Application of Optical Genome Mapping Technology in Detecting Complex Chromosomal Rearrangement / 中山大学学报(医学科学版)

ObjectiveTo investigate the application of optical genome mapping （OGM） technology in detecting complex chromosomal rearrangement. MethodsWe recruited five patients who were diagnosed as complex chromosomal rearrangement at the Reproductive Medicine Center of the Sixth Affiliated Hospital of Sun Yat-sen University from January 2022 to June 2023. They underwent OGM， nanopore sequencing and pre-implantation genetic testing （PGT）. The results were compared with the results of karyotype and chromosomal microarray analysis （CMA）/ copy number variation sequencing （CNV-Seq）. ResultsOGM could detect translocation， invert inversion， and triplet translocation， which were consistent with the results of OGM and CMA/ CNV-Seq. But OGM could not detect Robertsonian translocation. ConclusionBecause of its ultra-long reads， OGM realizes the detection across repetitive regions， and it has great advantages when applied in patients with complex chromosome rearrangement or uncertain karyotype analysis. It can accurately locate breakpoints.

Effect of Chuanxiong Rhizoma on the absorption and clearance of four coumarins in Angelicae Dahuricae Radix / 药学学报

To study the changes in the pharmacokinetic behavior of four coumarins (bergapten, oxypeucedanin, imperatorin and isoimperatorin) in rats before and after combinating Angelicae Dahuricae Radix with Chuanxiong Rhizoma. The plasma concentrations of the drugs were determined by ultra performance liquid chromatography-fluorescence detection (UPLC-FLD) for dose response and time dependent curves. The pharmacokinetic parameters were calculated by DAS 3.2.8, and SPSS 20.0 was used to analyze the differences of main pharmacokinetic parameters between the two groups. The result showed: comparing with Angelicae Dahuricae Radix group, the area under drug time curve (AUC0-24 h) of bergapten, oxypeucedanin and imperatorin increased by 177.2%, 97.14% and 54.43% respectively, AUC0-∞ increased by 282.3%, 104.2%, and 75.40% respectively, and clearance rate (CLZ/F) decreased by 68.26%, 51.08% and 43.98% respectively; the peak drug concentration (Cmax) of four coumarins was significantly increased; the distribution volume (VZ/F) of bergapten was significantly decreased. These data indicated that Chuanxiong Rhizoma can promote the absorption of coumarins in Angelicae Dahuricae Radix, slow down the elimination of coumarins, and increase their bioavailability in vivo. The animal experiment scheme in this study has been approved by the Experimental Animal Ethics Committee of Beijing University of Chinese Medicine (approval number: BUCM-4-2020083105-3072).

Ovarian Stimulation and In Vitro Maturation for Fertility Preservation in Female Cancer Patients / 中山大学学报(医学科学版)

@#【Objective】To determine the effect of ovarian stimulation or in vitro maturation for fertility preservation in female cancer patients. 【Methods】 A retrospective study was conducted in 27 females who underwent fertility preservation procedures in our center.【Results】Female patients were included in this study with an average age of 27.1. Patients spent on average for 6.8 d to retrieve oocytes since their attendance day. Total amount of Gn was on average 910 U per patient and for patients with breast cancer，the average estrogen level on trigger day reached 360 pg/mL. The maturation rate of oocytes from ovarian stimulation cycles was 82.6% ，which of that in emergency in- vitro maturation cycles was 38.1%.【Conclusion】The development capability of oocytes from cancer patients are comparable with those of other infertility patients. Peak estradiol levels were controlled by the administration of letrozole. In vitro maturation of oocytes performed at random time of the menstrual cycle may result in a lower maturation rate ，which is associated with the time limit of the follow- up cancer treatment. In conclusion，clinicians should consider a more holistic approach for female cancer patients，which focuses not only on the characteristic of the primary cancer but also on the phase of the menstrual cycle at their attendance day.

