Blastulation rate using fresh versus devitrified oocytes using homologous and heterologous sperm in an assisted reproduction clinic in a middle-income country

Background: Assisted reproduction has revolutionized the field of reproductive medicine, giving hope to many couples who face difficulties conceiving naturally. In this case, fresh oocytes and devitrified oocytes play a fundamental role in fertility treatments.Methods: Descriptive, retrospective and observational study in which the records of the HISPAREP reproduction clinic of the Spanish hospital were analyzed in the period from 2020 to 2022.Results: A total of 136 blastocysts that reached day 5 were studied compared to 8 embryos that were blocked before reaching blastulation. Of this, 76 oocytes were fresh, of which 97.4% reached day 5 (74) and the remaining two were blocked. Of the vitrified ones, a total of 68 were devitrified and of these 91.2% reached D5. Of the correctly fertilized oocytes, 96.5% (136) reached blastocysts compared to 5 that were blocked and 3 that were blocked from those that were not fertilized. Regarding the fertilization rate of fresh oocytes, a total of 76 was obtained, which is equivalent to 100%, and of the devitrified oocytes, 65 were fertilized (95.6%) and 3 were not fertilized (4.4%). With a variation of 6.2% in the blastocyst rate, between fresh and devitrified oocytes.Conclusions: There is no significant difference in the reproductive results of fresh and vitrified oocytes. It is necessary to try to carry out more studies with a larger sample size to obtain more conclusive results. The reproductive outcomes of fresh oocytes and oocytes devitrified using fresh sperm and frozen sperm are similar. With this, fresh oocytes have a slightly higher probability of implantation.

Relation between serum hormones and semen parameters in sub-fertile males: is 17-hydroxyprogesterone really a game changer?

Background: Semen analysis and serum hormone assessment remains an integral part of assessment of infertile males. 17-OHP has recently been demonstrated as a very good marker of intratesticular testosterone environment. Our study was designed to find relationship between individual serum hormone levels and semen parameters in sub fertile men, and to find whether 17-OHP fares better or worse in predicting baseline semen parameters compared to other routinely tested hormones.Methods: A retrospective analytical study was conducted on 74 patients, after matching inclusion and exclusion criteria, from July 2022 to December 2023. All included patients were investigated with Semen analysis and Serum biomarker levels (FSH, total testosterone, estradiol, total testosterone/estradiol (T/E2) ratio, and 17-hydroxyprogesterone).Results: Only 10 patients (13.51%) had normal semen analysis, while rest 64 (86.49%) had abnormality in at least 1 semen parameter. Serum testosterone and T/E2 ratio had significant difference (p<0.05) between the two groups. Comparing individual semen parameters against all hormones, FSH and total testosterone had significant association with sperm count, concentration and total motility. Total testosterone also had a significant relation with progressive motility, morphology and semen volume (p<0.05). T/E2 ratio had significant association with Sperm count, motility and morphology, and semen volume (p<0.05). But, 17-OHP and estradiol were not found to have any significant association with any baseline semen parameters in our study (p>0.05). But, serum 17OHP was found to be significantly associated with sexual dysfunction in males (p<0.05).Conclusions: Our study inferred that serum FSH, testosterone level and T/E2 ratio can be used to predict baseline semen parameters, but 17OHP did not have any association with baseline semen parameters. However, serum 17OHP can serve as a novel marker for male sexual dysfunction.

El laboratorio de fertilidad masculina, su evolución en los últimos 20 años / The male fertility laboratory, its evolution over the last 20 years / O laboratório de fertilidade masculina, sua evolução ao longo dos últimos 20 anos

Resumen La mayor accesibilidad a los tratamientos de reproducción asistida (RA) y los avances de la criobiología produjeron cambios en los laboratorios de andrología. El objetivo de este trabajo fue analizar la demanda y evolución de las variables seminales en las últimas dos décadas, caracterizar el laboratorio andrológico actual, evaluar el impacto de la incorporación del aseguramiento de la calidad y la inclusión de los sistemas computarizados (CASA). Se utilizaron datos de las medias mensuales del control de calidad interno (n=22 528) y encuestas a profesionales de laboratorios andrológicos (n=65) y a médicos especialistas en fertilidad (n=33). La demanda global se redujo significativamente con el aumento de las solicitudes de primera vez. El volumen y recuento, variables dependientes de andrógenos, disminuyeron con los años. El criterio estricto en morfología disminuyó el porcentaje de normales; la mitad de los médicos encuestados recibieron resultados entre 0 y 10% y el 40% consideró que ponía en riesgo el valor clínico de la variable. El sistema CASA permitió objetivar la cinética espermática e incrementar el porcentaje de progresivos rápidos, pero pocos laboratorios lo incorporaron. El 66% de los médicos resuelven el factor andrológico severo por tratamientos clínicos y el 95% utiliza técnicas de RA. El análisis de semen es ejecutado fundamentalmente por bioquímicos especializados, con baja adhesión a la automatización y acreditación del laboratorio, pero con participación en programas de evaluación externa de calidad. La demanda disminuyó como consecuencia del aumento del tratamiento por RA. La reducción del porcentaje de formas normales compromete su utilidad clínica.

