[Identification and visual analysis of ginsenosides in multi-steamed roots of Panax quinquefolium based on UPLC-Q-TOF-MS/MS and MALDI-MSI].

This study aims to investigate the component variations and spatial distribution of ginsenosides in Panax quinquefolium roots during repeated steaming and drying. Ultra performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) was employed to identify the ginsenosides in the root extract. Matrix-assisted laser desorption/ionization mass spectrometry imaging(MALDI-MSI) was employed to visualize the spatial distribution and spatiotemporal changes of prototype ginsenosides and metabolites in P. quinquefolium roots. The UPLC results showed that 90 ginsenosides were identified during the steaming process of the roots, and polar ginsenosides were converted into low polar or non-polar ginsenosides. The content of prototype ginsenosides decreased, while that of rare ginsenosides increased, which included 20(S/R)-ginsenoside Rg_3, 20(S/R)-ginsenoside Rh_2, and ginsenosides Rk_1, Rg_5, Rs_5, and Rs_4. MALDI-MSI results showed that ginsenosides were mainly distributed in the epidermis and phloem. As the steaming times increased, ginsenosides were transported to the xylem and medulla. This study provides fundamental information for revealing the changes of biological activity and pharmacological effect of P. quinquefolium roots that are caused by repeated steaming and drying and gives a reference for expanding the application scope of this herbal medicine.

Identification of raffinose family oligosaccharides in processed Rehmannia glutinosa Libosch using matrix-assisted laser desorption/ionization mass spectrometry image combined with machine learning.

RATIONALE: Currently, research on oligosaccharides primarily focuses on the physiological activity and function, with a few studies elaborating on the spatial distribution characterization and variation in the processing of Rehmannia glutinosa Libosch. Thus, imaging the spatial distributions and dynamic changes in oligosaccharides during the steaming process is significant for characterizing the metabolic networks of R. glutinosa. It will be beneficial to characterize the impact of steaming on the active ingredients and distribution patterns in different parts of the plant. METHODS: A highly sensitive matrix-assisted laser desorption/ionization mass spectrometry image (MALDI-MSI) method was used to visualize the spatial distribution of oligosaccharides in processed R. glutinosa. Furthermore, machine learning was used to distinguish the processed R. glutinosa samples obtained under different steaming conditions. RESULTS: Imaging results showed that the oligosaccharides in the fresh R. glutinosa were mainly distributed in the cortex and xylem. As steaming progressed, the tetra- and pentasaccharides were hydrolyzed and diffused gradually into the tissue section. MALDI-MS profiling combined with machine learning was used to identify the processed R. glutinosa samples accurately at different steaming intervals. Eight algorithms were used to build classification machine learning models, which were evaluated for accuracy, precision, recall, and F1 score. The linear discriminant analysis and random forest models performed the best, with prediction accuracies of 0.98 and 0.97, respectively, and thus can be considered for identifying the steaming durations of R. glutinosa. CONCLUSIONS: MALDI-MSI combined with machine learning can be used to visualize the distribution of oligosaccharides and identify the processed samples after steaming for different durations. This can enhance our understanding of the metabolic changes that occur during the steaming process of R. glutinosa; meanwhile, it is expected to provide a theoretical reference for the standardization and modernization of processing in the field of medicinal plants.

Adjuvant activity of tubeimosides by mediating the local immune microenvironment.

Rhizoma Bolbostemmatis, the dry tuber of Bolbostemma paniculatum, has being used for the treatment of acute mastitis and tumors in traditional Chinese medicine. In this study, tubeimoside (TBM) I, II, and III from this drug were investigated for the adjuvant activities, structure-activity relationships (SAR), and mechanisms of action. Three TBMs significantly boosted the antigen-specific humoral and cellular immune responses and elicited both Th1/Th2 and Tc1/Tc2 responses towards ovalbumin (OVA) in mice. TBM I also remarkably facilitated mRNA and protein expression of various chemokines and cytokines in the local muscle tissues. Flow cytometry revealed that TBM I promoted the recruitment and antigen uptake of immune cells in the injected muscles, and augmented the migration and antigen transport of immune cells to the draining lymph nodes. Gene expression microarray analysis manifested that TBM I modulated immune, chemotaxis, and inflammation-related genes. The integrated analysis of network pharmacology, transcriptomics, and molecular docking predicted that TBM I exerted adjuvant activity by interaction with SYK and LYN. Further investigation verified that SYK-STAT3 signaling axis was involved in the TBM I-induced inflammatory response in the C2C12 cells. Our results for the first time demonstrated that TBMs might be promising vaccine adjuvant candidates and exert the adjuvant activity through mediating the local immune microenvironment. SAR information contributes to developing the semisynthetic saponin derivatives with adjuvant activities.

