Based on cancer collateral theory， this paper proposes toxin-stasis accumulation in collaterals and yin deficiency with toxin hyperactivity as the core pathogenesis of postoperative recurrence of triple-negative breast cancer. Impaired qi transformation in collateral and depletion of yin interact， contributing to toxin-stasis binding， inadequate collateral nourishment， and unrestrained yang activity. The temporal dimension emphasizes yin deficiency with toxin hyperactivity and early reactivation of latent toxin； the spatial dimension emphasizes toxin-stasis accumulation and dissemination through collaterals. Treatment follows the principle of regulating qi and resolving collaterals. Before recurrence， emphasis is placed on nourishing collaterals to restrain latent toxin； after recurrence， on resolving binding while protecting collaterals. Herbal combinations are further tailored to disorders of qi， blood， and fluid collaterals and to the sites of dissemination， integrating treatment according to recurrence status and affected collateral systems. These theoretical propositions are intended to inform clinical reasoning and require clinical validation.

Objective:To investigate the changes of T lymphocyte subsets in patients with stage Ⅰ and Ⅱ cervical cancer after surgery and their relationship with postoperative lymph node metastasis according to the International Federation of Gynecology and Obstetrics (FIGO) stage (2014) .Methods:A total of 192 patients with FIGO stage ⅠA, ⅠB1, ⅠB2 and ⅡA1 who received radical cervical cancer resection and pelvic lymph node dissection in People's Hospital of Yuechi County of Sichuan Province and West China Guang'an Hospital of Sichuan University from November 2018 to November 2020 were selected for this study. According to FIGO stage, patients were divided into stage Ⅰ group ( n=85) and stage Ⅱ group ( n=107) . The dynamic changes of T lymphocytes subsets in patients with different FIGO stages were compared before and after surgery. Repeated measurement of variance was used to analyze the levels of T lymphocytes subsets in patients of different stages during treatment. Logistic regression was used to analyze the influencing factors of postoperative lymph node metastasis in patients with cervical cancer. Multivariate logistic regression was used to analyze the relationship between T lymphocytes subsets and postoperative lymph node metastasis. Receiver operator characteristic (ROC) curve was used to analyze the predictive efficacy of T lymphocytes level in postoperative lymph node metastasis. Results:The postoperative lymph node metastasis rate in stage Ⅱ patients [32.71% (35/107) ] was higher than that in stage Ⅰ patients [14.12% (12/85) ], with a statistically significant difference ( χ2=8.86, P=0.003) . Compared with the stage Ⅱ group, the levels of CD3 +, CD4 + T lymphocytes and CD4 +/CD8 + ratio were significantly higher in the stage Ⅰ group 1 day before surgery (all P<0.001) , and the level of CD8 + T lymphocytes was significantly lower ( P<0.001) . The levels of CD3 +, CD4 +, CD8 + T lymphocytes and the ratio of CD4 +/CD8 + showed dynamic changes at different stages after surgery. On 1, 7 and 30 days after surgery, the levels of CD3 +, CD4 + T lymphocytes and the ratio of CD4 +/CD8 + in stage Ⅰ group were higher than those in stage Ⅱ group (all P<0.001) , CD8 + T cell levels were lower than those in stage Ⅱ group (all P<0.001) . There were statistically significant differences in T lymphocytes subsets CD3 +, CD4 +, CD8 + and CD4 +/CD8 + time effect, intergroup effect and interaction effect between the two groups (all P<0.001) . Univariate analysis showed that the pathological type ( OR=1.85, 95% CI: 1.14-2.33, P=0.015) , differentiation degree ( OR=1.93, 95% CI: 1.18-2.67, P=0.024) , depth of myometrial invasion ( OR=2.08, 95% CI: 1.26-2.59, P=0.012) , tumor morphology ( OR=2.17, 95% CI: 1.57-2.63, P=0.009) , parametrial invasion ( OR=1.95, 95% CI: 1.43-2.76, P=0.036) and lymphovascular space invasion ( OR=2.03, 95% CI: 1.28-2.57, P=0.021) were the influencing factors for postoperative lymph node metastasis in patients with FIGO stage Ⅰ and Ⅱ cervical cancer. Multivariate analysis showed that the degree of differentiation ( OR=1.75, 95% CI: 1.08-2.03, P=0.015) , depth of myometrial invasion ( OR=2.30, 95% CI: 1.43-2.84, P=0.021) , parametrial invasion ( OR=2.50, 95% CI: 1.76-2.97, P=0.018) and lymphovascular space invasion ( OR=1.96, 95% CI: 1.03-2.51, P=0.033) were independent factors for postoperative lymph node metastasis in patients with FIGO stage Ⅰ and Ⅱ cervical cancer. Multivariate logistic regression analysis showed that the levels of CD3 +, CD4 +, CD8 + T cells and the ratio of CD4 +/CD8 + in patients with stage Ⅰ and stage Ⅱ cervical cancer 1 day before surgery were independent influencing factors for postoperative lymph node metastasis (all P<0.05) . ROC curve analysis showed that the areas under the curve of CD3 +, CD4 +, CD8 + T lymphocytes levels and the ratio of CD4 +/CD8 + in stage Ⅰ patients 1 day before surgery for predicting postoperative lymph node metastasis were 0.86, 0.82, 0.83, 0.89, respectively, and those in stage Ⅱ patients were 0.90, 0.93, 0.87, 0.95, respectively. CD4 +/CD8 + ratio was significantly more effective in predicting postoperative lymph node metastasis than other indexes (all P<0.001) . Conclusions:The levels of CD3 +, CD4 + T lymphocytes, and the CD4 +/CD8 + ratio in patients with FIGO stage Ⅰ and Ⅱ cervical cancer are significantly higher in 1-30 days after surgery than before, while the level of CD8 + T lymphocytes is significantly lower than before. There is a significant correlation between T lymphocytes subsets and lymph node metastasis after surgery. In addition, low differentiation, depth of myometrial invasion ≥1/2, parametrial invasion, and lymphovascular space invasion are independent risk factors for postoperative lymph node metastasis.

