Animals ; Atherosclerosis ; Humans ; Serine Proteinase Inhibitors ; Serpins

The aim of this study was to investigate the role of seminal plasma miR-210-3p in the impairment of semen quality caused by varicocele. This study included 102 patients whose semen quality was normal when they were diagnosed with varicocele. A 2-year follow-up for included patients was performed, and they were divided into Group A (semen quality became abnormal) and Group B (semen quality remained normal) according to the results of semen analysis during the follow-up. Semen parameters and seminal plasma miR-210-3p expression were investigated by semen analysis and quantitative real-time polymerase chain reaction, respectively. In vitro experiments with GC-2 cells were performed to explore the role of miR-210-3p in spermatogenic cells. The results of quantitative real-time polymerase chain reaction showed that the level of seminal plasma miR-210-3p in Group A was higher than that in Group B both after 2-year follow-up and when they were diagnosed with varicocele (both P < 0.01). Apoptosis and proliferation assays showed that miR-210-3p induces apoptosis of spermatogenic cells by promoting caspase-3 activation. In conclusion, our study indicated that seminal plasma miR-210-3p induces spermatogenic cell apoptosis by activating caspase-3 in patients with varicocele. Seminal plasma miR-210-3p may be a potential biomarker for predicting impaired semen quality caused by varicocele.

Objective To determine the impact of fluorine and aluminum,and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro and to explore its mechanisms.Methods The osteoclasts and bone marrow stromal cells (BMSCs) isolated from long bone of new born rats were cultured,respectively,in TC199 medium (containing 10％ fetal bovine serum) with fluoride,aluminum and fluoride combined with aluminum.The osteoclasts were inoculated in 96-well culture plate and ivory slice,BMSCs in 6-well culture plate,and culture medium was changed after 2 hours incubation.The cells were divided into control group,fluoride group,aluminum group and fluoride combined with aluminum group; the doses of sodium fluoride were 0,1.0 × 10-4,0,1.0 × 10-4 mol/L and the doses of aluminum chloride were 0,0,1.0 × 10-5,1.0 × 10-5 mol/L,respectively.Tartrate-resistant acid phosphatase (TRAP) staining positive cells were counted under light microscope after TRAP staining on the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue.The expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) was detected by real-time fluorescence quantitative PCR in BMSCs after 8 h treatment.Results ① Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the numbers of osteoclasts (F =7.15,6.56 and 7.98,respectively,all P ＜ 0.05).The numbers of osteoclasts in fluoride group,aluminum group and fluoride combined with aluminum group[(136.9 ± 22.99),(135.4 ± 23.5),(163.0 ± 24.4) per well] were higher than that in the control group[(92.5 ± 22.1) per well,all P ＜ 0.05].② Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the resorption pit area on ivory slices(F =10.47,12.64,14.29,respectively,all P ＜ 0.05).The resorption pit area on ivory slices in fluoride group,aluminum group and fluoride combined with aluminum group[(0.242 ± 0.031),(0.293 ± 0.026),(0.333 ± 0.016)mm2 per slice] was higher than that in the control group [(0.088 ± 0.030)mm2 per slice,all P ＜ 0.05].③Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the expression ratios of RANKL/OPG in BMSCs (F =8.15,15.38,23.59,respectively,all P ＜ 0.05).The expression ratios of RANKL/OPG in BMSCs in fluoride group,aluminum group and fluoride combined with aluminum group [(193.98 ± 137.93)％,(326.11 ± 176.78)％,(599.84 ± 275.82)％] were higher than that in the control group[(100.00 ± 56.02)％,all P ＜ 0.05].Conclusions Both fluoride and aluminum can cause increase in the number of osteoclasts in vitro and promote cell differentiation and bone resorption activity,which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.The stimulating effects of fluoride on osteoclasts differentiation and bone resorption is enhanced by aluminum.

Background Normal ultrastructure is the anatomical basis of retinal pigment epithelial(RPE) cells to perform normal physiological function.At present the precipitation method is often used to detect the ultrastructure of RPE cells with transmission electron microscopy(TEM).Objective The aim of this study was to explore a simple and feasible approach to examine the ultrastructure of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells.Methods hESCs were induced and differentiated into RPE cells by the spontaneous differentiation method,and the expressions of microphthalmia associated transcription factor MITF and paired-box gene 6 (PAX6),specific protein of RPE cells,in the cells were detected by immunofluorescence assay.hESC-RPE cells were inoculated into Transwell filter,and the ultrastructure of the cell sheet was examined under the TEM.Then the ultrastructure of the cell sheet specimens was compared with those of hESC-RPE cells from cell precipitation and RPE cell specimens of 90-day-old Long Evans rats.Results MITF and PAX6 were positively expressed in hESC-RPE cells.The normal ultrastructure were visible in the RPE cells of rats under the TEM,including apical microvilli,polarized melanin granules,cellular nucleus,basement membrane and intercellular junctions,and the ultrastructure of hESC-RPE cell sheet on Transwell was similar to the RPE cells in rats.However,only scatter melanin granules,nonpolar nucleus and scanty microvilli were observed under the TEM in the hESC-RPE cells by cell precipitation method.Conclusions Without digestion process,hESC-RPE cell sheet on Transwell can retain the normal ultrastructure of hESC-RPE cells under the TEM,with a more simple and reliable advantage.

