Prednisone may rebuild the immunologic homeostasis: Alteration of Th17 and Treg cells in the lymphocytes from rats' spleens after treated with prednisone-containing serum.

BACKGROUND: This study aimed to investigate alterations of T helper 17 (Th17), regulatory T (Treg) cells and relative cytokines after treating with prednisone-contained serum. Lymphocytes were isolated from female rats' spleens. METHODS: The splenic lymphocytes were divided into four groups: which were treated with normal rats' serum (control); prednisone-containing rats' serum (PDN); normal rats' serum and cytokines (CTK); cytokines and prednisone-containing rats' serum (PDN + CTK). The mRNA expression level of RORC, Foxp3 and interleukin-17 (IL-17) was examined by reverse transcription-polymerase chain reaction. The quantities of Th17 and Treg cells were tested by flow cytometry, and the concentrations of IL-17 and IL-10 were detected by enzyme-linked immunosorbent assay. RESULTS: Higher mRNA expression level of Foxp3, percentages of Treg/CD4+ , and concentrations of IL-10, lower mRNA expressions of RORC and IL-17, concentrations of IL-17 and percentages of Th17/CD4+ in PDN group were detected, compared with control group (all p < 0.01). Similar trend was detected in PDN + CTK group, compared with CTK group (all p < 0.01). CONCLUSION: Our results suggest that prednisone may rebuild the immunologic homeostasis and may be used in human diseases with changes in the imbalance immune system such as unexplained recurrent spontaneous abortion (URSA), hepatitis B infection, or other autoimmune diseases.

Prednisone may induce immunologic tolerance by activating the functions of decidual immune cells in early pregnancy.

The objective of this study was to investigate alterations in human first-trimester decidual immune cells (DICs) and relevant cytokines after treatment with prednisone. Decidual lymphocytes were treated with prednisone alone, cytokines alone or the combination of prednisone and cytokines. Levels of STAT3, STAT5, RORC and FOXP3 mRNA were assayed using quantitative real-time PCR, proportions of CD4+ T helper 17 (Th17) and CD4+ T regulatory (Treg) cells were measured using flow cytometry, and concentrations of interleukin (IL)-17A and IL-10 were determined using enzyme-linked immunosorbent assay. After treatment with prednisone alone, levels of STAT5 and FOXP3 mRNA were significantly higher than in untreated control cells (both P < 0.01), while levels of RORC mRNA were significantly lower than in controls (P < 0.05). Levels of STAT3 mRNA did not vary significantly with treatment. After treatment with prednisone alone, proportions of Th17/CD4+ cells and levels of IL-17A were significantly lower than in control cells, and proportions of Treg/CD4+ cells and levels of IL-10 significantly higher than in controls (all P < 0.01). Our results suggest that prednisone may improve pregnancy outcomes by restoring immunological homeostasis through up-regulation of STAT5 and FOXP3, induction of DIC differentiation into Treg cells, inhibition of DIC differentiation into Th17 cells, reduction of IL-17A secretion and induction of IL-10 secretion.

Proportion of Uterine Malignant Tumors in Patients with Laparoscopic Myomectomy: A National Multicenter Study in China.

BACKGROUND: The Food and Drug Administration recently announced that the use of morcellation may cause fibroids or pelvic dissemination and metastasis of uterine sarcoma; therefore, the use of morcellation is limited in the USA. A large sample study is necessary to assess the proportion of uterine malignant tumors found in patients with laparoscopic myomectomy. METHODS: A national multicenter study was performed in China. From 2002 to 2014, 33,723 cases were retrospectively selected. We calculated the prevalence and recorded the clinical characteristics of the patients with malignancy after morcellation application. A total of 62 cases were finally pathologically confirmed as malignant postoperatively. Additionally, the medical records of the 62 patients were analyzed in details. RESULTS: The proportion of postoperative malignancy after morcellation application was 0.18% (62/33,723) for patients who underwent laparoscopic myomectomy. Nearly 62.9% (39/62) of patients had demonstrated blood flow signals in the uterine fibroids before surgery. And, 23 (37.1%) patients showed rapid growth at the final preoperative ultrasound. With respect to the pathological types, 38 (61.3%) patients had detectable endometrial stromal sarcoma, 13 (21.0%) had detectable uterine leiomyosarcoma, only 3 (3.2%) had detectable carcinosarcoma, and 5 (8.1%) patients with leiomyoma had an undetermined malignant potential. CONCLUSIONS: The proportion of malignancy is low after using morcellation in patients who undergo laparoscopic myomectomy. Patients with fast-growing uterine fibroids and abnormal ultrasonic tumor blood flow should be considered for malignant potential, and morcellation should be avoided.

