). The recurrence rate of eyes with ulcer diameter ≥8.2 mm was 50.00% (5/10), which was significantly higher than 11.11% (2/18) of eyes with ulcer <8.2 mm ( P=0.036). The recurrent lesions began at the edge of implant bed accounted for 85.71% (6/7), and the recurrent lesions located below graft accounted for 14.29% (1/7). In 7 eyes with recurrent AK, 6 eyes were completely cured.Among recurrent AK eyes after LKP, 2 eyes were cured by long-term medical therapy, and 2 eyes were cured by extended-diameter LKP, and another 1 eye was cured by conjunctival flap covering surgery.One eye with recurrent AK after PKP was cured by extended-diameter PKP. Conclusions:The risk factors of recurrent AK after surgery are application of glucocorticoids before surgery and big lesions.Recurrent AK after surgery is curable by individualized therapy targeting to different clinical characteristics.

We studied the function and mechanism of miR-24 in regulating beta-like globin gene expression. We first detected the expression of miR-24 during erythroid differentiation and also detected the globin gene expression in miR-24 overexpressing K562 cells through q-PCR. Dual-luciferase reporter assay and Western blotting were used to identify target genes of miR-24. "Rescue experiment" was further used to investigate the regulation of miR-24 on globin gene expression whether depending on targeting Sp1 or not. We found that miR-24 increased during hemin-induced K562 cells and EPO-induced HPCs (hematopoietic progenitor cells) erythroid differentiation. Overexpression of miR-24 in K562 cells promoted the epsilon- and gamma-globin gene expression during hemin-induced erythroid differentiation through targeting the negative globin regulator Sp1. These results suggested that miR-24 can improve the expression of beta-like globin gene through targeting Sp1.
Cell Differentiation ; Gene Expression Regulation ; Hematopoietic Stem Cells ; metabolism ; Humans ; K562 Cells ; MicroRNAs ; genetics ; Sp1 Transcription Factor ; genetics ; epsilon-Globins ; genetics ; gamma-Globins ; genetics

Epitranscriptomics focuses on the RNA-modification-mediated post-transcriptional regulation of gene expression. The past decade has witnessed tremendous progress in our understanding of the landscapes and biological functions of RNA modifications, as prompted by the emergence of potent analytical approaches. The hematopoietic system provides a lifelong supply of blood cells, and gene expression is tightly controlled during the differentiation of hematopoietic stem cells (HSCs). The dysregulation of gene expression during hematopoiesis may lead to severe disorders, including acute myeloid leukemia (AML). Emerging evidence supports the involvement of the mRNA modification system in normal hematopoiesis and AML pathogenesis, which has led to the development of small-molecule inhibitors that target N6-methyladenosine (m 6A) modification machinery as treatments. Here, we summarize the latest findings and our most up-to-date information on the roles of m 6A and N7-methylguanine in both physiological and pathological conditions in the hematopoietic system. Furthermore, we will discuss the therapeutic potential and limitations of cancer treatments targeting m 6A.

Objective:To investigate the role of nicotinamide (NIC) in the differentiation of neural crest cells from human embryonic stem cells (hESCs), and lay the foundation for the induction of hESC-derived corneal endothelial cells.Methods:hESCs line H1 cultured for 5-7 days was used for induction.According to the different components of the neural crest induction medium, cells were assigned into different groups for 7-days induction, including group treated without NIC cultured in induction medium only, group treated with NIC cultured in induction medium containing 10 mmol/L NIC, NIC+ resveratrol (Res) group cultured in induction medium containing 10 mmol/L NIC and 10 μmol/L Res and Sirtinol group cultured in induction medium containing 10 μmol/L Sirtinol.Res and Sirtinol were used as SIRT1 activity agonist and inhibitor, respectively.The relative mRNA expression levels of hESCs and neural crest cell markers were detected by real-time fluorescence quantitative PCR at 1, 3, 5 and 7 days during the induction.The expression of neural crest cells markers after 7 days of induction was assayed by immunofluorescence staining.The induction efficiency of NIC and the effect of SIRT1 regulation on human natural killer 1 (HNK-1) positive cells expression were evaluated through flow cytometry analysis of percentages of nerve growth factor receptor (P75) and HNK-1 + cells. Results:Compared with the group treated without NIC, the mRNA expressions of totipotent genes octamer transcription factor 4 (OCT4) and homeodomain proteins (NANOG) were significantly decreased, and the mRNA expression levels of neural crest cell markers P75, HNK-1, SRY-related HMG box (SOX) 9 and SOX10 were significantly increased in the group treated with NIC after 5 days of induction (all at P<0.05). In the group treated without NIC, P75 was weakly expressed, and HNK-1 was sporadically expressed, and transcription factor AP-2β (AP-2β) and paired-like homeodomain transcription factor 2 (PITX2) were not detected.In the group treated with NIC, P75, HNK-1, AP-2β and PITX2 were strongly expressed.The proportion of P75 + HNK-1 + cells and P75 + cells were both significantly higher in the group treated with NIC than without NIC ( t=8.481, P=0.001; t=2.987, P=0.041). The percentage of HNK-1 + cells in groups treated without and with NIC, NIC+ Res group and Sirtinol group were (34.267±12.522)%, (89.633±1.358)%, (64.667±6.429)% and (86.300±3.460)%, respectively, with a statistically significant overall difference ( F=36.799, P<0.001). The proportion of HNK-1 + cells in NIC+ Res group was significantly lower than that in the groups treated with NIC and Sirtinol (all at P<0.05). Conclusions:NIC promotes the differentiation of hESCs-derived neural crest cells by inhibiting the activity of SIRT1 to enhance the expression of HNK-1.NIC treatment may provide a new strategy for source of seed cells in the treatment of neural crest cell-related diseases, such as corneal endothelial transplantation.

