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This paper introduces a new class of explanation structures, called robust counterfactual witnesses (RCWs), to provide robust, both counterfactual and factual explanations for graph neural networks. Given a graph neural network M, a robust counterfactual witness refers to the fraction of a graph G that are counterfactual and factual explanation of the results of M over G, but also remains so for any "disturbed" G by flipping up to k of its node pairs. We establish the hardness results, from tractable results to co-NP-hardness, for verifying and generating robust counterfactual witnesses. We study such structures for GNN-based node classification, and present efficient algorithms to verify and generate RCWs. We also provide a parallel algorithm to verify and generate RCWs for large graphs with scalability guarantees. We experimentally verify our explanation generation process for benchmark datasets, and showcase their applications., Comment: This paper has been accepted by ICDE 2024

Cryoprotectant-free vitrification is a common method for spermatozoa cryopreservation by direct plunging into liquid nitrogen. However, the commercial liquid nitrogen could be potentially contaminated by microorganisms. Warming temperature plays an essential role for quality of human spermatozoa after vitrification. This study aimed to evaluate comparatively a quality spermatozoa after vitrification in liquid nitrogen and clean liquid air as well as with two warming rates: at 42 degrees C and 45 degrees C. After performing of routine swim-up of normozoospermia samples, spermatozoa from the same ejaculate were divided into two groups: vitrified in liquid nitrogen (LN) and sterile liquid air (LA). Spermatozoa of LN group were warmed at 42 degrees C, and spermatozoa of LA groups were divided and warmed at 42 degrees C (LA42) and 45 degrees C (LA45). Then spermatozoa motility, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), reactive nitrogen species (RNS), and viability were assessed. It was no found significant differences in quality of spermatozoa from LN and LA groups in the motility, ROS, MMP, RNS rates after warming at 42 degrees C. A tendency to obtain better spermatozoa quality was found with using of warming by 42 degrees C in comparison with 45 degrees C. It was concluded that cryoprotectant-free vitrification by direct dropping of human spermatozoa into clean liquid air can be used as an alternative to cooling in liquid nitrogen. Warming of spermatozoa at 42 degrees C allows to preserve the spermatozoa physiological parameters.

Introduction: Spermatozoa cryopreservation is an important technique to preserve fertility for males. This study aimed at exploring the stability of epigenetics information in human spermatozoa, manipulated by two different technologies, freezing and vitrification. Methods: Spermatozoa samples were distributed into three groups: 1. Fresh spermatozoa (control group), 2. Frozen spermatozoa, 3. Vitrified spermatozoa. Epigenetic differences of fresh and cryopreserved spermatozoa were evaluated using high-throughput RNA sequencing. Results: Differentially expressed genes (DEGs) in frozen (1103 genes) and vitrified (333 genes) spermatozoa were evaluated. The bioinformatical analysis identified 8 and 15 significant pathways in groups of frozen and vitrified spermatozoa, respectively. The majority of these pathways are most relevant to immune and infectious diseases. The DEGs of the fertilization process are not detected during vitrification. The freezing process induces more down-regulation of genes and is relevant to apoptosis changes and immune response. Conclusion: Cryopreservation of human spermatozoa is an epigenetically safe method for male fertility preservation. Cryoprotectant-free vitrification can induce more minor biological changes in human spermatozoa, in comparison with conventional freezing.

Background: Gene set enrichment analysis (GSEA) was conducted on raw data, and alternative splicing (AS) events were found after mRNA sequencing of human spermatozoa. In this study, we aimed to compare unknown micro-epigenetics alternations in fresh and cryopreserved spermatozoa to evaluate the effectivity of cryopreservation protocols. Methods: Spermatozoa were divided into three groups: fresh spermatozoa (group 1), cryoprotectant-free vitrified spermatozoa (group 2), and conventionally frozen spermatozoa (group 3). Nine RNA samples (three replicates in each group) were detected and were used for library preparation with an Illumina compatible kit and sequencing by the Illumina platform. Results: Three Gene Ontology (GO) terms were found to be enriched in vitrified spermatozoa compared with fresh spermatozoa: mitochondrial tRNA aminoacylation, ATP-dependent microtubule motor activity, and male meiotic nuclear division. In alternative splicing analysis, a number of unknown AS events were found, including functional gene exon skipping (SE), alternative 5 ' splice sites (A5SS), alternative 3 ' splice sites (A3SS), mutually exclusive exon (MXE), and retained intron (RI). Conclusions: Cryopreservation of spermatozoa from some patients can agitate epigenetic instability, including increased alternative splicing events and changes in crucial mitochondrial functional activities. For fertilization of oocytes, for such patients, it is recommended to use fresh spermatozoa whenever possible; cryopreservation of sperm is recommended to be used only in uncontested situations.

Introduction: The in vitro culture of primordial follicles is the only available option for preserving fertility in prepubertal girls with malignant tumors. The cultivation of primordial follicles in scaffolds as artificial ovaries is a promising approach for this. Methods: Dissociated follicles were placed into an artificial ovarian scaffold composed of fibrinogen and thrombin. The follicles were cultured in a dish dedicated to live cell imaging and observed for growth using immunofluorescence and development via optical microscopy. The morphology of the follicles in the scaffold was three-dimensionally reconstructed using the Imaris software. Growth and development were also quantified. Results: The morphology of artificial ovaries began to degrade over time. Within approximately 7 days, primordial follicles were activated and grew into secondary follicles. A comparison of optical and confocal microscopy results revealed the superior detection of live cells using confocal microscopy. The three-dimensional reconstruction of the confocal microscopy data enabled the automatic enumeration and evaluation of the overall morphology of many follicles. Conclusions: The novel artificial ovary-enabled primordial follicles to enter the growth cycle after activation and grow into secondary follicles. The use of a fibrin scaffold as a carrier preserves the developmental potential of primordial germ cells and is a potentially effective method for preserving fertility in prepubertal girls.

