Relationship between semen parameters, serum InhB, and INSL-3 levels, and the degree of varicocele

Abstract Background Varicocele is an abnormal expansion of the pampininias venous plexus in the scrotum, resulting in impaired sperm production and reduced sperm quality. The exact pathophysiological mechanism leading to varicocele-related infertility has not been fully elucidated. Although treatable, varicocele may lead to male infertility. Objective To investigate the relationship between semen parameters, serum InhB and INSL-3 levels, and the degree of varicocele in male patients. Methods Serum InhB and INSL-3 were detected. To evaluate the relationship between semen parameters and serum InhB and INSL-3 levels. To evaluate the value of semen parameters and serum InhB and INSL-3 levels in distinguishing disease severity in patients with varicocele. Results Serum INSL-3 in patients with varicocele decreased with the severity of the disease. Serum INSL-3 was positively correlated with total sperm count and frequency of normal sperm morphology. There was a weak correlation between serum InhB and semen volume, concentration, and total sperm. Patients with different disease severity were similar within the groups, with partial overlap or similarity between varicocele Grade I and Grade II, and significant differences between Grade III and Grade I and II. Semen volume, concentration, total sperm, normal sperm morphology, and serum InhB and INSL-3 levels could distinguish the degree of varicocele. Conclusion Semen parameters and the combination of serum InhB and INSL-3 levels in patients with varicocele are closely related to the severity of the disease. Serum INSL-3 is expected to be a potential biomarker for early clinical intervention.

Analysis on Quality of Sojae Semen Praeparatum Based on Traditional Quality Evaluation / 中国实验方剂学杂志

ObjectiveBased on the quality evaluation experience of "it is better to have a fragrant and strong aroma" summarized by materia medica of past dynasties， the chemical components of Sojae Semen Nigrum（SSN） and Sojae Semen Praeparatum（SSP） were systematically compared and analyzed， and the main fermentation products in different fermentation time were quantitatively analyzed， so as to clarify the transformation law of internal components in the processing process and provide scientific basis for the modern quality control of SSP. MethodUltra performance liquid chromatography-quadrupole tandem time-of-flight mass spectrometry（UPLC-Q-TOF-MS） was used for the structural identification of the chemical constituents of SSN and SSP， and with the aid of Progenesis QI v2.3 software， the negative ion mode was employed for principal component analysis（PCA） pattern recognition， and the data were analyzed with the aid of orthogonal partial least squares-discriminant analysis（OPLS-DA） for two-dimensional data to obtain S-plot， and components with |P|>0.1 were selected as the differential constituents. The contents of isoflavonoids in SSP during fermentation was determined by UPLC， and the samples were taken every 8 h in the pre-fermentation period and every 2 d in the post-fermentation period， and the dynamic changes of isoflavonoid contents in different fermentation stages were analyzed. The contents of amino acids and nucleosides in SSP and SSN from different fermentation stages were quantitatively analyzed by phenyl isothiocyanate（PITC） pre-column derivatization and high performance liquid chromatography（HPLC） gradient elution， and the contribution of flavor substances to the "delicious" taste of SSP was discussed by taste intensity value（TAV）. ResultA total of 19 kinds of differential components were screened out， mainly soybean saponins and isoflavones， and their contents decreased significantly or even disappeared after fermentation. In the pre-fermentation process of SSP， glycoside bond hydrolysis mainly occurred， and isoflavone glycosides in SSN were degraded and converted into the corresponding aglycones， the content of flavor substances such as amino acids increased gradually. In the post-fermentation process， protein degradation mainly occurred， after 8 d of post-fermentation， the content of isoflavones was basically stable， while the total content of amino acids increased by 8-40 times on average. Different amino acids form the special flavor of SSP， such as the TAV of glutamate is always ahead of other flavor substances， and sweet substances such as alanine and valine have made relatively great contributions to SSP. ConclusionBased on the law of constituent transformation， combined with the traditional evaluation index of "fragrant and strong"， it is difficult to control the fermentation degree of SSP by the existing standards in the 2020 edition of Chinese Pharmacopoeia. It is suggested that description of the characteristics of SSP be refined and changed to "fragrant， delicious and slightly sweet"， and at the same time， the post-fermentation index compounds such as glutamic acid， alanine and valine should be added as the quality control indicators of SSP， so as to standardize the production process and improve the quality of SSP.