Abstract Increasing availability to assisted reproduction (AR) treatments in Argentina and advances in cryobiology resulted in changes in andrology laboratories. The aim of this study was to evaluate the demand and evolution of seminal variables in the last two decades, characterise the current andrology laboratory, evaluate the impact of the incorporation of quality assurance and the introduction of computer assisted semen analysis (CASA). Data were taken from internal quality control (IQC) monthly means (n=22 528) and professionals in charge of laboratories (n=65) and fertility physicians' (n=33) surveys. Overall demand decreased significantly while first-time orders increased. Sperm volume and sperm count -androgen dependent parameters- decreased over the years. Strict morphology criteria reduced the percentage of normal results; half of the physicians received results between 0 and 10% and 40% considered that it compromised the clinical value of the variable. The CASA system made it possible to objectify sperm kinetic, increasing the percentage of fast progressives, but few laboratories have incorporated it. Sixty-six percent of physicians resolve severe andrological factor by clinical treatments and 95% use AR techniques in those cases. Semen analysis is mainly performed by specialised biochemists, with low adherence to laboratory automatisation and accreditation, but with participation in external quality assessment programmes. The demand decreased because of the increase in AR treatment. The lower percentage of normal forms compromises their clinical utility.

Resumo O aumento do acesso aos tratamentos de reprodução assistida (RA) e os avanços na criobiologia levaram a mudanças nos laboratórios de andrologia. O objetivo deste trabalho foi analisar a demanda e a evolução das variáveis de sêmen nas últimas duas décadas, caracterizar o laboratório de andrologia atual, avaliar o impacto da incorporação da garantia da qualidade e a inclusão dos sistemas computadorizados (CASA). Foram utilizados dados das médias mensais do controle de qualidade interno (n= 22 528) e pesquisas a profissionais de laboratórios andrológicos e a médicos especialistas em fertilidade (n=33). A demanda geral diminuiu significativamente com o aumento das solicitações de primeira vez. O volume e a contagem de esperma, parâmetros dependentes de andrógenos, diminuíram ao longo dos anos. O critério morfológico rigoroso diminuiu a porcentagem de normais; metade dos médicos entrevistados recebeu resultados entre 0 e 10% e 40% considerou que isso comprometía o valor clínico do parâmetro. O sistema CASA, permitiu objetivar a cinética espermática e aumentar o percentual de progressões rápidas, mas poucos laboratórios o incorporaram. 66% dos médicos resolvem o fator andrológico grave por tratamentos clínicos e 95% utilizam técnicas de RA nesses casos. A análise do sêmen é realizada principalmente por bioquímicos especializados, com baixa aderência à automação e acreditação laboratorial, mas com participação em programas de avalação externa de qualidade. A demanda diminuiu como consequência do aumento do tratamento por RA. A diminuição em percentagem de formas normais compromete sua utilidade clínica.

Correlation of lipid profile of infertile men with abnormal semen parameters

Background: Male’s inability to impregnate a fertile female is referred as male factor infertility. It accounts for 40-50% of infertile couples and affects almost 7% of all men. Male comorbidities and conditions, which negatively affect men’s health, have been repeatedly associated with impaired reproductive functioning. Cholesterol is the main substrate for steroid synthesis and it plays crucial role in formation of sperm plasma membrane and thus in spermatogenesis, highlighting the role of serum lipids in male fecundity. Purpose of the study is to evaluate correlation lipid profile of male partners with abnormal semen parameters in infertile couples.Methods: A cross sectional study was carried out in the infertility clinic of a tertiary care hospital for over 18 months and 151 infertile men with abnormal semen parameters (as per WHO 2010 criteria) were enrolled. Evaluation of lipid profile was done and its relationship with abnormal semen parameters was analyzed.Results: Significant positive correlation was seen between LDL and Triglyceride with sperm concentration and total sperm count. However, significant negative correlation was seen between Triglyceride and sperm motility and TMSC. Total and progressive motility was found to be significantly associated with total cholesterol (p<0.05). Infertile men having higher total cholesterol were found to have better total and progressive motility compared to men with less total cholesterol.Conclusions: Our findings demonstrated that lipid profile has significant correlation with semen parameters, specifically sperm concentration, count and motility.

Low-dose prednisone is an effective rescue for deteriorating semen parameters following vasovasostomy

ABSTRACT Objective: This retrospective study aimed to evaluate the effectiveness of low-dose prednisone as a rescue therapy for patients with deteriorating semen parameters following vasovasostomy. Materials and Methods: Electronic medical records were queried at the University of Miami with documented CPT code 55400 (Bilateral Vasovasostomy) between January 2016 and April 2023. Records were then reviewed to identify patients who demonstrated ≥50% decrease in semen parameters, specifically sperm concentration, motility and total motile sperm count. Patients who were treated with 6 weeks of low-dose prednisone were identified, and baseline semen parameters and subsequent changes after prednisone therapy were assessed. A Mann-Whitney U Test was used to compare semen parameter changes before and after prednisone. Adverse effects associated with prednisone were monitored. Results: A total of 8 patients were identified with deteriorating semen parameters who were treated with 6 weeks of low-dose prednisone. Following prednisone therapy, all patients demonstrated improvements in total motile sperm count (TMSC), with a median improvement of 6 million. The median relative improvement in TMSC was 433%. Sperm concentration and motility also improved compared to post-operative baseline. No adverse effects were reported during the treatment period. Conclusions: Low-dose prednisone therapy appears to be a safe and effective intervention for managing deteriorating semen parameters following VV. The observed improvements in TMSC suggest the potential of prednisone to rescue patients with delayed failure after VV. Further research with larger sample sizes is warranted to confirm the safety and efficacy of low-dose prednisone as a rescue therapy in this specific patient population. Optimizing VV outcomes is crucial in male infertility, and further exploration of steroid therapy and innovative biotechnologies is warranted.