Cryoprotectant-free vitrification is a common method for spermatozoa cryopreservation by direct plunging into liquid nitrogen. However, the commercial liquid nitrogen could be potentially contaminated by microorganisms. Warming temperature plays an essential role for quality of human spermatozoa after vitrification. This study aimed to evaluate comparatively a quality spermatozoa after vitrification in liquid nitrogen and clean liquid air as well as with two warming rates: at 42 degrees C and 45 degrees C. After performing of routine swim-up of normozoospermia samples, spermatozoa from the same ejaculate were divided into two groups: vitrified in liquid nitrogen (LN) and sterile liquid air (LA). Spermatozoa of LN group were warmed at 42 degrees C, and spermatozoa of LA groups were divided and warmed at 42 degrees C (LA42) and 45 degrees C (LA45). Then spermatozoa motility, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), reactive nitrogen species (RNS), and viability were assessed. It was no found significant differences in quality of spermatozoa from LN and LA groups in the motility, ROS, MMP, RNS rates after warming at 42 degrees C. A tendency to obtain better spermatozoa quality was found with using of warming by 42 degrees C in comparison with 45 degrees C. It was concluded that cryoprotectant-free vitrification by direct dropping of human spermatozoa into clean liquid air can be used as an alternative to cooling in liquid nitrogen. Warming of spermatozoa at 42 degrees C allows to preserve the spermatozoa physiological parameters.

Mitochondria-targeted antioxidants have great potential to counterbalance the generated reactive oxygen species (ROS) because they cross the inner membrane of the mitochondria. Still, their use was not reported in vitrified human spermatozoa. Our laboratory has successfully vitrified spermatozoa without the use of permeable cryoprotectants, but subcellular-level evidence was missing. Therefore, this study aimed to improve spermatozoa vitrification using a mitochondria-targeted antioxidant (mitoquinone, MitoQ), reveal ultrastructural changes in the spermatozoa due to the use of a permeable cryoprotectant, and report alterations of functional proteins during the spermatozoa vitrification process. For this, each of 20 swim-up-prepared ejaculates was divided into seven aliquots and diluted with a vitrification medium supplemented with varying concentrations of MitoQ (0.02 and 0.2 mu M), glycerol (1, 4, and 6%), and a combination of MitoQ and glycerol. All aliquots were vitrified by the aseptic capillary method developed in our laboratory. The spermatozoa function assays revealed that the addition of either MitoQ (0.02 mu M), glycerol (1%), or a combination of MitoQ (0.02 mu M) and glycerol (1%) in the vitrification medium results in better or equivalent spermatozoa quality relative to the control. Transmission electron microscopy revealed that MitoQ protects the spermatozoa from undergoing ultrastructural alterations, but glycerol induced ultrastructural alterations during the vitrification process. Next, we performed label-free quantitative proteomics and identified 1,759 proteins, of which 69, 60, 90, and 81 were altered in the basal medium, 0.02 mu M MitoQ, 1% glycerol, and Mito-glycerol groups, respectively. Actin, tubulins, and outer dense fiber proteins were not affected during the vitrification process. Some of the identified ubiquitinating enzymes were affected during spermatozoa vitrification. Only a few proteins responsible for phosphorylation were altered dur

Objective: To investigate whether the ratio of the serum progesterone level on the day of human chorionic gonadotrophin (hCG) administration to the basal serum progesterone level (P-hCG/P-basal) is a predictor of pregnancy outcome during in vitro fertilization (IVF). Methods: A total of 12,708 cycles were performed in 9747 patients between 19 and 36 years of age who were undergoing controlled ovarian stimulation from October 2011 to July 2016 for their first or second attempts at IVF followed by fresh embryo transfer (ET). hCG was administered 36 h before oocyte retrieval to trigger final oocyte maturation. The serum progesterone level was measured on menstrual cycle days 2-4 (basal progesterone, P-basal) and on the day of hCG administration (P-hCG). P-hCG/P-basal was calculated. Live birth rates were compared among various ordinal P-hCG/P-basal intervals (< 0.5, 0.5-1.0, 1.0-1.5, 1.5-2.0, 2.0-2.5, > 2.5). Results: The average age of the patients recruited was 29.9 years. The average basal progesterone level was 0.8 ng/ml, while the average progesterone level on the day of hCG administration was 0.9 ng/ml. The live birth rates (according to the abovementioned ordinal P-hCG/P-basal intervals) were 47.3, 49.9, 47.8, 46.3, 45.5 and 44.0%, respectively. The live birth rates were significantly higher for patients with P-hCG/P-basal between 0.5-1.0 (OR = 1.14, 95% CI, 1.02-1.27, p = .02). Conclusions: P-hCG/P-basal between 0.5-1.0 predicts a higher live birth rate in IVF. Both P-hCG/P-basal and P on hCG day is less predictive value for predicting live birth rate.

Objective: Primordial follicles in premature ovarian failure (POF) patients are very difficult to be activated spontaneously, so that mature oocytes are difficult to be obtained for in vitro fertilization. The aim of our review is to analyze and to systematize the published data regarding effectiveness of different strategies for in vitro activation of cryopreserved ovarian tissue. Study design: According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a review of the literature was performed for all relevant full-text articles published in PubMed in English. Meta-analysis conducted using STATA 14.0. The random-effects model was used to combine 8 study results because the examination of heterogeneity was minimal. Results: One hundred and seventy seven patients after in vitro activation treatment (IVA) of ovarian tissue had accumulatively 26 pregnancies through IVF or natural pregnancy and then produced 18 live births. The random-effects model showed that the total clinical pregnancy and baby born rates reported in 8 studies evidence about effectiveness of IVA. Conclusion: In vitro activation of primordial follicles as a new potential treatment for ovarian disorder patients, can be a promising option for fertility preservation. Drug-free activation of ovarian tissue in comparison with drug-included activation seemed to be more efficient. (C) 2021 Elsevier B.V. All rights reserved.

The protocol of aseptic cryoprotectant-free vitrification on human spermatozoa is well documented. However, data about the effect of permeable cryoprotectants at this procedure is limited. Presented study aimed to test the aseptic capillary vitrification technologies using permeable cryoprotectant-included or cryopmtectant-free media. Thirty-two normal samples were included and analyzed after vitrification in three different media and thawing. Three treatment groups were formed: Group 1, basic medium; Group 2, basic medium with 0.25 M sucrose; Group 3, basic medium with glycerol. Before plunging into liquid nitrogen, capillaries were filled by 10 til of spermatozoa suspension and isolated from liquid nitrogen by location in hermetically closed 0.25 ml straws. Progressive motility, plasma membrane integrity, total motility/viability after 24, 48 and 72 h in vitro culture, apoptosis and mitochondrial membrane potential (Delta Psi m) were determined after thawing at 42 degrees C. Progressive motility of spermatozoa in groups 1, 2, 3 was 24.9 +/- 1.7%, 34.5 +/- 2.8% and 34.0 +/- 1.4%, respectively (P-1-2,P-3<0.05). The plasma membrane integrity of spermatozoa in groups 2 and 3 (48.4 +/- 2.9% and 45.5 +/- 3.9%, respectively) was higher than in Group 1 (33.3 +/- 2.1%, P < 0.05). After 24 h, 48 h and 72 h in vitro culture, the total motility and viability of spermatozoa in Group 1 was significantly lower than Group 2 and Group 3. The apoptosis rate in Group 3 (44.5 +/- 3.0%) and Group 2 (47.7 +/- 4.1%) were lower than in Group 1 (52.5 +/- 4.4%; P < 0.05). Delta Psi m rates in Group 3 and Group 2 were higher than in Group 1 (P < 0.05) with no statistical differences between this parameter in Group 2 and Group 3 (P > 0.1). In conclusion, supplementation of medium for aseptic capillary technology for cryoprotectant-free vitrification of human spermatozoa by permeable cryoprotectant does not improve the quality of spermatozoa after warming.

