ObjectiveTo explore the effect of Yishen Tongluo prescription on sperm DNA fragmentation index （DFI） and sperm mitochondrial membrane potential （MMP） in patients with asymptomatic idiopathic asthenospermia infertility. MethodsA total of 128 patients with asymptomatic idiopathic asthenospermia were randomly assigned to an experimental group （64 cases） and a control group （64 cases）. The experimental group received Yishen Tongluo prescription， while the control group was treated with Wuzi Yanzongwan combined with L-carnitine oral solution. One treatment course lasted 12 weeks. Spouse pregnancy rate， sperm progressive motility （PR）， total sperm motility （PR+NP）， sperm function （sperm tail hypotonic swelling rate， sperm acrosin activity）， sperm DFI， and sperm MMP were compared between the two groups before and after treatment. Adverse reactions were observed and recorded during the study， and clinical efficacy and safety were systematically evaluated. ResultsA total of 121 patients completed the study， including 61 in the experimental group and 60 in the control group. The spouse pregnancy rate in the experimental group was 14.75% （9/61）， higher than that in the control group at 6.67% （4/60）， though the difference was not statistically significant. Clinical efficacy in the experimental group was superior to that in the control group （P<0.05）. Compared with the results before treatment， sperm PR， PR + NP， sperm tail hypotonic swelling rate， sperm acrosin activity， sperm DFI， and sperm MMP were significantly improved in both groups after treatment （P<0.05）， with greater improvements in the experimental group （P<0.05）. However， there was no significant change in sperm concentration in either group after treatment. During the study， no abnormal safety indicators or significant adverse reactions occurred in either group. ConclusionThe kidney-tonifying and collateral-dredging method shows good clinical efficacy in the treatment of asymptomatic idiopathic asthenospermia infertility. Yishen Tongluo prescription can improve sperm motility， increase spouse pregnancy rate， enhance sperm function， and demonstrates good safety. Its mechanism may be related to reducing sperm DFI and increasing sperm MMP.

OBJECTIVE: To observe the clinical efficacy of heat-sensitive moxibustion combined with western medication for preconception intervention in prethrombotic state of recurrent spontaneous abortion (RSA) with kidney deficiency and blood stasis. METHODS: A total of 100 RSA patients of prethrombotic state with kidney deficiency and blood stasis were randomized into a combination group (50 cases, 5 cases were eliminated) and a medication group (50 cases, 5 cases were eliminated). In the medication group, the aspirin enteric-coated tablet was given orally at a dose of 75 mg a time, once daily. On the basis of the treatment in the medication group, in the combination group, heat-sensitive moxibustion was applied at the heat-sensitive points selected among the areas of Guanyuan (CV4), Shenque (CV8), and bilateral Sanyinjiao (SP6), Zusanli (ST36), Qihai (CV6), Taixi (KI3), Zigong (EX-CA1), Luanchao (Extra), Xuehai (SP10), and Yinlingquan (SP9), about 40 min a time, once every two days. Both groups were treated for 3 menstrual cycles continuously. Pregnancy success rate of 12 weeks was recorded in the two groups in follow-up of 3 months after treatment completion, during which conception was tried under the guidance of doctor. The TCM symptom score was observed and the coagulation-fibrinolysis indexes (activated partial thromboplastin time [APTT], prothrombin time [PT], platelet count [PLT], D-dimer [D-D], fibrinogen [FIB], protein S [PS], protein C [PC] and antithrombin Ⅲ [AT-Ⅲ]) were detected before and after treatment in the two groups. RESULTS: The pregnancy success rate of 12 weeks was 80.0% (32/40) in the combination group, which was higher than 54.3% (19/35) in the medication group (P<0.05). After treatment, the TCM symptom scores were decreased compared with those before treatment in the two groups (P<0.05), and the TCM symptom score in the combination group was lower than that in the medication group (P<0.05). Compared before treatment, the APTT and PT was prolonged (P<0.05), the levels of PLT, FIB and D-D were reduced (P<0.05), the activity of AT-Ⅲ, PS and PC was increased (P<0.05) after treatment in the two groups. After treatment, in the combination group, the APTT was longer (P<0.05), the levels of PLT, FIB and D-D were lower (P<0.05), the activity of AT-Ⅲ, PS and PC was higher (P<0.05) than those in the medication group. CONCLUSION Heat-sensitive moxibustion combined with western medication can effectively improve the prethrombotic state and TCM clinical symptoms in RSA patients with kidney deficiency and blood stasis, enhance pregnancy success rate, its mechanism may be related to ameliorating hypercoagulability.
Humans ; Female ; Moxibustion ; Adult ; Pregnancy ; Abortion, Habitual/blood* ; Young Adult ; Combined Modality Therapy ; Kidney/drug effects* ; Acupuncture Points ; Aspirin/administration & dosage*