Micro-dissection testicular sperm extraction for non-obstructive azoospermia patients with the history of secondary testicular injury / 中华男科学杂志

<p><b>Objective</b>To investigate the value of micro- dissection testicular sperm extraction (micro-TESE) in the treatment of non-obstructive azoospermia (NOA) in patients with the history of secondary testicular injury.</p><p><b>METHODS</b>Totally, 121 NOA patients with the history of secondary testicular injury underwent micro-TESE in our hospital from September 2014 to December 2017. We analyzed the correlation of the sperm retrieval rate with the causes of testicular injury and compared the outcomes of the ICSI cycles with the sperm retrieved from the NOA males by micro-TESE (the micro-TESE group) and those with the sperm ejaculated from severe oligospermia patients (sperm concentration <1×10⁶/ml, the ejaculate group). Comparisons were also made between the two groups in the female age, two-pronucleus (2PN) fertilization rate, transferrable embryos on day 3 (D3), D3 high- quality embryos, D14 blood HCG positive rate, embryo implantation rate, and clinical pregnancy rate.</p><p><b>RESULTS</b>Testicular sperm were successfully retrieved by micro-TESE in 86.0% of the patients (104/121), of whom 98.4% had the history of orchitis, 75.5% had been treated surgically for cryptorchidism, and 63.6% had received chemo- or radiotherapy. No statistically significant differences were observed between the micro-TESE and ejaculate groups in the 2PN fertilization rate (59.4% vs 69.3%, P > 0.05), D14 blood HCG positive rate (44.6% vs 57.9%, P > 0.05), embryo implantation rate (31.8 %% vs 32.6%, P > 0.05) and clinical pregnancy rate (41.5% vs 48.7%, P > 0.05). However, the rate D3 transferrable embryos was significantly lower in the micro-TESE than in the ejaculate group (40.5% vs 52.2%,P < 0.05), and so was that of D3 high-quality embryos (32.5% vs 42.1%, P < 0.05).</p><p><b>CONCLUSIONS</b>Micro-TESE can be applied as the first choice for NOA patients with the history of secondary testicular injury, but more effective strategies are to be explored for the improvement of ICSI outcomes with the sperm retrieved by micro- TESE.</p>

The effect of noise exposure on insulin sensitivity in mice may be mediated by the JNK/IRS1 pathway

BACKGROUND@#Epidemiological studies have suggested that noise exposure may increase the risk of type 2 diabetes mellitus (T2DM), and experimental studies have demonstrated that noise exposure can induce insulin resistance in rodents. The aim of the present study was to explore noise-induced processes underlying impaired insulin sensitivity in mice.@*METHODS@#Male ICR mice were randomly divided into four groups: a control group without noise exposure and three noise groups exposed to white noise at a 95-dB sound pressure level for 4 h/day for 1, 10, or 20 days (N1D, N10D, and N20D, respectively). Systemic insulin sensitivity was evaluated at 1 day, 1 week, and 1 month post-noise exposure (1DPN, 1WPN, and 1MPN) via insulin tolerance tests (ITTs). Several insulin-related processes, including the phosphorylation of Akt, IRS1, and JNK in the animals' skeletal muscles, were examined using standard immunoblots. Biomarkers of inflammation (circulating levels of TNF-α and IL-6) and oxidative stress (SOD and CAT activities and MDA levels in skeletal muscles) were measured via chemical analyses.@*RESULTS@#The data obtained in this study showed the following: (1) The impairment of systemic insulin sensitivity was transient in the N1D group but prolonged in the N10D and N20D groups. (2) Noise exposure led to enhanced JNK phosphorylation and IRS1 serine phosphorylation as well as reduced Akt phosphorylation in skeletal muscles in response to exogenous insulin stimulation. (3) Plasma levels of TNF-α and IL-6, CAT activity, and MDA concentrations in skeletal muscles were elevated after 20 days of noise exposure.@*CONCLUSIONS@#Impaired insulin sensitivity in noise-exposed mice might be mediated by an enhancement of the JNK/IRS1 pathway. Inflammation and oxidative stress might contribute to insulin resistance after chronic noise exposure.