Objective: To investigate whether the ratio of the serum progesterone level on the day of human chorionic gonadotrophin (hCG) administration to the basal serum progesterone level (P-hCG/P-basal) is a predictor of pregnancy outcome during in vitro fertilization (IVF). Methods: A total of 12,708 cycles were performed in 9747 patients between 19 and 36 years of age who were undergoing controlled ovarian stimulation from October 2011 to July 2016 for their first or second attempts at IVF followed by fresh embryo transfer (ET). hCG was administered 36 h before oocyte retrieval to trigger final oocyte maturation. The serum progesterone level was measured on menstrual cycle days 2-4 (basal progesterone, P-basal) and on the day of hCG administration (P-hCG). P-hCG/P-basal was calculated. Live birth rates were compared among various ordinal P-hCG/P-basal intervals (< 0.5, 0.5-1.0, 1.0-1.5, 1.5-2.0, 2.0-2.5, > 2.5). Results: The average age of the patients recruited was 29.9 years. The average basal progesterone level was 0.8 ng/ml, while the average progesterone level on the day of hCG administration was 0.9 ng/ml. The live birth rates (according to the abovementioned ordinal P-hCG/P-basal intervals) were 47.3, 49.9, 47.8, 46.3, 45.5 and 44.0%, respectively. The live birth rates were significantly higher for patients with P-hCG/P-basal between 0.5-1.0 (OR = 1.14, 95% CI, 1.02-1.27, p = .02). Conclusions: P-hCG/P-basal between 0.5-1.0 predicts a higher live birth rate in IVF. Both P-hCG/P-basal and P on hCG day is less predictive value for predicting live birth rate.

Mitochondria-targeted antioxidants have great potential to counterbalance the generated reactive oxygen species (ROS) because they cross the inner membrane of the mitochondria. Still, their use was not reported in vitrified human spermatozoa. Our laboratory has successfully vitrified spermatozoa without the use of permeable cryoprotectants, but subcellular-level evidence was missing. Therefore, this study aimed to improve spermatozoa vitrification using a mitochondria-targeted antioxidant (mitoquinone, MitoQ), reveal ultrastructural changes in the spermatozoa due to the use of a permeable cryoprotectant, and report alterations of functional proteins during the spermatozoa vitrification process. For this, each of 20 swim-up-prepared ejaculates was divided into seven aliquots and diluted with a vitrification medium supplemented with varying concentrations of MitoQ (0.02 and 0.2 mu M), glycerol (1, 4, and 6%), and a combination of MitoQ and glycerol. All aliquots were vitrified by the aseptic capillary method developed in our laboratory. The spermatozoa function assays revealed that the addition of either MitoQ (0.02 mu M), glycerol (1%), or a combination of MitoQ (0.02 mu M) and glycerol (1%) in the vitrification medium results in better or equivalent spermatozoa quality relative to the control. Transmission electron microscopy revealed that MitoQ protects the spermatozoa from undergoing ultrastructural alterations, but glycerol induced ultrastructural alterations during the vitrification process. Next, we performed label-free quantitative proteomics and identified 1,759 proteins, of which 69, 60, 90, and 81 were altered in the basal medium, 0.02 mu M MitoQ, 1% glycerol, and Mito-glycerol groups, respectively. Actin, tubulins, and outer dense fiber proteins were not affected during the vitrification process. Some of the identified ubiquitinating enzymes were affected during spermatozoa vitrification. Only a few proteins responsible for phosphorylation were altered dur

The protocol of aseptic cryoprotectant-free vitrification on human spermatozoa is well documented. However, data about the effect of permeable cryoprotectants at this procedure is limited. Presented study aimed to test the aseptic capillary vitrification technologies using permeable cryoprotectant-included or cryopmtectant-free media. Thirty-two normal samples were included and analyzed after vitrification in three different media and thawing. Three treatment groups were formed: Group 1, basic medium; Group 2, basic medium with 0.25 M sucrose; Group 3, basic medium with glycerol. Before plunging into liquid nitrogen, capillaries were filled by 10 til of spermatozoa suspension and isolated from liquid nitrogen by location in hermetically closed 0.25 ml straws. Progressive motility, plasma membrane integrity, total motility/viability after 24, 48 and 72 h in vitro culture, apoptosis and mitochondrial membrane potential (Delta Psi m) were determined after thawing at 42 degrees C. Progressive motility of spermatozoa in groups 1, 2, 3 was 24.9 +/- 1.7%, 34.5 +/- 2.8% and 34.0 +/- 1.4%, respectively (P-1-2,P-3<0.05). The plasma membrane integrity of spermatozoa in groups 2 and 3 (48.4 +/- 2.9% and 45.5 +/- 3.9%, respectively) was higher than in Group 1 (33.3 +/- 2.1%, P < 0.05). After 24 h, 48 h and 72 h in vitro culture, the total motility and viability of spermatozoa in Group 1 was significantly lower than Group 2 and Group 3. The apoptosis rate in Group 3 (44.5 +/- 3.0%) and Group 2 (47.7 +/- 4.1%) were lower than in Group 1 (52.5 +/- 4.4%; P < 0.05). Delta Psi m rates in Group 3 and Group 2 were higher than in Group 1 (P < 0.05) with no statistical differences between this parameter in Group 2 and Group 3 (P > 0.1). In conclusion, supplementation of medium for aseptic capillary technology for cryoprotectant-free vitrification of human spermatozoa by permeable cryoprotectant does not improve the quality of spermatozoa after warming.

