Etiological risk factors for subfertility among Palestinian women in Gaza

The inability to procreate is frequently considered a personal tragedy and a hardship for couples, impacting on the entire family and even the local community. In Gaza strip, Palestine, there has been no study on etiological risk factors for subfertility. The present study aimed to identify risk factors associated with subfertility among women in Gaza, Palestine. One hundred and sixty-nine women in the study group and 115 women in the control group were included. Cases were selected randomly from those referred to the A1 Basma Fertility Center, Gaza, Palestine. Data were collected through close-ended questionnaire, sonography, hormonal analysis and thrombophilia profile that included the methylenetetrahydrofolate reductase （MTHFR 677 C 〉 T）, factor V leiden （1691 G 〉 A） and pro- thrombin （20210 G 〉 A） genes. By using univariate analyses, the effects of different patient-related variables on the presence of subfertility were evaluated. A multiple logistic regression model was constructed, crude and adjusted odds ratios （OR） and 95% confidence intervals （95% CI） were calculated. The findings showed that 73.5 % （169/230） of the women referred to the A1 Basma Center sought treatment for subfertility. Different etiological risk factors were associated with subfertility, the most frequent of which in descending order were： thrombophilic disorders, fallopian tube problems, sex hormone abnormalities and polycystic ovary syndrome with an adjusted OR of 21.42, 13.63, 11.69 and 10.29, respectively. In conclusion, several etiological risk factors are responsible for subfertility among women in Gaza. Comprehensive evaluation of infertile women should be considered in the course of treatment; otherwise, the duration of sterility may be extended.

Short-term effects of organic amendments on soil fertility and root growth of rubber trees on Hainan Island, China

Rubber[Hevea brasiliensis(Willd.ex A.Juss.)Müll.Arg.]plantations are the largest cultivated forest type in tropical China.Returning organic materials to the soil will help to maintain the quality and growth of rubber trees.Although many studies have demonstrated that organic waste materials can be used to improve soil fertility and structure to promote root growth,few studies have studied the eff ects of organic amendments on soil fertility and root growth in rubber tree plantations.Here,bagasse,coconut husk or biochar were applied with a chemical fertilizer to test their eff ects on soil properties after 6 months and compared with the eff ects of only the chemical fertilizer.Results showed that the soil organic matter content,total nitrogen,available phosphorus and available potassium after the chemical fertilizer(F)treatment were all signifi cantly lower than after the chemical fertilizer+bagasse(Fba),chemical fertilizer+coconut husk(Fco)or chemical fertilizer+biochar(Fbi)(p<0.05).Soil pH in all organic amendments was higher than in the F treatment,but was only signifi cantly higher in the Fbi treatment.In contrast,soil bulk density in the F treatment was signifi cantly higher than in treatments with the organic amendments(p<0.05).When compared with the F treatment,soil root dry mass increased signifi-cantly by 190%,176%and 33%in Fba,Fco and Fbi treatments,respectively(p<0.05).Similar results were found for root activity,number of root tips,root length,root surface area and root volume.Conclusively,the application of bagasse,coconut husk and biochar increased soil fertility and promoted root growth of rubber trees in the short term.However,bagasse and coconut husk were more eff ective than biochar in improving root growth of rubber trees.

A Simple One-Step System Enhances the Availability of High-Quality Sperm for Assisted Reproductive Procedures

Over the last forty years, in vitro fertilization, which has expanded to assisted reproductive technologies (ART), has gone from an experimental procedure to the mainstay of infertility treatment. A technique that once made news with each birth is now responsible for 2% - 3% of the babies born in several nations of the world. This has happened due to significant advances in hormone therapies, culture techniques, and the specialization of equipment designed to support oocytes and embryos. However, for all the advances made to support female fertility, little has changed in male treatment since the advent of intracytoplasmic sperm injection in the early 1990’s. Recently, a number of authors have documented problems with sperm preparation techniques. Some report DNA damage, others membrane and organelle issues, all of which potentially hamper fertilization rates and possibly take-home baby rates. Further, as the clinical workload of ART has increased and staffing shortages have become critical, all labs are looking for simpler, more efficient ways to perform job functions. This study describes a simple, one-step method for preparing semen samples for ART. This new technique minimizes excessive manipulation of the sample compared to current standards and is less likely to cause cell damage. Preliminary results suggest a significant enhancement in recovered sample motility and an optimal sample for ART procedures with minimal sample manipulation.