Herbal Textual Research on Euryales Semen in Famous Classical Formulas / 中国实验方剂学杂志

This paper systematically combed and verified the name， origin， producing area， quality evaluation， harvesting， processing of Euryales Semen in famous classical formulas by consulting relevant ancient materia medica， medical books， prescription books and modern literature. The results showed that Euryales Semen was first collected by materia medica under the name of Jitoushi， and since the Ming dynasty， Qianshi has been used as a proper name and continues to this day， with other aliases such as Yanhuishi. Euryale ferox， a plant of the Nymphaeaceae family， is the same as that used in the past dynasties. However， due to long-term artificial domestication， the varieties vary with the origin， including Beiqian and Suqian. The medicinal part of Euryales Semen is mature seed kernel， its origin of ancient records mainly includes Shandong， Jiangsu， Henan and other places， since the Ming and Qing dynasties， Euryales Semen produced in Suzhou has been highly praised. Since modern times， it has gradually summarized and formed the best quality evaluation method of Euryales Semen with full grains， white cross-section， powdery enough and no broken powder. The harvesting time in the past dynasties was mainly August or in autumn. The main processing methods in the past dynasties included peeling for powder， pounding powder after steaming， drying and frying. Up to now， two mainstream processing methods of cleansing and stir-frying have been formed. Based on the research results， it is recommended that the mature seed kernel of E. ferox be used in famous classical formula Yihuangtang. Combined with the processing requirements of the original formula， it is suggested to refer to the stir-frying method in the general principles of processing of the current edition of Chinese Pharmacopoeia.

The Effect of Smoking on the Semen Quality in Male Infertile / 昆明医科大学学报

Objective To investigate the effect of smoking on the semen quality in infertile men.Methods A total of 360 male infertility patients were enrolled and divided into the smoking group(n=190)and non-smoking group(n=170)based on whether they smoked or not.Furthermore,the smoking group was subdivided into group A(≤10 sticks/d,n=63),group B(11～20 sticks/d,n=80),and group C(>20 sticks/d group,n=47)according to the amount of smoking.Semen volume,liquefaction time,sperm concentration,motility,DNA fragmentation rate and normal morphological rate were observed and compared between and within the groups.Results There were significant differences in semen volume,liquefaction time,sperm motility,normal morphological rate and DNA fragmentation rate between the smoking group and the non-smoking group(P<0.05).The semen volume,sperm motility and normal morphological rate of the smoking group were lower than those in the non-smoking group,and the DNA fragmentation rate and semen liquefaction time were higher than those in the non-smoking group.And with the increase of smoking volume,sperm motility and normal morphological rate decreased,semen liquefaction time and DNA fragmentation rate increased,and there was no significant difference in the sperm concentration between the smoking group and non-smoking group(P>0.05).There was no significant difference in the semen volume between the three groups with different smoking amounts(P>0.05).Conclusion Smoking has a negative impact on the sperm quality parameters such as semen volume,sperm motility,normal morphological rate,sperm motility,liquefaction time and DNA fragmentation,and the effect of heavy smoking is particularly obvious.We should strengthen the comprehensive health education,promote the healthy lifestyles and reduce smoking.

Dilution conditions and standardization study for semen samples using computer-aided sperm analysis / 临床检验杂志

Objective To explore the dilution conditions and standards in detecting the semen samples with high sperm concentration using computer-aided sperm analysis(CASA)systems.Methods CASA systems with 10 μm depth disposable counting chambers were used for the examination.The samples were divided into undiluted group(Group 1∶sperm concentration＜50×106/mL)and diluted groups(Group 2∶50×106/mL≤ sperm concentration＜100× 106/mL;Group 3∶sperm concentration≥100× 106/mL).When sperm concentration＜50×106/mL,no dilution was performed.When sperm concentration≥50× 106/mL,the samples were diluted with saline at 1∶n/(50×106)ratio(n=sperm concentration,n/[50×106]rounded down)to＜50×106/mL of sperm concentration.The sperm concentration,progressive motility(PR),non-progressive motility(NP),total motility(PR+NP)and immotile sperm percent-age(IM)were analyzed before and after dilution.The consistency of results pre-and post-dilution was compared.ROC curve was used to analyze the optimal dilution threshold.Results The differences in the parameters pre-and post-dilution gradually rosed with the increased sperm concentration.ROC curve analysis showed optimal dilution thresholds were 133.05 × 106/mL,101.75 × 106/mL,118.60×106/mL,90.90×106/mL,111.83×106/mL for the sperm concentration,PR+NP,PR,NP and IM respectively.Considering sperm concentration and NP were most affected the undiluted high concentration samples,the optimal comprehensive dilution threshold was determined as 125.08× 106/mL.Conclusion When sperm concentration exceeds 125×106/mL,it is recommended to dilute semen sample with normal saline.

