Effects of Jinkui Shenqi and Wuzi Yanzong pill on sperm motility and sperm DNA fragmentation rate in asthenospermia

This study was designed to compare the effects of Jinkui Shenqi and Wuzi Yanzong pill on sperm motility and sperm DNA fragmentation rate in patients with asthenospermia. 130 cases were randomly divided into an observation and control group (n=65). The control group was treated with the Wuzi Yanzong pill while the observation group with the Jinkui Shenqi pill. The sperm motility parameters rate (PR), semen concentration, sperm motility, DFI and α-glucosidase, fructose, seminal plasma zinc (Zn), acid phosphatase (ACP) in seminal plasma biochemistry and other indexes of were observed. The biochemical indexes of seminal plasma of α-glucosidase, fructose, Zn, ACP in two groups were significantly (p<0.05) improved after treatment. Compared with the control group, the indexes of the observation group improved more obviously after treatment. Pearson correlation analysis of DFI and PR indexes in 130 patients before treatment showed that sperm DFI and PR percentage were negatively correlated in asthenospermia patients (r =-0.572, P<0.05). There was no significant difference in DFI, semen concentration, PR, and sperm motility between the two groups before treatment. The DFI, semen concentration, PR and sperm viability of the two groups showed a tendency to improve after treatment, and the effect of the observation group was less significant than that of the control group (p<0.05). Two groups of patients have completed treatment successfully, no adverse events occurred during treatment. Jinkui Shenqi pill can effectively treat asthenospermia, which can effectively improve the effect of sperm motility in patients. It has less adverse reactions, safe and reliable, and is worthy of promotion.

The combined effect of lifestyle intervention and antioxidant therapy on sperm DNA fragmentation and seminal oxidative stress in IVF patients: a pilot study

ABSTRACT Purpose: Sperm DNA fragmentation (SDF) and seminal oxidative stress are emerging measurable factors in male factor infertility, which interventions could potentially reduce. We evaluated (i) the impact of lifestyle changes combined with oral antioxidant intake on sperm DNA fragmentation index (DFI) and static oxidation-reduction potential (sORP), and (ii) the correlation between DFI and sORP. Materials and Methods: We conducted a prospective study involving 93 infertile males with a history of failed IVF/ICSI. Ten healthy male volunteers served as controls. Semen analysis was carried out according to 2010 WHO manual, whereas seminal sORP was measured using the MiOXSYS platform. SDF was assessed by sperm chromatin structure assay. Participants with DFI >15% underwent a three-month lifestyle intervention program, primarily based on diet and exercise, combined with oral antioxidant therapy using multivitamins, coenzyme Q10, omega-3, and oligo-elements. We assessed changes in semen parameters, DFI, and sORP, and compared DFI results to those of volunteers obtained two weeks apart. Spearman rank correlation tests were computed for sORP and DFI results. Results: Thirty-eight (40.8%) patients had DFI >15%, of whom 31 participated in the intervention program. A significant decrease in median DFI from 25.8% to 18.0% was seen after the intervention (P <0.0001). The mean DFI decrease was 7.2% (95% CI: 4.8-9.5%; P <0.0001), whereas it was 0.42% (95%CI; -4.8 to 5.6%) in volunteers (P <0.00001). No differences were observed in sperm parameters and sORP. Based on paired sORP and DFI data from 86 patients, no correlation was observed between sORP and DFI values (rho=0.03). Conclusion: A 3-month lifestyle intervention program combined with antioxidant therapy reduced DFI in infertile men with elevated SDF and a history of failed IVF/ICSI. A personalized lifestyle and antioxidant intervention could improve fertility of subfertile couples through a reduction in DFI, albeit controlled trials evaluating reproductive outcomes are needed before firm conclusions can be made. Trial registration number and date: clinicaltrials.gov NCT03898752, April 2, 2019.