OBJECTIVE: To compare the clinical efficacy of herbal cake-separated moxibustion at Baliao acupoints combined with western medication versus western medication alone in treating erectile dysfunction (ED) with kidney deficiency and blood stasis. METHODS: A total of 108 ED patients were randomly divided into a combination group (54 cases, 2 cases dropped out) and a western medication group (54 cases, 1 case was eliminated, 2 cases dropped out). The western medication group received oral tadalafil tablets 5 mg a time, once daily. The combination group received additional herbal cake-separated moxibustion at Baliao acupoints (Shangliao [BL31], Ciliao [BL32], Zhongliao [BL33], Xialiao [BL34]) twice weekly (administered on Tuesdays and Fridays). Both groups were treated for 4 weeks. The 5-question international index of erectile function (IIEF-5) score, erectile hardness assessment (EHS) score, TCM syndrome score, serum sex hormone levels (follicle-stimulating hormone [FSH], luteinizing hormone [LH], prolactin [PRL], testosterone [T]), penile hemodynamic parameters [peak systolic velocity (PSV) of penile cavernosal artery, end-diastolic velocity (EDV), resistance index (RI)] were compared before and after treatment between the two groups, and the safety and clinical efficacy were evaluated. RESULTS: After treatment, the IIEF-5 scores, EHS scores, serum T levels, PSV and RI of penile cavernosal artery were increased compared with those before treatment in the two groups (P<0.05). Except for RI, the combination group exhibited significantly higher improvements in the above indexes than those in the western medication group (P<0.05). After treatment, the TCM syndrome scores, the serum FSH, LH levels, and EDV in the two groups and the serum PRL level in the combination group were decreased compared with those before treatment (P<0.05), while the above indexes in the combination group were lower than those in the western medication group (P<0.05). The total effective rate of the combination group was 88.5% (46/52), which was superior to 72.5% (37/51) in the western medication group (P<0.05). No significant adverse reactions occurred in either group. CONCLUSION Herbal cake-separated moxibustion at Baliao acupoints combined with western medicine can regulate sex hormone levels, improve penile blood supply, enhance erectile function and hardness, and is superior to western medication alone, with no observed adverse reactions.
Humans ; Male ; Moxibustion ; Erectile Dysfunction/drug therapy* ; Middle Aged ; Adult ; Acupuncture Points ; Drugs, Chinese Herbal/administration & dosage* ; Kidney/physiopathology* ; Aged ; Follicle Stimulating Hormone/blood* ; Luteinizing Hormone/blood* ; Treatment Outcome

AIM:To investigate the protective effect of Yishen-Tongluo prescription(YTP)against oxidative stress-induced senescence of renal tubular epithelial cells via phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB/AKT)/mammalian target of rapamycin(mTOR)pathway in vitro,and to explore its potential mechanism.METHODS:Human renal tubular epithelial HK-2 cells were cultured in vitro and exposed to 400 μmol/L H2O2 to establish a cellular se-nescence model.The cells were treated with 1 g/L YTP and/or 1 μmol/L 740 Y-P(PI3K activator)for 48 h,and were di-vided into control group,H2O2 group,YTP group,YTP+H2O2 group,740 Y-P+H2O2 group and 740 Y-P+YTP+H2O2 group.Senescent cells were analyzed by senescence-associated β-galactosidase(SA-β-Gal)staining.The protein levels of P21,P16,γ-H2AX,microtubule-associated protein 1 light chain 3B(LC3B),P62,PI3K,p-PI3K,AKT,p-AKT,mTOR,p-mTOR,P70S6K and p-P70S6K were determined by Western blot.The proliferation of the cells was detected by EdU staining.Intracellular level of reactive oxygen species(ROS)was assessed by DCFH-DA probe,and laser confocal microscopy was used to observe autophagic flux in HK-2 cells.Each experiment was repeated 3 times.RESULTS:(1)Compared with control group,the SA-β-Gal-positive cells in H2O2 group were significantly increased,the EdU-positive cells were significantly reduced,and the protein levels of P21,P16 and γ-H2AX were significantly elevated(P<0.01).The ratios of p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR and p-P70S6K/P70S6K were significantly increased(P<0.05).The protein level of LC3B-Ⅱ was incresed,the P62 expression was reduced(P<0.01),and the ratio of autolyso-somes to autophagosomes increased(P<0.05).(2)Compared with H2O2 group,the intracellular ROS level in YTP+H2O2 group significantly decreased(P<0.01).The SA-β-Gal-positive cells were significantly reduced,the EdU-positive cells were significantly increased,and the protein levels of P21,P16 and γ-H2AX were decreased(P<0.05).The ratios of p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR and p-P70S6K/P70S6K were significantly reduced(P<0.05).The protein lev-el of LC3B-Ⅱ was increased,the P62 expression was reduced(P<0.05),and the ratio of autolysosomes to autophago-somes decreased(P<0.05).(3)Compared with YTP+H2O2 group,the ratios of p-PI3K/PI3K,p-AKT/AKT,p-mTOR/mTOR and p-P70S6K/P70S6K,and the protein level of LC3B-Ⅱ were decreased in 740 Y-P+YTP+H2O2 group(P<0.01),while the P62 expression increased(P<0.05).CONCLUSION:The YTP can clear accumulated intracellular ROS,res-cue autophagic flux by inhibiting the PI3K/AKT/mTOR signaling pathway,thus alleviating oxidative stress-induced HK-2 cell senescence.