Objective: Primordial follicles in premature ovarian failure (POF) patients are very difficult to be activated spontaneously, so that mature oocytes are difficult to be obtained for in vitro fertilization. The aim of our review is to analyze and to systematize the published data regarding effectiveness of different strategies for in vitro activation of cryopreserved ovarian tissue. Study design: According to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, a review of the literature was performed for all relevant full-text articles published in PubMed in English. Meta-analysis conducted using STATA 14.0. The random-effects model was used to combine 8 study results because the examination of heterogeneity was minimal. Results: One hundred and seventy seven patients after in vitro activation treatment (IVA) of ovarian tissue had accumulatively 26 pregnancies through IVF or natural pregnancy and then produced 18 live births. The random-effects model showed that the total clinical pregnancy and baby born rates reported in 8 studies evidence about effectiveness of IVA. Conclusion: In vitro activation of primordial follicles as a new potential treatment for ovarian disorder patients, can be a promising option for fertility preservation. Drug-free activation of ovarian tissue in comparison with drug-included activation seemed to be more efficient. (C) 2021 Elsevier B.V. All rights reserved.

AIMS: Little is known about the relation between the long-term joint exposure to various ambient air pollutants and the incidence of heart failure (HF). We aimed to assess the joint association of various air pollutants with HF risk and examine the modification effect of the genetic susceptibility. METHODS AND RESULTS: This study included 432 530 participants free of HF, atrial fibrillation, or coronary heart disease in the UK Biobank study. All participants were enrolled from 2006 to 2010 and followed up to 2018. The information on particulate matter (PM) with diameters ≤2.5 µm (PM2.5), ≤10 µm (PM10), and between 2.5 and 10 µm (PM2.5-10) as well as nitrogen oxides (NO2 and NOx) was collected. We newly proposed an air pollution score to assess the joint exposure to the five air pollutants through summing each pollutant concentration weighted by the regression coefficients with HF from single-pollutant models. We also calculated the weighted genetic risk score of HF. During a median of 10.1 years (4 346 642 person-years) of follow-up, we documented 4201 incident HF. The hazard ratios (HRs) [95% confidence interval (CI)] of HF for a 10 µg/m3 increase in PM2.5, PM10, PM2.5-10, NO2, and NOx were 1.85 (1.34-2.55), 1.61 (1.30-2.00), 1.13 (0.80-1.59), 1.10 (1.04-1.15), and 1.04 (1.02-1.06), respectively. We found that the air pollution score was associated with an increased risk of incident HF in a dose-response fashion. The HRs (95% CI) of HF were 1.16 (1.05-1.28), 1.19 (1.08-1.32), 1.21 (1.09-1.35), and 1.31 (1.17-1.48) in higher quintile groups compared with the lowest quintile of the air pollution score (P trend <0.001). In addition, we observed that the elevated risk of HF associated with a higher air pollution score was strengthened by the genetic susceptibility to HF. CONCLUSION: Our results indicate that the long-term joint exposure to various air pollutants including PM2.5, PM10, PM2.5-10, NO2, and NOx is associated with an elevated risk of incident HF in an addit

This study aimed to compare the quality of human spermatozoa vitrified by direct plunging into liquid nitrogen vs. liquid air. Spermatozoa were divided into three groups: fresh spermatozoa (Group F) were used as a control. Spermatozoa suspension (20 mu l) was vitrified in open granules by direct dropping into liquid nitrogen (Group LN) or clean liquid air (Group LA). After warming at 37 degrees C, the progressive motility rate of Group F was reduced from 65.9 +/- 2.5% to 34.0 +/- 1.9% (Group LN) and 38.1 +/- 2.3% (Group LA), respectively (P-1-2,P-3 < 0.05). The reductions in viability were 65.6 +/- 2.2%, 29.0 +/- 1.8%, and 36.6 +/- 2.6% for Groups F, LN, and LA, respectively (P-1-2,P-3 < 0.05). Comparing spermatozoa vitrified in liquid nitrogen vs. liquid air, no significant differences were detected in motility (34.0 +/- 1.9% vs. 38.1 +/- 2.3%), viability (29.0 +/- 1.8% vs. 36.6 +/- 2.6%), early apoptosis (13.8 +/- 1.5% vs. 14.3 +/- 1.8%), late apoptosis (45.5 +/- 1.8% vs. 43.7 +/- 2.2%), and necrosis (19.5 +/- 2.0% vs. 15.0 +/- 1.8%; p>0.01 for all respective differences). There was a statistical tendency for increasing rates of progressive motility and viability and decreasing rates of apoptosis and necrosis when comparing spermatozoa vitrified in liquid air vs. liquid nitrogen. It is concluded that cryoprotectant-free vitrification by the direct dropping of human spermatozoa in a clean cooling agent (liquid air) is a good alternative to the use of nonsterile liquid nitrogen and can be used to cool cells while minimising the risk of microbial contamination.

Purpose High progesterone is associated with low implantation rate. Our previous study demonstrated that DNA methylation in endometrium was increased in women with high progesterone in IVF cycles. However, the DNA methylation status is still not yet confirmed, and how it affects endometrial receptivity in high progesterone is still unknown. Current study investigated the effects of high progesterone on DNA methylation and gene expression of adhesion molecules on endometrium during implantation window. Methods A cohort study included 20 women with high progesterone (HP) and 20 with normal progesterone (NP) on the day of human chorionic gonadotropin (hCG) administration after controlled ovarian hyperstimulation in IVF cycle. Endometrial tissues were collected on the 7th day after hCG administration. Immunohistochemical staining of DNA methyltransferases (DNMT1 and DNMT3B) and adhesion molecules (MUC1, CDH1 and CTNNB1) were performed. Methylation of MUC1, CDH1, and CTNNB1 promoter regions was detected by Sequenom MassARRAY or bisulfite sequencing PCR. RT-qPCR was used to quantify mRNA expression levels, and correlation of methylation and gene expression level of the adhesion molecules were determined. Results DNMT3B, but not DNMT1, in nucleus of luminal and glandular epithelial cells in HP group was significantly higher than that in NP group. Promoter regions of CDH1 and CTNNB1, but not MUC1, in endometrium of HP group were hypermethylated. Protein and mRNA expression of MUC1, CDH1, and CTNNB1 in endometrium of HP group was significantly lower than that in NP group. Level of DNA methylation was negatively correlated with the gene expression of CDH1 and CTNNB1, but not MUC1. Conclusions DNA hypermethylation and low expression of adhesion molecules on endometrium were associated with high progesterone during implantation window, which may contribute to the underlying epigenetic mechanism in the failure of IVF treatment.