Actualización sobre COVID-19 e histofisiología del sistema reproductor masculino / Update on COVID-19 and histophysiology of the male reproductive system

RESUMEN: La reciente pandemia de la COVID-19 ha sacudido a la sociedad teniendo una importante repercusión en el campo de la salud y de la investigación. Dada su relevancia, se han llevado a cabo estudios sobre los efectos del SARS-CoV-2 en la fisiología humana. En concreto, sobre la posible presencia y transmisión del virus a través del sistema reproductor masculino y su posible efecto en el éxito reproductivo. Conocer si la presencia del virus altera los órganos responsables del desarrollo y maduración de las células de la serie espermatogénica podría revelarnos su implicación en la calidad seminal. Por ello, nos planteamos esta revisión, con el fin de analizar las principales evidencias científicas sobre los efectos del SARS-CoV-2 en la histofisiología del sistema reproductor masculino y sobre la capacidad fecundante de los espermatozoides.

SUMMARY: The recent COVID-19 pandemic has shaken up society, having a significant impact on the field of health and research. Given its relevance, studies have been performed on the effects of SARS-CoV-2 on human physiology. In particular, the possible presence and transmission of the virus through the male reproductive system could affect reproductive success. Knowing if the presence of the virus disrupts the organs responsible for the development and maturation of the cell lines involved in spermatogenesis could reveal its implications in sperm quality. For that reason, we proposed this review, in order to analyze the main scientific evidence on the effects of SARS-CoV-2 on the histophysiology of the male reproductive system and sperm fertilizing capacity.

Sperm DNA damage compromises embryo development, but not oocyte fertilisation in pigs

BACKGROUND: The assessment of sperm DNA integrity has been proposed as a complementary test to conventional mammalian semen analysis. In this sense, single-strand (SSB) and double-strand (DSB) DNA breaks, the two types of sperm DNA fragmentation (SDF), have been reported to have different aetiologies and to be associated to different fertility outcomes in bovine and humans. Considering that no studies in porcine have addressed how SDF may affect sperm quality and fertility outcomes, the present work aimed to determine the impact of global DNA damage, SSB and DSB on sperm quality and in vitro fertilising ability. To this end, 24 ejaculates (one per boar) were split into three aliquots: the first was used to assess sperm quality parameters through a computer-assisted sperm analysis (CASA) system and flow cytometry; the second was used to perform in vitro fertilisation, and the third, to evaluate sperm DNA integrity using alkaline and neutral Comet assays. RESULTS: The results showed that global DNA damage negatively correlates (P 0.05). CONCLUSION: Considering all these findings, this work sets a useful model to study how SDF negatively influences fertility.

Influence of the sperm DNA fragmentation index on the outcome of rescue ICSI and the clinical value of rescue ICSI / 中南大学学报(医学版)

OBJECTIVES@#As a remedy for the failure of in vitro fertilization (IVF), rescue intracytoplasmic sperm injection (R-ICSI) has been widely carried out, but it has failed to significantly improve the fertilization rate and clinical pregnancy rate. Sperm DNA fragmentation index (DFI) was highly correlated with pregnancy outcome of artificial assisted reproduction. This study aims to investigate the effect of the sperm DFI on the outcome of R-ICSI and the clinical value of R-ICSI.@*METHODS@#This retrospective analysis was conducted among 140 infertile couples receiving R-ICSI in from January 2014 to December 2019. The subjects were assigned into a total fertilization failure (TFF)+low DFI group (R-ICSI after TFF and DFI<30%) (n=63), a TFF+high DFI group (R-ICSI after TFF and DFI≥30%) (n=16), a partial fertilization failure (PFF)+low DFI group (R-ICSI after PFF and DFI<30%) (n=52), a PFF+high DFI group (R-ICSI after PFF and DFI≥30%) (n=9). All transferred embryos were come from R-ICSI. The general clinical data [infertility duration, male age, female age, basal serum level of follicle stimulating hormone (FSH), basal serum level of luteinizing hormone (LH), antral follicle count, endometrial thickness of human chorionic gonadotropin (HCG) day, and eggs] and R-ICSI cycle outcomes (fertilization rate, normal fertilization rate, cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate and live birth rate) were analyzed. In addition, the effect of R-ICSI on the fertilization outcome of conventional IVF total fertilization failure and partial fertilization failure was explored.@*RESULTS@#There was no significant difference in the general clinical data and R-ICSI cycle outcome between the TFF+low DFI group and the TFF+high DFI group (all P>0.05). There was no significant difference in the general clinical data between the PFF+low DFI group and the PFF+high DFI group (all P>0.05). The fertilization rate and normal fertilization rate in the PFF+low DFI group were significantly higher than those in the PFF+high DFI group (85.40% vs 72.41%, 71.90% vs 58.62%, respectively; both P<0.05). However, there was no significant difference in cleavage rate, good embryo rate, implantation rate, clinical pregnancy rate, and live birth rate between the 2 groups (all P>0.05). The R-ICSI cycle of TFF: A total of 79 fresh cycles, 57 fresh transplant cycles, a total of 761 unfertilized oocytes, and 584 M II oocytes were treated with R-ICSI, the fertilization rate was 83.22%, the normal fertilization rate was 75.51%, the cleavage rate was 98.15%, the good embryo rate was 40.74%, the implantation rate was 30.56%, and the clinical pregnancy rate was 43.86%; 29 live births were obtained. The R-ICSI cycle of PFF: A total of 61 fresh cycles, 31 fresh transplant cycles, a total of 721 unfertilized oocytes, and 546 M II oocytes were treated with R-ICSI; the fertilization rate was 83.33%, the normal fertilization rate was 69.78%, the cleavage rate was 97.36%, the good embryo rate was 44.39%, the implantation rate was 25.42%, and the clinical pregnancy rate was 45.16%; 12 live births were obtained.@*CONCLUSIONS@#In the case of partial fertilization failure of IVF, the sperm DFI affects the fertilization rate and normal fertilization rate of R-ICSI; whether it is a TFF of IVF or PFF of IVF, ICSI can be used as an effective remedy way.