Environmental pollutants generated by industrialization,characterized by microplastics,are increasingly impairing male fertility in the form of"environmental hidden toxins".Although the concept of environmental toxicity has been proposed for a long time,its practical application remains limited.Therefore,this study innovatively introduces and elaborates on the concept of"environmental hidden toxins",highlighting its pathogenic characteristics,including latency,turbidity,consumptive nature,concurrent nature,and transmutation.Environmental hidden toxin can be classified into five categories based on the properties:stagnation toxins,scorching toxins,turbid toxins,desiccating toxins,and latent toxins.The core pathogenesis of environmental hidden toxins-induced male infertility involves three stages:firstly,invading the lungs and spleen,leading to the gradual depletion of healthy qi;secondly,forming toxin-stasis complexes that obstruct the spermatic pathway;and lastly,penetrating deeply into the seminal chamber,directly damaging the genuine essence.The treatment principle emphasizes"strengthening the foundation and clearing the source,"advocating dual strategies of detoxification and strengthening vital qi.This includes enhancing clarity and lowering turbidity,fortifying the earth to generate metal to replenish healthy qi;resolving stasis,unblocking collaterals,and dispelling toxins to eliminate pathogenic factors;and nourishing the kidneys,replenishing essence,and expelling toxins to preserve the genuine essence.All the above approaches form a holistic traditional Chinese medicine(TCM)strategy that treating both manifestation and root cause of disease.The study provides theoretical foundations and offers novel clinical insights into TCM interventions for male infertility caused by environmental pollutants.

Objective : To explore the effect and mechanism of total saponins of astragalus membranaceus combined with bone marrow mesenchymal stem cells (BMSCs) regulating the stromal cell-derived factor-1 (SDF-1) / chemo- kine receptor 4 ( CXCR4) pathway in the treatment of autoimmune premature ovarian failure ( POF) model rats． Methods : Rats were randomly divided into POF group，total saponins of astragalus membranaceus group，BMSCs group ，combination group ，combination + SDF-1 / CXCR4 pathway inhibitor ( AMD3100 ) group ，and control group，with 12 rats in each group．Except for the control group，all other groups of rats needed to construct a POF model．After successful modeling，treatment would begin．The drug injected into the tail vein once a week，and the drug injected into the abdominal cavity once a day for 4 weeks．ELISA was applied to detect the levels of anti ovari- an antibodies (AOAB) ，β-endorphin ( β-EP) ，vascular endothelial growth factor (VEGF) ，tumor necrosis factor- α (TNF-α) ，and interferon-γ ( INF-γ) in serum．Changes in ovarian index were detected．HE staining was ap- plied to detect the number of mature follicles，developing follicles，and blocked follicles in the ovary．Flow cytome- try was applied to detect the proportion of BMSCs in the ovaries．QRT-PCR was applied to detect the mRNA ex- pression of mesenchymal transition factor (cMET) ，hepatocyte growth factor (HGF) ，SDF-1，and CXCR4 in ovar- ian tissue．Western blot was applied to detect SDF-1 and CXCR4 proteins in ovarian tissue． Results : Compared with the control group，the serum levels of AOAB，TNF-α , INF-γ , and the number of blocked follicles in the ova- ries were higher，while the serum levels of β-EP，VEGF，ovarian index，the numbers of mature follicles and devel- oping follicles in the ovary，the expression of SDF-1，CXCR4 mRNA and protein were lower in the POF group (P < 0. 05) ．Compared with the POF group，the serum levels of AOAB，TNF-α , INF-γ , and the number of blocked follicles in the ovaries were lower，while the serum levels of β-EP，VEGF，ovarian index，the numbers of mature follicles and developing follicles in the ovary，the proportion of BMSCs，the expression of cMET ，HGF，SDF-1， CXCR4 mRNA and SDF-1，CXCR4 protein were higher in the total saponins of astragalus membranaceus group， BMSCs group，and combination group，and the corresponding indicators in the combination group showed the most obvious change trend (P＜0. 05) ．AMD3100 reversed the effect of total saponins of astragalus membranaceus com- bined with BMSCs on POF rats． Conclusion The combination of total saponins of astragalus membranaceus and BMSCs may inhibit inflammation in POF rats，promote follicular development，increase the proportion of BMSCs in the ovaries，and promote their homing by activating the SDF-1 / CXCR4 pathway，thereby improving ovarian func- tion．