Response of Tomato on Calcareous Soils to Different Seedbed Phosphorus Application Rates

Field experiments were conducted with five rates (0, 75, 150, 225, and 450 kg P2O5 ha-1) of seedbed P fertilizer application to investigate the yield of tomato in response to fertilizer P rate on calcareous soils with widely different levels of Olsen P (13-142 mg kg-1) at 15 sites in some suburban counties of Beijing in 1999. Under the condition of no P fertilizer application, tomato yield generally increased with an increase in soil test P levels, and the agronomic level for soil testing P measured with Olsen method was 50 or 82 mg kg-1 soil to achieve 85% or 95% of maximum tomato yield, respectively. With regard to marketable yield, in the fields where Olsen-P levels were < 50 mg kg-1, noticeable responses to applied P were observed. On the basis of a linear plateau regression, the optimum seedbed P application rate in the P-insufficient fields was 125 kg P2O5 ha-1 or about 1.5-2 times the P removal from harvested tomato plants. In contrast, in fields with moderate (50 < Olsen P < 90 mg kg-1) or high (Olsen P > 90 mg kg-1) available P, there was no marked effect on tomato fruit yield. Field survey data indicated that in most fields with conventional P management, a P surplus typically occurred. Thus, once the soil test P level reached the optimum for crop yield, it was recommended that P fertilizer application be restricted or eliminated to minimize negative environmental effects.

Mitigating N2O and NO Emissions from Direct-Seeded Rice with Nitrification Inhibitor and Urea Deep Placement

Soil-emitted nitrous oxide(N2O) and nitric oxide(NO) in crop production are harmful nitrogen(N) emissions that may contribute both directly and indirectly to global warming. Application of nitrification inhibitors, such as dicyandiamide(DCD), and urea deep placement(UDP), are considered effective approaches to reduce these emissions. This study investigated the effects of DCD and UDP, compared to urea and potassium nitrate, on emissions, nitrogen use efficiency and grain yields under direct-seeded rice. High-frequency measurements of N2O and NO emissions were conducted using the automated closed chamber method throughout the crop-growing season and during the ratoon crop. Both UDP and DCD were effective in reducing N2O emissions by 95% and 73%, respectively. The highest emission factor(1.53% of applied N) was observed in urea, while the lowest was in UDP(0.08%). Emission peaks were mainly associated with fertilization events and appeared within one to two weeks of fertilization. Those emission peaks contributed to 65%–98% of the total seasonal emissions. Residual effects of fertilizer treatments on the N2O emissions from the ratoon crop were not significant;however, the urea treatment contributed 2%, whereas UDP contributed to 44% of the total annual emissions. On the other hand, cumulative NO emissions were not significant in either the rice or ratoon crops. UDP and DCD increased grain yields by 16%–19% and N recovery efficiency by 30%–40% over urea. The results suggested that the use of DCD and UDP could mitigate N2O emissions and increase grain yields and nitrogen use efficiency under direct-seeded rice condition.

