Zhibai Dihuang Pill Alleviates Ureaplasma urealyticum-Induced Spermatogenic Failure and Testicular Dysfunction via MAPK Signaling Pathway.

The testicles and sperm are extremely susceptible to inflammation and oxidative stress. Although Zhibai Dihuang Pill (ZDP) has been reported to treat various infertilities including male infertility induced by Ureaplasma urealyticum (UU) infection, its mechanism is still poorly understood. This study is aimed at clarifying the underlying mechanism of ZDP to protect against UU-infected male infertility. We found that UU-infected infertile rats exhibited weight loss, reduced food intake, and decreased sperm count and vitality. The administration of ZDP improved the general state and sperm motility of rats. In addition, UU infection led to spermatogenesis disorders, impaired secretory function and blood-testis barrier (BTB) of Sertoli cells, and elevated inflammation and oxidative stress. As expected, ZDP suppressed inflammation and oxidative stress to alleviate spermatogenesis disorders. Our research showed that ZDP could improve spermatogenesis disorders and testicular function primarily through the mitogen-activated protein kinase (MAPK) signaling pathway. ZDP exerts its anti-inflammatory and antioxidant effects via the MAPK signaling pathway, thus playing an important role in ameliorating spermatogenesis failure and testicular dysfunction.

[Impact of Ureaplasma urealyticum infection on the MRPS22 protein expression in rat spermatogenic cells and intervening effect of Zhibai Dihuang Decoction].

OBJECTIVE: To investigate the impact of Ureaplasma urealyticum (UU) infection (UUI) on the expression of the mitochondrial ribosomal protein S22 (MRPS22) in rat spermatogenic cells and the intervening effect of Zhibai Dihuang Decoction (ZBDH). METHODS: Forty-five SD male rats were randomly divided into four groups of equal number: normal control, UUI model control, ZBDH and azithromycin, and the UUI model was made by bladder injection of the standard UU strain in the latter three groups. After modeling, the rats in the ZBDH and azithromycin groups were treated intragastrically with ZBDH at 1 g/kg/d and azithromycin at 0.105 g/kg/d respectively, while those in the normal and UUI model control groups with normal saline at 1 ml/kg/d. At 21 days after intervention, all the animals were sacrificed and their testes harvested for observation of the apoptosis and mitochondrial ultrastructure of the spermatogenic cells, measurement of the mitochondrial membrane potential (MMP) by flow cytometry, and determination of the mRNA and protein expressions of MRPS22 by RT-PCR and Western blot, respectively. RESULTS: The apoptotic index of the rat sperma-togenic cells was significantly higher in the UUI model control than in the ZBDH and azithromycin groups (ï¼»11.23 ± 1.65ï¼½ % vs ï¼»6.62 ± 0.49ï¼½ % and ï¼»7.82 ± 0.81ï¼½ %, P < 0.01), but lower in the ZBDH than in the azithromycin group (P < 0.05). The mitochondrial ultrastructure of the spermatogenic cells was markedly improved in the ZBDH and azithromycin groups as compared with that in the model control. The MMP level was remarkably lower in the model control than in the normal control (ï¼»8.77 ± 1.73ï¼½ % vs ï¼»22.33 ± 1.66ï¼½ %, P < 0.01), but higher in the ZBDH (ï¼»18.26 ± 1.32ï¼½ %) than in the model control (P < 0.01) and the azithromycin group (ï¼»15.91 ± 1.69ï¼½ %) (P < 0.01). The mRNA and protein expressions of MRPS22 were significantly lower in the model control (8.02 ± 3.21 and 22.65 ± 5.31) than in the normal control (15.43 ± 2.54 and 33.31 ± 7.09), ZBDH (11.26 ± 3.82 and 33.35 ± 3.96), and azithromycin group (8.79 ± 2.03 and 28.11 ± 4.13) (all P < 0.01), but both higher in the ZBDH than in the azithromycin group (P < 0.01). There was a positive correlation between the MRPS22 protein expression and MMP (r = 0.639, P < 0.01). CONCLUSIONS: UU infection induces the apoptosis of rat spermatogenic cells by inhibiting the mRNA and protein expressions of MRPS22 and reducing the mitochondrial membrane potential, while ZBDH can decrease the apoptosis of spermatogenic cells by improving the mRNA and protein expressions of MRPS22 and enhancing the mitochondrial membrane potential.