The effect of age and abstinence time on semen quality: a retrospective study / 亚洲男科学杂志(英文版)

This study analyzed the effects of male age and abstinence time on semen quality and explored the best abstinence time for Chinese males among different age groups. Semen parameters, including sperm kinetics, morphology, and DNA fragmentation index (DFI), were reviewed from 2952 men. Samples were divided into six age groups (≤25 years, 26-30 years, 31-35 years, 36-40 years, 41-45 years, and >45 years) and were divided into six groups according to different abstinence time (2 days, 3 days, 4 days, 5 days, 6 days, and 7 days). The differences in semen quality between the groups were compared, and the effect of age and abstinence time on semen quality was analyzed. Significant differences were observed in semen volume, progressive motility (PR), and DFI among the age groups (all P < 0.05), and no significant differences were observed in sperm morphological parameters (all P > 0.05). There were significant differences in semen volume, PR, and DFI among different abstinence time groups (all P < 0.05) and no significant differences in sperm morphological parameters (all P > 0.05). Pearson analysis showed that male age and abstinence time were both significantly correlated with sperm kinetics and DFI (both P < 0.05), while no significant correlation was found with sperm morphological parameters (all P > 0.05). The box plots and histograms of men's age, abstinence time, and semen quality show that most semen quality parameters differ significantly between the 2 days and 7 days abstinence groups and other groups at different ages. Except for the sperm morphology parameters, sperm kinetic parameters and sperm DFI are linearly related to male age and abstinence time.

Local renin angiotensin system and sperm DNA fragmentation / 亚洲男科学杂志(英文版)

The renin angiotensin system (RAS) appears to influence male fertility at multiple levels. In this work, we analyzed the relationship between the RAS and DNA integrity. Fifty male volunteers were divided into two groups (25 each): control (DNA fragmentation ≤20%) and pathological (DNA fragmentation >20%) cases. Activities of five peptidases controlling RAS were measured fluorometrically: prolyl endopeptidase (which converts angiotensin [A] I and A II to A 1-7), neutral endopeptidase (NEP/CD10: A I to A 1-7), aminopeptidase N (APN/CD13: A III to A IV), aminopeptidase A (A II to A III) and aminopeptidase B (A III to A IV). Angiotensin-converting enzyme (A I to A II), APN/CD13 and NEP/CD10 were also assessed by semiquantitative cytometry and quantitative flow cytometry assays, as were the receptors of all RAS components: A II receptor type 1 (AT1R), A II receptor type 2 (AT2R), A IV receptor (AT4R or insulin-regulated aminopeptidase [IRAP]), (pro)renin receptor (PRR) and A 1-7 receptor or Mas receptor (MasR) None of the enzymes that regulate levels of RAS components, except for APN/CD13 (decrease in fragmented cells), showed significant differences between both groups. Micrographs of RAS receptors revealed no significant differences in immunolabeling patterns between normozoospermic and fragmented cells. Labeling of AT1R (94.3% normozoospermic vs 84.1% fragmented), AT4R (96.2% vs 95.3%) and MasR (97.4% vs 87.2%) was similar between the groups. AT2R (87.4% normozoospermic vs 63.1% fragmented) and PRR (96.4% vs 48.2%) were higher in non-fragmented spermatozoa. These findings suggest that fragmented DNA spermatozoa have a lower capacity to respond to bioactive RAS peptides.