Current situation and improvement strategies of semen testing in hospitals without developing assisted reproductive technolo-gy in Hunan province / 临床检验杂志

Objective To investigate the current status of semen testing in the hospitals without assisted reproductive technology in Hu-nan Province,propose improvement strategies to enhance the diagnosis of male infertility and promote reproductive health services in these hospitals.Methods Questionnaire surveys and on-site investigations were conducted to examine the semen testing status in 67 hospitals without assisted reproductive technology in Hunan Province.The existing problems were summarized,and improvement strate-gies were suggested.Results Among the 67 hospitals,59.7％(40/67)performed semen testing.Of these,45％(18/40)possessed dedicated personnel,60％(24/40)possessed staff received relevant training,and 22.5％(9/40)held certificates.Only 30％(12/40)conducted sperm morphology testing,among them 20％(8/40)able to stain and interpret following WHO standards.The hospitals of 37.5％(15/40)used computer-assisted sperm analysis(CASA)systems,and 22.5％(9/40)were equipped with phase-contrast microscopes.30％(12/40)had dedicated examination areas,and 32.5％(13/40)had independent ejaculation rooms,17.5％(7/40)used disposable slides to observe sperm concentration.In 2022,the hospitals of 57.5％(23/40)had an average daily sample volume＜1.The hospital of 67.5％(27/40)performed standard operating procedures,40％(16/40)followed the WHO 5th la-boratory manual for routine testing,15％(6/40)conducted internal quality control,and 12.5％(5/40)participated in external quali-ty assessment(EQA).Another 55％(22/40)wished to participate in EQA.Conclusion The semen testing capacity in the hospitals,in which assisted reproductive technology is not yet carried out currently,urgently requiring multifaceted improvements.The proposed strategies include emphasizing semen testing,establishing reproductive medicine consortia for comprehensive support,establishing a provincial quality control center for EQA,founding a professional committee for a learning platform,offering training to enhance staff expertise,and including the pre-pregnancy semen testing in public health programs.

Effects of Diosmine on semen quality and sex hormone concentration in patients with spermatovenous liga-tion / 实用医学杂志

Objective To investigate the effect of diosmine on semen quality and sex hormone concentra-tion in patients with spermatovenous ligation.Methods Clinical data were retrospectively collected from January 2019 to January 2022 on patients with varicocele admitted to Department of Urology,General Hospital of Southern Theater Command,divided into a control group and a study group based on whether they had a history of Diosmine use after surgery.Clinical data from both groups were matched using propensity score matching.Sperm quality and sex hormone tests were then compared before surgery,three months after surgery and six months after surgery.Results There were 73 in the control group and 73 in the study group.Both groups were balanced in terms of age,BMI,seminal fluid volume and testosterone(P＞0.05).Sperm motility,forward motility and testosterone were all increased 3 and 6 months after surgery,and sperm malformations,sperm DNA fragment index,follicle stimulating hormone and luteinizing hormone were all reduced significantly(P＜0.05).The results were also better than those in the control group for total sperm motility,forward motility,testosterone,sperm DNA fragment index,follicle stimulating hormone and luteinizing hormone(P＜0.05).Conclusions Diosmine can help to restore semen quality and improve sex hormone concentration in patients with varicocele after Laparoscopic varicocelectomy.

The relationship between semen quality and trace element levels in seminal plasma and miR-184 levels in seminal vesicles of male infertility patients / 实用医学杂志

Objective Explore the relationship between semen quality and trace element levels in seminal plasma,as well as miR-184 levels in exosomes of male infertility patients.Methods 155 male infertility patients in our hospital from January 2022 to July 2023 were selected as the infertility group,and another 155 male patients with normal physical examination during the same period were selected as the normal group.Compare the general information,semen quality parameters,trace elements,and miR-184 levels in exosomes between two groups,and analyze the correlation between male infertility patients and the above laboratory indicators through Spearman rank correlation analysis.Results The sperm concentration,total sperm motility(PR + NP),forward motility sperm(PR),average curve velocity,average linear velocity,average path velocity,and average lateral amplitude of sperm head(ALH)in the infertile group were lower than those in the normal group(P<0.05);The levels of Zn,Mg,and Ca in the infertile group were lower than those in the normal group,while the levels of Cu,Pb,and relative expression of miR-184 were higher than those in the normal group(P<0.05);The levels of Zn and Cu elements in male infertility patients are positively and negatively correlated with sperm concentration,total sperm motility(PR+NP),and forward motility sperm(PR),respectively.The levels of Pb elements are negatively correlated with the average lateral amplitude of sperm head(ALH),and P<0.05.The relative expression level of miR-184 in male infertility patients is negatively correlated with sperm concentration,total sperm motility(PR + NP),forward motility sperm(PR),and average sperm curve velocity,with P<0.05.Conclusion The quality of semen in male infertility patients is closely related to the levels of trace elements in semen and miR-184 levels in exosomes.