Testicular and epididymal dysfunctions: searching a new index for the differential diagnosis / Disfunciones testiculares y epididimarias: búsqueda de un nuevo índice para el diagnóstico diferencial

Objective: This proof-of-principle aims to develop an index to aid the differential diagnosis of disorders affecting testicular and/or epididymis. A total of 202 subject data were evaluated in two groups: fertile men with children naturally conceived within 1 year of unprotected intercourse (n = 36) and infertile men (n = 166) who had attempted a pregnancy more than 1 year with unprotected intercourse. Materials and methods: Semen parameters (sperm count, vitality, motility, morphology, and hypoosmotic swelling test [HOST]) were evaluated. The index was calculated by dividing the percentage HOST by the percentage of sperm progressive motility in the fertile group (n = 36). Results: A normal range from 1.23 to 1.53 was determined. Using this index, the outcomes of semen analysis from infertile men were grouped in three study groups: below 1.23 (n = 24), normal (n = 44), and higher than 1.53 (n = 98). These parameters were significantly decreased in semen with normal range (p < 0.01) and in indexes higher than 1.53 (p < 0.0001). Receiver operating characteristic curves compared progressive motility and morphology in infertile men with indexes higher than 1.53 shows that semen samples with normal sperm progressive motility and morphology did not suggest dysfunctions in testis and epididymis. Semen samples with asthenozoospermia suggested epididymal dysfunction (area under the curve [AUC] 0.889, confidence interval [CI] 0.783-1), whereas semen samples with teratoasthenozoospermia suggested dysfunction in both testicles and epididymis (AUC 0.891, CI 0.77-1). Conclusions: The current index proof-of-principle of the success of such a strategy provides valuable information about whether a disorder individually affects testicular and/or epididymal function.

Objetivo: Esta prueba de principio tiene como objetivo desarrollar un índice que ayude al diagnóstico diferencial de los trastornos testiculares y/o epidídimales. Métodos: Se evaluaron 202 individuos divididos en dos grupos: hombres fértiles con hijos concebidos de forma natural en el plazo no mayor a un año (n = 36) y hombres infértiles (n = 166), los cuales habían intentado un embarazo por más de un año. Se evaluaron los parámetros seminales (concentración, viabilidad, movilidad, morfología y prueba de hinchazón hipoosmótica [HOST]). El índice se calculó dividiendo el porcentaje de HOST por el porcentaje de movilidad espermática progresiva en el grupo fértil (n = 36). Resultados: Se determinó un rango normal de 1,23 a 1,53. Utilizando este índice, los resultados del análisis del semen de los hombres infértiles se agruparon en tres grupos de estudio: por debajo de 1,23 (n = 24), normal (n = 44) y superior a 1,53 (n = 98). En contraste, estos parámetros disminuyeron significativamente en el semen de rango normal (p < 0,01) y en los índices superiores a 1,53 (p < 0,0001). Las curvas ROC comparadas con la movilidad espermática progresiva y la morfología en los hombres infértiles con índices superiores a 1,53 muestran que las muestras de semen con movilidad progresiva y morfología normales no sugieren disfunciones en los testículos y epidídimos. Las muestras de semen con astenozoospermia sugerirían una disfunción del epidídimo (AUC 0,889, IC 0,783-1), mientras que las muestra de semen que presentaban teratoastenozoospermia sugerirían una disfunción tanto en los testículos como en el epidídimo (AUC 0,891, IC 0,77-1).

Dimethyl acetamide and dimethyl sulfoxide associated at glucose and egg yolk for cryopreservation of Pseudoplatystoma corruscans semen

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

Therapeutic Effect of Ziziphi Spinosae Semen-Albiziae Flos on Depression in Rats / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Ziziphi Spinosae Semen-Albiziae Flos on the nucleotide-binding oligomerization domain，NOD-like receptor thermal protein domain associated protein 1 （NLRP1）/chemokine ligand 1 （CXCL1）/chemokine receptor 2 （CXCR2） pathway in the hippocampus of the rat model of chronic unpredictable mild stress （CUMS）-induced depression. MethodA total of 120 male SD rats were randomized into blank，CUMS，CUMS + low-，medium-，and high-dose （4，8，16 g·kg-1） Ziziphi Spinosae Semen-Albiziae Flos，and CUMS + venlafaxine hydrochloride （0.008 g·kg-1） groups，with 20 rats in each group.The rat model of depression was established by solitary feeding combined with CUMS.The behaviors and spatial learning and memory abilities of rats were examined by sugar water consumption test，tail suspension test，forced swimming test，and Morris water maze test.Quantitative real-time PCR （Real-time PCR） and Western blot were employed to determine the expression of factors associated with the NLRP1/CXCL1/CXCR2 pathway in the hippocampus.Enzyme-linked immunosorbent assay was employed to determine the levels of tumor necrosis factor-α （TNF-α），interleukin （IL）-18，IL-1β，and IL-6 in the hippocampus.The immunofluorescence assay was used to measure the levels of reactive oxygen species （ROS） in the hippocampus. ResultCompared with the blank group，the CUMS group showed decreased preference to sugar water and times of crossing the platform （P<0.01），and increased immobility time of tail suspension，forced swimming floating time，and escape latency （P<0.01）.Compared with the CUMS group，the administration of Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride alleviated the effects of CUMS on the above-mentioned behaviors and spatial learning and memory abilities of the rats （P<0.05，P<0.01）.Compared with the blank group，the CUMS group showed up-regulated protein levels of NLRP1，CXCL1，and CXCR2 （P<0.01） and elevated levels of IL-18，IL-1β，TNF-α，and IL-6 （P<0.01） in the hippocampus.The treatment with Ziziphi Spinosae Semen-Albiziae Flos and venlafaxine hydrochloride attenuated the activation of NLRP1/CXCL1/CXCR2 signaling pathway and lowered the levels of inflammatory cytokines in the hippocampus of CUMS rats （P<0.05，P<0.01）.In addition，Ziziphi Spinosae Semen-Albiziae Flos lowered the level of ROS in the hippocampus （P<0.05，P<0.01）. ConclusionZiziphi Spinosae Semen-Albiziae Flos can mitigate the depressive behaviors of the rat model of CUMS-induced depression by inhibiting the activation of NLRP1/CXCL1/CXCR2 signaling pathway.