Evaluation of the sperm DNA fragmentation index in infertile Japanese men by in-house flow cytometric analysis / 亚洲男科学杂志(英文版)

Semen analysis has long been used to evaluate male fertility. Recently, several sperm function tests have been developed. Of those, the sperm DNA fragmentation index (DFI), which describes the status of the sperm DNA, is thought to be a suitable parameter for evaluating male fertility. However, there have been no large-scale studies on the sperm DFI of Japanese men. Therefore, we investigated the feasibility of using an in-house flow cytometry-based sperm DFI analysis based on the sperm DNA fragmentation test of sperm chromatin structure assay (SCSA) to assess male fertility in Japan. This study enrolled 743 infertile and 20 fertile Japanese men. To evaluate reproducibility, inter- and intraobserver precision was analyzed. A receiver operating characteristic curve analysis was used to set a cutoff value for the sperm DFI to identify men who could father children by timed intercourse or intrauterine insemination. The variability of the sperm DFI among fertile volunteers was determined. The relationship between semen parameters and the sperm DFI was assessed by Spearman's rho test. A precision analysis revealed good reproducibility of the sperm DFI. The cutoff value of sperm DNA fragmentation in infertile men was 24.0%. Semen volume had no relationship with the sperm DFI. Sperm concentration, sperm motility, total motile sperm count, and percentage of normal-shaped sperm were significantly and negatively correlated with the sperm DFI. The median sperm DFI was smaller in fertile volunteers (7.7%) than that in infertile men (19.4%). Sperm DNA fragmentation analysis can be used to assess sperm functions that cannot be evaluated by ordinary semen analysis.

Random sperm DNA fragmentation index is not associated with clinical outcomes in day-3 frozen embryo transfer / 亚洲男科学杂志(英文版)

Damage to sperm DNA was proposed to play an important role in embryonic development. Previous studies focused on outcomes after fresh embryo transfer, whereas this study investigated the influence of sperm DNA fragmentation index (DFI) on laboratory and clinical outcomes after frozen embryo transfer (FET). This retrospective study examined 381 couples using cleavage-stage FET. Sperm used for intracytoplasmic sperm injection (ICSI) or in vitro fertilization (IVF) underwent density gradient centrifugation and swim up processing. Sperm DFI had a negative correlation with sperm motility (r = -0.640, P < 0.01), sperm concentration (r = -0.289, P < 0.01), and fertilization rate of IVF cycles (r = -0.247, P < 0.01). Sperm DFI examined before and after density gradient centrifugation/swim up processing was markedly decreased after processing (17.1% vs 2.4%, P < 0.01; 65 randomly picked couples). Sperm progressive motility was significantly reduced in high DFI group compared with low DFI group for both IVF and ICSI (IVF: 46.9% ± 12.4% vs 38.5% ± 12.6%, respectively; ICSI: 37.6% ± 14.1% vs 22.3% ± 17.8%, respectively; both P < 0.01). The fertilization rate was significantly lower in high ( ≥25%) DFI group compared with low (<25%) DFI group using IVF (73.3% ± 23.9% vs 53.2% ± 33.6%, respectively; P < 0.01) but was equivalent in high and low DFI groups using ICSI. Embryonic development and clinical outcomes after FET were equivalent for low and high DFI groups using ICSI or IVF. In this study, sperm DFI did not provide sufficient information regarding embryo development or clinical outcomes for infertile couples using FET.