Dietary exposure and risk assessment of chlorinated paraffins in roots and rhizomes of traditional Chinese medicine herbs.

Traditional Chinese medicine (TCM) provides therapeutic and health care effects through dietary intake. Owing to the susceptibility of plants to contaminations, a risk assessment system is urgently needed to ensure the safe use of TCMs. In this study, the contamination levels and risks associated with the dietary intake of short-chain chlorinated paraffins (SCCPs) and medium-chain chlorinated paraffins (MCCPs) were investigated in six kinds of frequently-used TCM herbs. The concentrations varied from 144.4 to 1527.8 ng·g-1 dw for SCCPs and non-detect to 1214.1 ng·g-1 dw for MCCPs, with mean values of 551.5 and 259.8 ng·g-1 dw, respectively. A geographic distribution analysis indicated that the concentrations of CPs in TCMs were mainly associated with their levels of contamination in the ambient environment. Carbon atom-chlorine congener profiles of CPs were dominated by C10Cl7-8 and C14Cl7-8 congeners, accounting for 20.1% and 32.4% of the total SCCP and MCCP concentrations, respectively. Principal component analysis indicated that the TCM species might be the main factor influencing the accumulation of SCCPs congeners. Finally, a risk assessment reveals that the estimated daily intake and margin of exposure were far below levels that might pose a health risk, indicating an acceptable dietary intake of SCCPs and MCCPs in the studied TCMs. This is the first report of CPs in the TCM herbs and the obtained results are expected to aid in future evaluation of the quality of TCMs and ensuring diet and drug safety.

Analysis of dietary exposure and risk assessment of pesticide residues in roots and rhizomes of Chinese herbs.

Medicine food homologous (MFH) plants provide therapeutic and health care effects through diet. Thus, a risk assessment system for hazardous ingredient residues is urgently required to ensure their safe use. In this study, the pesticide contamination of six root and rhizome Chinese herbs, Ginseng Radix et Rhizoma, Panacis Quinquefolii Radix, Pseudostellariae Radix, Salviae Miltiorrhizae Radix et Rhizoma, Codonopsis Radix, and Glehniae Radix, and the risks associated with their intake were investigated. A total of 420 MFH plant samples collected from 22 provinces in China were tested, and 61 pesticides were detected in 413 samples. Multiple pesticide residues were detected in each MFH sample, with contents ranging from 0.0002 to 3.010 mg/kg dry weight. Carbendazim (≥47.14%) and propham (≥40%) were the most frequently detected pesticides. Risk assessment determined by hazard quotients indicated that the risks were acceptable, with no short- or long-term adverse health effects. However, considering the high incidence of residues and the detection of unregistered or even prohibited pesticides, strict supervision of soil quality and pesticide application (particularly cadusafos) in MFH plant cultivation are recommended to aid in monitoring MFH plant quality and ensuring diet and drug safety. PRACTICAL APPLICATION: Ensure the diet and drug safety of Chinese herbs.

Mass spectrometry investigation of nucleoside adducts of fatty acid hydroperoxides from oxidation of linolenic and linoleic acids.

The widespread presence of lipid hydroperoxides in foodstuffs and biological samples has aroused great attentions in recent years, while it remains challenging for analysis of the fragility of O - O bond linkage of peroxides. In this present study, we explored the utility of electrospray ionization mass spectrometry (ESI-MS) for characterization of two fatty acid hydroperoxides from oxidation of linoleic acid and α-linolenic acid, which are the essential fatty acids abundant in many seeds and vegetable oils. The results indicated that in-source fragmentation occurred in the detection of the two fatty acid hydroperoxides in both positive and negative ion modes, which yielded characteristic fragments for ESI-MS analysis. In addition, the genotoxicity of fatty acid hydroperoxides for generation of nucleoside adducts was investigated. It was found that a variety of nucleoside adducts were formed from the reactions of fatty acid hydroperoxides and nucleosides. Furthermore, the decomposition products of the fatty acid hydroperoxides were determined, which provided evidence to elucidate the reaction mechanism for formation of nucleoside adducts.