Effects of different frequencies of pricking-bloodletting at auricular points plus auricular point sticking therapy on the serum levels of related factors in patients with acne vulgaris / 针灸推拿医学（英文版）

Objective: To observe the effects of different frequencies of pricking-bloodletting at auricular points plus auricular point sticking therapy on the levels of the serum immunoglobulin G (IgG), immunoglobulin M (IgM), testosterone (T), and insulin-like growth factor-1 (IGF-1) in patients with acne vulgaris (AV). Methods: Ninety patients with AV were randomly assigned to treatment group 1, treatment group 2, and treatment group 3 according to the random number remainder grouping method, with 30 cases in each group. All three groups received pricking-bloodletting at auricular points plus auricular point sticking therapy. Treatment group 1 was treated once a week, treatment group 2 was treated twice a week, and treatment group 3 was treated 3 times a week. Four-week treatment was taken as 1 treatment course, and 3 treatment courses were observed. On the day before the start of the study and on the day next to the end of each course, the global acne grading system (GAGS) score was recorded, and 3 mL of blood from the median cubital vein was collected to test the serum levels of IgG, IgM, T, and IGF-1. After 3 courses of treatment, the efficacy index was calculated according to the GAGS score, and the serum indicators in patients with effective treatment (efficacy index ≥20％) and complete sampling were analyzed. Results: A total of 60 patients were included in the final blood indicator analysis. After 3 courses of treatment, the intra-group comparisons showed that the serum IgG level increased significantly in patients in treatment group 1 (P<0.01); the serum T level decreased significantly in the female patients in treatment group 2 (P<0.05); the IGF-1 level significantly decreased in the patients in all three groups (P<0.05 or P<0.01). There were no significant differences in the serum levels of IgG, IgM, T, and IGF-1 among the three groups at each time point (P>0.05). Conclusion: Pricking-bloodletting at auricular points plus auricular point sticking therapy can affect the levels of serum IgG, T, and IGF-1 in AV patients. The level of the serum IGF-1 can be reduced by treatment once a week, twice a week, or 3 times a week. Treatment once a week can increase the patients' serum IgG level, and treatment twice a week can significantly decrease the serum T level in female patients. Reducing the serum IGF-1 level may be one of the action mechanisms of pricking-bloodletting at auricular points plus auricular point sticking therapy in treating AV.

Isolation and identification of two new compounds from Semen Persicae / 药学学报

Five compounds were isolated from the ethyl acetate fraction of Semen Persicae by using various chromatographic methods, including ODS, Sephadex LH-20, HPLC and semipreparative HPLC. Their structures were identified by 1D-NMR, 2D-NMR, HR-ESI-MS, UV, IR, circular dichroism (CD) and ECD calculation techniques: (2R,3R)-5,7,4′-trihydroxy-3′-methoxy-3-formylflavan-3-ol-5-O-β-D-glucopyranoside (1), (7R,8S)-dihydrodehydrodiconiferyl 6″-benzoyl alcohol-9-O-β-D-glucopyranoside (2), (7R,8S)-dihydrodehydrodiconiferyl alcohol-9-β-O-D-glucopyranosid (3), 2-methoxy-4-(2-propenyl)-phenyl-O-β-D-glucopyranoside (4), 2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]-propane-1,3-diol (5). Compound 1 and 2 are new compounds, and compounds 3-5 were obtained from Prunus davidiana (Carr.) Franch. for the first time.