Objective:To investigate the alterations in the brain structural network properties related to cognitive emotion regulation strategies in young, first-episode and drug-naive (FEDN) patients with major depressive disorder (MDD).Methods:One hundred and twelve young patients who were diagnosed with FEDN MDD were enrolled in the Second Xiangya Hospital of Central South University between October 2019 and October 2023, and 143 healthy controls (HC) were recruited for reference during the same period. Clinical data and T 1 structural images of high-resolution magnetic resonance imaging were collected from all participants, and the cognitive emotion regulation was assessed using the Chinese version of the Cognitive Emotion Regulation Questionnaire (CERQ-C). The brain structural alterations related to cognitive emotion regulation strategies were explored using whole-brain voxel-based morphometry (VBM) group analysis and individual differential structural covariance network (IDSCN) analysis. Partial correlation analysis was performed between the IDSCN anomalous structural concatenated edge and subdimensions of the CERQ-C. Results:The scores on the sub-dimension of negative cognitive emotion regulation (self-condemnation, rumination, catastrophization, blaming others) in the young FEDN MDD patients in CERQ-C were significantly higher than those in HC (14.28±2.30 vs 12.45±2.00, t=6.501, P<0.001;14.40±2.97 vs 11.31±2.93, t=7.934, P<0.001;12.19±3.75 vs 7.59±2.58, t=10.553, P<0.001;10.26±3.00 vs 8.74±2.89, t=3.916, P<0.001; respectively). There was no statistically significant difference in the VBM group analysis (all P>0.05). However, IDSCN single sample t test results showed significant group differences between the left orbitofrontal inferior gyrus and the left supplementary motor area, the left orbitofrontal middle gyrus and the bilateral precuneus, and the left superior temporal gyrus and the bilateral hippocampus (all P<0.05). With controlling for age, sex, and other factors, partial correlation analysis results showed that the left orbital inferior frontal gyrus-left supplementary motor area connection in patients with MDD was positively correlated with the self-condemnation and acceptance scores of the CERQ-C ( r=0.226, P=0.027; r=0.216, P=0.035), while negatively correlated with the score of blaming others ( r=-0.252, P=0.013). Additionally, connections between the right hippocampus and left superior temporal gyrus were significantly associated with the scores on catastrophization subdimension of the CERQ-C ( r=0.229, P=0.025). Conclusion:Due to its sensitivity to individualized analyses, the IDSCN analysis found that abnormal topological property changes were mainly manifested in the limbic system, the sensorimotor system, and the default network in young FEDN MDD patients, and were linked to their negative cognitive emotion regulation strategies, even though the VBM group analysis did not yield any significant results.

