OBJECTIVETo evaluate the results of requesting a second consecutive sperm ejaculate in order to reduce ICSI cycles by PESA or TESE on the day of oocyte pick-up in assisted reproductive technology (ART).

METHODSWe collected 68 semen samples as a second consecutive ejaculate from 34 men, compared the semen volume and sperm concentration, motility and total count between the first and the second ejaculation, and analyzed the laboratory results and clinical outcomes of fertilization with the mixed sperm.

RESULTSThe 34 males ejaculated twice within 4 hours by masturbation, with an interval of 26-183 (94.9 +/- 39.8) minutes between the first and second ejaculation. The volume of the first ejaculate was (2.0 +/- 1.4) ml, significantly higher than that of the second ([1.5 +/- 0.9] ml) (P = 0.007), although the numbers of motile sperm and grade a + b sperm of the first ([40.8 +/- 25.3]% and [30.9 +/- 22.4]%) were significantly lower than those of the second ([52.2 +/- 21.1]% and [39.9 +/- 17.5]%) (P < 0.05). There were no statistically significant differences in the sperm concentration or total sperm count between the two ejaculates (P > 0.05). The ICSI, IVF + ICSI, and IVF cycles were 3, 3 and 28 respectively among the 34 couples undergoing ART. The number of retrieved oocytes, normal fertilization rate, high quality embryo rate and frozen cycles/fresh transfer cycles ratio were 15.5 +/- 8.7, 57.0% (247/433), 58.7% (145/247) and 20/24 for the IVF cycle, 21.7 +/- 8.3, 61.5% (40/65), 67.5% (27/40) and 3/2 for the ICSI cycle, and 10.0 +/- 2.6, 72.4% (21/29), 66.7% (14/21) and 3/3 for the IVF + ICSI cycle. Fourteen live births were achieved out of the 18 pregnancies, including 6 healthy boys and 9 healthy girls.

CONCLUSIONA clinical pregnancy rate of >30% can be achieved by requesting a second consecutive sperm ejaculate on the day of oocyte pick-up in order to collect more sperm and/or increase the total number of motile sperm for ART. And this method can avoid other invasive sperm processing techniques and the need of unnecessary micromanipulative fertilization.

Adult ; Female ; Humans ; Male ; Middle Aged ; Ovulation Induction ; Pregnancy ; Reproductive Techniques, Assisted ; Sperm Retrieval ; Young Adult

Objective Long-chain triacylglycerol (LCT) by three producers,each mixed with the same medium-chain triacylglycerol (MCT),were compared with the brand MCT/LCT in causing focal necrosis of hepatocytes in beagle dogs (a bioequivalence evaluation).Methods 21 beagle dogs (male,0.7-1.5 years old,10-15 kg) were used in this study.According to the sources of the LCT,the animals were divided into Group A (LCT made in China),Group B (LCT made in Japan),Group C (LCT made in Germany),and the control group (the brand 10％ MCT/LCT).Central venous port was placed via the lumber vein of the animals under general anesthesia.After 2 weeks of rehabilitation,MCT/LCT was administered through this port for 28 days at 9 g/ (kg · d) [while the routine dose used clinically was 1 g/ (kg · d)].The laboratory indexes and the pathomorphism of the liver and kidney were studied single blindly.Results Laboratory tests,including liver and kidney function,blood coagulation function and lipid metabolism,did not identify differences among emulsions with different sources of LCT.Liver biopsy at day 28 showed no focal necrosis in Group C and the control group;there was minor damage in Group B;and Group A had obvious liver necrosis.and the pathological findings of other organs are similar.No significant difference was observed in biopsies of other organs.Conclusions Emulsions with different sources of LCT varied in their damage to the liver.Generics with LCT of higher quality were equivalent to the brand MCT/LCT in terms of safety.