Comparison of contents variation of six isoflavones in black beans, semifinished and finished Sojae Semen Praeparatum / 药学实践杂志

Objective To compare the contents variation of six flavonoids includingdaidzin，glycitin, genistin, daidzein, glycitein and genisteinin black beans, semifinished and finished Sojae Semen Praeparatum．Methods The contents of flavonoids were determined by HPLC，the condition were Diamonsil C18 column (4.6×250 mm, 5 μm) , column temperature 30 ℃, detection wavelength 260 nm, mobile phase 0.2% acetic acid water (A) - methanol (B), gradient elution, flow rate 1.0 ml/min．Results The linearity of this method to determine 6 isoflavones was good (r≥0.9993) within the determination range, and the recovery rate met the requirements. The RSD of precision, repeatability and stability experiment was less than 4%, 3%and 3%. The results of HPLC showed that the contents of six flavonoidsin Sojae Semen Praeparatum increased significantly compared with black beans. And, the contents of six flavonoids in finished Sojae Semen Praeparatum were slightly more than those in semifinished Sojae Semen Praeparatum. Conclusion The HPLC method established in this study could accurately determine the content of 6 isoflavones in Sojae Semen Praeparatum. The content of six isoflavones in black beans could be increased by the fermentation, and the combined isoflavones were transformed into free isoflavones during the fermentation process.

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

This article has systematically sorted out and verified the name， origin， producing area， quality evaluation， harvesting and processing of Pruni Semen by consulting ancient materia medica， medical books， prescription books and modern literature， in order to provide a basis for the development of famous classical formulas containing Pruni Semen. The results showed that Pruni Semen， as a medicinal material， has been widely used in medical literature of past dynasties since it was collected in Shennong Bencaojing， and also included under the names such as Yuhe， Yuzi and Yuli， and aliases such as Jueli， Queli and Chexiali. The primordial plants mentioned in the past dynasties involve about 12 species of Rosaceae， but with Prunus humilis， P. japonica and P. glandulosa as mainstream varieties used in the past dynasties， while the 2020 edition of Chinese Pharmacopoeia stipulates that the basal plants are P. humilis， P. japonica and P. pedunculata. Most of the ancient records for the origin of Pruni Semen are found everywhere in high mountains， valleys and hills， modern literature records that its origin varies according to its base， for example， P. humilis and P. japonica are mainly produced in Hebei， eastern Inner Mongolia， Liaoning， Shandong and other regions of China， and P. pedunculata is mainly produced in Inner Mongolia. Modern literature summarizes its quality as faint yellow， full and fulfilling， neat and not broken， and non-oiling， and the small Pruni Semen is better than the big Pruni Semen. The ancient processing methods of Pruni Semen mainly include blanching and peeling， blanching and peeling followed by frying， and blanching and peeling followed by pounding， with the common feature of blanching and peeling. The successive editions of Chinese Pharmacopoeia stipulate that it should be pounded when used. Based on the results of the herbal textual research and the writing time of Bianzhenglu， and combined with the market survey of Pruni Semen， it is suggested that P. humilis or P. japonica should be used as the origin of Pruni Semen in Sanpiantang， and it is harvested when the fruits are ripe， the kernels are collected by removing the stones， and processed by blanching， peeling and pounding consulting the decoction method in the current edition of Chinese Pharmacopoeia.

Study on the tissue distribution of eight effective components of Ziziphi Spinosae Semen aqueous extract by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry / 药学学报

The aim is to study the tissue distribution characteristics of eight effective components in normal rats after oral administration of Ziziphi Spinosae Semen (ZSS) aqueous extract. An ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) analysis method was developed and validated for the determination of four flavonoids and four saponins in rat tissue using puerarin and ginsenoside Re as the internal standard (IS), respectively. Tissue samples including the heart, liver, spleen, lung, kidney, muscle, brain, small intestine, and serum, were collected from each rat at 0.5 h, 1.0 h, and 2.0 h after oral administration of ZSS aqueous extract (15 g·kg-1). All calibration curves exhibited good linearity (r > 0.994 6) over a wide concentration range for all components. The intra-day and inter-day precisions (RSD) at four different levels were both less than 19.77%, and the accuracies (RE) ranged from -19.68% to 19.46%; The extraction recoveries of the eight components ranged from 86.70% to 114.29%, and the matrix effects were from 82.14% to 114.57%. The validated method was successfully applied to the tissue distribution study of the eight components. The levels of swertisin, spinosin, 6‴-feruloylspinosin, and kaempferol-3-O-rutinoside in the small intestine were highest, then followed by the kidney, heart, and liver. Meanwhile, the levels of jujuboside A (JuA), jujuboside B (JuB), and jujuboside A1 (JuA1) in the small intestine were highest, then followed by the lung, spleen, and kidney. The concentrations of betulinic acid in the small intestine were higher than heart, lung, kidney, and liver. The flavonoids and saponins of ZSS with extremely low content could pass through the blood-brain barrier. The research results will provide an experimental basis for explaining the mechanism of nourishing the heart and tranquilizing the mind of ZSS. The animal experimental operations involved in this study followed the regulations of the Animal Ethics Committee of Shanxi University of Chinese Medicine and passed the animal experimental ethical review (No. 2021DW172).