Background::The conversion of adenosine (A) to inosine (I) through deamination is the prevailing form of RNA editing, impacting numerous nuclear and cytoplasmic transcripts across various eukaryotic species. Millions of high-confidence RNA editing sites have been identified and integrated into various RNA databases, providing a convenient platform for the rapid identification of key drivers of cancer and potential therapeutic targets. However, the available database for integration of RNA editing in hematopoietic cells and hematopoietic malignancies is still lacking.Methods::We downloaded RNA sequencing (RNA-seq) data of 29 leukemia patients and 19 healthy donors from National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database, and RNA-seq data of 12 mouse hematopoietic cell populations obtained from our previous research were also used. We performed sequence alignment, identified RNA editing sites, and obtained characteristic editing sites related to normal hematopoietic development and abnormal editing sites associated with hematologic diseases.Results::We established a new database, "REDH", represents RNA editome in hematopoietic differentiation and malignancy. REDH is a curated database of associations between RNA editome and hematopoiesis. REDH integrates 30,796 editing sites from 12 murine adult hematopoietic cell populations and systematically characterizes more than 400,000 edited events in malignant hematopoietic samples from 48 cohorts (human). Through the Differentiation, Disease, Enrichment, and knowledge modules, each A-to-I editing site is systematically integrated, including its distribution throughout the genome, its clinical information (human sample), and functional editing sites under physiological and pathological conditions. Furthermore, REDH compares the similarities and differences of editing sites between different hematologic malignancies and healthy control.Conclusions::REDH is accessible at http://www.redhdatabase.com/. This user-friendly database would aid in understanding the mechanisms of RNA editing in hematopoietic differentiation and malignancies. It provides a set of data related to the maintenance of hematopoietic homeostasis and identifying potential therapeutic targets in malignancies.

Objective To investigate the relationship of miRNA gene polymorphisms with blood pressure(BP)responses to the sodium and potassium diet intervention.Methods In 2004,we recruited 514 participants from 124 families in seven villages of Baoji,Shaanxi Province,China.All subjects were given a three-day normal diet,followed by a seven-day low-salt diet,a seven-day high-salt diet,and finally a seven-day high-salt and potassium supplementation.A total of 19 miRNA single nucleotide polymorphisms(SNPs)were selected for analysis.Results Throughout the sodium-potassium dietary intervention,the BP of the subjects fluctuated across all phases,showing a decrease during the low-salt period and an increase during the high-salt period,followed by a reduction in BP subsequent to potassium supplementation during the high-salt diet.MiR-210-3p SNP rs 12364149 was significantly associated with systolic BP(SBP),diastolic BP(DBP)and mean arterial pressure(MAP)responses to low-salt diet.MiR-4638-3p SNP rs6601178 was significantly associated with SBP while miR-26b-3p SNP rs115254818 was significantly associated with MAP responses to low-salt intervention.In addition,miR-26b-3p SNP rs115254818 was significantly correlated with SBP,DBP and MAP responses to high-salt intervention.MiR-1307-5p SNPs rs1 1191676 and rs2292807 were associated with SBP and MAP responses to high-salt diet.MiR-4638-3p SNP rs6601178,miR-210-3p SNP rs12364149,miR-382-5p SNP rs4906032 and rs4143957 were significantly associated with SBP response to high-salt diet.In addition,miR-26b-3p SNP rs115254818 was significantly associated with SBP,DBP and MAP responses to potassium supplementation.MiR-1307-5p SNPs rs11191676,rs2292807,and miR-19a-3p SNP rs4284505 were significantly associated with SBP responses to high-salt and potassium supplementation.Conclusion miRNA gene polymorphisms are associated with BP response to sodium and potassium,suggesting that miRNA genes may be involved in the pathophysiological process of salt sensitivity and potassium sensitivity.