Objective To explore clinical value of serum total bile acid(TBA) and cholinesterase(CHE) in infantile hepatopathy.Methods Sixty infants diagnosed respectively with breast milk jaundice,cytomegalovirus hepatitis and congenital bile duct teratism were included.Twenty healthy infants were as control.The infants of cytomegalovirus hepatitis were self-compared in acute stage and convalescence stage.Serum TBA and CHE in every group were measured and compared.Results Serum TBA in control group,breast milk jaundice group,cytomegalovirus hepatitis group and congenital bile duct teratism group were(6.47?3.25),(8.71?1.79),(45.86?40.85),(119.50?31.73) ?mol/L,respectively;and CHE were(11295.50?1731.91),(10762.89?1237.30),(9771.32?860.27),(7967.304?31.58) U/L,respectively.Compared with other groups,the level of serum TBA and CHE in cytomegalovirus hepatitis group and the congenital bile duct teratism group were significantly different(all P

OBJECTIVE: To assess the effect of disinfectant (Cavicide) with benzethon chloramine and isopropanol as main active ingredients disinfectant on dental impression accuracy. METHODS: The effect of Cavicide on three impression materials (alginate, polyether and vinylpolysiloxane) were assessed using a standard model. The standard model was digitized by an extraoral scanner (IScan D103i, Imetric). For each kind of impression materials, thirty impressions were taken following the manufactures' instruction in the same conditions. Subsequently, the impressions were randomly divided into three groups, with ten impressions in each group. After the impression taking was completed, the three groups underwent pure water rinse for 1 min (blank control, BC), 2% glutaraldehyde solution immersion disinfection for 30 min (glutaraldehyde, GD), and Cavicide solution spray disinfection for 5 min (Cavicide, CC), respectively. All the impressions were digitized by the extraoral scanner (IScan D103i, Imetric) after disinfection and exported to a dedicated three-dimensional analysis software (Geomagic Qualify 2014, Geomagic, USA). In the software, the digital models of the impressions were trimmed to teeth and then superimposed with the digitized standard model via best-fit alignment. Root mean square (RMS) was used to evaluate the deviations between the impression and the standard model. The deviation in the anterior and posterior regions was evaluated respectively. One-way ANOVA test and the LSD post-hoc test were used to compare the deviations between the three groups (P < 0.05). The color map of each superimposition was saved for visual analysis. RESULTS: For the polyether and vinylpolysiloxane materials, the difference between the three groups was not statistically significant (P=0.933, P=0.827). For the alginate material, the difference in posterior region between group GD and group BC, as well as group GD and group CC were statistically significant (GD vs. BC, P=0.001; GD vs. CC, P=0.002), while the difference between group BC and group CC was not statistically significant (P=0.854). The visual analysis showed an obvious deviation in the buccal-lingual direction in group GD. CONCLUSION Disinfectant (Cavicide) with benzethon chloramine and isopropanol as main active ingredients using spray disinfection has no effect on the accuracy of the alginate, polyether and vinylpolysiloxane impressions.
2-Propanol ; Chloramines ; Dental Impression Materials ; Dental Impression Technique ; Disinfectants ; Disinfection ; Models, Dental

Animals ; Arsenic ; toxicity ; Comet Assay ; DNA Damage ; drug effects ; Lymphocytes ; drug effects ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Wistar ; Tobacco Smoke Pollution ; adverse effects

Biomarkers ; blood ; Dimethylformamide ; pharmacokinetics ; Humans ; Occupational Exposure