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

In this paper， the name， origin， quality evaluation， producing area and processing methods of Lablab Semen Album in the famous classical formulas were researched by reviewing the ancient materia medica， medical books， prescription books and modern literature. The results showed that the name of Lablab Semen Album in the past dynasties was mostly derived from its shape and color， called Biandou and Baibiandou. The mainstream origin used in the past dynasties was Lablab purpureus， the medicinal parts were mainly white mature seeds， with the addition of the leaves in the Song dynasty and the flowers in the Ming dynasty. Since modern times， the authentic producing areas of Lablab Semen Album are Suzhou， Zhejiang and other places， and now mainly produced in Chuxiong and Xinping， Yunnan and Panzhihua， Sichuan. The traditional quality evaluation of Lablab Semen Album is evaluated as large， solid， full and white. The harvesting time of this herb is recorded from the eighth to the ninth lunar month in related literature， the pods are picked when the seeds are ripe， and the seeds are dried in the sun. In ancient times， the processing of Lablab Semen Album mainly consisted of frying the seeds with skin and then pulverizing for use， or soaking and peeling seeds for raw use. Based on the conclusion of the textual research， it is recommended that the seeds or flowers of the white flowering plants of L. purpureus， a member of the leguminosae， should be used in the famous classical formulas， and the dried seeds or dried flowers of Lablab Semen Album can be used as medicine if the formula did not clearly indicate processing requirements.

Associations of mobile phone use with male semen quality and sex hormones / 环境与职业医学

Background Experimental studies have shown that radiofrequency electromagnetic waves emitted by mobile phones can cause adverse effects on male reproductive health, including decreased semen quality and altered sex hormones. However, the results of epidemiological studies on the relationship between mobile phone use and male semen quality are inconsistent. Furthermore, there are few epidemiological studies on the association of mobile phone use with sex hormones. Objective To explore the associations of mobile phone use with male semen quality and sex hormones. Methods A total of 2045 men visited the reproductive medicine center of a hospital in Wuhan and ordered infertility examination were recruited from December 2018 to January 2020. Information on mobile phone use was obtained using a questionnaire. Among them, 1232 and 1694 men were eligible for semen quality analyses and sex hormone analyses, respectively. Multiple linear and logistic regression models were used to analyze the associations of mobile phone use with male semen quality and sex hormones. Results After adjusting for potential confounders, there was no statistically significant associations of mobile phone use with sperm progressive motility, sperm total motility, sperm concentration, sperm count, or serum luteinizing hormone (P>0.05). However, serum total testosterone showed a declined tendency with increasing daily duration of mobile phone use (Ptrend=0.08). Compared with men with daily mobile phone use of 0-2 h, men with daily mobile phone use of 2.1-5, 5.1-8, and >8 h showed decreased serum total testosterone concentrations by 6.29% (95%CI: 0.40%-11.84%), 6.01% (95%CI: 0.60%-12.19%), and 7.87% (95%CI: 0.40%-14.79%), respectively. Conclusion Mobile phone use is not associated with male semen quality and serum luteinizing hormone, but increasing daily duration of mobile phone use is potentially associated with a tendency to lower male serum total testosterone.

Screening of active ingredients for estrogenic effect of total flavonoids from Cuscutae Semen / 中国药房

OBJECTIVE To screen the active ingredient with estrogenic effect from total flavonoids of Cuscutae Semen. METHODS The estrogenic effect of total flavonoids from 10 batches of Cuscutae Semen was evaluated with mouse uterus coefficient and endometrial thickness as evaluation indexes， establish its fingerprint and calibrate the common peak. Common peak and spectrum-effect relationship of the above two indicators were analyzed by bivariate relationship analysis and grey correlation analysis to screen active components with estrogenic effect. UPLC-Q-TOF-MS technology was used to characterize the active components. RESULTS The estrogenic effect of total flavonoids from 10 batches of Cuscutae Semen was good. Twenty-eight and thirty-three common peaks of total flavonoids in Cuscutae Semen were obtained in the positive and negative ion modes respectively. The constituents represented by peaks 7，10，12-16，26 in positive ion mode and peaks 2，5，8，9，12，16，19，22-26 in negative ion mode were highly correlated with the estrogenic effect of total flavonoids from Cuscutae Semen. Further identification showed that the active substances with estrogenic effect from the total flavonoids of Cuscutae Semen were 5，7，3′， 4′-tetramethoxyflavone， 6- O-（trans） p-coumarin-furanfructose-（2→1）-glucopyranoside， rutin， kaempferol-3，7-diglucoside， apigenin-7-O-glucoside， hyperoside， baicalin， quercitin， quercetin， apigenin， kaempferol， isorhamnetin， rhododendron， isoquercetin， kaempferol-3-furan arabinoside， 2，6-octadecanediacetylic acid. CONCLUSIONS A total of 16 chemical components with estrogenic effect are screened from total flavonoids of Cuscutae Semen in the study， which can provide reference for the development of phytoestrogens.