比较临床应用第5天和第6天囊胚的玻璃化冷冻和传统的低温冷藏

Objective: To evaluate implantation of day 5 and day 6 vitrified and slow-frozen blastocysts. Design: Retrospective analysis comparing two cryopreservation techniques. Setting: Private IVF clinic. Patient(s): Five hundred eight cryopreserved embryo transfer candidates. Intervention(s): Supernumerary day 5 and day 6 blastocysts were vitrified or slow-frozen and transfered after warming or thawing. Main Outcome Measure(s): Comparison of two cryopreservation techniques with respect to survival rate, implantation, and pregnancy rates of day 5 and day 6 blastocysts. Result(s): In 254 vitrified transfer cycles, survival, embryonic implantation, and clinical pregnancy rates for day 5 blastocysts were 95.9% , 33.4% , 48.7% , respectively, and for day 6 blastocysts 97.5% , 25.9% , 42.8% . In 254 slow-frozen transfer cycles, survival, embryonic implantation, and clinical pregnancy rates for day 5 blastocysts were 91.4% , 29.6% , 42.8% , respectively, and for day 6 blastocysts 94.8% , 28.2% , 43.1% . Overall there was a slightly, but not significantly, higher outcome regarding implantation and clinical pregnancy with the use of day 5 blastocysts (31.3% and 45.4% , respectively) versus day 6 blastocysts (26.7, and 42.9% , respectively). Conclusion(s): Vitrification technique yields the same implantation and pregnancy rate as slow-frozen blastocyst transfers. Slow growing embryos can be cryopreserved on day 6, because they yield a satisfactory survival, implantation, and pregnancy rate.

The Association between Mycoplasma Species Infection and Tubal Obstruction

Objectives: Numerous factors can cause infertility. Tubal factor accounts for approximately 30% of infertility in females. This study was conducted to evaluate the rate of asymptomatic Mycoplasma hominis and Ureaplasma urealyticum infection in females diagnosed with tubal obstruction. Methods: This is a retrospective case-control study conducted at New Life Fertility Centre. We have identified and reviewed the health records of 167 subfertile women that had HSG and an endocervical swab for Mycoplasma hominis and Ureaplasma urealyticum done. Results: We compared the endocervical swab results of ureaplasma and mycoplasma in the patients with tubal obstruction (group 1) to the patients with normal patent tubes (group 2). Diagnosis of tubal patency was based on the HSG results. Our results show that there is a significantly higher rate of Ureaplasma urealyticum/Mycoplasma hominis infection in group 1 compared to group 2. Conclusion: Our data shows that there is a high rate of U. urealyticum and M. hominis infection in patients diagnosed with tubal factor of infertility and it can be a marker in the prediction of the tubal pathology.

FSH/LH Ratio as a Predictor of the IVF Outcome in Young Women

In Vitro Fertilization (IVF) is the treatment for many causes of infertility. Many studies were done to investigate different factors that can affect the success rate. This study was conducted to evaluate if cycle day 3 (CD3) follicle-stimulating hormone (FSH)/luteinizing hormone (LH) ratio can be a predictor for the IVF outcome in young sub-fertile females ≤ 35 years with normal baseline FSH. This is a retrospective case control study conducted at the Centre of Fertility and Andrology Care (CFAC) in Egypt where 235 sub-fertile women underwent IVF. Patients were grouped based on CD3 FSH/LH ratio. Group A consisted of ≤35-year-old women with FSH/LH ratio ≤35-year-old women with FSH/LH ratio ≥ 2. The primary outcomes include the fertilization rate, implantation rate and the clinical pregnancy rate. The secondary outcomes include duration and the total dose of gonadotrophin used. We found that, there was no significant difference in the total dose of gonadotropin used during the IVF cycle. Also, there was no significant difference in the number of retrieved and fertilized oocytes and the number of good embryos. Clinical pregnancy rate was the same in both groups. In conclusion, in patients younger than 35 years, CD3 FSH/LH ratio is not a predictor for IVF outcome.