Objective:To study the effect of Wenjing Huayu Zhitong therapy （Xiangyan Zhitong prescription， XZP） on the mitogen activated protein kinase （MAPK）/extracelluar regulated protein kinase （ERK） signaling pathway of primary dysmenorrhea （PD） rats， and explore the pathogenesis of PD and the mechanism of action of Wenjing Huayu Zhitong therapy. Method:Forty-eight female SPF-grade Wistar rats were randomly divided into blank group，model group， western medicine group， low-dose XZP group， medium-dose XZP group， and high-dose XZP group， with 8 rats in each group. In addition to the blank group， dysmenorrhea rat model with cold coagulation and blood stasis syndrome was established by cold stimulation combined with estradiol benzoate and oxytocin. The rats in the blank group，model group，western medicine group， low-dose XZP group， medium-dose XZP group， and high-dose XZP group were given distilled water， distilled water，0.06 g·kg^-1 ibuprofen， 6.55 g·kg^-1 XZP， 13.09 g·kg^-1 XZP， and 26.18 g·kg^-1 XZP， respectively， by gavage for 6 days. The writhing latency and writhing frequency of rats were recorded within 30 min after oxytocin injection.Western blot was used to detect the protein expression of B-Raf， mitogen activates extracellular regulated kinases1/2 （MEK1/2）， extracellular regulated kinases1/2 （ERK1/2）， p-MEK1/2， p-ERK1/2， c-Jun， and cyclooxygenase-2 （COX-2） in rat uterus. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was employed to detect the mRNA expression of B-Raf， MEK1， MEK2， ERK1， ERK2， c-Jun， and cyclooxygenase-2 （COX-2） in rat uterus. Result:Compared with the model group，the treatment groups showed insignificantly prolonged writhing latency and significantly reduced writhing frequency （P<0.01）. On the 6^th day of modeling， there was no significant difference in the quantitative scores of symptoms and signs among the treatment groups. On the 12^th day of modeling， the scores changed little in the western medicine group and the low-dose XZP group and decreased significantly in the medium- and high-dose XZP groups （P<0.01） compared with those in the model group. Compared with those in the blank group， the protein and mRNA levels of p-MEK1/2， p-ERK， B-Raf， c-Jun， and COX-2 in the model group were significantly up-regulated （P<0.01）. Compared with those in the model group， the protein and mRNA levels of p-MEK1/2， p-ERK1/2， B-Raf， c-Jun， and COX-2 in the western medicine group， medium-dose XZP group， and high-dose XZP group were significantly down-regulated （P<0.05，P<0.01）. Conclusion:The mechanism of Wenjing Huayu Zhitong therapy in treating PD with cold coagulation and blood stasis syndrome may be related to the down-regulation of MAPK/ERK signaling pathway.

Objective:To observe the clinical efficacy of Wenjing Huayu Zhitong therapy in treating primary dysmenorrhea (PD) with cold coagulation and blood stasis, and to explore its immune mechanisms on PD. Method:The 108 PD patients with cold coagulation and blood stasis syndrome were collected and randomly divided into traditional Chinese medicine (TCM) group, ibuprofen group and placebo group according to the random number table method, with 36 cases in each group. All patients received corresponding medicines three days before menstruation. The patients in TCM group were treated with TCM and ibuprofen sustained release capsule simulator. The patients in ibuprofen group were treated with ibuprofen sustained-release capsule and TCM simulator. The patients in placebo group were treated with TCM simulator and ibuprofen sustained-release capsule simulator. Treatment lasted for 6 consecutive days for three menstrual cycles, and follow-up was conducted for three menstrual cycles after drug withdrawal. The visual analogue score (VAS), total time of abdominal pain and TCM symptom scores in each menstrual cycle were recorded. The levels of CD3⁺, CD4⁺, CD8⁺ and the ratio of CD4⁺/CD8⁺ in peripheral blood before and after treatment were detected by flow cytometry. Result:After treatment for three menstrual cycles, both the TCM group and ibuprofen group were better than placebo group in reducing VAS score and reducing total abdominal pain time (P<0.01). The long-term follow-up effect after drug withdrawal in TCM group was significantly better than that in ibuprofen group (P<0.01). The total effective rate was 91.43% (32/35) in TCM group, 66.67% (10/33) in ibuprofen groups, and 30.30% (10/33) in placebo group . The efficacy of the TCM group was better than that of the ibuprofen group (χ²=-2.971, P<0.01), and the efficacy of the ibuprofen group was better than that of the placebo group (χ²=-2.371, P<0.05). After treatment, the levels of CD3⁺, CD4⁺ and CD4⁺/CD8⁺ in TCM group were significantly increased and the levels of CD8⁺ were decreased significantly as compared with those before treatment (P<0.01). After treatment, the levels of CD4⁺ and CD4⁺/CD8⁺ in TCM group were higher (P<0.05，P<0.01)，while the levels of CD8⁺ were significantly lower than those in ibuprofen group and placebo group (P<0.01). Conclusion:Wenjing Huayu Zhitong therapy can reduce the VAS score and accumulative time of abdominal pain, and improve the dysmenorrhea symptoms in patients with PD. Reversal of the T cell subsets disorder may be one of its mechanisms in treating PD with cold coagulation and blood stasis.