Oxidative origin of sperm DNA fragmentation in the adult varicocele

ABSTRACT Purpose: Sperm DNA fragmentation is a major cellular mechanism underlying varicocele-related male infertility. However, the type of DNA fragmentation - whether oxidative or of another nature - remains unknown. Thus, the aim of this study was to evaluate single- and double-stranded sperm DNA fragmentation, and oxidative-induced sperm DNA damage in men with varicocele. Materials and Methods: A cross-sectional study was performed, including 94 normozoospermic adults, of which 39 men without varicocele (controls) and 55 men with varicocele grades II or III, uni- or bilaterally. All men collected semen by masturbation. After semen analysis, the remaining volume was used for evaluation of three types of sperm DNA damage: (i) total DNA fragmentation, using an alkaline comet assay, (ii) double-stranded DNA fragmentation, using a neutral comet assay, and (iii) oxidative DNA damage, using an alkaline comet assay associated with the DNA glycosylase formamidopyrimidine enzyme. In each assay, percentage of sperm with any degree of DNA fragmentation, and with high DNA fragmentation were compared between the groups using an unpaired Student's t test or a Mann-Whitney test. Results: The varicocele group presented a higher rate of sperm with fragmented DNA (both any and high DNA fragmentation), considering single-stranded DNA fragmentation, double-stranded DNA fragmentation, or a combination of both, as well as oxidative- induced DNA fragmentation. Conclusions: Patients with varicocele have an increase in sperm DNA fragmentation levels, particularly in oxidative stress-induced sperm DNA damage.

Effects of sperm DNA fragmentation index on semen parameters and pregnancy outcomes in intrauterine insemination / 中华男科学杂志

Objective@#To analyze the correlation of the sperm DNA fragmentation index (DFI) level with semen parameters and pregnancy outcomes of artificial insemination of the husband (AIH) in the cycle of intrauterine insemination (IUI).@*METHODS@#We collected the clinical data on 777 cases of IUI, including female clinical indicators, male semen parameters, sperm DFI and pregnancy outcomes. According to the DFI level, we divided the patients into three groups: DFI < 15%, 15% ≤ DFI < 30% and DFI ≥ 30%.@*RESULTS@#The sperm DFI level was significantly elevated with the increased age of the males (P = 0.002) and closely related to the total number of motile sperm (P = 0.002) and total sperm motility (P = 0.000) before treatment, as well as to sperm concentration (P = 0.000), total sperm motility (P = 0.001) and total number of progressively motile sperm (P = 0.000) after density gradient centrifugation. The rate of clinical pregnancy was decreased in the DFI ≥ 30% group. There were no statistically significant differences between sperm DFI and the rates of clinical pregnancy and abortion.@*CONCLUSIONS@#Male age significantly affects the sperm DFI level. Sperm DFI is closely related to sperm motility and total number of progressively motile sperm, but not to the rates of clinical pregnancy and abortion in patients undergoing IUI. IUI can be used as an effective method of assisted reproduction for male infertility./.

Assessment of heavy metals and its impact on DNA fragmentation in different fish species / Avaliação de metais pesados e seu impacto na fragmentação de DNA em diferentes espécies de peixe

Abstract This study was conducted to assess water pollution by examining DNA fragmentation in selected fish organs (kidney, liver, gills, and muscle tissue) from Wallago attu, Sperata sarwari, Vulgaris vulgaris, and Labeo rohita collected from a known polluted section of the Chenab River, Pakistan, and from a control site. The fish were caught using a gill net and were assigned to three different weight groups (W1, W2, and W3) to study the degree of variation in DNA fragmentation in relation to body weight. In fish from the polluted site, DNA fragmentation was higher in kidney, liver, gills, and muscles, compared to the control. No significant DNA fragmentation was observed in fish collected from the control site. Highly significant (P < 0.01) relationship between body weight and DNA fragmentation was found in the organs of fish procured at the contaminated site. DNA fragmentation in body organs was found to be affected by the concentrations of lead, copper, nickel, and cadmium in W. attu, S. sarwari, L. rohita, and V. vulgarus harvested from Chenab River. DNA fragmentation in different freshwater fish species is therefore a reliable biomarker of water pollution.