Recovery of Essential Plant Nutrients from Biofuel Residual

Essential plant nutrients contained in residues and wastes generated during biofuel processing can be recovered for further production of bioenergy biomass. The objective of this study was to determine the relative agronomic efficiency of “processed” biofuel residual (PBR). Liquid biofuel residual was “processed” by precipitating phosphate and ammonium in the residual with magnesium into a struvite-like material. Then, in a series of greenhouse experiments, we evaluated the fertility potential of PBR, using sweet sorghum (Sorghum bicolor (L.) Moench), as a test bioenergy crop. We compared the agronomic effectiveness of PBR to inorganic commercial fertilizers, biosolids, and poultry manure as nutrient sources. The sources were either applied alone or in combination with supplemental essential plant nutrients (S, K, Mg, and micronutrients). In each of the greenhouse experiments, the crop was grown for 12 wk on soil of minimal native fertility. After each harvest, sufficient water was applied to the soil in each pot over a 6-wk period to yield ~2 L (~one pore volume) of leachate to assess potential total N and soluble reactive phosphorus (SRP) losses. Dry matter yields from the PBR treatment applied alone were significantly greater than yields from inorganic fertilizers, biosolids, and poultry manure treatments applied alone, and similar to yields obtained when the supplemental essential plant nutrients were added to the inorganic fertilizer, biosolids, and manure treatments. Leachate N and SRP concentrations from the PBR treatment were significantly lower than in the treatments with inorganic fertilizers, poultry manure, and biosolids. We conclude that PBR can substitute for inorganic fertilizers and other organic sources of plant nutrients to produce bioenergy biomass cheaply, without causing offsite N and P losses in vulnerable soils.

Maternal low protein diet and fetal programming of lean type 2 diabetes

Maternal nutrition is found to be the key factor that determines fetal health in utero and metabolic health during adulthood.Metabolic diseases have been primarily attributed to impaired maternal nutrition during pregnancy,and impaired nutrition has been an immense issue across the globe.In recent years,type 2 diabetes(T2D)has reached epidemic proportion and is a severe public health problem in many countries.Although plenty of research has already been conducted to tackle T2D which is associated with obesity,little is known regarding the etiology and pathophysiology of lean T2D,a variant of T2D.Recent studies have focused on the effects of epigenetic variation on the contribution of in utero origins of lean T2D,although other mechanisms might also contribute to the pathology.Observational studies in humans and experiments in animals strongly suggest an association between maternal low protein diet and lean T2D phenotype.In addition,clear sex-specific disease prevalence was observed in different studies.Consequently,more research is essential for the understanding of the etiology and pathophysiology of lean T2D,which might help to develop better disease prevention and treatment strategies.This review examines the role of protein insufficiency in the maternal diet as the central driver of the developmental programming of lean T2D.

Is bigger better: The association between follicle size and livebirth rate following IVF?

Purpose: To analyze the correlation between lead follicle size at hCG trigger and IVF outcome. Methods: A review of all patients undergoing their first IVF cycle between 1/2005-12/2009 was performed. Four groups were evaluated (<18 mm, 18 - 18.9mm, 19 - 19.9 mm and ≥20 mm), based on the mean diameter of the two largest follicles on day of ovulation trigger (OT);cycle parameters and outcomes were analyzed. Results: 1577 cycles were reviewed. There were no significant differences noted for cycle parameters or outcomes for the 4 groups. However, although LBR was not significantly different, there was a decline noted as lead follicle size increased. Conclusions: Delaying the administration of OT to enhance follicular growth does not appear to improve IVF outcome. Larger lead follicles do not yield a higher percentage of mature oocytes, embryos available for transfer or LBR. A misguided zest to achieve a higher quantity of fertilizable oocytes may impair oocyte and embryo quality.

Evaluation of Sperm DNA Fragmentation amongst Infertile Black Africans. A Nigerian Study