ObjectiveTo evaluate the clinical effects of Yishen Tongluo Prescription in treating patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction， as well as the effects of this formula on biochemical indices and oxidative stress of seminal plasma. MethodsPatients who fulfilled the inclusion criteria were randomly allocated by the random number table method into an observation group （61 patients， including 2 dropped out and 59 completed） and a control group （61 patients， including 2 dropped out， 1 excluded， and 58 completed）. The observation group took Yishen Tongluo Prescription orally， 1 dose/day， and the control group took levocarnitine oral solution， 10 mL/time， 3 times/day. Both groups received a treatment course of 12 weeks and were followed up for 6 weeks. The percentage of progressive motile sperms （PR）， percentage of sperms with normal morphology， seminal plasma oxidative stress indicators ［malondialdehyde （MDA） and superoxide dismutase （SOD）］， seminal plasma biochemical indicators ［acid phosphatase （ACP）， zinc （Zn）， fructose （Fru）， and alpha-glucosidase （α-Glu）］， and traditional Chinese medicine （TCM） symptom scores were recorded for both groups before and after treatment. ResultsThe total response rate in the observation group was 76.3% （45/59）， which was higher than that （44.8%， 26/58） in the control group （χ²=12.120， P<0.05）. After treatment， both groups of patients showed increases in PR， percentage of sperms with normal morphology， and seminal plasma SOD level （P<0.05）， which were higher in the observation group than in the control group （P<0.05）. The MDA level and TCM symptom scores decreased in both groups after treatment （P<0.05）， and the decreases were more obvious in the observation group than in the control group （P<0.05）. After treatment， the seminal plasma levels of ACP， Zn， Fru， and α-Glu in the observation group （P<0.05） and that of α-Glu in the control group （P<0.05） rose. Moreover， the observation group had higher seminal plasma levels of ACP， Zn， Fru， and α-Glu than the control group （P<0.05）. Neither group showed obvious adverse reactions. ConclusionIn summaryYishen Tongluo prescription is able to improve the sperm vitality and reduce the sperm abnormality rate in infertile patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction. It may exert therapeutic effects by regulating the microenvironments of the epididymis， prostate， and seminal vesicles， thereby recovering the seminal plasma biochemical levels， enhancing the antioxidant capacity of seminal plasma， clearing excessive reactive oxygen species， and alleviating oxidative stress.