Resumo Este estudo foi conduzido para avaliar a poluição da água examinando a fragmentação do DNA em órgãos de peixes selecionados (rim, fígado, brânquias e tecido muscular) de Wallago attu, Sperata sarwari, Vulgaris vulgaris e Labeo rohita coletados de uma conhecida área poluída do rio Chenab, Paquistão e de um local de controle. Os peixes foram capturados usando uma rede branquial e foram divididos em três grupos de pesos diferentes (W1, W2 e W3) para estudar o grau de variação na fragmentação do DNA em relação ao peso corporal. Nos peixes do local poluído, a fragmentação do DNA foi maior nos rins, fígado, brânquias e músculos, em comparação ao controle. Não foi observada fragmentação significativa do DNA em peixes coletados no local de controle. Relação altamente significativa (P <0,01) entre o peso corporal e a fragmentação do DNA foi encontrada nos órgãos dos peixes adquiridos no local contaminado. Verificou-se que a fragmentação do DNA nos órgãos do corpo é afetada pelas concentrações de chumbo, cobre, níquel e cádmio em W. attu, S. sarwari, L. rohita e V. vulgarus colhidos no rio Chenab. A fragmentação do DNA em diferentes espécies de peixes de água doce é, portanto, um biomarcador confiável da poluição da água.

Distribution of TCM syndrome types and their correlation semen parameters in infertility patients with varicocele / 中华男科学杂志

Objective@#To explore the distribution of Traditional Chinese Medicine (TCM) syndrome types and their relationship with semen parameters in infertility male patients with varicocele (VC).@*METHODS@#Using Questionnaire on Clinical Symptoms of Varicocele-Caused Male Infertility, we made an investigation among 147 infertility male patients with VC, determined the types of their TCM syndromes, obtained their semen parameters, and analyzed the distribution of the TCM syndrome types and their correlation with semen parameters.@*RESULTS@#Of the TCM syndrome types identified, kidney deficiency and stagnated heat constituted the largest proportion (34.7%), and the mixed type accounted for a significantly higher percentage than the simple type (P < 0.05). The patients with kidney deficiency and stagnated heat, compared with those with other syndrome types, had a dramatically lower sperm concentration (［21.62 ± 9.25］ vs ［28.88 ± 12.92］ ×10⁶/ml, P < 0.01), but a higher percentage of morphologically abnormal sperm (［98.33 ± 0.15］% vs ［96.27 ± 0.18］%, P < 0.05) and DNA fragmentation index (［19.72 ± 3.17］% vs ［10.96 ± 3.82］%, P < 0.01). No statistically significant differences were observed in the percentage of progressively motile sperm among different TCM syndrome types.@*CONCLUSIONS@#Kidney deficiency and stagnated heat is a main TCM syndrome type in infertility male patients with varicocele and correlated with sperm concentration, the percentage of morphologically abnormal sperm and DNA fragmentation index.

Correlation of Mycoplasma genitalium infection with semen parameters and sperm DNA integrity in male infertility patients / 中华男科学杂志

Objective@#To analyze the relationship of Mycoplasma genitalium (MG) infection with routine semen parameters and sperm DNA integrity in male infertility patients.@*METHODS@#Totally, 114 semen samples, 34 MG-positive and 80 MG-negative, were collected from male infertility patients and subjected to routine semen analysis with the computer-assisted sperm analysis system, Papanicolaou staining for observation of sperm morphology, and sperm chromatin diffusion (SCD) test for detection of sperm DNA integrity. Semen parameters and DNA integrity were compared between the MG-positive and MG-negative groups with SPSS 21.0 statistical software and the relationship between the semen parameters and DNA integrity analyzed by Pearson correlation analysis.@*RESULTS@#The MG-positive samples, compared with the MG-negative ones, showed significantly decreased semen volume (［2.87 ± 0.37］ vs ［3.86 ± 0.43］ ml, P 0.05).@*CONCLUSIONS@#MG infection may be an important factor affecting sperm quality in male infertility patients. Active prevention and treatment of MG infection can help prevent male infertility.