Background: Male infertility is approaching an epidemic proportion. Almost 50% of all cases of infertility may be associated with a male factor. The diagnostic usefulness of sperm DNA integrity is now accessible as an additional tool to Seminal Fluid Analysis. Objective: To assess sperm DNA fragmentation index (SDFI) in male infertility and its relationship with obesity, alcohol consumption and cigarette smoking among infertile Nigerians. Patients and Methods: Patients who presented for infertility at three health facilities of Nordica Fertility Center in Lagos, Asaba and Abuja cities in Nigeria. STATA 13 was used for student’s t-test to compare the means of continuous variables among smokers and non-smokers and among alcohol consumers and non-consumers. Linear regression analysis was employed to assess the correlation between SDFI as dependent variable and some independent variables. Results: There was no significant difference in the SDFI of men aged &#8722;1.80, P-value = 0.04) than that of obese men (30.2%). Obese men were 2.12 times as likely to have SDFI ≥25% compared to normal weight men (χ2&#45;2.16, P-value = 0.14, OR = 2.12, 95% CI: 0.77, 5.80). Mean SDFI of men who consume alcohol (37.1%) was significantly higher (t = &#8722;1.97, P-value = 0.03) than that of those who did not consume alcohol. Although Pearson’s correlation matrix (r) indicated that sperm DNA fragmentation index was positively correlated with history of infertility (r = 0.01), groin surgery (r = 0.04), mumps (r = 0.04) and sexually transmitted illness (r = 0.04), however the degree of correlation was not significant (P-value ≥ 0.5) in each case. Conclusion: This is the first report in Black Africa that describes a correlation between sperm DNA integrity, as measured by the halo test and age, BMI and alcohol consumption. Men with normal BMI were more likely to have excellent to good SDFI and hence good fertility potential. Data from this study indicate that the infertile men had significantly higher sperm DNA fragmentation. Obese men and those engaged in alcohol consumption also had higher sperm DNA fragmentation indices.

Pyospermia in Men Seeking Assisted Reproduction Technology

Introduction: Pyospermia is often detected in semen, especially when investigating for male infertility. Pyospermia, in a significantly large number is often correlated with inferior sperm parameters and reduced fertility. Pyospermia may be a factor of a fundamental genitourinary infection. This study assessed the relationship between pus cells and semen parameters in infertile Nigerian males. Subjects and Methods: A cross-sectional descriptive study was carried out at Nordica Fertility Center, Lagos, Nigeria, from 2004 to 2009. A total of 907 subjects were analyzed. Subjects ages (years) were categorized into ≤30, 30.1 - 35, 35.1 - 40, 40.1 - 45, 45.1 - 50 and >50, BMI into Results: Leucocytopenia of ≥3 per high power field (/hpf) occurred in 67 (7.4%) men. The overall means (±sd) of pus cells was 1.4 (0.9), 1.4 (0.7), 1.4 (0.7) and 1.6 (1.6) per high power field in all, normospermia, oligoospermic and azoospermic clusters of patients respectively. Mean (±sd) pus cells in semen was 6.0 (4.0), the highest, among obese (BMI ≥ 30) azoospermic patients. The proportion of subjects with pyospermia of 1 - 2 was highest (323/346, 93.3%) among oligoospermic patients while that with pyospermia of ≥3 was highest (10/109, 9.2%) among azoospermic patients. Obese azoospermic patients were 1.8 times as likely to have ≥3/hpf pyospermia compared to normal normospermic patients (x2 = 0.05, P-value = 0.51, OR = 1.82, 95% CI: 0.30, 11.02). Mean progressive motility (%) was significantly reduced (t = 1.95, P-value = 0.03) among patients with ≥3/hpf pyospermia (33.75 ± 19.23) compared to those with 1 - 2/hpf pyospermia (38.58 ± 21.63). Patients with sperm oval head ≤ 30% had significantly higher pyospermia (t = 2.22, P-value = 0.013) than those with sperm oval head of >30%. Conclusion: Elevated pus cell counts were observed among obese azoospermic men, those with reduced sperm progressive motility and those with lower count of sperms with oval head morphology. Clinicians should consider pyospermia relative to body mass index and mean progressive motility in their management of male infertility.

Human Sperm Freezing: Mini Update

Sperm freezing is widely used in ART clinics around the globe. Very little has actually changed with respect to cryopreservation protocols and methodology of freezing over the last 50 years. The aim of this paper is to briefly review the basic principles that underlie freezing and ice crystal formation and also provide a brief overview of newer sperm freezing techniques like sperm vitrification and freeze drying of sperm.