[Efficacy and safety of № I Empirical Prescription for Chronic Prostatitis in the treatment of type ⠢ refractory chronic prostatitis].

OBJECTIVE: To evaluate the efficiency and safety of № I Empirical Prescription for Chronic Prostatitis (№ I EPCP) in the treatment of type Ⅲ refractory chronic prostatitis. METHODS: We randomly assigned 53 cases of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis to an experimental and a control group to receive № I EPCP at 1 dose per day and saw palmetto extract at 160 mg bid), respectively, all for 8 weeks. Before and after 4 and 8 weeks of treatment, we obtained The National Institute of Health Chronic Prostatitis Symptom Index (NIH-CPSI) scores, Traditional Chinese Medicine Syndrome Scores (TCMSS), maximum urinary flow rate (Qmax), average urinary flow rate (Qavg), Hamilton Depression Rating Scale (HAMD) scores and Hamilton Anxiety Rating Scale (HAMA) scores, and compared them between the two groups of patients. RESULTS: Totally 48 of the patients completed the medication and follow-up, 25 in the experimental and 23 in the control group. Compared with the baseline, the NIH-CPSI scores after 8 weeks of treatment were significantly decreased in the experimental (27.82 ± 7.25 vs 15.46 ± 4.77, P <0.05) and the control group (25.98 ± 6.47 vs 21.06 ± 5.74, P <0.05), and so were the TCMSSs (24.64 ± 9.82 vs 16.42 ± 6.33 and 9.15 ± 3.74, P <0.05, and 23.67 ± 8.73 vs 18.55 ± 5.92 and 13.48 ± 4.45, P <0.05); the Qmax at 8 weeks were dramatically increased in the experimental group (ï¼»18.45 ± 7.81ï¼½ vs ï¼»23.44 ± 8.73ï¼½ ml/s, P <0.05) and the control (ï¼»17.58 ± 6.92ï¼½ vs ï¼»21.26 ± 8.32ï¼½ ml/s, P <0.05), and so was the Qavg (ï¼»11.27 ± 5.33ï¼½ vs ï¼»16.51 ± 7.36ï¼½ ml/s, P <0.05 and ï¼»10.66 ± 5.82ï¼½ vs ï¼»13.44 ± 6.16ï¼½ ml/s, P <0.05); the HAMD scores were remarkably reduced in the experimental group (22.74 ± 6.37 vs 17.62 ± 5.71 and 12.54 ± 5.22, P <0.05) and the control (23.55 ± 7.14 vs 22.34 ± 6.88 and 21.62 ± 5.63, P <0.05), and so were the HAMA scores (21.37 ± 7.15 vs 18.42 ± 6.35 and 14.63 ± 7.11, P <0.05 and 20.54 ± 6.77 vs 19.87 ± 6.24 and 19.42 ± 7.04, P <0.05). No obvious adverse reactions were observed in either of the two groups during the medication. CONCLUSIONS: № I EPCP deserves promotion and clinical application for its definite effectiveness and safety in the treatment of type Ⅲ refractory chronic prostatitis with damp-heat and blood stasis.

[Effects of Zhibai Dihuang Decoction on mitochondrial cytochrome oxidase in the spermatogenic cells of rats with ureaplasma urealyticum infection].