OBJECTIVE TNF- related apoptosis- inducing ligand(TRAIL)is a promising cancer therapeutic agent due to its minimal toxicity to normal tissues and remarkable apoptotic activity in tumors. However, most breast cancer cells are resistant to TRAIL- induced apoptosis. Our objectives are to investigate the underlying molecular mechanisms and to develop strategies to overcome such resistance. METHODS To identify modulators of TRAIL-induced apoptosis, we carried out a genome wide siRNA screen. To validate the screening result, we either silenced or overexpressed the identified genes in various breast cancer cells and changes in growth and TRAIL-induced cell apoptosis were determined in vitro and in an orthotopic xenograft mouse model. Finally, we investigated whether small molecules targeting the identified genes improve the effectiveness of TRAIL-therapy. RESULTS We unexpectedly identified androgen receptor (AR) to be responsible for TRAIL resistance. While AR is classically viewed as the key factor in prostate cancer progression, we found that AR expression levels were markedly elevated in human invasive breast cancer specimens including triple- negative breast cancers (TNBC) that are highly aggressive with poor prognosis. Importantly, breast cancer cell lines express different levels of AR that correlated with their TRAIL resistance. AR overexpression in MDA- MB- 231 and MDA- MB- 436 cells suppressed the TRAIL sensitivity whereas knockdown of AR rendered MCF-7 and MDA-MB-453 cells sensitive to TRAIL-induced apoptosis. AR overexpression also induced TRAIL resistance in breast tumors in vivo. Further, we observed an upregulation of the TRAIL receptor, death receptor 5 (DR5) in breast cancer cells, following the removal or inhibition of AR by its antagonists Casodex and MDV3100. Treatment with AR antagonists also enhanced TRAIL- induced breast cancer cell apoptosis. CONCLUSION AR signaling suppresses TRAIL-induced breast cancer cell apoptosis, in part, by suppressing DR5 expression, and a combination of AR antagonists together with TRAIL may be a novel and effective therapy for TNBC.

Objective To study the effect of selenium on peripheral and splentic T-cell subset, apoptosis of spleen cells in fluorosis chicken and its mechanism. Methods One hundred and eighty 8-day Hailanhe chicks were randomly divided into 3 groups(each 60): ①control group: 195 mg/kg fluoride and 0.08 mg/kg of selenium; ②fluorine group : 1000 mg/kg fluoride and 0.08 mg/kg of selenium ;③selenium antagonism group : 1000 mg/kg On 30~(th), 60~(th), 90~(th) day, peripheral and splentic CD4~+, CD8~+ T-cell subset analyses underwent flow cytometry and apoptosis of spleen cells were detected by TUNEL for study subjects. Results Compared with control group, the CD4~+ T-cell subset of peripheral in fluorine group was decreased obviously in 30,60,90 days[ (35.36± 4.27)% vs (24.29 ± 2.96)%, (47.65 ± 5.42)% vs (41.62 ± 3.96)%, (49.58 ± 3.98) % vs (42.35 ± 6.03 )%, P < 0.05 or < 0.01 ], CD4~+/CD8~+ ratio also was decreased obviously [ ( 1.701 ± 0.145 )% vs (1.393 ± 0.163)%,(2.712 ± 0.345)% vs (1.781 ± 0.201)%,(2.438 ± 0.356)% vs (1.973 ± 0.229)%, P< 0.05 or < 0.01]. Compared with fluorine group, the CD4~+ T-cell subset of peripheral in selenium antagonism group [ (29.40 ± 3.38)%, (45.40 ± 6.01 )%, (46.85 ± 5.25)%, P < 0.05 or < 0.01 ] was increased obviously in 30,60,90 days,CD4~+/CD8~+ ratio in 60,90 days[(2.004 ±0.314)%,(2.211±0.229)%,all P<0.01]also was increased obviously.Compared with control group,the CD4~+ T-cell subset of spleen cells in fluorine group was decreased obviously in 30,60,90 days[(47.33±5.35)% vs(41.91±4.83)%,(49.28±5.24)% vs(41.26 ±4.56)%,(34.31±4.15)%vs(29.33±2.89)%,all P<0.01],CD4~+/CD8~+ ratio also was decreased obviously[(1.927 ±0.244)% vs(1.525 ±0.265)%,(1.847±0.224)% vs(1.640±0.198)%.(1.265±0.174)% vs(0.878±0.092)%,P<0.05 or<0.01].Compared with fluorine group,the CD4~+ T-cell subset of spleen cells in selenium antagonism group in 60,90 days[(44.87±5.43)%,(32.62±3.37)%,all P<0.05]was increased obviously,CD4~+/CD8~+ ratio in 30,60, 90 days[(1.703 ±0.201)%,(1.772±0.215)%,(0.991±0.124)%,P<0.05 or<0.01]also was increased obviously. The apoptosis ratio of spleen cells in fluorine group in 30,60,90 days[(2.31±0.36)%,(2.76±0.22)%,(3.04± 0.29)%]was higher than that in control group[(1.14±0.21)%,(1.23±0.23)%,(1.29±0.20)%,P<0.01].The apoptosis ratio of spleen cells in selenium antagonism group in 60,90 days[(2.42 ±0.32)%,(2.73±0.39)%]was lower than that in fluorine group(P<0.05 or<0.01).Conclusion A certain concentration of selenium can antagonize the immunity inhibition of fluorine by decreasing apoptosis and improving the unbalance of T-cell subset.