Clinical and neonatal outcomes of using a modified micro cryotube for cryopreservation of small numbers of spermatozoa for TESA-ICSI cycles.

Cryopreservation of a small number of human spermatozoa is still a major challenge for embryologists. The aim of this study was to evaluate the clinical pregnancy and neonatal outcomes of intracytoplasmic sperm injection (ICSI) using a modified micro cryotube as freezing carrier for freezing small numbers of human spermatozoa collected by testicular sperm aspiration (TESA). We conducted a retrospective study to analyses the ICSI outcomes of using frozen-thawed few testicular spermatozoa in males with obstructive azoospermia (OA) from June 2017 to June 2021. Of 155 ICSI treatment cycles, 79 cycles were allocated to frozen sperm group and a modified micro cryotube was used for freezing testicular sperm, 76 cycles were allocated as fresh sperm group. No significant differences were observed in fertilization rate, good quality embryo rate, and blastocyst rate between the frozen sperm group and fresh sperm group (P > 0.05). Similarly, in the fresh embryo transfer cycles plus the first frozen-thawed embryo transfer cycles, the total clinical pregnancy rate (54.43% vs 57.89%), implantation rate (46.08% vs 49.47%), miscarriage rate (13.95% vs 13.64%) and live birth rate (45.57% vs 48.68%) were not statistically different between the frozen and fresh sperm groups (P > 0.05). In addition, there was no statistical differences in the mean gestational age (38.33weeks ± 1.74 vs 37.89weeks ± 1.87), preterm delivery rate (5.56% vs 10.81%), mean birth weight at delivery (3026.50 g ± 577.64 vs 2977.56 g ± 528.93), and low birth weight (12.50% vs 19.51%) between the two groups (P > 0.05 in all cases). Modified micro cryotube for cryopreservation of rare testicula rretrieved spermatozoa did not negatively affect the pregnancy and neonatal outcomes in TESA-ICSI cycles. The presented method may be a useful alternative for cryopreservation of small numbers of human spermatozoa in clinical setting.

Synthesis of symmetrical secondary oligoethylene glycolated amines from diethanolamine.

Monodisperse oligoethylene glycols (M-OEGs)-containing symmetrical secondary amines are highly valuable synthetic intermediates in drug development and materials sciences. Scalable three-step synthesis of M-OEGs secondary amines with flexible M-OEGs and/or alkyl chains is described herein. Through reduction amination of diethanolamine, Williamson ether synthesis, and subsequent deprotection, a series of M-OEGs secondary amines with diverse and fine-tunable chemical structures were conveniently prepared. The presented strategy is attractive with readily available starting materials, simple catalytic systems, scalable synthesis, and avoids the use of explosive sodium azide.

Tannic acid-Fe complex derivative-modified electrode with pH regulating function for environmental remediation by electro-Fenton process.

A heterogeneous electro-Fenton (hetero-EF) system can effectively broaden the applicable pH range, although the decreased electrogeneration efficiency of H2O2 at elevated pH (especially neutral conditions) is unfavorable for the efficient removal of organic pollutants. Herein, a tannic acid-Fe complex derivative-modified carbon felt (TFD@CF) cathode was prepared for hetero-EF treatment of organic pollutants over a wide pH range. Interestingly, the as-prepared hetero-EF cathode could act as a pH regulator that acidified the solution over a wide pH range. As expected, the TFD@CF cathode exhibited excellent hetero-EF activity for the removal of diverse organic pollutants (such as methyl orange, methylene blue, sulfamerazine, bisphenol A and 2,4-dichlorophenoxyacetic acid) at neutral and even alkaline pH (removal efficiency >90 %). A total of 2.98 kWh kg-1 COD-1 with 83.2 % COD removal could be achieved by the TFD@CF cathode for the treatment of actual textile dyeing secondary wastewater. Electrochemical characterizations proved that the TFD@CF cathode had excellent electrochemical properties with improved electron transfer ability and a well-pronounced Fe(III) electroreductive response. Meanwhile, more acidic groups were newly generated during the electrochemical reaction (an increase of 30.1 %), thus dissociating more H+ into solution. The identification of reactive oxygen species suggested that OH and 1O2 could be responsible for the removal of organic pollutants in the TFD@CF EF system. These interesting findings may provide new insights into the design of multifunctional hetero-EF cathodes for the removal of refractory organic pollutants.

Synergistic effects of oxidation, coagulation and adsorption in the integrated fenton-based process for wastewater treatment: A review.