Correlation Analysis of Baseline Antimullerian Hormone Level and Live Birth Rate of Fresh Embryo Transfer Cycles and Frozen Embryo Transfer Cycles / 中山大学学报(医学科学版)

[Objective]To investigate the relationship of baseline antimullerian hormone(AMH)and live birth rate of IVF/ICSI and further explore the prognostic effect of AMH on live birth rate.[Methods]All non-polycystic ovary patients who underwent their first embryo transfers in our unit and had basal serum AMH evaluated between 2010 and 2015 were evaluated in this retrospective study. Patients were grouped according to their AMH level,i.e. low AMH group with AMH less than 1.1 ng/mL(n = 485),middle AMH group with AMH between 1.1 ng/mL and 7.0 ng/mL (n = 1 989),and high AMH group with AMH higher than 7.0 ng/mL (n=468). For age subgroup analysis,patients were stratified as follow:group A(age≤29 years),group B(30~34 years),group C(35~39 years)and group D(over 40 years). We compared clinical outcomes between AMH groups in different age groups usingunivariate and multivariate analysis. ROC analysis was utilized to assess predictive value of AMH on live birth rate.[Results](1)In both fresh and frozen embryo transfers,baseline AMH was significantly related to clinical outcomes. The lower AMH was,the lower implantation rate,clinical pregnancy rate,and live birth rate. However,higher miscarriage rate was observed. All difference reached statistically significant.(2)In age subgroup analysis,we demonstrated AMH was related to live birth rate in patients in group A,B, and C,regardless of fresh or frozen embryo transferred. In those over 40 years,AMH was related to live birth rate in frozen cycles (P < 0.05)but not fresh cycles(P = 0.092). The further multivariate analysis confirmed the above results after controlling po?tential confounding variables.(3)The AUC of ROC analysis for AMH predicting live birth rate were 0.647,0.633 for fresh and fro?zen cycles respectively.[Conclusion]Baseline AMH as one of excellent ovarian reserve markers ,was significantly related to live birth rate in fresh or frozen cycles. Baseline AMH was an independent prognostic factor of live birth rate,but its predictive value on live birth rate was of limited clinical value.

Application Analysis of All-blastocyst-culture in Elderly Infertile Patients with Low Ovarian Reserve / 中山大学学报(医学科学版)

[Objective]To determine whether all-blastocyst-culture can benefit elderly infertile patients with low ovarian reserve.[Methods]To retrospectively analyze elderly patients with low ovarian reserve undergoing IVF/ICSI in Reproductive Medicine Re?search Center of the Sixth Affiliated Hospital of Sun Yat-sen University from May 2016 to April 2017.We divide them into group A (All-blastocyst-culture)and group B(None-blastocyst-culture)based on different culture approach of D3 transferrable embryo. To compare the cumulative biochemical pregnancy rate ,cumulative clinical pregnancy rate between them.[Results]A total of 231 eligi?ble patients are included,with130 in group A and 101 in group B. The basic conditions of the two groups show no difference. Cumula?tive biochemical pregnancy rate/clinical pregnancy rate in group A is higher than that of group B ,though the difference is not statisti?cally significant(P>0.05). After removing patients who did not undergo embryo transfer due to failure in blastocyst culture ,the cumu?lative biochemical pregnancy rate/clinical pregnancy rate in group A is significantly higher than group B(P<0.05). Multivariate analy?sis of the patient′s basic condition and the outcome of blastocyst culture showed that the number of D3 transferrable embryo was a risk factor for the failure of blastocyst culture(OR=0.277,95%CI:0.103~0.744,P<0.05).[Conclusion]All-blastocyst-culture will not adversely affect the pregnancy outcome of elderly infertile patients with low ovarian reserve. On the contrary ,once they obtain transfer?rable embryos, pregnancy outcome in All-Blastocyst-Culture group are better than None-Blastocyst-Culture group. A small number of D3 transferrable embryo is a risk factor for failure to culture blastocyst. If the patients were fully informed consent ,we can consider implementing all-blastocyst-culture for elderly infertile patients with low ovarian reserve.

Predictive Value of Early Serum Beta-Human Chorionic Gonadotropin Measurements After Cleavage or Blastocyst Embryo Transfers / 中山大学学报(医学科学版)