Semen cuscutae in the treatment of osteoarthritis:network pharmacology analysis and experimental validation / 中国组织工程研究

BACKGROUND:Semen cuscutae has the effect of tonifying the liver and kidney system and benefiting the essence.The main pathogenesis of osteoarthritis is deficiency of the liver and kidneys.Therefore,it is hypothesized that there is a link between semen cuscutae and osteoarthritis. OBJECTIVE:To explore the potential relationship between osteoarthritis and semen cuscutae and validate the mechanism of semen cuscutae based on the network pharmacology and molecular docking analysis. METHODS:First,the active ingredients and targets of semen cuscutae were screened in TCMSP,and the genes related to osteoarthritis were collected in the disease databases GeneCard's,OMIM and TTD.The intersected genes were taken and then subjected to a series of analyses and screened for hub genes.Through the enrichment analysis of hub genes,the pathway of semen cuscutae acting on osteoarthritis was selected.The role of hub genes was verified by molecular docking.Therefore,the appropriate active ingredients of semen cuscutae were selected for experimental verification. RESULTS AND CONCLUSION:There were 11 active ingredients of semen cuscutae,66 intersection target genes of semen cuscutae and osteoarthritis,and 12 hub genes,including tumor necrosis factor,interleukin 1B,TP53,RAC-alpha serine/threonine protein kinase(AKT1),vascular endothelial growth factor A,matrix metalloproteinase 9,prostaglandin peroxidase 2,cystatinase 3,epidermal growth factor,peroxisome proliferator-activated receptor gamma,interleukin 10,vascular cell adhesion factor 1.After the enrichment analysis of the hub genes,the classical inflammatory pathway,nuclear factor-κB signaling pathway,was selected for subsequent validation of semen cuscutae to alleviate osteoarthritic inflammation.Through the results obtained after molecular docking of each active ingredient and the hub gene of the pathway prostaglandin peroxidase 2,sesamin with the highest affinity was selected for subsequent cell experiments,and the experimental results confirmed that sesamin,the active ingredient of semen cuscutae,could reduce the expression of cyclooxygenase 2 by inhibiting the nuclear factor-κB signaling pathway induced by interleukin-1β.To conclude,sesamin,the active ingredient of semen cuscutae,reduces the expression of cyclooxygenase 2 by inhibiting the nuclear factor-κB signaling pathway induced by interleukin-1β,thereby improving inflammation in osteoarthritis and expanding the therapeutic effect of semen cuscutae in osteoarthritis.

Study on the relationship between semen quality and bacterial infection in infertile men in Guangdong province / 现代医院

Objective To study the correlation between semen quality and bacterial infection in men with abnormal fer-tility,and provide clinical basis for guiding the reproductive health of men with abnormal fertility.Methods 200 male semen samples with abnormal fertility were collected,and then separated and cultured for 48 hours.According to the culture results,they were divided into three groups:the non-pathogenic group,the pathogenic group,and the sterile group.The bacterial resist-ance analysis of the pathogenic group was conducted,and the semen quality between each group was compared.Results After 48 hours of isolation and cultivation,200 semen samples had been tested,non-pathogenic bacteria was detected in 163 semen samples,accounting for 81.5% ;pathogenic bacteria was detected in 33 semen samples,accounting for 16.5% ;and bacteria was not detected in 4 semen samples,accounting for 2.0% .The top three strains of pathogenic bacteria in 33 cases were Escherichia coli,Streptococcus agalactiae,and Enterococcus faecalis,with drug resistance rates of 80.0% ,87.5% ,and 100.0% ,respec-tively.Conclusion The detection rate of bacterial culture in semen of men with abnormal fertility is relatively high,and patho-genic bacteria can affect semen quality.

Effects of Ziziphi Spinosae Semen-Albiziae Flos on the IRE1α/ASK1/JNK Pathway of Endoplasmic Reticulum Stress in Hippocampus of Depression Model Rats / 中药新药与临床药理