OBJECTIVE: To explore the effects of Zhibai Dihuang Decoction (ZDD) on mitochondrial cytochrome oxidase (COX) in the spermatogenic cells of rats with ureaplasma urealyticum (UU) infection. METHODS: From forty 4－5 months old SD rats, 30 were randomly selected for the establishment of the model of testicular UU infection by inoculating the bladder with UU suspension and the other 10 injected with normal saline as controls (group A). At 7 days after inoculation, the rat models of testicular UU infection were treated orally with normal saline (group B), ZDD at 1 g per kg of the body weight per day (group C), and azithromycin at 0.105 g per kg of the body weight per day (group D), respectively, once daily for 21 days. Then all the animals were sacrificed and the epididymal and testicular tissues collected for examination of sperm motility with the color sperm dynamic detection system, measurement of the COX activity with the immunohistochemical DAB method, and determination of the mRNA expressions of COXⅠ and COXⅡ by RT-PCR. RESULTS: Compared with group A, group B showed significant decreases in such sperm parameters as grade a sperm (ï¼»1.03 ± 0.09ï¼½ vs ï¼»0.07 ± 0.03ï¼½ %, P<0.01), grade b sperm (ï¼»2.07 ± 0.52ï¼½ vs ï¼»0.35 ± 0.13ï¼½ %, P<0.01), straight line velocity (VSL) (ï¼»10.95 ± 0.98ï¼½ vs ï¼»6.78 ± 1.05ï¼½ µm/s, P<0.01), curvilinear velocity (VCL) (ï¼»42.03 ± 1.35ï¼½ vs ï¼»38.10 ± 7.65ï¼½ µm/s, P>0.05), average path velocity (VAP) (ï¼»16.22 ± 1.52ï¼½ vs ï¼»10.05 ± 1.80ï¼½ µm/s, P<0.01), and the mRNA expressions of COX Ⅰ (ï¼»2.25 ± 0.24ï¼½ vs ï¼»0.93 ± 0.10ï¼½ %, P<0.01) and Ⅱ (ï¼»6.72 ± 0.37ï¼½ vs ï¼»2.95 ± 0.78ï¼½ %, P<0.01). After treatment, all the parameters were remarkably increased in groups C and D (grade a sperm: ï¼»1.11 ± 0.30ï¼½ and ï¼»0.60 ± 0.19ï¼½%; grade b sperm: ï¼»2.40 ± 0.59ï¼½ and ï¼»1.32 ± 0.27ï¼½ %; VSL: ï¼»12.11 ± 1.62ï¼½ and ï¼»11.47 ± 1.21ï¼½ µm/s; VCL: ï¼»54.30 ± 2.35ï¼½ and ï¼»45.75 ± 1.64ï¼½ µm/s; VAP ï¼»18.40 ± 1.27ï¼½ and ï¼»16.69 ± 1.02ï¼½ µm/s; expression of COXⅠ mRNA: ï¼»1.86 ± 0.30ï¼½ and ï¼»1.74 ± 0.17ï¼½ %) as compared with those in group B (P<0.05or P<0.01) except the COX activity and the expression of COX Ⅱ mRNA (P>0.05), and all the parameters were significantly higher in group C than in D (P<0.05or P<0.01). CONCLUSIONS: UU infection can reduce grades a and b sperm, linear, curvilinear and mean sperm velocities, and the mRNA expressions of COX Ⅰ and Ⅱ while ZDD can improve these parameters. The improvement of sperm motility may not be associated with the activity of COX, and the COX activity may be related to the mRNA expression of COX II but not that of COXⅠ.

[Effect of Zhibai Dihuang Decoction on Expressions of TRPV1 and TRPV5 in Spermatogenic Cells of UU-infected Rats].

OBJECTIVE: To observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD. METHODS: Totally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot. RESULTS: The UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01). CONCLUSION: ZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

[Combination of acupuncture, cupping and medicine for treatment of fibromyalgia syndrome: a multi-central randomized controlled trial].