[Objective]To compare early serum beta-human chorionic gonadotropin (β-hCG) levels after cleavage or blastocyst embryo transfers (ET) in predicting pregnancy outcome.[Methods]A total of 2421 IVF-ET cycles in our center performed from June 2010 to May 2015 and resulted in clinical intrauterine pregnancies were analyzed retrospectively. The predictive value was compared betweenβ-hCG on day 14 after cleavage ET andβ-hCG on day 12 after blastocyst ET.[Results]Serumβ-hCG levels of patients re?sulted in clinical intrauterine pregnancies were significantly higher with blastocyst ET compared with cleavage ET. This significant dif?ference was also existed in patients resulted in miscarriage, ongoing pregnancy (OP) or live birth (LB). However, this significant differ?ence was only existed in frozen embryo transfers. For a frozen cleavage ET, the cut-off value was 475 U/L (sensitivity 79%, specificity 61.3%) in predicting LB. For a frozen blastocyst ET, the cut-off value was 575 U/L (sensitivity 74.9%, specificity 59.2%) in predicting LB.[Conclusion]In frozen embryo transfers, early serumβ-hCG level after blastocyst ET is higher than cleavage ET. The cut-off val?ue in predicting pregnancy outcome is different according to the stage embryo transferred. Early serum β-hCG can effectively predict live birth after blastocyst or cleavage ET.

Array comparative genomic hybridization analysis of early-stage arrested human embryos / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate chromosomal euploidies in early-stage arrested human embryos.</p><p><b>METHODS</b>To determine the euploidy status of the 24 chromosomes, 13 embryos were analyzed, which included 5 arrested at 4-cell stage, 4 arrested at 8-cell stage, and 4 embryos at blastocyst stage regardless of their morphological scores. All embryos were subjected to biopsy, whole genome amplification, and array comparative genome hybridization analysis.</p><p><b>RESULTS</b>Chromosome euploidies of the arrested embryos can be normal, aberrant and chaotic. Mosaicism is prevalent in early stage cleavage, whilst most of the blastocysts, even with poor morphology, are normal diploid.</p><p><b>CONCLUSION</b>Arrested embryo may have normal chromosomes euploidy. Mosaicism is common in cleavage stage embryos. Early stage embryo arrest may not be solely attributable to chromosomal aneuploidies and needs further research.</p>

Effects of M II stage oocytes zona pellucida birefringence on pregnancy outcome

OBJECTIVE@#To explore the effects of different M II stage oocytes zona pellucida birefringence on pregnancy outcome.@*METHODS@#A total of 46 couples with infertile which induced by single cause received in-vitro fertilization treatment were analyzed retrospectively, and randomly divided into the high zona birefringence (HZB)/HZB group, HZB/low zona birefringence (LZB) group and LZB/LZB group according to different oocytes zona pellucida birefringence. Intracytoplasmic sperm injection outcome was analyzed and compared.@*RESULTS@#The proportion of HZB oocytes, implantation rate and the pregnancy rate were decreased in three groups (HZB/HZB group>HZB/LZB group>LZB/LZB group) (P0.05). Logistic regression analysis showed that factors affect M II stage oocytes zona pellucida birefringence were age, basal FSH level and the LH level on the day of HCG injection. Age and FSH levels were negatively correlated with the single oocyte zona pellucida birefringence; While the LH level on the day of hCG injection was positively correlated with the single oocyte zona pellucida birefringence.@*CONCLUSIONS@#The primary influence factors on M II stage oocytes zona pellucida are age, basal FSH level and the LH level on the day of hCG injection. The birefringence value of zona pellucida can affect the pregnancy outcome.

Change and significance of growth differentiation factor 9 and bone morphogenetic protein expression during oocyte maturation in polycystic ovary syndrome patients with ovarian stimulation / 中华妇产科杂志