Fragmentación del ADN espermático en parejas tratadas por fertilización in vitro / Sperm DNA fragmentation in couplestreated by in vitro fertilization

RESUMEN Introducción: Se estudian los factores que afectan el ADN espermático en la actualidad y en qué medida influyen en la aplicación de las técnicas de reproducción asistida de alta tecnología. La fertilización in vitro es un tema de interés que ha motivado a muchos investigadores. Objetivo: Determinar la relación que existe entre el índice de fragmentación del ADN de la cromatina espermática y los resultados de la técnica de fertilización in vitro en parejas infértiles. Métodos: Se realizó un estudio descriptivo transversal en 107 parejas infértiles remitidas del Programa Nacional de Atención a la pareja infértil, donde se precisó la asociación entre el potencial fertilizante y las variables de resultados: tasa de fertilización, calidad embrionaria y embarazo. Se estudiaron, además, algunos factores que pueden influir en el grado de fragmentación del ADN. Resultados: El potencial fertilizante no se relacionó con la tasa de fertilización, ni con el embarazo clínico, pero sí con la calidad embrionaria (P= 0.036). La edad paterna resultó ser estadísticamente significativa en relación con el potencial fertilizante (P= 0.032). La presencia de varicocele se asoció con el bajo potencial fertilizante (OR= 5,27; IC 95 por ciento [1.34 - 24.09]). Conclusiones: El índice de fragmentación del ADN de la cromatina espermática afecta negativamente la calidad de los embriones a transferir. La edad y el varicocele se relacionan positivamente con el grado de fragmentación del ADN espermático. El consumo de alcohol, el hábito de fumar y la exposición a agentes físicos y químicos no se relacionaron con el índice de fragmentación del ADN espermático en este estudio(AU)

ABSTRACT Introduction: The factors that affect the spermatic DNA are being studied at present and in what extent they affect the implementation of high technology assisted reproduction techniques. In vitro fertilization is a topic of interest that has motivated many researchers. Objective: To determine the relationship between the rate of DNA fragmentation of sperm chromatin and the results of in vitro fertilization technique in infertile couples. Methods: A descriptive cross-sectional study was conducted in 107 infertile couples referred to the National Program of Attention to the Infertile Couple, where it was specified the association between the potential fertilizer and outcome variables: rate of fertilization, embryo quality and pregnancy. In addition, there were studied some factors that may influence the degree of DNA fragmentation. Results: The fertilizing potential was not related to the rate of fertilization, or with the clinical pregnancy, but it did with the embryo quality (p= 0.036). The paternal age proved to be statistically significant in relation to the fertilizing potential (p= 0.032). The presence of varicocele was associated with the low fertilizing potential (OR = 5.27; CI 95 percent [1.34 - 24.09]). Conclusions: The rate of DNA fragmentation of sperm chromatin negatively affects the quality of the embryos to be transferred. The age and varicocele are positively related with the degree of sperm DNA fragmentation. The consumption of alcohol, the smoking habit and the exposure to chemical and physical agents were not associated with the rate of sperm DNA fragmentation in this study(AU)

Shikonin Exerts Cytotoxic Effects in Human Colon Cancers by Inducing Apoptotic Cell Death via the Endoplasmic Reticulum and Mitochondria-Mediated Pathways

The apoptotic effects of shikonin (5,8-dihydroxy-2-[(1R)-1-hydroxy-4-methylpent-3-enyl]naphthalene-1,4-dione) on the human colon cancer cell line SNU-407 were investigated in this study. Shikonin showed dose-dependent cytotoxic activity against SNU-407 cells, with an estimated IC50 value of 3 µM after 48 h of treatment. Shikonin induced apoptosis, as evidenced by apoptotic body formation, sub-G1 phase cells, and DNA fragmentation. Shikonin induced apoptotic cell death by activating mitogen-activated protein kinase family members, and the apoptotic process was mediated by the activation of endoplasmic reticulum (ER) stress, leading to activation of the PERK/elF2α/CHOP apoptotic pathway, and mitochondrial Ca2+ accumulation. Shikonin increased mitochondrial membrane depolarization and altered the levels of apoptosis-related proteins, with a decrease in B cell lymphoma (Bcl)-2 and an increase in Bcl-2-associated X protein, and subsequently, increased expression of cleaved forms of caspase-9 and -3. Taken together, we suggest that these mechanisms, including MAPK signaling and the ER-and mitochondria-mediated pathways, may underlie shikonin-induced apoptosis related to its anticancer effect.