Objective Based on the endoplasmic reticulum stress(ERS)inositol-requiring enzyme-1α(IRE1α)/apoptosis signal-regulated kinase 1(ASK1)/c-Jun N-terminal kinase(JNK)pathway to investigate the intervention effect of Ziziphi Spinosae Semen(ZSS)-Albiziae Flos(AF)on the depression model of rats,which were prepared by solitary rearing combined with chronic unpredictable mild stress(CUMS).Methods A total of 144 rats were randomly divided into normal group,model group,high-,medium-and low-dose groups of ZSS-AF,and Venlafaxine group.In addition to the normal group,the rats in other groups were isolated for 21 days combined with CUMS to prepare the depression model.The behavioral changes of rats were observed by open field test and Morris water maze test.The ultrastructural changes of hippocampal neurons were observed by transmission electron microscope.TUNEL staining was used to observe the apoptosis of nerve cells in hippocampus.The protein expression levels of IRE1α,phosphorylated(P)-IRE1α,ASK1,P-ASK1,JNK,P-JNK,B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and cysteme aspartate specific protease-3(Caspase-3)in hippocampus were detected by Western Blot.The mRNA expression levels of IRE1α,ASK1,JNK,Bax,Bcl-2 and Caspase-3 in hippocampus were detected by Real-Time PCR.Results Compared with the normal group,the scores of horizontal movement and vertical movement in the open field test of rats in the model group were decreased(P＜0.01).In the water maze test,the escape latency was increased and the number of crossing the original platform was decreased(P＜0.01).The number of hippocampal apoptosis was increased(P＜0.01).The protein expression levels of P-IRE1 α/IRE1 α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 and mRNA expressions of IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were increased,while the protein and mRNA expression levels of Bcl-2 were decreased(P＜0.01).Compared with model group,the scores of horizontal movement and vertical movement in the open field test of rats in each dose ZSS-AF groups and Venlafaxine group were increased(P＜0.01).In the water maze test,the escape latency was decreased and the times of crossing the original platform was increased(P＜0.01).The number of hippocampal apoptosis was decreased(P＜0.01).The mRNA expression levels of P-IRE1α/IRE1α,P-ASK1/ASK1,P-JNK/JNK,Bax,Caspase-3 protein and IRE1α,ASK1,JNK,Bax,Caspase-3 in hippocampus were decreased,while the protein and mRNA expression levels of Bcl-2 were increased(P＜0.05,P＜0.01).The effect of medium-dose ZSS-AF group was better than that of high-and low-dose groups.Conclusion ZSS-AF may play an antidepressant role by regulating IRE1α/ASK1/JNK pathway of endoplasmic reticulum stress.

Inspiration of The Inner Canon of Yellow Emperor （《黄帝内经》） to the Construction of the Theory of "Middle Jiao Deficiency Causes Seminal Changes" / 中医杂志

Based on the theory of spleen-stomach in The Inner Canon of Yellow Emperor （《黄帝内经》）， the theory of "middle jiao deficiency causes seminal changes" focuses on the ways in which the spleen-stomach deficiency affect male seminal disorders， as well as the ideas and methods of treating seminal disorders from the perspective of the spleen and stomach. Based on the discussion of spleen and stomach in The Inner Canon of Yellow Emperor， the theoretical connotations of “middle jiao deficiency causes seminal changes” are clarified in terms of the scope of the disease， the etiology of the spleen deficiency， the pathogenesis changes， and the treatment of the disease. It is believed that the connotation of "semen" includes the semen itself and the semen chamber as a whole， and that the core pathogenesis of "seminal changes" is the deficiency of spleen and stomach， which also involves kidney deficiency. Taking the theory of "middle jiao deficiency causes seminal changes" as a guide， the treatment idea of "regulating middle jiao and rectifying essence" was established， with the key of tonifying the middle jiao and regulating transportation and transformation of spleen and stomach， and the aim of regulating essence chamber and promoting the metabolism of turbid substances and the production of essence. It is divided into three methods， including tonifying， transporting， and clearing the middle jiao to rectify semen， so as to providing new ideas for the treatment of male seminal disorders from the spleen and stomach.

Analysis of semen quality parameters among 155 sperm donors / 预防医学

Objective@#To analyze the semen quality parameters of sperm donor volunteers, so as to provide insights into male infertility control and related research.@*Methods@#A total of 155 sperm donors were recruited from the Human Sperm Bank of Zhejiang Province using the convenience sampling method from January to March 2021. Demographic information were collected through questionnaire surveys. Semen were collected, and parameters including semen volume, sperm concentration, total number of sperm, forward motility rate and total sperm viability were measured. Semen quality was evaluated according to the WHO Laboratory Manual for the Examination and Processing of Human Semen.@*Results@#@*Conclusion@#There were 20.65% of the 155 sperm donors with unqualified semen, and the unqualified rates of forward motility rate and total sperm viability were relatively high.

Screening Effective Sites of Momordicae Semen-Epimedii Folium and Anti-lung Cancer Mechanism of Its Prescription / 中国实验方剂学杂志