Objective To investigate the expression pattern and significance of two importantoocyte-secreted factors:growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)during oocyte maturation in women with polycystic ovary syndrome(PCOS)and infertile women due to husband factors.Methods Total of 25 oocytes[9 at germinal vesicle GV stage,9 at M Ⅰ stage and 7 at M Ⅱ stage]were obtained from 12 patients with PCOS and 82 oocytes(29 at GV stage,26 at M Ⅰ stage and 27 at M Ⅱ stage)were from 56 controls.The nested quantitative real time(RT)PCR was uscd to detect the abundance of GDF9 and BMP15 mRNA in each oocyte.Results(1)The expression level of GDF9 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 44.8(4.2-529.0),27.6(9.8-172.7)and 49.0(0.2-65.9)respectively,the expression in were 149.9(55.4-387.9),29.9(2.5-205.8)and 657.8(149.4-1376.2)in control group,respectively.The expression of GDF9 mRNA at M Ⅱ stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences didn't reach statistical significant at GV or M Ⅰ stage between the two groups(P ＞ 0.05).The expression of GDF9 mRNA displayed some changes at different maturation stage in controls(P ＜ 0.05,P ＜ 0.01),however,the expression didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).(2)The expression level BMP15 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 0.1(0.1-22.0),3.2(0.6-55.0)and 6.4(3.2-8.5),respectively,the expression were 41.6(6.5-96.1),4.0(2.0-10.4)and 49.7(2.3-139.5)in control group,respectively.The expression of BMP15 mRNA at GV stage was significantly lower in PCOS group than in controls(P ＜ 0.01),however,the differences were not significant at M Ⅰ or M Ⅱ stage between the two groups(P ＞0.05).The expression of BMP15 mRNA also displayed some changes at different maturation stage in controls(P ＜ 0.05),however,the level didn't demonstrate any dynamic changes in PCOS group(P ＞ 0.05).Conclusion It was suggested that the low expression of oocyte secreted factors in mature oocytes from PCOS patients might be associated with impaired oocyte quality and developmental competence in PCOS.

Evaluation of the fidelity of multiple displacement amplification from small number of cells / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the fidelity of multiple displacement amplification (MDA) from small number of cells (1-10 cells) by 10K 2.0 SNP mapping array.</p><p><b>METHODS</b>A fibroblast cell line (Tri-18; GM02732, 47, XY, +18) was used as the template, and 6 groups were set up in the study. Groups A and B were positive and negative control, respectively; groups C-F were experimental groups involving the MDA products from 1, 2, 5 and 10 cells respectively. In combination of single nucleotide polymorphism (SNP) array, the product of each group was assessed based on the genome coverage, loss of heterozygosity (LOH) rate and allele dropout (ADO) rate.</p><p><b>RESULTS</b>The nonspecific product of negative control presented an average call rate of 3.2%. The genome coverage of the MDA product increased from 86.4% to 96.4% with the increasing number of template from 1 to 10 cells, while the LOH rate and ADO rate decreased significantly (P<0.05).</p><p><b>CONCLUSION</b>MDA is a highly efficient and reliable method for whole genome amplification. The fidelity of MDA will be improved significantly with the increasing number of template cells. 10K 2.0 SNP mapping array is a quick, accurate and comprehensive method to evaluate the fidelity of amplified DNA products, but the ADO SNPs should be distinguished from those of preferential amplification from the LOH loci to avoid errors.</p>

Detection of Aneuploidy from Single Cells by Array-Comparative Genetic Hybridization / 中山大学学报(医学科学版)

[Objective] To set up an optimized protocol for aneuploidy detection from single cells through Array- Comparative Genetic Hybridization (CGH).[Method] Two cell lines,trisomy 18 (Tri-18;GM02732,47,XY,+18) and chromosome 4 segment deletion [sDel-4;GM00343,46,XY,4(del) (qter ＞ p14)],were used in the study.In combination of 10 k 2.0 SNP mapping array platform and multiple displacement amplification (MDA),the diagnostic accurate rates of MDA product from single cells of the two cell lines using gDNA and single-cell MDA product as reference were compared.[Result] An extremely lower call rate (3.2 ± 1.2)% in the negative control group was observed compared to the experiment groups.When the single-cell MDA product was used as reference,the standard deviations of Log2 (signal intensity ratio) were significantly decreased in both groups,compared with when the gDNA as reference (P ＝ 0.004).Through CNAT analytic software,some specific chromosomes (16,17,19,and 22) presented obvious preferential amplification (PA) when the gDNA was used as reference,but this PA could be eliminated when single-cell MDA product was used as reference.[Conclusion] 10 k 2.0 SNP mapping array in combination with MDA could be a quick,highly efficient and accurate method to detect aneuploidy in single cells.

