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Objective To study the effect of Ureaplasma urealyticum on the level of DCXR expression and apoptosis in human spermatozoa. Methods 145 cases were in infertile group. Semen routine analysis was referred to the WHO standard method. Western blot was used to detect the level of DCXR expression in spermatozoa. The localization of the protein in human spermatozoa was determined by indirect immunofluorescent staining. The sperm apoptosis was assayed by TUNEL. Analysis the indicators of change before and after treatment with sensitive antibiotics for Uu infection. Results The expression of DCXR protein and the percentage of DCXR-positive rate were lowerin UU positive group than those in UU negative group. The expression of DCXR protein and the percentage of DCXR-positive rate were higher after treatment, than before therapy in Uu positive group (P < 0.05). The sperm apoptosis rate in Uu positive group were higher than those in Uu negative. The sperm apoptosis rate was lower after treatment than before therapy in Uu positive group (P < 0.05). Conclusion Ureaplasma urealyticum infection may affect the level of DCXR protein expression, the percentage of the DCXR-positive rate and sperm apoptosis in human spermatozoa. After effective treatment of Uu infection, the sperm protein DCXR expression and sperm apoptosis would be obviously improved.
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Objective To compare the effects of different ways of blood purification on inflammatory cytokines clearance in patients with maintenance hemodialysis.Methods The clinical data of 50 patients with maintenance hemodialysis were retrospectively analyzed.The low flux dialysis,high flux dialysis were used respectively,and the changes of hypersensitive C-reactive protein (hs-CRP),tumor necrosis factor alpha (TNF-α),interleukin 1 (IL-1),interleukin 6 (IL-6) after dialysis were compared between the two methods.Results After low flux dialysis,the hs-CRP,TNF-α,IL-1,IL-6 levels were (6.58 ± 3.69) mg/L,(1.54 ± 0.42) g/L,(105.46 ±8.20) ng/L,(27.58 ± 6.83) ng/L,respectively,which after high flux dialysis were (2.93 ± 1.85) mg/L,(1.13 ±0.27) g/L,(97.21 ± 7.l4.) ng/L,(20.35 ± 5.76) ng/L,respectively,the differences were statistically significant (t =6.253,5.806,5.365,5.806,all P ＜ 0.05).Conclusion Compared with the low flux dialysis,high flux dialysis is more effective in inflammatory cytokines clearance in patients with maintenance hemodialysis.
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Objective To study the expression of vascular endothelial cell growth factor (VEGF) and its receptor 2 (VEGFR2) and apoptosis in the epididymis of rats with chronic arsenic poisoning.Methods Forty male Sprague-Dawley rats,weighting 160-200 g,were selected and randomly divided into four groups:high dose [60.0 mg/L sodium arsenite (NaAsO2)],medium dose (12.0 mg/L NaAsO2),low dose arsenic infected groups (2.4 mg/L NaAsO2),and control group (distilled water).The rats were treated with arsenic through drinking water for 6 consecutive months.At the end of the experiment,all rats were killed and epididymis was separated rapidly,apoptosis cells of rat epididymis were determined via the TUNEL method,the expressions of VEGF and VEGFR2 were observed through immunohistochemistry.Results The apoptosis of the epididymis tubules epithelial cells were compared in control,low,medium,and high dose groups (124.68 ± 6.59,138.96 ± 8.48,152.59 ± 10.79,170.69 ±16.60),the differences were statistically significant (F =10.562,P ＜ 0.05),with increased exposure doses of arsenic,apoptosis cells were increased.VEGF expression levels of the body,head and tail in epididymis were compared in control,low,medium,and high dose groups (148.50 ± 0.60,134.20 ± 0.85,98.23 ± 0.80;136.70 ± 0.95,128.20 ±0.72,90.30 ± 0.12;127.80 ± 0.62,117.60 ± 0.72,84.51 ± 0.60;120.26 ± 0.46,90.12 ± 0.36,66.43 ± 0.90),the differences were statistically significant (F =46.445,45.867,41.381,P ＜ 0.05),and VEGF expression levels in arsenic infected groups were lower than that in control group (P ＜ 0.05);VEGFR2 expression levels of the head and tail (130.70 ± 0.89,128.80 ± 0.70;126.30 ± 0.65,121.42 ± 0.62;120.26 ± 0.12,117.84 ± 0.55;102.60 ±0.78,104.92 ± 1.15) were compared,the differences were statistically significant (F =3.452,2.701,P ＜ 0.05),and VEGFR2 expression levels in arsenic infected groups were lower than that in control group (P ＜ 0.05).Conclusion Arsenism has inhibitory effect on the expression of VEGF and VEGFR2 in the epididymis,leading to epididymis epithelial cell apoptosis,which indicates it may play an important role in male infertility.
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AIM:To investigate the effects of P34H gene silencing on the expression of P34H and activity of hyaluronidase (HYD) in mouse sperm.METHODS:The recombinant plasmid series of P34H targeted short hairpin RNA (shRNA) were constructed by GV248 plasmids vector.These recombinant plasmids were transformed into DH 5αcompetent cells, and the plasmids were taken from DNA sequencing analysis .The HEK293T cells were co-transfected with shRNA and lentiviral packaging plasmids .The 3 kinds of recombinant lentiviruses and negative control lentiviruses were used to in -ject into the mouse epididymis and the expression of P 34H at mRNA and protein levels was detected by real-time PCR and Western blot, respectively.The location of P34H protein on the mouse spermatozoa was determined by indirect immunofluo-rescent staining using P34H antibody.The positive rate and activity intensity of HYD was detected by modified sodium hya-luronate-gelatin membrane.RESULTS:DNA sequencing analysis confirmed that the 3 P34H-shRNA sequences were suc-cessfully inserted into the lentiviral vectors .P34H expression in epididymis tissue was significantly decreased at both mR-NA and protein levels compared with those of the non-transfected and normal control vectors (P<0.05).The GV-P34H-shRNA-1 played a significant role in reducing the percentage of P 34H positive rate and the activity of HYD in mouse sperm.The silencing effect did not significantly differ between the non-transfected and normal control vectors .CONCLU-SION:Silencing of P34H significantly inhibits the percentage of P 34H positive rate and the activity of hyaluronidase in mouse sperm.