Integrated Proteomics and Metabolomics Assessment Indicated Metabolic Alterations in Hypothalamus of Mice Exposed to Triclosan.

Triclosan (TCS) is a ubiquitous antimicrobial used in many daily consumer products. It has been reported to induce endocrine disrupting effects at low doses in mammals, disturbing sex hormone function and thyroid function. The hypothalamus plays a crucial role in the maintenance of neuroendocrine function and energy homeostasis. We speculated that the adverse effects of TCS might be related to the disturbance of metabolic processes in hypothalamus. The present study aimed at investigating the effects of TCS exposure on the protein and metabolite profiles in hypothalamus of mice. Male C57BL/6 mice were orally exposed to TCS at the dosage of 10 mg/kg/d for 13 weeks. The hypothalamus was isolated and processed for mass spectrometry (MS)-based proteomics and metabolomics analyses. The results showed that a 10.6% decrease (P = 0.066) in body weight gain was observed in the TCS exposure group compared with vehicle control group. Differential analysis defined 52 proteins and 57 metabolites that delineated TCS exposed mice from vehicle controls. Among the differential features, multiple proteins and metabolites were found to play vital roles in neuronal signaling and function. Bioinformatics analysis revealed that these differentially expressed proteins and metabolites were involved in four major biological processes, including glucose metabolism, purine metabolism, neurotransmitter release, and neural plasticity, suggesting the disturbance of homeostasis in energy metabolism, mitochondria function, neurotransmitter system, and neuronal function. Our results may provide insights into the neurotoxicity of TCS and extend our understanding of the biological effects induced by TCS exposure.

Metabolomic approach for rapid differentiation of Fritillaria bulbs by matrix-assisted laser desorption/ionization mass spectrometry and multivariate statistical analysis.

The bulbs of Fritillaria have been used for centuries as food and medicinal products in many Asian countries. Different Fritillaria species have distinct pharmacological effects despite of their similar appearances. Effective differentiation of Fritillaria species can avoid adulteration and is crucial to its clinical uses. In this paper, a hybrid method of matrix assisted laser desorption/ionization mass spectrometry and multivariate statistical analysis was developed for the rapid and reliable differentiation of Fritillaria species for the first time. Significantly different patterns for five Fritillaria species were obtained by MALDI-MS after instant sample extractions. Different groups of Fritillaria were confidently differentiated via an orthogonal partial least square model. In addition, a metabolomic taxonomy of five Fritillaria species was obtained based on MALDI-MS data.

Activation of RAW264.7 macrophages by active fraction of Albizia julibrissin saponin via Ca2+-ERK1/2-CREB-lncRNA pathways.

The saponin active fraction from the stem bark of Albizia julibrissin (AJSAF) is an ideal vaccine adjuvant, but its mechanism of action remains unclear. The recent evidences indicate that long noncoding RNAs (lncRNAs) play essential roles in regulating the activation and function of macrophages. The current experiments were designed to investigate the effects of AJSAF on the activation of RAW264.7 macrophages and to explore its intracellular molecular mechanisms using a global gene expression microarray. AJSAF could significantly enhance phagocytic activity, induce reactive oxygen species (ROS), promote surface molecule expression, and up-regulate the mRNA and protein expression of cytokines and chemokines in RAW264.7 cells. AJSAF induced the differential expression of 223 mRNAs and 103 lncRNAs in RAW264.7 cells. Bioinformatics were used to predict the potential target mRNAs and function of up-regulated lncRNA A_30_P01018532 in RAW264.7 cells induced by AJSAF. The total 99 co-expressed mRNAs were classified as putative target genes of A_30_P01018532. A_30_P01018532 was associated with the inflammatory and immune response. AJSAF significantly increased the intracellular free Ca2+ levels and induced the phosphorylation of ERK1/2 and CREB in RAW264.7 cells. Moreover, Ca2+ chelator BAPTA-AM, ERK1/2 inhibitor PD98059 and CREB inhibitor KG-501 significantly inhibited the up-regulation of TNF-α, CCL2, CXCL2, CCL22, and A_30_P01018532 in RAW264.7 cells induced by AJSAF. These results suggested that AJSAF could activate RAW264.7 cells via Ca2+-ERK1/2-CREB pathways and that A_30_P01018532 might be an important regulator of mRNA expression in AJSAF-activated macrophage. This study may provide insights into the molecular mechanisms of action of AJSAF.