ObjectiveTo preliminarily confirm the effective anti-lung cancer sites of Momordicae Semen and Epimedii Folium and study their mechanism of action. MethodOn the basis of preliminary research， the extraction method of Momordicae Semen and Epimedii Folium was optimized and the effective parts were screened under the guidance of pharmacological effects. Different ethanol elution and water elution sites of Momordicae Semen and Epimedii Folium were obtained through adsorption and elution with D101 macroporous resin. The methylthiazolyldiphenyl-tetrazolium bromide （MTT） colorimetric assay was used to detect the effects of total drug extracts and different elution sites on the proliferation of various tumor cell lines， and to screen for the optimal elution site and tumor sensitive strains. Flow cytometry was used to detect the effect of the elution sites of Momordicae Semen and Epimedii Folium on intracellular reactive oxygen species （ROS） and apoptosis in A549 cells. Western blot was used to compare the expressions of tumor protein 53 （p53）， Bcl-2-associated X protein （Bax）， cysteinyl aspartate specific proteinase-3 and 9 （Caspase-3 and Caspase-9） proteins in A549 cells. ResultThe inhibitory effect of Momordicae Semen on the proliferation of A549 cells was better than the kernel of Momordicae Semen， with 50% inhibitory concentration （IC50） being （86.83±2.88） mg·L-1 and （95.10±18.13） mg·L-1， respectively. The effect of total extracts of Epimedii Folium on A549 anti proliferation IC50 value was （4.71±0.81） mg·L-1. The IC50 values of the 40%， 60%， and 80% ethanol and anhydrous ethanol eluted macroporous resins of the total extracts of Momordicae Semen and Epimedii Folium inhibiting A549 proliferation were （45.32±4.38）、 （14.95±0.73）、 （17.07±1.76）、 （14.46±2.35）、 （51.7±2.26）、 （12.37±0.67）、 （20.29±0.93）、 and （3.43±0.91） mg·L-1， respectively. Compared with the normal group， the 1∶1 combination of Momordicae Semen and Epimedii Folium inhibited A549 cell proliferation in a time-dependent and concentration-dependent manner. Compared with the normal group， 50 mg·L-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased intracellular ROS expression （P<0.01）. Compared with the normal group， 12.5， 25， 50 mg·L-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of A549 cell apoptosis （P<0.01）. Compared with the normal group， 25， 50 mg·L-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of p53 in A549 cells （P<0.01）. Compared with the normal group， 12.5， 25， 50 mg·L-1 of the combination of Momordicae Semen and Epimedii Folium significantly increased the expression of Bax （P<0.01）. Compared with the normal group， 50 mg·L-1 of the combination of Momordicae Semen and Epimedii Folium significantly reduced the expressions of Caspase-3 and Caspase-9 （P<0.01）. ConclusionThe anti-tumor effect of Momordicae Semen is better than that of the kernel of Momordicae Semen. The anti-tumor substances of Momordicae Semen and Epimedii Folium mainly concentrate in the 60% ethanol to anhydrous ethanol elution site. A549 cells are sensitive to the 1∶1 combination of Momordicae Semen and Epimedii Folium， which can effectively inhibit the cell proliferation. The mechanism may be related to increasing the generation of ROS in A549 cells， promoting their apoptosis， increasing the expressions of apoptotic proteins such as p53 and Bax， and reducing the expressions of Caspase-3 and Caspase-9.

Exploring Detoxication Mechanism of Dioscoreae Bulbiferae Rhizoma Processed with Phaseoli Radiati Semen Juice Based on Target Organ Ferroptosis / 中国实验方剂学杂志

ObjectiveTo investigate the attenuating effect of Dioscoreae Bulbiferae Rhizoma（DBR） processed with Phaseoli Radiati Semen（PRS） juice， and explore the attenuating mechanism based on ferroptosis of the main toxic target organ. MethodSixty male ICR mice were randomly divided into blank group， DBR group， water roasted DBR group（hereinafter referred to as water group）， PRS juice-roasted DBR group 1（DBR-PRS 10∶1， stuffy moistening for 40 min， stir-fried at 130 ℃ for 18 min， hereinafter referred to as group 1）， PRS juice-roasted DBR group 2（DBR-PRS 10∶1， stuffy moistening for 80 min， stir-fried at 100 ℃ for 14 min， hereinafter referred to as group 2）， PRS juice-roasted DBR group 3（DBR-PRS=20∶3， stuffy moistening for 40 min， stir-fried at 160 ℃ for 14 min， hereinafter referred to as group 3）. The raw and processed groups of DBR were gavaged with their corresponding 95% ethanol extract at a dose of 3 g·kg-1·d-1， while the blank group was gavaged with an equal volume of 0.5% sodium carboxymethyl cellulose， once a day for 14 consecutive days. Hematoxylin-eosin（HE） staining was used to observe the histopathological changes of mouse liver. Alanine aminotransferase（ALT） and aspartate aminotransferase（AST） levels in serum， as well as malondialdehyde（MDA）， ferrous ions（Fe2+）， reduced glutathione（GSH） and superoxide dismutase（SOD） levels in liver tissue were detected by the biochemical detection. Western blot was used to detect the expression of iron key proteins such as ferritin heavy chain 1（FTH1） and glutathione peroxidase 4（GPX4）. ResultHE staining results showed that the liver tissue structure of the blank group was clear， the morphology of hepatocytes was normal， the cytoplasms of hepatocytes in the DBR group and water group were loose and vacuolar， with obvious pathological damages， and the pathologic damages of mice in the group 1-3 were significantly improved. Compared with the blank group， the levels of ALT， AST， MDA and Fe2+ in mice from the DBR group were significantly increased（P<0.01）， while GSH and SOD levels were significantly reduced（P<0.01）， and the protein expression levels of FTH1 and GPX4 were significantly decreased（P<0.01）. Compared with the DBR group， the ALT， AST，MDA and Fe2+ levels of mice in the group 1-3 were significantly reduced（P<0.05， P<0.01）， the GSH and SOD levels and the protein expression levels of FTH1 and GPX4 were significantly increased（P<0.01）. Compared with the water group， the AST and MDA levels of mice in the group 1-3 were significantly reduced（P<0.05， P<0.01）， the SOD level significantly increased（P<0.05， P<0.01）， the FTH1 protein expression significantly increased（P<0.01）， and the serum ALT level of mice in the group 2-3 significantly reduce（P<0.01）， Fe2+ level significantly reduced（P<0.01）， GSH level significantly increased（P<0.05， P<0.01）， and GPX4 protein expression significantly increased（P<0.05， P<0.01）. Among the group 1-3， the group 3 had the best detoxification effect. ConclutionProcessing with PRS juice can reduce the liver injury induced by DBR， and the mechanism may be related to the inhibition of ferroptosis in the liver.