OBJECTIVE: To observe the effects of electroacupuncture (EA) with different frequencies on spermatogenic function, testicular morphology and oxidative stress in oligoasthenospermia (OAT) rats, and to explore the mechanism and the optimal parameters of EA for OAT. METHODS: Sixty SPF-grade male SD rats were randomly divided into a solvent control group, a model group, a 2 Hz EA group, a 100 Hz EA group and a 2 Hz/100 Hz EA group, with 12 rats in each group. Except for the solvent control group, the other 4 groups were administered ornidazole suspension (800 mg·kg^-1·d^-1) by gavage for 28 d to establish the OAT model. Starting from the 1st of modeling, EA was applied at "Guanyuan" (CV4), "Qihai" (CV6) and bilateral "Sanyinjiao" (SP6) and "Zusanli" (ST36) in the 3 EA groups, continuous wave of 2 Hz, continuous wave of 100 Hz, and disperse-dense wave of 2 Hz/100 Hz were used in the 2 Hz EA group, the 100 Hz EA group, and the 2 Hz/100 Hz EA group, respectively, with current intensity of 1-3 mA, 30 min a time, once every other day, for 28 consecutive days. After intervention, the testicular index was calculated, epididymal sperm quality was assessed, and the fertility ability was observed; morphology of testicular tissue was observed by HE staining, and the Johnson score was calculated; the positive expression of reactive oxygen species (ROS) in testicular tissue was detected by immunofluorescence; the activity of superoxide dismutase (SOD) and catalase (CAT), as well as the level of malondialdehyde (MDA) in testicular tissue were measured by ELISA; the protein expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) in testicular tissue was detected by Western blot. RESULTS: Compared with the solvent control group, in the model group, the testicular index, sperm concentration, sperm motility and the number of offspring were decreased (P<0.01), the seminiferous tubules atrophied and the Johnson score decreased (P<0.01); the activity of SOD and CAT, as well as the protein expression of Nrf2 and HO-1 in testicular tissue were decreased (P<0.01); the sperm deformity rate, the positive expression of ROS and the MDA level in testicular tissue were increased (P<0.01). Compared with the model group, in the 2 Hz EA group, the 100 Hz EA group and the 2 Hz/100 Hz EA group, the testicular index, sperm concentration, sperm motility and the number of offspring were increased (P<0.05, P<0.01), the pathological morphology of testicular tissue improved and the Johnson scores increased (P<0.01); the activity of SOD and CAT, as well as the protein expression of Nrf2 and HO-1 in testicular tissue were increased (P<0.05, P<0.01); the sperm deformity rate, the positive expression of ROS and the MDA level in testicular tissue were decreased (P<0.05, P<0.01). Compared with the 2 Hz EA group, in the 2 Hz/100 Hz EA group, the testicular index, sperm concentration, sperm motility, as well as the CAT activity and HO-1 protein expression in testicular tissue were increased (P<0.01, P<0.05); the positive expression of ROS was decreased (P<0.01). Compared with the 100 Hz EA group, in the 2 Hz/100 Hz EA group, the testicular index was increased (P<0.01), the positive expression of ROS in testicular tissue was decreased (P<0.01). CONCLUSION EA with 2 Hz continuous wave, 100 Hz continuous wave, and 2 Hz/100 Hz disperse-dense wave can all improve the spermatogenic arrest and reduce the level of oxidative stress in testicular tissue in OAT rats, the mechanism may be related to up-regulating the protein expression of Nrf2 and HO-1 and improving oxidative stress. EA with disperse-dense wave of 2 Hz/100 Hz shows the optimal effect.
Male ; Animals ; Electroacupuncture ; Oxidative Stress ; Rats ; Rats, Sprague-Dawley ; Spermatogenesis ; Oligospermia/genetics* ; Humans ; Testis/metabolism* ; Superoxide Dismutase/metabolism* ; Asthenozoospermia/genetics* ; Acupuncture Points ; Malondialdehyde/metabolism*

Objective:To investigate the alterations in the brain structural network properties related to cognitive emotion regulation strategies in young, first-episode and drug-naive (FEDN) patients with major depressive disorder (MDD).Methods:One hundred and twelve young patients who were diagnosed with FEDN MDD were enrolled in the Second Xiangya Hospital of Central South University between October 2019 and October 2023, and 143 healthy controls (HC) were recruited for reference during the same period. Clinical data and T 1 structural images of high-resolution magnetic resonance imaging were collected from all participants, and the cognitive emotion regulation was assessed using the Chinese version of the Cognitive Emotion Regulation Questionnaire (CERQ-C). The brain structural alterations related to cognitive emotion regulation strategies were explored using whole-brain voxel-based morphometry (VBM) group analysis and individual differential structural covariance network (IDSCN) analysis. Partial correlation analysis was performed between the IDSCN anomalous structural concatenated edge and subdimensions of the CERQ-C. Results:The scores on the sub-dimension of negative cognitive emotion regulation (self-condemnation, rumination, catastrophization, blaming others) in the young FEDN MDD patients in CERQ-C were significantly higher than those in HC (14.28±2.30 vs 12.45±2.00, t=6.501, P<0.001;14.40±2.97 vs 11.31±2.93, t=7.934, P<0.001;12.19±3.75 vs 7.59±2.58, t=10.553, P<0.001;10.26±3.00 vs 8.74±2.89, t=3.916, P<0.001; respectively). There was no statistically significant difference in the VBM group analysis (all P>0.05). However, IDSCN single sample t test results showed significant group differences between the left orbitofrontal inferior gyrus and the left supplementary motor area, the left orbitofrontal middle gyrus and the bilateral precuneus, and the left superior temporal gyrus and the bilateral hippocampus (all P<0.05). With controlling for age, sex, and other factors, partial correlation analysis results showed that the left orbital inferior frontal gyrus-left supplementary motor area connection in patients with MDD was positively correlated with the self-condemnation and acceptance scores of the CERQ-C ( r=0.226, P=0.027; r=0.216, P=0.035), while negatively correlated with the score of blaming others ( r=-0.252, P=0.013). Additionally, connections between the right hippocampus and left superior temporal gyrus were significantly associated with the scores on catastrophization subdimension of the CERQ-C ( r=0.229, P=0.025). Conclusion:Due to its sensitivity to individualized analyses, the IDSCN analysis found that abnormal topological property changes were mainly manifested in the limbic system, the sensorimotor system, and the default network in young FEDN MDD patients, and were linked to their negative cognitive emotion regulation strategies, even though the VBM group analysis did not yield any significant results.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.