Correlation between methylenetetrahydrofolate reductase gene polymorphism and oligoasthenospermia and the effects of folic acid supplementation on semen quality.

BACKGROUND: The present study investigated the correlation between 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism and oligoasthenospermia, as well as the effects of folic acid supplementation on semen quality. METHODS: The present study was a case control study. The PCR-chip assay was applied to analyze the distribution characteristics of the frequencies and genotypes of the MTHFR C677T allele in 167 Han Chinese patients with idiopathic male infertility (including 86 patients with oligospermia and 81 patients with asthenospermia) and in 78 males with normal semen parameters. Moreover, homocysteine (Hcy) levels were assessed for the different groups. Semen quality was measured following three months of folic acid supplementation for the oligospermia and asthenospermia groups. RESULTS: The cytosine-thymine (CT) genotype (50% vs. 39.5%) and the thymine-thymine (TT) genotype (51.2% vs. 7.7%) carriers in the oligospermia group exhibited significantly higher percentages compared with those of the control group. The percentage of the CT genotype carriers in the asthenospermia group was significantly higher compared with that of the control group (59.3% vs. 50%), while the frequency of the TT genotype was significantly increased (22.2% vs. 7.7%). Furthermore, serum Hcy levels in the oligospermia and asthenospermia groups were significantly higher compared with those of the control group. The data also demonstrated that sperm density increased significantly following three months of folic acid supplementation to patients with oligospermia or asthenospermia. In these patients, the highest increase was noted for the subjects carrying the TT genotype. CONCLUSIONS: The MTHFR C677T mutation and the elevated Hcy levels are important risk factors for the development of oligoasthenospermia. Folic acid supplementation can significantly improve sperm density.

Role of ginsenosides in reactive oxygen species-mediated anticancer therapy.

Cancer is still a global public health problem, which is the leading cause of death in most countries. Ginseng has been used for centuries all over the world as a panacea that promotes longevity. As the king of herb plants, ginseng holds great promise as a new treatment option which is used either by itself or in combination with other medicinal ingredients that is widely accepted as complementary and alternative medicine in cancer therapy. Ginsenosides, the major pharmacologically active ingredients of ginseng, have been shown to have multiple medicinal effects including prominent anticancer activity. The purpose of this review is to give our perspective about the roles of ginsenosides in reactive oxygen species (ROS)-mediated anticancer therapy. Additionally, to provide new sheds light for further improvement and carry out pre-clinical and clinical trials to develop it successfully into a potential anticancer agent. Panax herbs and their derivate/metabolites ginsenosides exert beneficial effects for treating various types of cancers. The mechanism of ROS-mediated anticancer activities of ginsenosides varies depending on the specific type of cancer cells involved. Ginsenosides may suppress cancer cell proliferation through anti-oxidation on tumor initiation and induce apoptosis, paraptosis or autophagy via generation of ROS on tumor progression, promotion, angiogenesis, invasion and metastasis by various signaling pathways e.g., activation of AMPK, MEK, ASK-1/JNK, ESR2-NCF1-ROS, ER-dependent PI3K/Akt/Nrf2, P53-CHOP, ROS-JNK-autophagy, and/or inhibition of PI3K/Akt signaling pathways. These multiple effects rather than a single may play a crucial role in emerging ginsenosides as a successful anticancer drug.

Pathway analysis of global gene expression change in dendritic cells induced by the polysaccharide from the roots of Actinidia eriantha.