Effect and Mechanism of Cuscutae Semen Total Flavonoids Combined with Tripterygium wilfordii Polyglycoside Tablets on Ovarian Germline Stem Cells of Physiological Mice by Notch Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Cuscutae Semen total flavonoids combined with Tripterygium wilfordii polyglycoside tablets （TWPT） on ovarian germline stem cells of female physiological mice through neurogenic locus notch homolog （Notch） signaling pathway. MethodSixty female Kunming mice （5 weeks old） were randomly divided into normal group， Tripterygium wilfordii polyglycoside tablets low-， high-dose groups （13.65 mg·kg-1·d-1 and 27.3 mg·kg-1·d-1， 1 and 2 times clinical equivalent dose）， Cuscutae Semen total flavonoids low- and high-dose groups （150 mg·kg-1·d-1 and 300 mg·kg-1·d-1）， and combination group （13.65 mg·kg-1·d-1 TWPT and 150 mg·kg-1·d-1 Cuscutae Semen total flavonoids）， with 10 in each group. After 3 weeks of continuous administration， the uterus/brain and ovarian/brain indexes were calculated， and the pathological changes of ovarian tissue were observed under light microscope. The content of estradiol in serum was determined by enzyme linked immunosorbent assay （ELISA）. Immunofluorescence assay was performed to observe the expressions of germline stem cell markers in ovarian epithelium， including mouse vasa homologue （Mvh）， octamer-binding transcription factor 4 （Oct4）， tyrosine-protein kinase receptor （c-kit）， Nanog， Notch signaling pathway molecules， neurogenic locus notch homolog protein 1 （Notch1）， hes family BHLH transcription factor 1（Hes1）， and jagged canonical Notch ligand 1 （JAG1）. ResultCompared with the normal group， low and high doses of TWPT had no significant effect on the uterus/brain and ovary/brain indexes and the uterus and ovary morphologies of mice， while only the number of atretic follicles was increased （P<0.01）. The expressions of ovarian germline stem cell markers and Notch signaling pathway molecules had a decreasing trend in TWPT low-dose group， while the expressions of Mvh， c-kit， and Nanog were down-regulated （P<0.05， P<0.01） and the expressions of Notch1 and Hes1 were also reduced （P<0.01） in TWPT high-dose group. However， the above indexes were increased in Cuscutae Semen total flavonoids low-dose group （P<0.05， P<0.01）. Compared with the low does of TWPT group， the combination group had a decrease in the increased number of atretic follicles （P<0.01）， an improvement in the down-regulated expressions of Mvh and Nanog （P<0.01）， and an increase in the expressions of Notch1 and Hes1 （P<0.05， P<0.01）. ConclusionOvarian germline stem cells are the source target of the reproductive toxicity of TWPT. Cuscutae Semen total flavonoids participate in the regulation of the germline stem cell pathways to alleviate the reproductive toxicity caused by TWPT， and its mechanism of action may be related to the Notch signaling pathway.

Effects of the COVID-19 pandemic on semen quality in male partners of infertile couples: a hospital-based observational study / 亚洲男科学杂志(英文版)

The effects of the coronavirus disease 2019 (COVID-19) pandemic on male fertility have received considerable attention because human testes contain high levels of angiotensin-converting enzyme-2 receptors, through which severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) can enter. Early studies showed decreases in semen quality during and after recovery from COVID-19. However, no semen quality studies have examined the effects of widespread subclinical and mild disease, as well as changes in lifestyle, psychosocial behavior, intake of dietary supplements, and stress. This cross-sectional study compared semen quality parameters in male partners of infertile couples between men who underwent semen analysis before the COVID-19 pandemic (prepandemic group) and men who underwent semen analysis during the pandemic period (pandemic group); the analysis sought to clarify the overall effects of the pandemic. No participants in the pandemic group had experienced clinically overt disease. Among the 239 participants, mean body weight (P = 0.001), mean body mass index (P < 0.001), median sperm concentration (P = 0.014), total sperm count (P = 0.006), and total percentages of motile (P = 0.013) and abnormal cells (P < 0.001) were significantly greater in the pandemic group (n = 137) than those in the prepandemic group (n = 102). Among abnormal cells, the percentages of cells with excess residual cytoplasm (P < 0.001), head defects (P < 0.001), and tail defects (P = 0.015) were significantly greater in the pandemic group than those in the prepandemic group. With the exception of morphology, the overall semenogram results were better in the pandemic group than those in the prepandemic group.