Objective To prepare a mouse model of asthenozoospermia(AZS)with high sperm DNA fragmentation(SDF)using corticosterone(CORT)and sodium benzoate(NaB).Methods Fifty 3-week-old male ICR mice were divided randomly into CORT-treated(n=30)and NaB-treated(n=20)groups.The CORT group was further divided into the following six groups(n=5 per group):high CORT(500 μg/mL),medium CORT(200 μg/mL),and low CORT(10 μg/mL)drinking water group,drinking water control group,CORT injection(40 mg/kg)group,and injection control group(normal saline).The animals were modeled continuously for 50 d.Mice in the NaB group were further divided into four groups(n=5 per group):high NaB(500 mg/kg),medium NaB(300 mg/kg),and low NaB(100 mg/kg)gavage groups,and control group(normal saline).The animals were modeled continuously for 50 d.The physiological state of the mice in each group was observed and mass changes were recorded continuously.The sperm motility capacity and DNA fragmentation index(DFI)of the sperm were observed from the tail of the epididymis after the end of the modeling.Results The rate of mass change in the CORT-injection moding group showed a downward trend.There was no significant difference(P＞0.05)in the high NaB gavage group,and the rate of body mass change in the high NaB gavage group was significantly decreased compared with the control group(P＜0.05).The percentages of forward motility sperm were significantly decreased in the CORT injection group(P＞0.05)and the percentage in the high NaB gavage group(P＜0.05),compared with the control group.The DFI was increased in the CORT injection group compared with the control group,but the difference was not significant(P＞0.05),and the DFI in the high NaB gavage group was significantly increased compared with the control group(P＜0.05).Conclusions Intragastric gavage with NaB 500 mg/(kg·d)for 50 d is an ideal method for constructing an animal model of AZS with high SDF.

Environmental pollutants generated by industrialization,characterized by microplastics,are increasingly impairing male fertility in the form of"environmental hidden toxins".Although the concept of environmental toxicity has been proposed for a long time,its practical application remains limited.Therefore,this study innovatively introduces and elaborates on the concept of"environmental hidden toxins",highlighting its pathogenic characteristics,including latency,turbidity,consumptive nature,concurrent nature,and transmutation.Environmental hidden toxin can be classified into five categories based on the properties:stagnation toxins,scorching toxins,turbid toxins,desiccating toxins,and latent toxins.The core pathogenesis of environmental hidden toxins-induced male infertility involves three stages:firstly,invading the lungs and spleen,leading to the gradual depletion of healthy qi;secondly,forming toxin-stasis complexes that obstruct the spermatic pathway;and lastly,penetrating deeply into the seminal chamber,directly damaging the genuine essence.The treatment principle emphasizes"strengthening the foundation and clearing the source,"advocating dual strategies of detoxification and strengthening vital qi.This includes enhancing clarity and lowering turbidity,fortifying the earth to generate metal to replenish healthy qi;resolving stasis,unblocking collaterals,and dispelling toxins to eliminate pathogenic factors;and nourishing the kidneys,replenishing essence,and expelling toxins to preserve the genuine essence.All the above approaches form a holistic traditional Chinese medicine(TCM)strategy that treating both manifestation and root cause of disease.The study provides theoretical foundations and offers novel clinical insights into TCM interventions for male infertility caused by environmental pollutants.

Objective To prepare a mouse model of asthenozoospermia(AZS)with high sperm DNA fragmentation(SDF)using corticosterone(CORT)and sodium benzoate(NaB).Methods Fifty 3-week-old male ICR mice were divided randomly into CORT-treated(n=30)and NaB-treated(n=20)groups.The CORT group was further divided into the following six groups(n=5 per group):high CORT(500 μg/mL),medium CORT(200 μg/mL),and low CORT(10 μg/mL)drinking water group,drinking water control group,CORT injection(40 mg/kg)group,and injection control group(normal saline).The animals were modeled continuously for 50 d.Mice in the NaB group were further divided into four groups(n=5 per group):high NaB(500 mg/kg),medium NaB(300 mg/kg),and low NaB(100 mg/kg)gavage groups,and control group(normal saline).The animals were modeled continuously for 50 d.The physiological state of the mice in each group was observed and mass changes were recorded continuously.The sperm motility capacity and DNA fragmentation index(DFI)of the sperm were observed from the tail of the epididymis after the end of the modeling.Results The rate of mass change in the CORT-injection moding group showed a downward trend.There was no significant difference(P＞0.05)in the high NaB gavage group,and the rate of body mass change in the high NaB gavage group was significantly decreased compared with the control group(P＜0.05).The percentages of forward motility sperm were significantly decreased in the CORT injection group(P＞0.05)and the percentage in the high NaB gavage group(P＜0.05),compared with the control group.The DFI was increased in the CORT injection group compared with the control group,but the difference was not significant(P＞0.05),and the DFI in the high NaB gavage group was significantly increased compared with the control group(P＜0.05).Conclusions Intragastric gavage with NaB 500 mg/(kg·d)for 50 d is an ideal method for constructing an animal model of AZS with high SDF.