This study aimed to compare the quality of human spermatozoa vitrified by direct plunging into liquid nitrogen vs. liquid air. Spermatozoa were divided into three groups: fresh spermatozoa (Group F) were used as a control. Spermatozoa suspension (20 mu l) was vitrified in open granules by direct dropping into liquid nitrogen (Group LN) or clean liquid air (Group LA). After warming at 37 degrees C, the progressive motility rate of Group F was reduced from 65.9 +/- 2.5% to 34.0 +/- 1.9% (Group LN) and 38.1 +/- 2.3% (Group LA), respectively (P-1-2,P-3 < 0.05). The reductions in viability were 65.6 +/- 2.2%, 29.0 +/- 1.8%, and 36.6 +/- 2.6% for Groups F, LN, and LA, respectively (P-1-2,P-3 < 0.05). Comparing spermatozoa vitrified in liquid nitrogen vs. liquid air, no significant differences were detected in motility (34.0 +/- 1.9% vs. 38.1 +/- 2.3%), viability (29.0 +/- 1.8% vs. 36.6 +/- 2.6%), early apoptosis (13.8 +/- 1.5% vs. 14.3 +/- 1.8%), late apoptosis (45.5 +/- 1.8% vs. 43.7 +/- 2.2%), and necrosis (19.5 +/- 2.0% vs. 15.0 +/- 1.8%; p>0.01 for all respective differences). There was a statistical tendency for increasing rates of progressive motility and viability and decreasing rates of apoptosis and necrosis when comparing spermatozoa vitrified in liquid air vs. liquid nitrogen. It is concluded that cryoprotectant-free vitrification by the direct dropping of human spermatozoa in a clean cooling agent (liquid air) is a good alternative to the use of nonsterile liquid nitrogen and can be used to cool cells while minimising the risk of microbial contamination.

Purpose High progesterone is associated with low implantation rate. Our previous study demonstrated that DNA methylation in endometrium was increased in women with high progesterone in IVF cycles. However, the DNA methylation status is still not yet confirmed, and how it affects endometrial receptivity in high progesterone is still unknown. Current study investigated the effects of high progesterone on DNA methylation and gene expression of adhesion molecules on endometrium during implantation window. Methods A cohort study included 20 women with high progesterone (HP) and 20 with normal progesterone (NP) on the day of human chorionic gonadotropin (hCG) administration after controlled ovarian hyperstimulation in IVF cycle. Endometrial tissues were collected on the 7th day after hCG administration. Immunohistochemical staining of DNA methyltransferases (DNMT1 and DNMT3B) and adhesion molecules (MUC1, CDH1 and CTNNB1) were performed. Methylation of MUC1, CDH1, and CTNNB1 promoter regions was detected by Sequenom MassARRAY or bisulfite sequencing PCR. RT-qPCR was used to quantify mRNA expression levels, and correlation of methylation and gene expression level of the adhesion molecules were determined. Results DNMT3B, but not DNMT1, in nucleus of luminal and glandular epithelial cells in HP group was significantly higher than that in NP group. Promoter regions of CDH1 and CTNNB1, but not MUC1, in endometrium of HP group were hypermethylated. Protein and mRNA expression of MUC1, CDH1, and CTNNB1 in endometrium of HP group was significantly lower than that in NP group. Level of DNA methylation was negatively correlated with the gene expression of CDH1 and CTNNB1, but not MUC1. Conclusions DNA hypermethylation and low expression of adhesion molecules on endometrium were associated with high progesterone during implantation window, which may contribute to the underlying epigenetic mechanism in the failure of IVF treatment.

Non-PR, IFPRI4; 3 Building Inclusive and Efficient Markets, Trade Systems, and Food Industry; 4 Transforming Agricultural and Rural Economies; 5 Strengthening Institutions and Governance; Capacity Strengthening, MTID

Kashin-Beck disease (KBD) is an endemic degenerative osteoarthropathy of uncertain etiology. Our study sought to identify a correlation between small proteoglycans decorin and biglycan expression and Kashin-Beck Disease. Immunohistochemistry was used to assess the decorin and biglycan levels in cartilage specimens from both child KBD patients, and rats fed with T-2 toxin under a selenium-deficient condition. Real-time PCR and Western blot were used to assess mRNA and protein levels of decorin and biglycan in rat cartilages, as well as in C28/I2 chondrocytes stimulated by T-2 toxin and selenium in vitro. The result showed that decorin was reduced in all zones of KBD articular cartilage, while the expression of biglycan was prominently increased in KBD cartilage samples. Increased expression of biglycan and reduced expression of decorin were observed at mRNA and protein levels in the cartilage of rats fed with T-2 toxin and selenium- deficiency plus T-2 toxin diet, when compared with the normal diet group. Moreover, In vitro stimulation of C28/I2 cells with T-2 toxin resulted in an upregulation of biglycan and downregulation of decorin, T-2 toxin induction of biglycan and decorin levels were partly rescued by selenium supplement. This study highlights the focal nature of the degenerative changes that occur in KBD cartilage and may suggest that the altered expression pattern of decorin and biglycan have an important role in the onset and pathogenesis of KBD.

Zinc cobaltite (ZnCo 2 O 4 ) is a promising material for supercapacitors with appreciable theoretical capacitance. However, it suffers from poor electrical conductivity and large volume expansion during charge-discharge processes. In this work, a novel material, ZnCo 2 O 4 /C microspheres in a yolk-shell structure, is fabricated by a facile refluxing process in combination with a calcination treatment. Carbon is in situ formed via the pyrolysis of polyvinyl pyrrolidone and uniformly dispersed both in the core and the shell. Benefiting from the unique structure and the synergistic effect of the two components, this material exhibits remarkable electrochemical properties such as high specific capacitance (1821 F g -1 at 5 A g -1 ), excellent rate performance, and superior cycling stability (no capacitance loss over 9000 cycles). The assembled asymmetric supercapacitor coupled with an active carbon anode can deliver a high energy density of 45.9 W h kg -1 at a power density of 700 W kg -1 with an excellent cycling stability (i.e. a capacitance retention rate over 95% after 9000 cycles).