Will male semen quality improve with environmental quality? / 亚洲男科学杂志(英文版)

Wenzhou has improved its environmental quality because of comprehensive environmental remediation; nevertheless, the semen quality of infertile males remains unclear. This study determined whether better environmental quality improved semen quality in this region. We recorded semen quality data from 22 962 infertile males from January 2014 to November 2019 at the Center for Reproductive Health of The First Affiliated Hospital of Wenzhou Medical University (Wenzhou, China). Patients were predominantly 30-35 years old (33.1%) and workers (82.0%), with high school education or lower (77.6%); more than a half of the patients (52.6%) were Wenzhou household registration; and most patients (77.5%) had abnormal semen quality. Patients who were older than 40 years and workers, and those with Wenzhou household registration, had significantly worse semen quality (all P < 0.05). From 2014 to 2019, progressive sperm motility, total sperm motility, and semen volume showed increasing linear trends in all patients (P = 0.021, 0.030, and 0.005, respectively), yet normal sperm morphology showed a linearly decreasing trend (P = 0.046). Sensitivity analyses for subgroups yielded similar results. In conclusion, the improvement of environmental quality and better function of the accessory glands are associated with progressive sperm motility, total sperm motility, and semen volume. Normal sperm morphology is influenced by occupational exposures and personal lifestyle and does not improve with environmental quality.

Predictors of microsurgical varicocelectomy efficacy in male infertility treatment: critical assessment and systematization / 亚洲男科学杂志(英文版)

In this review, we tried to systematize all the evidence (from PubMed [MEDLINE], Scopus, Cochrane Library, EBSCO, Embase, and Google Scholar) from 1993 to 2021 on the predictors of microsurgical varicocelectomy efficacy in male infertility treatment. Regarding the outcomes of varicocele repair, we considered semen improvement and pregnancy and analyzed them separately. Based on the 2011 Oxford CEBM Levels of Evidence, we assigned a score to each trial that studied the role of the predictor. We systematized the studied predictors based on the total points, which were, in turn, calculated based on the number and quality of studies that confirmed or rejected the studied predictor as significant, into three levels of significance: predictors of high, moderate, and low clinical significance. Preoperative total motile sperm count (TMSC) coupled with sperm concentration can be a significant predictor of semen improvement and pregnancy after varicocelectomy. In addition, for semen improvement alone, scrotal Doppler ultrasound (DUS) parameters, sperm DNA fragmentation index (DFI), and bilateral varicocelectomy are reliable predictors of microsurgical varicocelectomy efficacy.

Contamination status and exposure risk of mycotoxins in Coicis Semen / 中国中药杂志

By investigating the contamination status and predicting the exposure risk of mycotoxin in Coicis Semen, we aim to provide guidance for the safety supervision of Chinese medicinal materials and the formulation(revision) of mycotoxin limit standards. The content of 14 mycotoxins in the 100 Coicis Semen samples collected from five major markets of Chinese medicinal materials in China was determined by UPLC-MS/MS. The probability evaluation model based on Monte Carlo simulation method was established after Chi-square test and One-way ANOVA of the sample contamination data. Health risk assessment was performed on the basis of margin of exposure(MOE) and margin of safety(MOS). The results showed that zearalenone(ZEN), aflatoxin B_1(AFB_1), deoxynivalenol(DON), sterigmatocystin(ST), and aflatoxin B_2(AFB_2) in the Coicis Semen samples had the detection rates of 84%, 75%, 36%, 19%, and 18%, and the mean contamination levels of 117.42, 4.78, 61.16, 6.61, and 2.13 μg·kg~(-1), respectively. According to the limit standards in the Chinese Pharmacopoeia(2020 edition), AFB_1, AFs and ZEN exceeded the standards to certain extents, with the over-standard rates of 12.0%, 9.0%, and 6.0%, respectively. The exposure risks of Coicis Semen to AFB_1, AFB2, ST, DON, and ZEN were low, while 86% of the samples were contaminated with two or more toxins, which needs more attention. It is suggested that the research on the combined toxicity of different mycotoxins should be strengthened to accelerate the cumulative exposure assessment of mixed contaminations and the formulation(revision) of toxin limit standards.