The impact of atosiban on pregnancy outcomes in women undergoing in vitro fertilization-embryo transfer: A meta-analysis.

BACKGROUND: Atosiban is administered to women undergoing in vitro fertilization-embryo transfer (IVF-ET) to improve pregnancy outcomes. However, the results of this treatment were controversial. We conducted this meta-analysis to investigate whether atosiban improves pregnancy outcomes in the women undergoing in vitro fertilization (IVF). METHODS: Databases of PubMed, EMBASE, Web of Science, China BioMedicine, and Google Scholar were systematically searched. Meta-analyses were performed to investigate whether atosiban improves pregnancy outcomes in the women undergoing IVF. RESULTS: Our results showed that atosiban was associated with higher implantation (OR = 1.63, 95% CI: 1.17-2.27; P = 0.004) and clinical pregnancy (OR = 1.84, 95% CI: 1.31-2.57; P < 0.001) rates. However, atosiban showed no significant association with the miscarriage, live birth, multiple pregnancy or ectopic pregnancy rates. When a further subgroup analysis was performed in the women undergoing repeated implantation failure (RIF), implantation (OR = 1.93, 95% CI: 1.45-2.57; P < 0.001), clinical pregnancy (OR = 2.48, 95% CI: 1.70-3.64; P <0.001) and the live birth (OR = 2.89, 95% CI: 1.78-4.67; P < 0.001) rates were significantly higher in the case group. Nevertheless, no significant difference was detected in the miscarriage and multiple pregnancy rates between the case and control groups. CONCLUSION: Atosiban may be more appropriate for women undergoing RIF and play only a limited role in improving pregnancy outcomes in the general population of women undergoing IVF. These conclusions should be verified in large and well-designed studies.

Interleukin 23 regulates the functions of human decidual immune cells during early pregnancy.

BACKGROUND: This study investigated the effects of interleukin 23 (IL-23) on the production of cytokines (IL-1, IL-4, IL-10, and IL-17), the differentiation of Treg/Th17 and STAT3 (i.e., signal transducer and activator of transcription 3) in human decidual immune cells (DICs) during early pregnancy. METHODS: DICs were treated with recombinant human IL-23 and an antibody against IL-23 subunit p19. The differentiation of Treg and Th17 cells was detected by flow cytometry. Levels of IL-23 receptor (IL-23R), STAT3, and phosphorylated STAT3 (pSTAT3) was examined by Western blot. The production of IL1, IL4, IL10, and IL-17 in DICs was measured by ELISA. RESULTS: Exogenous recombinant human IL-23 significantly promoted the differentiation of Th17 cells from DICs, while anti-IL-23 antibody significantly promoted the differentiation of Treg cells from DICs. Consistent with the differentiation of Th17 and Tregs cells, levels of IL-1ß and IL-17 correlated positively with IL-23 treatment, and anti-IL-23 antibody increased the secretion of IL-4 and IL-10 from DICs. Levels of pSTAT3, but not STAT3 or IL-23R, were significantly elevated by recombinant IL-23 treatment; anti-IL-23 antibody significantly decreased the levels of pSTAT3 and IL-23R in DICs. CONCLUSIONS: IL-23 mediates the differentiation of Th17 and Treg cells and the production of associated cytokines in DICs. The potential mechanism likely involves the STAT3 pathway.

The use of glyburide in the management of gestational diabetes mellitus: a meta-analysis.