Zinc cobaltite (ZnCo 2 O 4 ) is a promising material for supercapacitors with appreciable theoretical capacitance. However, it suffers from poor electrical conductivity and large volume expansion during charge-discharge processes. In this work, a novel material, ZnCo 2 O 4 /C microspheres in a yolk-shell structure, is fabricated by a facile refluxing process in combination with a calcination treatment. Carbon is in situ formed via the pyrolysis of polyvinyl pyrrolidone and uniformly dispersed both in the core and the shell. Benefiting from the unique structure and the synergistic effect of the two components, this material exhibits remarkable electrochemical properties such as high specific capacitance (1821 F g -1 at 5 A g -1 ), excellent rate performance, and superior cycling stability (no capacitance loss over 9000 cycles). The assembled asymmetric supercapacitor coupled with an active carbon anode can deliver a high energy density of 45.9 W h kg -1 at a power density of 700 W kg -1 with an excellent cycling stability (i.e. a capacitance retention rate over 95% after 9000 cycles).

Zinc cobaltite (ZnCo 2 O 4 ) is a promising material for supercapacitors with appreciable theoretical capacitance. However, it suffers from poor electrical conductivity and large volume expansion during charge-discharge processes. In this work, a novel material, ZnCo 2 O 4 /C microspheres in a yolk-shell structure, is fabricated by a facile refluxing process in combination with a calcination treatment. Carbon is in situ formed via the pyrolysis of polyvinyl pyrrolidone and uniformly dispersed both in the core and the shell. Benefiting from the unique structure and the synergistic effect of the two components, this material exhibits remarkable electrochemical properties such as high specific capacitance (1821 F g -1 at 5 A g -1 ), excellent rate performance, and superior cycling stability (no capacitance loss over 9000 cycles). The assembled asymmetric supercapacitor coupled with an active carbon anode can deliver a high energy density of 45.9 W h kg -1 at a power density of 700 W kg -1 with an excellent cycling stability (i.e. a capacitance retention rate over 95% after 9000 cycles).

Zinc cobaltite (ZnCo 2 O 4 ) is a promising material for supercapacitors with appreciable theoretical capacitance. However, it suffers from poor electrical conductivity and large volume expansion during charge-discharge processes. In this work, a novel material, ZnCo 2 O 4 /C microspheres in a yolk-shell structure, is fabricated by a facile refluxing process in combination with a calcination treatment. Carbon is in situ formed via the pyrolysis of polyvinyl pyrrolidone and uniformly dispersed both in the core and the shell. Benefiting from the unique structure and the synergistic effect of the two components, this material exhibits remarkable electrochemical properties such as high specific capacitance (1821 F g -1 at 5 A g -1 ), excellent rate performance, and superior cycling stability (no capacitance loss over 9000 cycles). The assembled asymmetric supercapacitor coupled with an active carbon anode can deliver a high energy density of 45.9 W h kg -1 at a power density of 700 W kg -1 with an excellent cycling stability (i.e. a capacitance retention rate over 95% after 9000 cycles).

In deep-sea sediment, the microbes present in anhydrite crystals are potential markers of the past environment. In the Atlantis II Deep, anhydrite veins were produced by mild mixture of calcium-rich hydrothermal solutions and sulfate in the bottom water, which had probably preserved microbial inhabitants in the past seafloor of the Red Sea. In this study, this hypothesis was tested by analyzing the metagenome of an anhydrite crystal sample from the Atlantis II Deep. The estimated age of the anhydrite layer was between 750 and 770 years, which might span the event of hydrothermal eruption into the benthic floor. The 16S/18S rRNA genes in the metagenome were assigned to bacteria, archaea, fungi and even invertebrate species. The dominant species in the crystals was an oil-degrading Alcanivorax borkumensis bacterium, which was not detected in the adjacent sediment layer. Fluorescence microscopy using 16S rRNA and marker gene probes revealed intact cells of the Alcanivorax bacterium in the crystals. A draft genome of A. borkumensis was binned from the metagenome. It contained all functional genes for alkane utilization and the reduction of nitrogen oxides. Moreover, the metagenomes of the anhydrites and control sediment contained aromatic degradation pathways, which were mostly derived from Ochrobactrum sp. Altogether, these results indicate an oxic, oil-spilling benthic environment in the Atlantis II basin of the Red Sea in approximately the 14th century. The original microbial inhabitants probably underwent a dramatic selection process via drastic environmental changes following the formation of an overlying anoxic brine pool in the basin due to hydrothermal activities. © 2016 Author(s).

In deep-sea sediment, the microbes present in anhydrite crystals are potential markers of the past environment. In the Atlantis II Deep, anhydrite veins were produced by mild mixture of calcium-rich hydrothermal solutions and sulfate in the bottom water, which had probably preserved microbial inhabitants in the past seafloor of the Red Sea. In this study, this hypothesis was tested by analyzing the metagenome of an anhydrite crystal sample from the Atlantis II Deep. The estimated age of the anhydrite layer was between 750 and 770 years, which might span the event of hydrothermal eruption into the benthic floor. The 16S/18S rRNA genes in the metagenome were assigned to bacteria, archaea, fungi and even invertebrate species. The dominant species in the crystals was an oil-degrading Alcanivorax borkumensis bacterium, which was not detected in the adjacent sediment layer. Fluorescence microscopy using 16S rRNA and marker gene probes revealed intact cells of the Alcanivorax bacterium in the crystals. A draft genome of A. borkumensis was binned from the metagenome. It contained all functional genes for alkane utilization and the reduction of nitrogen oxides. Moreover, the metagenomes of the anhydrites and control sediment contained aromatic degradation pathways, which were mostly derived from Ochrobactrum sp. Altogether, these results indicate an oxic, oil-spilling benthic environment in the Atlantis II basin of the Red Sea in approximately the 14th century. The original microbial inhabitants probably underwent a dramatic selection process via drastic environmental changes following the formation of an overlying anoxic brine pool in the basin due to hydrothermal activities. © 2016 Author(s).

In deep-sea sediment, the microbes present in anhydrite crystals are potential markers of the past environment. In the Atlantis II Deep, anhydrite veins were produced by mild mixture of calcium-rich hydrothermal solutions and sulfate in the bottom water, which had probably preserved microbial inhabitants in the past seafloor of the Red Sea. In this study, this hypothesis was tested by analyzing the metagenome of an anhydrite crystal sample from the Atlantis II Deep. The estimated age of the anhydrite layer was between 750 and 770 years, which might span the event of hydrothermal eruption into the benthic floor. The 16S/18S rRNA genes in the metagenome were assigned to bacteria, archaea, fungi and even invertebrate species. The dominant species in the crystals was an oil-degrading Alcanivorax borkumensis bacterium, which was not detected in the adjacent sediment layer. Fluorescence microscopy using 16S rRNA and marker gene probes revealed intact cells of the Alcanivorax bacterium in the crystals. A draft genome of A. borkumensis was binned from the metagenome. It contained all functional genes for alkane utilization and the reduction of nitrogen oxides. Moreover, the metagenomes of the anhydrites and control sediment contained aromatic degradation pathways, which were mostly derived from Ochrobactrum sp. Altogether, these results indicate an oxic, oil-spilling benthic environment in the Atlantis II basin of the Red Sea in approximately the 14th century. The original microbial inhabitants probably underwent a dramatic selection process via drastic environmental changes following the formation of an overlying anoxic brine pool in the basin due to hydrothermal activities. © 2016 Author(s).