Adjuvant acupuncture reduces first trimester pregnancy loss after IVF

Background: The objective of this study was to determine the effects of acupuncture treatment as an adjunctive therapy on the outcome of in vitro fertilization (IVF)/Intracytoplasmic Sperm Injection (ICSI). Material/Methods: We conducted a retrospective study of 238 patients who underwent conventional IVF/ ICSI alone or in conjunction with acupuncture over a course of 2 years. Patients in the two treatment groups were matched in terms of age and diagnosis. Acupuncture was administered in two sessions 5 to 7 days prior to and on the day of embryo transfer. Results: There were no differences between the two groups in terms of fertilization rate, pregnancy or implantation rates, and endometrial thickness. The number of oocytes retrieved (P < 0.005) and the number of first trimester miscarriages were significantly lower in the group receiving acupuncture (P < 0.001). Conclusions: Acupuncture reduces miscar-riage rates in patients undergoing IVF/ICSI possibly secondary to stress relief.

Pregnancy Rate after Embryo Transfer on Day 5 and Day 6 in IVF. A Systematic Review and Meta-Analysis

Study Question: The question: is there any difference in pregnancy rate between embryo transfer day 5 and day 6 in IVF? What Is Known Already? Blastocyst transfer is increasingly popular in assisted reproductive technology (ART) centers today. Very few articles concentrate on comparing Day 5 and Day 6 embryo transfer with conflict results. Objective, Study Design: Systematic review and meta-analysis of published controlled studies. Searches conducted from 2001-2020 on PubMed. Medline, EMBASE, and ISI Web of Science Electronic database is used to collect data, using the following search terms: blastocyst, Day 5, Day 6, embryo transfer (E.T.) and pregnancy rate. Materials, Setting, Methods: A total of 6 full-text articles preselected from 211 references, based on title and abstract. Two independent reviewers performed data selection and extraction according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis statement (PRISMA). This systematic review was conducted according to (PICO) standard. Random-effect meta-analysis performed on all data (overall analysis). Results and the Role of Chance: Data from 6 relevant articles were extracted and integrated into the meta-analysis that reported clinical pregnancy rate (CPR) as an outcome in 5640 cycles, 2274 cycle had embryo transfer at day 6 and the control was 3366 cycle had embryo transfer day 5 showed a significantly higher clinical pregnancy rate following Day 5 embryo transfer compared with Day 6 embryo transfer with odd ratio and 95% confidence limit 0.73 (0.66 - 0.82) p value < 0.000. Sensitivity analysis led to similar results and conclusions. Limitations, Reasons for Caution: The validity of meta-analysis results depends mainly on the quality and the number of published studies available. Indeed, this meta-analysis included no randomized controlled trial (RCT). Wider Implications of the Findings: In regards to the results of this original meta-analysis, ART practitioners should preferably transfer D5 rather than D6 blastocysts. Further RCTs are needed to address the question of whether D6 embryos should be transported.

Demographic and Assisted Reproduction Related Factors Associated with Dichorionic Triplet Gestations

Our objective was to evaluate which demographic factors or assisted reproductive technologies were associated with IVF triplet gestations where one of the embryos split, resulting in a dichorionic triplet gestation. This was a case-control study of dichorionic versus trichorionic triplet gestations that underwent assisted reproductive technology over the last 5 years at our fertility center. There were 53 cases of dichorionic triamniotic triplet gestations compared to 119 trichorionic triplet controls. There were no significant demographic differences between the cases and controls. 51/53 dichorionic triplets and 86/119 trichorionic triplets were conceived through IVF, the remaining utilized intrauterine insemination. ICSI was performed in virtually all patients that underwent IVF. Of the potential risk factors studied, hatching was used in 70.6% of dichorionic compared to 89.5% of trichorionic IVF triplets (p = 0.005);embryo transfer was performed on Day 5 or 6 compared to Day 3 in 88.0% dichorionic vs 71.8% trichorionic (p = 0.028). Frozen sperm was utilized more frequently with dichorionic than with trichorionic triplets, 26.0% vs 10.9% (p < 0.011). Only 4 (7.5%) of the IVF cases underwent pre-implantation genetics. Certain assisted reproductive technologies appear to be associated with embryo splitting and a dichorionic triplet gestation. More research is needed in this area to further elucidate these findings.