ETHNOPHARMACOLOGICAL RELEVANCE: The roots of Actinidia eriantha Benth have been used clinically to treat a variety of cancers in traditional Chinese medicine. The polysaccharide from this drug (AEPS) was previously reported to be a potential antitumor agent with immunomodulatory activity. However, the mechanisms of its antitumor action in immunomodulation have not yet been well-defined. AIM OF THE STUDY: To investigate the effects of AEPS on the phenotypic and functional maturation of dendritic cells and to explore the intracellular signaling mechanisms of its antitumor action in the immunomodulation. MATERIALS AND METHODS: The effects of AEPS on the phagocytic activity, expression of surface molecules, mRNA and protein expression levels of cytokines and chemokines in mouse bone-marrow derived dendritic cells (BMDCs) were determined by flow cytometry, qRT-PCR and ELISA, respectively. The transcriptional profile induced by AEPS was established using oligonucleotide microarray, and Ingenuity Pathway Analysis (IPA) was used to identify potential signaling pathways. Western blotting, neutralization experiments and inhibition assay were performed to confirm signaling pathway involved in maturation of DCs induced by AEPS. Furthermore, we discussed the downstream effects of the action of AEPS using clustering, network and pathway mapping approaches. RESULTS: AEPS could significantly reduced phagocytic activity, promoted expression of accessory and co-stimulatory molecules, and up-regulated the mRNA and protein expression levels of cytokines and chemokines in BMDCs. Microarray assay revealed that AEPS induced significantly differential expression of 452 genes including up-regulated cytokines (IL-6, IL-1ß, TNF-α, IL-10, IL-12p40, IFN-ß and IFN-γ), chemokines (MIP-1α, MIP-1ß, CCL5, MDC and MCP-1), transcription factors (STAT1, STAT2, STAT5b, IRF1 and IRF7) and pattern recognition receptors (TLR3, DDX58, DHX58 and IFIH1) in the BMDCs. AEPS-induced production of TNF-α and IL-12p40 from BMDCs was inhibited by antibodies against TLR2 and TLR4. Furthermore, AEPS induced the phosphorylation of NF-κB p65 in a time-dependent manner, and BAY 11-7082, an inhibitor of NF-κB, remarkably suppressed the production of cytokines induced by AEPS. CONCLUSION: AEPS triggers the phenotypic and functional maturation of DCs via TLR2/4 and NF-κB signaling pathway, resulting in augmented antitumor immune responses. Our results suggested that AEPS might be helpful in potentiating the efficiency of DC-based cancer immunotherapy. This study further expanded current knowledge on the mechanisms of antitumor action of AEPS.

Activation of RAW264.7 macrophages by the polysaccharide from the roots of Actinidia eriantha and its molecular mechanisms.

The polysaccharide from the roots of Actinidia eriantha (AEPS), a potent antitumor agent and immunological adjuvant, was investigated for the immunomodulatory effects on RAW264.7 macrophages and its molecular mechanisms. AEPS could significantly enhance the pinocytic and phagocytic activity, induce the production of NO, TNF-α, IL-10, IL-1ß and IL-6, and promote the expression of accessory and costimulatory molecules in RAW264.7 cells. PCR array assay revealed that AEPS up-regulated 28 genes including TLRs (TLR2, TLR8, TLR9), proinflammatory factors (IL-1ß, G-CSF, IL-1α, GM-CSF, IL-6, COX-2, TNF-α, IFN-ß, CXCL10, CCL2, TNF-ß, IL-10), and the genes involved in NF-κB signaling pathway, and down-regulated 6 genes such as TLR3, TLR4, PGLYRP1, EIF2αK2, MAP3K1 and IRF1. AEPS was further showed to promote cytoplasmic IκB-α degradation and increase nuclear NF-κB p65 levels in RAW264.7 cells. These results suggested that AEPS activated RAW264.7 macrophages and elicited a M1 and M2 response through TLRs/NF-κB signaling pathway.

The Inhibitory Effect of 3 ß -Hydroxy-12-oleanen-27-oic Acid on Growth and Motility of Human Hepatoma HepG2 Cells through JNK and Akt Signaling Pathway.

3 ß -Hydroxy-12-oleanen-27-oic acid (ATA) was a main antitumor active triterpene from the rhizomes of Astilbe chinensis. In this study, we investigated its effects on growth, apoptosis, cell cycle, motility/invasion, and metatasis in human hepatoma HepG2 cells in vitro and antimetastasis of B16-F10 melanoma in mice in vivo, as well as its molecular mechanisms of action using a high-throughput Cancer Pathway Finder PCR Array. ATA could not only induce tumor cells into apoptosis through the activation of both extrinsic and intrinsic pathways, arrest HepG2 cells in G2/M phase, but also suppress the invasion and metastasis abilities of HepG2 cells and the lung metastasis of B16-F10 melanoma in mice. PCR array assay revealed that ATA upregulated 9 genes including CDKN1A, MDM2, CFLAR (CASPER), TNFRSF10B (DR5), c-Jun, IL-8, THBS1, SERPINB5 (maspin), and TNF and downregulated 8 genes such as CCNE1, AKT, ANGPT1, TEK, TGFBR1, MMP9, U-PA, and S100A4. These results indicate that ATA could exert antitumor effects through activating JNK/MAPK and suppressing AKT signal transduction pathways and that ATA might be a potent anticancer agent.