PURPOSE: Glyburide has been used for managing gestational diabetes mellitus (GDM) in a number of countries. It is rather inexpensive. However, its efficacy and safety remain controversial. With this meta-analysis, we evaluated glyburide in comparison with insulin. MATERIAL/METHODS: With a systematic literature search strategy, a total of 93 randomized controlled trials (RCTs) with insulin and glyburide comparison were identified. Based on the revised Consolidated Standards of Reporting Trials (CONSORT) checklist, five of them met the inclusion criteria and were included in this meta-analysis. RESULTS: Six hundred and seventy four subjects were included in these five RCTs. When compared with insulin, glyburide had an increased relative risk (RR) for neonatal hypoglycemia (RR: 1.98; 95% confidence interval [CI]: 1.17, 3.36). Estimation of standard mean differences (SMD) showed that both fetal birth weight and incidence of macrosomia were higher in subjects receiving glyburide than in those receiving insulin (SMD: 0.21; 95% CI: 0.06, 0.36; RR: 2.22; 95% CI: 1.07, 4.61 respectively). There were no significant differences in maternal glucose control, glycated hemoglobin, the rate of Cesarean section, large-for-gestational age, neonatal hypocalcemia, length of stay for neonatal ICU admissions, preterm birth, or congenital anomalies. CONCLUSIONS: Our study suggested that in women with GDM, glyburide is as effective as insulin, but the risks of neonatal hypoglycemia, high fetal birth weight, and macrosomia were higher.

[Effects of strontium chloride activation on the cleavage rate and somatic cell nuclear transfer embryos in mice].

OBJECTIVE: To establish a suitable protocol for activating mouse somatic cell nuclear transfer embryos with strontium chloride (SrCl2). METHODS: We constructed and identified mouse nuclear transfer (NT) embryos. After nuclear injection, we activated the NT embryos using the following chemical activation methods: exposing the NT embryos to 5 and 10 mmol/L SrCl2 strontium for 1 -8 h, activating the NT embryos with 1-20 mmol/L SrCl2 strontium at 4 and 6 h, treating the NT embryos with 10 mmol/L SrCl2 strontium in different activating media, and exposing the NT embryos to 10 mmol/L SrCl2 strontium combined with 6-dimethylaminopurine (6-DMAP) and cytochalasin B (CB). After activation, the NT embryos were cultured in vitro in the cleavage medium. RESULTS: When the NT embryos were treated with SrCl2 at the concentration of 5 mmol/L, the cleavage rate was remarkably higher at 6 h (38.9%) than at 1 h (6.7%), 2 h (22.8%), 3 h (22.8%) and 4 h (25.6%) (P < 0.05), but with no significant differences from those at 5 h (28.9%), 7 h (34.4%) and 8 h (28.9%) (P > 0.05). When the NT embryos were treated with SrCl2 for 6 h, the rates of cleavage and blastulation were 68.9% and 7.2% at 10 mmol/L, markedly higher than at 1 mmol/L (28.3% and 0%), 2.5 mmol/L (35.6% and 0%), 5 mmol/L (37.8% and 1.1%), 7.5 mmol/L (60.6% and 2.2%), 15 mmol/L (51.7% and 1.1%), and 20 mmol/L (41.7% and 1.1%) (P < 0.05). The cleavage rate of the NT embryos cultured in the Ca2+ and Mg2+ KSOM medium was 27.8%, significantly lower than in the Ca(2+)-free KSOM (69.4%), Ca2+/Mg(2+)-free KSOM (66.1%), and Ca2+/Mg(2+)-free + EDTA KSOM (68.3%) (P < 0.05). The total cell blastocyst number was significantly larger in the NT embryos treated with SrCl2 + CB (45.40 +/- 2.23) than in those treated with SrCl2 (30.15 +/- 1.12), 6-DMAP (34.95 +/- 1.38), and 6-DMAP + CB (37.45 +/- 1.43) (P < 0.05). CONCLUSION: Six-hour treatment with 10 mmol/L SrCl2 in Ca2+ alone or in combination with CB can well activate NT embryos in mice.

Not every subseptate uterus requires surgical correction to reduce poor reproductive outcome.