We develop a timed calculus for Mobile Ad Hoc Networks embodying the peculiarities of local broadcast, node mobility and communication interference. We present a Reduction Semantics and a Labelled Transition Semantics and prove the equivalence between them. We then apply our calculus to model and study some MAC-layer protocols with special emphasis on node mobility and communication interference. A main purpose of the semantics is to describe the various forms of interference while nodes change their locations in the network. Such interference only occurs when a node is simultaneously reached by more than one ongoing transmission over the same channel., Comment: In Proceedings FTSCS 2012, arXiv:1212.6574

LAGE is a systematic framework developed in Java. The motivation of LAGE is to provide a scalable and parallel solution to reconstruct Gene Regulatory Networks (GRNs) from continuous gene expression data for very large amount of genes. The basic idea of our framework is motivated by the philosophy of divideand-conquer. Specifically, LAGE recursively partitions genes into multiple overlapping communities with much smaller sizes, learns intra-community GRNs respectively before merge them altogether. Besides, the complete information of overlapping communities serves as the byproduct, which could be used to mine meaningful functional modules in biological networks., Comment: 2 pages

The motivation for this paper is to apply Bayesian structure learning using Model Averaging in large-scale networks. Currently, Bayesian model averaging algorithm is applicable to networks with only tens of variables, restrained by its super-exponential complexity. We present a novel framework, called LSBN(Large-Scale Bayesian Network), making it possible to handle networks with infinite size by following the principle of divide-and-conquer. The method of LSBN comprises three steps. In general, LSBN first performs the partition by using a second-order partition strategy, which achieves more robust results. LSBN conducts sampling and structure learning within each overlapping community after the community is isolated from other variables by Markov Blanket. Finally LSBN employs an efficient algorithm, to merge structures of overlapping communities into a whole. In comparison with other four state-of-art large-scale network structure learning algorithms such as ARACNE, PC, Greedy Search and MMHC, LSBN shows comparable results in five common benchmark datasets, evaluated by precision, recall and f-score. What's more, LSBN makes it possible to learn large-scale Bayesian structure by Model Averaging which used to be intractable. In summary, LSBN provides an scalable and parallel framework for the reconstruction of network structures. Besides, the complete information of overlapping communities serves as the byproduct, which could be used to mine meaningful clusters in biological networks, such as protein-protein-interaction network or gene regulatory network, as well as in social network., Comment: 13 pages, 6 figures

王士雄纂 ; 呂慶熊, 林植梅仝校., 綫裝., 框15.5x11.6公分, 10行22字. 黑口, 左右雙邊, 單黑魚尾. 版心中鐫"霍亂"及小題, 下鐫葉次., 書名頁刻"光緖丁亥仲秋重鐫, 四明汲綆書莊藏版", 《中國中醫古籍總目》06832著錄., 鈐"莊兆祥印"朱, 白文各一方., Xian zhuang., Kuang 15.5 x 11.6 gong fen, 10 hang 22 zi. Hei kou, zuo you shuang bian, dan hei yu wei. Ban xin zhong juan "Huo luan" ji xiao ti, xia juan ye ci., Detailed notes in vernacular field only., Wang Shixiong zuan ; Lü Qingxiong, Lin Zhimei tong jiao., Qian "Zhuang Zhaoxiang yin" zhu, bai wen ge yi fang.

V.1. 王氏醫案 : 二卷 / 王士雄著 ; 周鑅輯錄 -- v.2-4. 王氏醫案續編 : 八卷 / 王士雄醫案 ; 張鴻...[et al.]輯., V.1. Wang shi yi an : er juan / Wang Shixiong zhu ; Zhou Hengji lu -- v.2-4. Wang shi yi an xu bian : ba juan / Wang Shixiong yi an ; Zhang Hong ...[et al.] ji., 王孟英著., 綫裝., 框17.9x13.3公分, 10行24字, 無界行. 白口, 四周雙邊, 單黑魚尾. 版心上鐫題名, 中鐫卷次, 下鐫葉次. 眉端行間均刻評點., 題名據叢書書名頁., 書名頁刻"海寧王孟英著, 醫案, 溫熱經緯, 霍亂論, 續醫案, 飮食譜" ; 書名背頁牌記刻"光緖壬辰年重刻上海醉六堂藏板錢炳書耑", 《中國叢書綜錄》p.725收錄., 《王氏醫案》原名《回春錄》 ;《王氏醫案續編》原名《仁術志》, 鈐"莊兆祥印"朱, 白文各一方., Xian zhuang., Kuang 17.9 x 13.3 gong fen, 10 hang 24 zi, wu jie hang. Bai kou, si zhou shuang bian, dan hei yu wei. Ban xin shang juan ti ming, zhong juan juan ci, xia juan ye ci. Mei duan xing jian jun ke ping dian., Ti ming ju cong shu shu ming ye., Detailed notes in vernacular field only., "Wang shi yi an" yuan ming "Hui chun lu" ; "Wang shi yi an xu bian" yuan ming "Ren shu zhi", Wang Mengying zhu., Qian "Zhuang Zhaoxiang yin" zhu, bai wen ge yi fang.

沈堯封輯 ; 王士雄參 ; 徐政杰補註 ; [沈]之藩校., 綫裝., 框18.2x13公分, 10行20字. 白口, 四周雙邊, 單黑魚尾. 版心上鐫題名, 中鐫卷次, 下鐫葉次., 分上, 下卷., 書名牌記刻"同治元年重鐫", 卷上卷端題"沈堯封先生輯, 前塘後學婿王士雄孟英參, 徐政傑靄輝補註, 男之藩友珊校"., 《中國中醫古籍總目》07436著錄., 鈐"哲如陳慶保藏書", "莊兆祥印", "莊兆祥"., Xian zhuang., Kuang 18.2 x 13 gong fen, 10 hang 20 zi. Bai kou, si zhou shuang bian, dan hei yu wei. Ban xin shang juan ti ming, zhong juan juan ci, xia juan ye ci., Fen shang, xia juan., Shu ming pai ji ke "Tongzhi yuan nian chong juan", Detailed notes in vernacular field only., Shen Yaofeng ji ; Wang Shixiong can ; Xu Zhengjie bu zhu ; [Shen] Zhifan jiao., Qian "Zheru Chen Qingbao cang shu", "Zhuang Zhaoxiang yin", "Zhuang Zhaoxiang".