Optimization of ethanol reflux extraction process of Ziziphi Spinosae Semen- Schisandrae Sphenantherae Fructus based on network pharmacology combined with response surface methodology / 中国中药杂志

The present study optimized the ethanol extraction process of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus drug pair by network pharmacology and Box-Behnken method. Network pharmacology and molecular docking were used to screen out and verify the potential active components of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus, and the process evaluation indexes were determined in light of the components of the content determination under Ziziphi Spinosae Semen and Schisandrae Sphenantherae Fructus in the Chinese Pharmacopoeia(2020 edition). The analytic hierarchy process(AHP) was used to determine the weight coefficient of each component, and the comprehensive score was calculated as the process evaluation index. The ethanol extraction process of Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus was optimized by the Box-Behnken method. The core components of the Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus drug pair were screened out as spinosin, jujuboside A, jujuboside B, schisandrin, schisandrol, schisandrin A, and schisandrin B. The optimal extraction conditions obtained by using the Box-Behnken method were listed below: extraction time of 90 min, ethanol volume fraction of 85%, and two times of extraction. Through network pharmacology and molecular docking, the process evaluation indexes were determined, and the optimized process was stable, which could provide an experimental basis for the production of preparations containing Ziziphi Spinosae Semen-Schisandrae Sphenantherae Fructus.

Protective Effect of Flos Puerariae，Hoveniae Semen， and Their Compatibility on Acute Alcoholic Gastric Mucosal Injury in Mice and Mechanism / 中国实验方剂学杂志

ObjectiveTo explore the improvement effect of Flos Puerariae， Hoveniae Semen， and their compatibility on acute alcoholic gastric mucosal injury， and lay a foundation for further development of Flos Puerariae， Hoveniae Semen， and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury. MethodThe acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor （15 mL·kg-1）. A total of 120 male ICR mice were randomly divided into 8 groups， namely， the blank group， model group， omeprazole group （0.026 g·kg-1）， Flos Puerariae-Hoveniae Semen （compatibility） high， medium， and low-dose groups （29.2，14.6， 7.3 g·kg-1）， Flos Puerariae group （19.5 g·kg-1）， and Hoveniae Semen group （19.5 g·kg-1）， with 15 mice in each group. After one week of adaptive feeding， the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg-1 for 3 d. From the 4th day， after 1 h of administration， Erguotou liquid was administrated at the rate of 15 mL·kg-1 and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration， the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph， and the activity of ethanol dehydrogenase （ADH） in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of nuclear transcription factor-κB （NF-κB） p65 and NF-κB inhibitory protein α （IκBα） were detected by real-time polymerase chain reaction （Real-time PCR）. ResultAs compared with the normal group， the content of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） in serum of mice in the model group was increased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues was increased （P<0.01）， and the mRNA expression of IκBα was decreased （P<0.01）. As compared with the model group， the drunkenness time of the omeprazole group， high and medium-dose compatibility groups， and Flos Puerariae group was prolonged （P<0.05）， the sober up time of the high and medium-dose compatibility groups was shortened （P<0.05）， the ethanol concentration in the serum of the high-dose compatibility group was decreased （P<0.05）， the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased （P<0.05）， the macroscopic injury score of the high， medium， and low-dose compatibility groups and Flos Puerariae group was decreased （P<0.05）， the score of pathological injury in the omeprazole group， high， medium， and low-dose compatibility groups， and Flos Puerariae group was decreased （P<0.01）， the expression of IL-6 in serum of all drug groups was decreased （P<0.05）， the expression of IL-1β in serum of the omeprazole group， high， medium， and low-dose Flos Puerariae groups， and Hoveniae Semen group was decreased （P<0.05）， the expression of TNF-α in serum of high and medium-dose groups was decreased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues of all drug groups was decreased （P<0.05）， and the mRNA expression of IκBα in gastric mucosa tissues of the omeprazole group and high， medium， and low-dose compatibility groups was increased （P<0.05）. As compared with the high-dose compatibility group， the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened （P<0.01）， the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged （P<0.01）， the concentration of ethanol in the serum of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group increased （P<0.05）， the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased （P<0.05）， the pathological injury score of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， the content of IL-1β in serum of low-dose compatibility group， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， and the mRNA expression of IκBα in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased （P<0.05）. As compared with the medium-dose compatibility group， the drunkenness time in the Hoveniae Semen group was shortened （P<0.05）， the sober up time in the Flos Puerariae group was prolonged （P<0.05）， the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased （P<0.01）， and the content of IL-1β in serum of the low-dose compatibility group， the Flos Puerariae group， and Hoveniae Semen group was increased （P<0.05）. As compared with the low-dose compatibility group， the pathological injury score of the Hoveniae Semen group was increased （P<0.05）. ConclusionFlos Puerariae， Hoveniae Semen， and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice， which may be related to the inhibition of the expression of NF-κB signal pathway in gastric mucosa， and the high-dose compatibility group has the optimal effect.