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Objective To investigate the effects of subchronic arsenic exposure on caspase-3 expression in Leydig cells and serum testosterone level in rats. Methods Forty SD rats were divided into four groups based on body mass (160-220 g) by random number table, 10 in each group and treated with As2O3 through drinking water at the doses of 0.0 (distilled water), 2.0, 12.0 and 60.0 mg/L for 14 weeks. Blood sample from heart was collected, serum testosterone was measured by enzyme linked immunosorbent assay (ELISA); the testicular tissue of rats was taken, the expression of caspase-3 was assayed by immunohistochemical staining, and its average gray value was analyzed with image analysis software (Biomias). Results There were statistically significant differences of the serum testosterone levels between groups (F= 37.99, all P< 0.05). Compared with the control group [(3.082 3 ± 0.653 0) ng/L], serum testosterone levels in middle-dose and high-dose groups [(1.694 6 ± 0.255 4), (1.350 5 ± 0.281 9)ng/L] were significantly lower (all P< 0.05). The numbers of positive cells of caspase-3 were 8.10 ± 1.91, 9.80 ± 2.15,10.00 ± 1.83 and 10.90 ± 2.38 in each vision in the 4 groups, there were statistically significant differences between groups (F=3.17, all P<0.05), compared with the control group , the middle-dose and high-dose groups were significantly higher (all P<0.05);the average gray values of caspase-3 were 135.10 ± 6.89, 130.00 ± 3.30, 117.10 ± 4.28, and 113.10 ± 5.38, there were statistically significant differences between groups (F = 41.09, P < 0.05), compared with the control group, the middle-and high-dose groups were decreased (all P<0.05). Conclusions One possible mechanism of As2O3 on male germ cell toxicity may be through up-regulation of the expression of caspase-3 in Leydig cells of SD rats and initiating the pathway of the apoptosis signal, which can lead to increased apoptosis of Leydig cells, decreased concentrations of testosterone, and damaged reproductive function in rats.
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Objective To investigate the effects of sub-chronic arsenic poisoning on hyaluronidase activity of rat sperm.Methods Forty Sprague-Dawley (SD) rats in cleanliness grade,weighing 100-200 g,were randomly divided into four groups:control group,low-dose,middle-dose and high-dose arsenic exposure groups,10 in each group,and allotted to cages.Rats of arsenic groups were exposed to arsenic through drinking water (As2O3 was dissolved in distilled water).The concentrations of As2O3 in low-dose,middle-dose and high-dose groups were 25,50,and 100 mg/L,respectively.Body weight of each rat was weighed once a week.The rats in control group drank distilled water freely.All rats were fed for three months.After the experiment,all rats were killed and their sperms were collected.The positive rates and hyaluronidase activity intensities of the sperms in different groups were examined by the mature improved fixed-substrate film method.Results The positive rate of sperm hyaluronidase in control,low-dose,middle-dose and high-dose groups were (90.8 ± 13.2)％,(79.1 ± 15.6)％,(59.7 ± 20.3)％ and (33.8 ± 14.5)％,respectively.Compared with the control group,the positive rates of the middledose and the high-dose groups were significantly reduced(all P ＜ 0.05).The activity intensities of hyaluronidase in the sperms in control,low-dose,middle-dose and high-dose groups were (45.1 ± 24.0),(31.1 ± 21.3),(19.4 ± 15.3) and (9.8 ± 3.1)μm,respectively.Compared with the control group,the activity intensities of hyaluronidase of the middle-dose and the high-dose groups were significantly reduced(all P ＜ 0.05).Conclusion Sub-chronic arsenic poisoning reduces the activity intensities and positive rates of sperm hyaluronidase of rat,resulting in reduced sperm fertilizing capacity.
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Objective To evaluate the effects of chronic arsenic poisoning on serum inhibin-B (INH-B) and testosterone of male rats.Methods Forty Sprague-Dawley rats in cleanliness grad were randomly divided into four groups according to body weight:high-dose,middle-dose,low-dose arsenic exposure groups and control group,10 animals in each group.Rats in high-dose,middle-dose,and low-dose groups were administered with arsenic in concentrations of 10.0,5.0,2.5,0.0 mg/kg in distilled water,respectively.Rats in control group were fed with distilled water freely.After 9 months,serum INH-B and testosterone were tested using an ELISA Kit.Results Serum INH-B and testosterone levels in the above mentioned four groups,respectively,were (66.31 ± 37.21),(75.13 ± 29.88),(119.02 ± 36.10),(133.04 ± 38.67)ng/L and (1.292 ± 0.715),(1.757 ± 0.649),(2.359 ± 0.675),(2.817 ± 0.890)μg/L,and the differences between different groups were statistically significant (F=84.88,8.214,all P ＜ 0.05).Serum INH-B and testosterone levels in high-dose and middle-dose groups were significantly lower than that of control group(all P＜ 0.05),whereas no significant difference was found between low-dose group and control group (all P ＞ 0.05).Conclusion Chronic arsenic poisoning has led to decreased concentration of serum INH-B and testosterone in male rats.