Biological Basis of Premature Ejaculation Based on Theory of "Brain-heart-kidney-semen Chamber" Axis / 中国实验方剂学杂志

The theory of "brain-heart-kidney-semen chamber" axis is proposed based on the basic theories of traditional Chinese medicine， the modern physiological characteristics of men's diseases， and clinical practice. According to this theory， dysfunctions of the brain， heart， kidney， and semen chamber are the core mechanisms for the occurrence of premature ejaculation， and the loss of control of the opening and closing of the seminal orifices due to the dysfunction of the semen chamber is the final link in the occurrence of premature ejaculation. The treatment of premature ejaculation based on the theory of "brain-heart-kidney-essence chamber" axis highlights the overall regulation of the Zang-fu organs involved in the disease， while focusing on the simultaneous treatment of the mind and body. By exploring the biological basis of the "brain-heart-kidney-essence chamber" axis and premature ejaculation， we propose that the biological basis of premature ejaculation and the axis is mainly related to the function decline of the local brain area， neuromodulation malfunction， central neurotransmitter imbalance， endocrine disorders， and enhanced sensory afferents of the penis. This study aims at providing a new approach for the prevention and treatment of premature ejaculation by traditional Chinese medicine and a scientific basis for the development of more effective therapeutic methods.

Mechanism of Cuscutae Semen to Control Premature Ovarian Failure Based on Lipid and Atherosclerosis Pathways / 中国实验方剂学杂志

ObjectiveThe active ingredients， action targets， and signaling pathways of Cuscutae Semen to control premature ovarian failure were initially predicted by network pharmacology and molecular docking techniques， and an animal model of premature ovarian failure was constructed to explore the mechanism of Cuscutae Semen based on lipid and atherosclerosis signaling pathways. MethodThe effective components and corresponding targets of drugs were obtained from Traditional Chinese Medicines Systems Pharmacology Platform （TCMSP）， Swiss Target Prediction， Pharmmapper， and other databases. GeneCards database was used to collect disease-related targets. Venny2.1.0 online tool was used to screen out the intersection targets of drugs and diseases， and STRING database and Cytoscape v3.7.2 software were used to construct the network diagram of "drug-component-target" and protein-protein interaction （PPI）. The gene ontology （GO） and the Kyoto encyclopedia of genes and genomes （KEGG） enrichment analyses of the intersection targets were performed by running the R language script. The molecular docking technology was utilized to dock drug components with targets and visualize some of the docking results. The mice were randomly divided into a blank group， a model group， a Cuscutae Semen group， and an estradiol valerate group， and the ovarian premature failure model was prepared by chronic stress. The blank group and the model group were gavaged with the same amount of normal saline， and the Cuscutae Semen group was given a Cuscutae Semen decoction of 2.6 g·kg-1·d-1. The estradiol valerate group was given an estradiol valerate solution of 0.13 mg·kg-1·d-1. After four weeks， samples were collected， and hematoxylin-eosin （HE） staining was performed to observe the histopathological changes in the ovary. Serum levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， estradiol （E2）， Muller's tube inhibitor/anti-Muller's tube hormone （AMH）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， and low-density lipoprotein cholesterol （LDL-C） were determined by enzyme-linked immunosorbent assay （ELISA）. The expression levels of extracellular regulatory protein kinase （ERK）， nuclear transcription factor-κB p65 （NF-κB p65）， nuclear transcription factor-κB suppressor α （IκBα）， interleukin-1β （IL-1β）， IL-6， and tumor necrosis factor-α （TNF-α） were measured by Western blot. ResultA total of 171 targets of Cuscutae Semen for the prevention and treatment of premature ovarian failure were screened， mainly including tumor protein p53 （TP53）， protein kinase B1 （Akt1）， sarcoma （SRC）， tumor necrosis factor （TNF）， epidermal growth factor receptor （EGFR）， etc. KEGG pathway enrichment analysis predicts that Cuscutae Semen is mainly involved in lipid and atherosclerosis， TNF signaling pathway， and TP53 signaling pathway to control premature ovarian failure. The animal experiments show that compared with the premature ovarian failure model group， the Cuscutae Semen group can significantly upregulate AMH， E2， and HDL-C （P<0.05， P<0.01）， significantly downregulate LH， TC， and LDL-C （P<0.01）， greatly reduce IL-1β， IL-6， and TNF-α protein levels， as well as ERK， NF-κB p65， and their phosphorylation levels （P<0.01）. ConclusionCuscutae Semen can regulate hormone levels and improve ovarian function through a multi-component， multi-target， and multi-pathway approach， and the mechanism may be related to the regulation of lipid and atherosclerosis signaling pathways.