Antioxidant activity of ethanol extract of Lycium barbarum's leaf with removal of chlorophyll / 한국영양학회지

PURPOSE: The aim of this study was to estimate the antioxidant activities of 50%, 70%, and 100% ethanol extracts of Lycium barbarum leaf and chlorophyll removal extract. METHODS: The antioxidant activities were estimated by measuring total polyphenol content and by assays of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfate) (ABTS) radical scavenging activities and ferric reducing antioxidant power (FRAP). In addition, reactive oxygen species (ROS) production, DNA fragmentation, and antioxidant enzyme (superoxide dismutase and catalase) activities of the extracts were measured in hydrogen peroxide (H2O2)-stressed HepG2 cells. RESULTS: The total polyphenol content, DPPH and ABTS radical scavenging activities, and FRAP value of the extracts increased in an ethanol concentration-dependent manner. The antioxidant activities of the chlorophyll-removal extracts were much higher than those of the chlorophyll-containing extracts. Cytotoxicity was not observed in HepG2 cells with extracts up to 1,000 µg/mL. All extracts inhibited ROS production in a concentration-dependent manner from 31.3 µg/mL and inhibited DNA damage at 250 µg/mL. The SOD and catalase activities of cell lines treated with the extracts and H2O2 were similar to those of normal cells, indicating a strong protective effect. CONCLUSION: Lycium barbarum leaf extracts had high antioxidant activities and protected H2O2-stressed HepG2 cells. Since the chlorophyll-removal extract exhibited higher antioxidant activities than the chlorophyll-containing ones and the cytoprotective effect was similar, chlorophyll removal extract of Lycium barbarum leaf could be developed as ingredients of functional food and cosmetics.

Los parámetros espermáticos funcionales como factor pronóstico en ICSI: Experiencia de la ciudad de Medellín / Functional Seminal Parameters as a Prognostic Factor in ICSI: Medellín City Experience

Introducción La infertilidad por factor masculino afecta al 30% de las parejas infértiles y la evaluación seminal es crítica en las determinaciones que conllevan a un posible tratamiento con el fin de tener un resultado exitoso. Objetivo El objetivo de este estudio fue evaluar la relación que existe entre los parámetros seminales convencionales y funcionales, con las tasas de fecundación, desarrollo embrionario y embarazo obtenidas después de inyección intracitoplasmática de espermatozoides (ICSI). Métodos 36 muestras seminales de parejas que se sometieron a ICSI (18 usando oocitos propios y 18 de donante), fueron evaluadas de manera convencional, posteriormente se seleccionaron los espermatozoides, se realizó ICSI y una alícuota se utilizó para cuantificar las siguientes pruebas funcionales: potencial de membrana mitocondrial, integridad de la membrana, detección de especies reactivas de oxígeno e índice de fragmentación del ADN. Resultados No se encontraron diferencias significativas en cuanto a los parámetros convencionales y funcionales en los dos grupos, como tampoco se encontró una relación significativa entre los parámetros evaluados y los resultados de ICSI. Sólo se observó que la tasa de embarazo fue mayor en el grupo de oocitos donados (p < 0,0001). Conclusiones Los datos obtenidos en este estudio sugieren que no existe correlación entre los parámetros evaluados y los resultados de ICSI. Eso se debe probablemente, a que la selección de los espermatozoides tanto por gradientes de densidad como la posterior selección durante el procedimiento del ICSI, tiene un bajo poder predictivo sumado a la capacidad que tiene el oocito de reparar los daños presentes en el espermatozoide.

Background Male infertility affects 30% of infertile couples and seminal evaluation is critical in the determinations that lead to a possible treatment in order to have a successful outcome. Objective The objective of this study was to evaluate the relationship between conventional and functional seminal parameters, with the rates of fertilization, embryo development and pregnancy obtained after ICSI. Methods 36 semen samples of couples that underwent ICSI (18 using own oocytes and 18 from donors) were conventionally evaluated, spermatozoa were subsequently selected, ICSI was performed and an aliquot was used to quantify the following functional tests: mitochondrial membrane potential, membrane integrity, reactive oxygen species detection and DNA fragmentation index. Results There were no significant differences in the conventional and functional parameters in the two groups, nor was there a significant relationship between the parameters evaluated and the ICSI results. It was only observed that the pregnancy rate was higher in the group of donated oocytes (p < 0.0001). Conclusions The data obtained in this study suggest that there is no correlation between the parameters evaluated and the ICSI outcome. This is probably because the selection of spermatozoa by density gradients in addition to the subsequent selection during ICSI has a low predictive power and also the ability of the oocyte to repair the damage present in the spermatozoa.