徐大椿著 ; 王士雄編., 綫裝., 框17.4x12公分, 9行21字. 黑口, 左右雙邊, 單黑魚尾. 版心中鐫題名, 下鐫葉次., 內封背頁牌記鐫"咸豐七年[1857]海昌蔣氏衍芬草堂校梓", 《中國中醫古籍總目》11354著錄., 本書疑為叢書零種(第四冊), 叢書名不詳, 叢書各子目版式不一., 鈐"莊兆祥印", "莊兆祥"., Xian zhuang., Kuang 17.4 x 12 gong fen, 9 hang 21 zi. Hei kou, zuo you shuang bian, dan hei yu wei. Ban xin zhong juan ti ming, xia juan ye ci., Nei feng bei ye pai ji juan "Xianfeng qi nian [1857] Haichang Jiang shi Yan fen cao tang jiao zi", Detailed notes in vernacular field only., Ben shu yi wei cong shu ling zhong (di si ce), cong shu ming bu xiang, cong shu ge zi mu ban shi bu yi., Xu Dachun zhu ; Wang Shixiong bian., Qian "Zhuang Zhaoxiang yin", "Zhuang Zhaoxiang".

王士雄[纂] ; 楊照藜, 汪曰楨評 ; 沈宗淦參., 綫裝 ., 框17.2x13.4公分,10行22字, 小字雙行同. 黑口, 左右雙邊, 對黑魚尾. 版心中鐫題名及卷次, 下鐫葉次., 書名頁刻"溫熱經緯, 海寧王孟英先生纂, 褎海徐三庚署". 書名背頁牌記刻"光緖壬辰[1892]孟秋學院前璧經堂昌記藏版", 前有咸豐二年趙夢齡序, 咸豐五年楊照藜序., 《中國中醫古籍總目》(06428)著錄., 鈐"莊兆祥印", "莊兆祥"., Xian zhuang ., Kuang 17.2 x 13.4 gong fen, 10 hang 22 zi, xiao zi shuang hang tong. Hei kou, zuo you shuang bian, dui hei yu wei. Ban xin zhong juan ti ming ji juan ci, xia juan ye ci., Detailed notes in vernacular field only., Wang Shixiong [zuan] ; Yang Zhaoli, Wang Yuezhen ping ; Shen Zonggan can., Qian "Zhuang Zhaoxiang yin", "Zhuang Zhaoxiang".

王士雄纂 ; 楊照藜, 汪曰楨評 ; 沈宗淦參., 綫裝., 框17.7x13.3公分, 10行22字, 小字雙行同. 黑口, 左右雙邊, 對黑魚尾. 版心中鐫題名及卷次, 下鐫葉次., 書名頁刻"溫熱經緯, 海寧王孟英先生纂, 褎海徐三庚署", 前有咸豐二年[1852]趙夢齡序, 咸豐五年[1855]楊照藜序., 《中國中醫古籍總目》(06428)著錄., 鈐"莊兆祥印", "莊兆祥"., Xian zhuang., Kuang 17.7 x 13.3 gong fen, 10 hang 22 zi, xiao zi shuang hang tong. Hei kou, zuo you shuang bian, dui hei yu wei. Ban xin zhong juan ti ming ji juan ci, xia juan ye ci., Detailed notes in vernacular field only., Wang Shixiong zuan ; Yang Zhaoli, Wang Yuezhen ping ; Shen Zonggan can., Qian "Zhuang Zhaoxiang yin", "Zhuang Zhaoxiang".

Background: A functional artificial ovary is a promising strategy to recover fertility and restore endocrine function in cancer patients. The aim of this study is to optimize the follicle isolation protocol for cryopreserved human ovarian tissues.Methods: Each of the cryopreserved human ovarian cortex pieces (OCPs) from 10 patients was cut into two equal parts and randomly distributed into two treatment groups. Group 1: OCPs digested with Tumor Dissociation Enzyme (TDE); Group 2: OCPs digested with Liberase Dispase High (DH). The efficiency of both groups were evaluated in terms of yield, viability, morphology, and a short-term in vitro culture (IVC) in alginate scaffolds.Results: The TDE can isolate more primordial follicles and smaller diameter of follicles than Liberase DH. The TDE also enabled the isolation of more bright red follicles, higher percent of viable follicles, more morphologically normal follicles, and lower oxidative stress levels compared with Liberase DH. After eight days of IVC, follicles in the TDE group had a higher growth rate from Day 0 to Day 8, and higher viability on Day 8 than the Liberase DH Group.Conclusion: The TDE can be considered an alternative to Liberase DH, enables the isolation of a higher number of healthy follicles from human OCPs, and improves follicle survival after IVC in contrast to Liberase DH.

Background: A functional artificial ovary is a promising strategy to recover fertility and restore endocrine function in cancer patients. The aim of this study is to optimize the follicle isolation protocol for cryopreserved human ovarian tissues.Methods: Each of the cryopreserved human ovarian cortex pieces (OCPs) from 10 patients was cut into two equal parts and randomly distributed into two treatment groups. Group 1: OCPs digested with Tumor Dissociation Enzyme (TDE); Group 2: OCPs digested with Liberase Dispase High (DH). The efficiency of both groups were evaluated in terms of yield, viability, morphology, and a short-term in vitro culture (IVC) in alginate scaffolds.Results: The TDE can isolate more primordial follicles and smaller diameter of follicles than Liberase DH. The TDE also enabled the isolation of more bright red follicles, higher percent of viable follicles, more morphologically normal follicles, and lower oxidative stress levels compared with Liberase DH. After eight days of IVC, follicles in the TDE group had a higher growth rate from Day 0 to Day 8, and higher viability on Day 8 than the Liberase DH Group.Conclusion: The TDE can be considered an alternative to Liberase DH, enables the isolation of a higher number of healthy follicles from human OCPs, and improves follicle survival after IVC in contrast to Liberase DH.