Comparing Results with Immature Germinal Vesicle Oocytes From Different In-vitro Fertilization Cycles / 中山大学学报(医学科学版)

[Objective] This study compared outcomes of in vitro maturation (IVM) and in vitro fertilization (IVF) intracytoplasmic sperm injection (ICSI) cycles after IVM of immature germinal vesicle (GV) oocytes.[Methods] ICSI was performed on metaphase II (MII) oocytes retrieved in 163 IVF-ICSI cycles (group I;n ＝ 987) or matured from GV stage oocytes in IVF-ICSI ( group II;n ＝ 132) and 37 IVM cycles ( group III;n ＝ 235).Fertilization and cleavage rates and embryo quality were compared among the three groups.[Results] The fertilization rate,cleavage rate and top quality embryos rate were higher in group I than group II and group III (84.9%,98.1%,and 61.6%;72.0%,90.5% and 22.1%l;75.3%,94.4%,and 25.1%,respectively).Blastomere numbers and morphology scores were highest in group I (P ＜ 0.05),but no significant differences existed between group II and group III.[Conclusion] The morphology of embryos developed from in vivo MII oocytes was superior to those from in vitro matured MII oocytes.No significant difference was observed in embryo morphology from immature GV oocytes in IVF and IVM cycles.

Efficiency comparison between two preimplantation genetic diagnostic methods for chromosomal translocation carriers / 中华妇产科杂志

Objective To compare the diagnostic efficiency between blastomere preimplantation genetic diagnosis (PGD) and polar body PGD for chromosomal translocation carriers. Methods Group A had 8 cycles using whole painting probes for the first polar body diagnosis, while group B had 29 cycles using two subtelomeric probes and one centromeric probe for the blastomere diagnosis. Results The fertilization rate of group A was significantly lower than group B [66. 1% (72/109) vs 85.2% (304/357) , P < 0.05]. There was no significant difference in the successful biopsy rate between two groups. However, group A had a significantly higher loss rate during fixation and higher no signal rate after fluorescence in situ hybridization [ FISH, 9. 6% (12/104) vs 1.6% (4/252), 11.2% (10/89) vs 3.0% (7/233) ]. Totally, the diagnostic efficiency in group A (72. 5% ,79/109 ) was significantly lower than that in group B( 89. 8%, 230/256, P < 0. 05 ). Although both the clinical pregnancy rate( 3/7 ) and implantation rate( 22. 2% ,4/18 ) of group A were higher, the differences were not statistically significant ( P > 0.05 ). Conclusion Both methods can be used efficiently in the PGD for chromosomal translocation carriers. Blastomere PGD has a higher diagnostic rate.

Study of acute lung Injury in rats with acute carbon monoxid poisoning(ACOP) / 中华急诊医学杂志

Objective To structure the model of acute carbon monoxid poisoning(ACOP)in rats. Evaluate the effectiveness of the poisoning on the pulmonary function and the significance of carbon monoxide hemoglobin(HbCO)and oxygenation index in diagnosis of acute lung injury(ALI)/acute respiratory distress syndrome(ARDS).Method Eighty healthy adult male Wistar rats were randomized into 4 groups.According to the concentration of CO,poisoning group was randomized into three groups(each group=20),group A,group B,group C.After poisoned,arterial blood was collected rapidly for arterial blood gas analysis.According to the pathological changes,the models were divided into ALI/ARDS group and non-ALI/ARDS group.Results Compared with control group,the incident rate of ALI/ARDS in group B(25%)and group C(55%)were significantly higher(P

Clinical analysis of intracytoplasmic sperm injection in patients with previous fertilization failure after conventional IVF / 中华男科学杂志

<p><b>OBJECTIVE</b>To analyze the clinical outcome of intracytoplasmic sperm injection (ICSI) in patients with previous fertilization failure after conventional IVF.</p><p><b>METHODS</b>Data from 20 ICSI cases (22 ICSI cycles) with previous complete failure of fertilization or with fertilization rate < or = 20% between January 2002 and December 2004 were retrospectively analyzed. The control group consisted of 100 consecutive ICSI cycles for male factor infertility in the same period.</p><p><b>RESULTS</b>The fertilization rate dramatically increased from 5.4% after conventional IVF to 76.9% after ICSI treatment (chi-squared = 264.66, P < 0.001). However, the fertilization rate in the subgroup with previous low fertilization was significantly lower than those in the control and in the subgroup without previous fertilization (67.9% vs 77.5%, 67.9% vs 84.2%). Compared with the control group, the subgroup without previous fertilization had a higher pregnancy rate and implantation rate, but only the difference in the implantation rate was statistically significant (40.5% vs 18.9%).</p><p><b>CONCLUSION</b>ICSI can overcome previous fertilization failure with conventional in vitro fertilization and thus improve the clinical outcome.</p>