OBJECTIVE: To evaluate whether hysteroscopic septoplasty should be performed in all women diagnosed with subseptate uterus. METHODS: In a prospective study, 138 patients diagnosed with subseptate uterus at the First Affiliated Hospital of Guangxi Medical University, Nanning, China, were enrolled between January 1, 2006, and March 1, 2011, and reproductive outcomes were compared among women who did and those who did not undergo hysteroscopic resection. Women were divided in 2 groups: group A comprised women with a history of recurrent spontaneous abortion (RSA), and was subdivided into control (A1) and surgery (A2) groups; group B comprised women with no history of poor reproductive outcomes, and was subdivided into control (B1) and surgery (B2) groups. RESULTS: The rates of pregnancy and term delivery were higher in group A2 than in group A1 (P<0.05). The incidence of RSA and preterm delivery was higher in group A1 than in group A2 (P<0.05). There was no difference in pregnancy rate, incidence of RSA, or preterm or term delivery between group B1 and group B2. CONCLUSION: Hysteroscopic septoplasty significantly improved pregnancy outcomes in women with a history of RSA, but did not influence reproductive outcomes in women with no history of poor pregnancy outcomes.

[Results of requesting a second consecutive sperm ejaculate on the day of oocyte pick-up in assisted reproductive technology].

OBJECTIVE: To 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). METHODS: We 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. RESULTS: The 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. CONCLUSION: A 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.

[Prenatal gene diagnosis of paternally inherited alpha-thalassemia by detecting fetal DNA in maternal plasma].

OBJECTIVE: To evaluate the value of diagnosis of alpha-thalassemia by analyzing fetal DNA in maternal plasma. METHODS: Ten families were screened, the husbands being alpha-thalassemia Southeast Asia deletion (SEA alpha-thalassemia-1) heterozygotes and the pregnant women being alpha-thalassemia-2 heterozygotes. Fluorescent polymerase chain reaction (PCR) and gene scanning were used to detect the paternally inherited genotypes of SEA alpha-thalassemia-1 gene mutation and short tandem repeats (STRs) in the maternal plasma fetal DNA. The results were compared to those of conventional prenatal diagnosis of fetal DNA in amniotic fluid, chorionic villus or cord blood. RESULTS: Paternally derived STR genotypes were detected in all specimens of plasma fetal DNA. Paternally inherited SEA alpha-thalassemia-1 gene mutation was detected in 4 cases, while the other 6 cases did not inherit the paternal mutation. The results were completely concordant with those of the conventional prenatal diagnosis. CONCLUSION: Noninvasive prenatal diagnostic method, the technique using fluorescent PCR and gene scanning to detect the fetal DNA and paternally inherited SEA alpha-thalassemia-1 gene mutation in maternal plasma helps exclude the fetuses with hemoglobin H diseases.

[Effect of platelet-activating factor on sperm function].

Platelet-activating factor (PAF) is a unique and novel signaling phospholipid that has pleiotropic biologic properties in addition to platelet activation. Recent studies show that this novel compound plays a significant role in male reproduction and sperm function. PAF binds surface special receptors inducing the formation of inositol triphosphate (IP3) and diacylglycerol (DAG) and increasing intracellular calcium. The concentrations of sperm-derived PAF may help predict sperm motility and fertilization potential, which may serve as a valuable marker for assessing male fertility. Exogenous PAF can improve sperm motility, acrosome reaction and pregnancy rates in human intrauterine inseminations.

[Identification of G6PD mutations in Guangxi by combination of denaturing high performance liquid chromatography, DNA sequencing and restriction endonuclease analysis].

OBJECTIVE: To explore new genotypes of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Guangxi. METHODS: G6PD mutations were identified by combination of denaturing high performance liquid chromatography (DHPLC), DNA sequencing and restriction endonuclease assay. RESULTS: Three cases (10.0%) of G6PD Viangchan (871G-->A, 1311C-->T) and one case (3.3%) of G6PD Union (1360C -->T) were first identified in China mainland. G6PD Ganton (1376G-->T, 30.0%) was the commonest mutation, followed by Kaiping (1388G-->A, 26.7%), and Gaohe (95A-->G, 23.3%). CONCLUSION: Besides G6PD Ganton, Kaiping and Gaohe, there are G6PD Viangchan and Union in Guangxi. G6PD Viangchan and Union is reported for the first time in China mainland.