OBJECTIVE: To evaluate the clinical effect of combination of acupuncture, cupping and medicine for treatment of fibromyalgia syndrome. METHODS: By using multi-central randomized controlled method, 186 cases were randomly divided into an acupuncture combined with cupping and western medicine group (group A), an acupuncture combined with cupping group (group B) and a western medicine group (group C) and treated continuously for 4 weeks. The treatment of acupuncture combined with cupping was produced by acupuncture at five mental points and moving cupping on the Hechelu of the back, once evrey other day, thrice each week, and the western medicine therapy by oral administration of Amitriptyline, once each day. The scores of McGill Pain Questionnaire (MPQ), the amount of tenderness point and the time of producing effect were compared and the therapeutic effects were assessed with the Hamilton Depression Scale (HAMD). RESULTS: The cured and markedly effective rate was 65.0% (39/60) in the group A, which was superior to 15.9% (10/63) in the group B and 16.1% (9/56) in the group C (both P < 0.001). After treatment, the scores of MPQ and HAMD and the amount of tenderness point all decreased in the three groups, group A being significantly better than group B and group C, and the time of producing effect in the group A was more earlier than those in the group B and the group C. CONCLUSION: The therapeutic effect of combination of acupuncture, cupping and medicine on fibromyalgia syndrome is superior to that of the simple acupuncture combined with cupping or the simple medicine.

Chronic fatigue syndrome treated by acupuncture and moxibustion in combination with psychological approaches in 310 cases.

OBJECTIVE: To observe clinical therapeutic effect of acupuncture and moxibustion combined with a psychological approach on chronic fatigue syndrome (CFS). METHODS: The treatment was given by acupuncture plus moxibustion combined with a psychological approach based on differentiation of symptoms and signs in 310 cases. RESULTS: Of 310 cases observed, 275 cases (88.7%) were clinically cured, 28 cases (9%) improved, and 7 cases (2.3%) failed. CONCLUSION: Acupuncture plus moxibustion combined with a psychological approach is an effective therapy for CFS.

[Determination of periplocin in different parts of Periploca sepium].

OBJECTIVE: To determinate the content of periplocin in different parts of Periploca sepium Bunge. METHODS: HPLC were carried out on ODS column, acetonitril: water (27: 73) as mobile phase, detection wavelength at 220nm. RESULTS: The content of periplocin in root bark, stem bark, xylem of root, xylem of stem are 1.03%, 0.65%, 0.26%, 0.39% respectively. No periplocin was detected in leaves and fruit. CONCLUSION: The plant of Periploca sepium Bunge should be multiply utilized.

[Controlled observation on electroacupuncture combined with cake-separated moxibustion for treatment of tennis elbow].

OBJECTIVE: To explore an effective method for treatment of tennis elbow. METHODS: One hundred and twenty-eight cases were randomly divided into a treatment group and a control group, 64 cases in each group. The treatment group were treated by electroacupuncture at Hegu (LI 4) and Taichong (LR 3), combined with cake-seperated moxibustion at Ashi points and Shousanli (LI 10) in the affected elbow, and the control group by routine block treatment of 1 mL lidocane and 25 mg prednisone. RESULTS: The cured rate and the effective rate were 40.6% and 93.7% in the treatment group, and 25.0% and 78.1% in the control group, respectively, with a significant difference between the two groups (P < 0.05). CONCLUSION: Electroacupuncture combined with cake-separated moxibustion is an effective method for tennis elbow.

[Quantification of metallothionein induced with selenium by using competitive ELISA].

Metallothionein induced by selenium was assayed with competitive ELISA. It was found that in control mice, the liver contained (2.47+/-0.90) microgram MT/g wet weight tissue, while the mice administered with ZnSO(4) contained (8.15+/-2.20) microgram MT/g wet weight liver tissue and the mice given with selenium malt contained (12.80+/-1.44) microgram MT/g wet weight liver. A significant difference was found between the control and the latter two groups (P<0.05, P<0.05). The MT content of selenium group was significantly higher than the zinc group. It is suggested that the selenium was more potent in induction of MT and was a stronger inducer than ZnSO(4) in its effective dose range, therefore selenium may be developed into a novel and safe MT inducer.