Closely linked polymorphic marker: successful application in preimplantation genetic diagnosis for beta-thalassemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To evaluate the applicability of the polymorphic marker closely linked with beta-globin gene for the preimplantation genetic diagnosis (PGD) in couples at risk of having child with beta-thalassemia.</p><p><b>METHODS</b>Single cell multiplex nested PCR which coamplifies the beta-globin gene and the closely linked polymorphic marker, HumTHO1 gene, was applied in six clinical PGD cycles for four couples with beta-thalassemia.</p><p><b>RESULTS</b>In six clinical PGD cycles, a total of 44 embryos were biopsied and 44 blastomeres were obtained. Forty-one blastomeres were amplified and thirty-five embryos were given definite diagnoses. Fourteen embryos were transferred back to the uterus of the patients and one pregnancy went on well and ended with one live healthy birth, which confirmed the results of PGD. The average amplification efficiency of single blastomere was 89.7% and the average allele drop-out(ADO) rate was 14.4%. The coamplification of HumTHO1 could help to detect the existence of ADO and contamination.</p><p><b>CONCLUSION</b>This is the first report on unaffected pregnancy resulting from PGD using multiplex nested PCR in China. The simultaneous amplification of polymorphic marker closely linked to beta-globin gene(HumTHO1) could help to resist the risk of misdiagnosis in PGD caused by ADO and contamination.</p>

Outcome of repeated epididymal sperm aspiration or testicular sperm extraction in azoospermic patients / 中华男科学杂志

<p><b>OBJECTIVE</b>To review the outcome of repeated percutaneous sperm aspiration (PESA) and testicular sperm extraction (TESE) for intracytoplasmic sperm injection (ICSI).</p><p><b>METHODS</b>Forty-three cycles of 31 cases of azoospermic patients which underwent at least two PESA or TESE for ICSI from January 2001 to December 2002 were collected. The sperm retrieval, fertilization, implantation and clinical pregnancy were analyzed.</p><p><b>RESULTS</b>Twenty-four cases underwent PESA and 7 cases underwent TESE. There were not any complications in these patients. Compared with the first cycle of 154 cases, the fertilization rate, implantation rate and clinical pregnancy rate were 78.39% vs 73.64%, 19.68% vs 18.38% and 34.88% vs 37.91%, respectively(P > 0.05).</p><p><b>CONCLUSIONS</b>Repeated PESA or TESE is safe and well tolerated in azoospermic patients. Compared with the first cycle, the differences of repeated PESA or TESE cycles in fertilization rate, implantation rate and clinical pregnancy rate were not statistically significant.</p>

Sperm retrieval methods and pregnancy outcome of 100 azoospermia patients / 中华男科学杂志

<p><b>OBJECTIVES</b>To review the retrospective treatment results of the azoospermia patients during January 2001 to January 2002 in the fertility center.</p><p><b>METHODS</b>One hundred males attempted intracytoplasmic sperm injection (ICSI) cycle for treatment of azoospermia. All patients were undergone sperm retrieval by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) while their wives received conventional ovarian hyperstimulation. The hormone levels, testicular histology, the rates of sperm retrieval, fertilization, implantation and pregnancy were analysed and evaluated.</p><p><b>RESULTS</b>Sperm were retrieved by PESA in 76 of 100 (76%) and by TESE in 23 of 100 (23%) men of azoospermia. The fertilization rate, implantation rate and clinical pregnancy rate were 71.3%, 20.35% and 42.11% respectively in PESA group, and 75.18%, 22.05% and 41.60% respectively in TESA group. Thirty-two clinical pregnancies were achieved with 15 ongoing pregnancies and subsequent live delivery for 15 cases in PESA group, and 2 cases of miscarriage, while 10 clinical pregnancies were achieved with 6 ongoing pregnancies, 2 cases of live delivery and 2 cases of miscarriage in TESA group. One case failed to retrieve sperm by TESE and canceled.</p><p><b>CONCLUSIONS</b>Hormonal levels and testicular histology are unable to predict which men with azoospermia will have sperm retrieved by PESA and TESE. PESA and TESE with ICSI are effective methods to treat azoospermia. There were no significant differences in fertilization, implantation and clinical pregnancy rate between two groups.</p>