Elective Single Embryo Transfer: Future Forward

Notwithstanding the most noteworthy verifiable live birth achievement rates for couples experiencing in vitro fertilization (IVF), there has been a scourge of iatrogenic twin and higher gestation development considered from this treatment. While some expanded peril is natural for the barren populace requiring treatment, the act of multiple embryo transfer adds to these inconveniences and results, particularly concerning its part in higher number pregnancies. Improvement in cryopreservation procedures has permitted conservation of supernumerary embryos for use in future cycles, and refinements in culture frameworks and embryo selection have brought about the transfer of elective single embryos while keeping up positive pregnancy rates. The willful exchange of a single top notch quality elective single embryo transfer e-SET (Blastocyst) has fundamentally lessened the multiple gestation rates and boosted the rate of singleton pregnancy without compromising the global success rates. We acknowledge that in high-risk situations (e.g. previous history of preterm gestation and poor maternal wellbeing), double-embryo transfer (DET) or triple embryo transfer (TET) ought to be disallowed because of unsuitably high perils. Be that as it may, we contend that ordering e-SET for every single young lady can be viewed as an unsatisfactory break of patient self-governance, particularly since DET/TET offers certain ladies money related and social focal points.

Colonization of neonate mouse spermatogonial stem cells co-culture with Sertoli cells in the presence and absence soft agar

Objective:To investigate the effects of soft agar on in-vitro proliferation of neonate mouse spermatogonial stem cells co-cultured with Sertoli cells.Methods:Tissues of neonate NMRI male mice testes were used for harvesting spermatogonial stem cells and Sertoli cells.After cell harvest,flow cytometry using promyelocytic leukemia zinc-finger(PLZF)protein antibody was used to assess the purity of the cells.The isolated testicular cells were cultured in the absence(the control group)or presence of soft agar-coated dishes(the experimental group)supplemented with leukemia inhibitory factor and glia cell line–derived neurotrophic factor for two weeks.Alkaline phosphatase activity was assessed in the colonies formed after two weeks of culture by alkaline phosphatase staining.On day 14 of culture,the expression levels of DNA-binding protein inhibitor(ID-4)and PLZF genes in the undifferentiated cells were evaluated by the detection of PLZF protein antibody using real-time PCR and immunocytochemistry techniques.The number and diameter of the colonies of spermatogonial stem cells were assessed by ImageJ software.Results:In the experimental group,the number and the diameter of colonies significantly increased as compared with those in the control group(P<0.05,P<0.01,respectively).In addition,the level of expression of ID-4 and PLZF genes in the undifferentiated cells significantly increased in the experimental group as compared with the control group(P<0.01).However,the expression of tyrosine-protein kinase kit(c-kit)gene in differentiated cells decreased in the experimental group as compared with the control group,but there was no significant difference between the two groups.Conclusions:Spermatogonial stem cells can be efficiently proliferated by culturing the stem cells in soft agar-coated dishes.The new protocol used in this study can be a valuable method for future studies.

Fertility preservation for female patients with childhood and adolescent cancer

With the significant cancer treatment in the past decades,>85%of children with cancer now survives into adulthood,and fertility preservation has become an important quality-of-life technology for them.1 Cancer treatment may include surgery,chemotherapy,radiotherapy,and/or hematopoietic stem cell transplantation(HSCT).2 Except for non-pelvic surgery,these treatments may affect ovarian function and consequently cause varying degrees of gonadal toxicity.3 Patients cured by anti-cancer treatment have a very high risk of premature ovarian insufficiency(POI).