Ex vivo relaxation effect of Cuscuta chinensis extract on rabbit corpus cavernosum.

The effect of Cuscuta chinensis extract on the rabbit penile corpus cavernosum (PCC) was evaluated in the present study. Penises obtained from healthy male New Zealand white rabbits (2.5-3.0 kg) were precontracted with phenylephrine (Phe, 10 µmol l(-1)) and then treated with various concentrations of Cuscuta chinensis extract (1, 2, 3, 4 and 5 mg ml(-1)). The change in penile tension was recorded, and cyclic nucleotides in the PCC were measured by radioimmunoassay (RIA). The interaction between Cuscuta chinensis and sildenafil was also evaluated. The result indicated that the PCC relaxation induced by Cuscuta chinensis extract was concentration-dependent. Pre-treatment with an nitric oxide synthase (NOS) inhibitor (Nω nitro-L-arginine-methyl ester, L-NAME), a guanylyl cyclase inhibitor (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, ODQ), or a protein kinase A inhibitor (KT 5720) did not completely inhibit the relaxation. Incubation of penile cavernous tissue with the Cuscuta chinensis extract significantly increased cyclic guanosine monophosphate (cGMP) and cyclic adenosine monophosphate (cAMP) in the PCC. Moreover, the Cuscuta chinensis extract significantly enhanced sildenafil-induced PCC relaxation. In conclusion, the Cuscuta chinensis extract exerts a relaxing effect on penile cavernous tissue in part by activating the NO-cGMP pathway, and it may improve erectile dysfunction (ED), which does not completely respond to sildenafil citrate.

[Efficacy of natural vitamin E on oligospermia and asthenospermia: a prospective multi-centered randomized controlled study of 106 cases].

OBJECTIVE: To explore the therapeutic effect of natural vitamin E (VitE) on oligospermia and asthenospermia in in- fertile men. METHODS: We conducted a prospective multi-centered randomized controlled study on 64 infertile men with oligospermia (31 as controls treated with Tamoxifen 10 mg bid and 33 as experimental cases treated with Tamoxifen 10 mg bid + VitE 100 mg tid) and 42 cases of asthenospermia (20 as controls treated with Levocarnitine oral solution 1 bottle bid and 22 as experimental cases treated with Levocarnitine oral solution 1 bottle bid + VitE 100 mg tid). We compared the control and experimental groups in sperm concentration and percentage of progressively motile sperm before and 3 months after medication, as well as the rate of clinical pregnancy and adverse events. RESULTS: Among the oligospermia patients, the average sperm concentrations in the control and experimental groups were 8.00 x 10(6)/ml and 10.66 x 10(6)/ml before medication (P > 0.05). After medication, the numbers of cases evaluated as with no, slight, moderate and marked improvement in sperm concentration were 10 and 9 (P > 0.05), 16 and 14 (P > 0.05), 5 and 4 (P > 0.05) and 0 and 0 (P >0.05); and the numbers of natural pregnancies were 0 and 6 in the control and experimental groups (P < 0.01). Among the asthenospermia patients, the average rates of progressively motile sperm were 17.00% and 18.10% in the control and experimental groups before medication (P > 0.05). After medication, the numbers of cases evaluated as with no, slight, moderate and marked improvement in the percentage of progressively motile sperm were 7 and 2 (P < 0.01), 4 and 8 (P < 0.01), 3 and 2 (P > 0.05) and 1 and 1 (P > 0.05), and the numbers of natural pregnancies were 5 and 9 in the two groups (P < 0.01), but no adverse events were observed. CONCLUSION: As a safe and effective adjuvant agent for the treatment of oligospermia and asthenospermia, vitamin E can improve sperm concentration, the percentage of progressively motile sperm, and finally the rate of natural pregnancy.