Fenton process is the most popular for wastewater treatment among all available advanced oxidation processes (AOPs). Numerous endeavors have been devoted to improving the oxidation efficiency of Fenton reaction in terms of promoting ·OH generation, accelerating iron redox cycle and extending applicable pH range. However, in addition to oxidation, coagulation and adsorption also simultaneously occur in the Fenton process, which play important role in the removal of pollutants. Rapid progress has revealed the synergistic effects of oxidation, coagulation and adsorption in the Fenton process, providing new ideas for the treatment of complex and refractory wastewater. Based on available studies, this review is the first to systematically summarize the research progress regarding the synergistic effects of oxidation, coagulation and adsorption in the integrated Fenton-based processes for wastewater treatment. The involved mechanism of the synergistic effects in different Fenton processes (homogeneous Fenton, heterogeneous Fenton and physical field-assistant Fenton coupling process) are critically reviewed. Furthermore, special attention has been paid to the representative applications of the synergistic effects in wastewater treatment (such as industrial organic wastewater, landfill leachate and heavy metal-organic complexes, etc.), particularly focusing on the operation parameters and removal performance. Finally, a conclusion of the review and subsequently, perspectives are given for possible research directions. We believe this review can provide useful information for researchers and end-users involved in the development and application of the Fenton process in wastewater treatment.

Laser-assisted selection of immotile spermatozoa has no effect on obstetric and neonatal outcomes of TESA-ICSI pregnancies.

BACKGROUND: Azoospermic patients have benefited from both epididymal and testicular spermatozoa intracytoplasmic sperm injection (ICSI) treatment and lasers have been used to identify viable, immotile spermatozoa before the procedure. There are limited studies on the safety of laser-assisted selection of immotile spermatozoa. The aim of this study was to investigate the impact of laser-assisted selection of immotile spermatozoa on the obstetric and neonatal outcomes after ICSI. METHODS: A retrospective comparative study was conducted on outcomes of ICSI cycles with testicular spermatozoa from June 2014 to June 2018. Of 132 cycles, 33 were allocated to the test group and oocytes were injected with immotile spermatozoa selected by laser, 99 cycles were allocated as control group. RESULTS: Compared with the control group, no significant differences were found in the pregnancy, implantation, miscarriage and live birth rates in the test group in either fresh or frozen transfer cycles. The cumulative live birth rate in the test group was 69.70%, which was slightly higher than in the control group (60.61%), but this was not statistically different. There were no differences in the average gestational age, premature birth rate, neonatal birth weight, and the malformation rate between the test and control groups (P > 0.05). In addition, the obstetric outcome between the two groups were not different (P > 0.05). CONCLUSIONS: No negative effect on perinatal and neonatal outcomes was seen by using laser-assisted selection of immotile spermatozoa for TESA-ICSI. This study endorses the use of laser-assisted selection of viable spermatozoa for ICSI cycles.

Does longer storage of blastocysts with equal grades in a cryopreserved state affect the perinatal outcomes?

AIM: Although mammalian embryos could be preserved in liquid nitrogen for thousands of years in theoretical models, the viability of cryopreserved blastocyst with varying grades remains to be speculated. In this study, we aimed to determine whether the longer storage time of blastocysts with equal grades could negatively affect the perinatal outcomes. MATERIALS AND METHODS: Single vitrified-warmed blastocyst was divided into four grades (AA, AB/BA, BB, BC/CB) according to the blastocyst score when freezing, and each grade of blastocyst was categorized into four storage duration categories: 28 days-1 year, 1-3 years, 3-5 years, and ≥5 years. Then the perinatal outcomes with different storage time were analyzed. RESULTS: Our results revealed that for blastocysts with the same grade, the length of storage time had no statistical effect on blastocyst survival rate, clinical pregnancy/implantation rate, live birth rate, and abortion rate. In addition, more advanced developmental blastocyst could obtain better pregnancy outcomes regardless of the cryopreservation length. Similar neonatal outcomes were obtained over time. CONCLUSIONS: Cryopreservation time could not negatively affect the perinatal outcomes of blastocysts with equal grades. Efficient blastocyst cryopreservation technology by vitrification can help older women obtain high-quality embryos at a young age.

Constructing a Robust Fluorescent DNA-Stabilized Silver Nanocluster Probe Module by Attaching a Duplex Moiety.

Fluorescent DNA-templated silver nanoclusters (DNA-Ag NCs) have served as excellent luminescent probes and operation units in various applications. However, the fluorescence property of DNA-Ag NCs is very sensitive to elongation or modification of the DNA template, limiting the breadth of applications. In this work, we propose a strategy for constructing a robust fluorescent DNA-Ag NCs probe module by attaching a duplex moiety to the nanocluster-bearing sequence. The fluorescence intensity of the DNA-Ag NCs can be enhanced 90-fold upon hybridization of the elongated moiety. Adenine in the linker sequence has a further enhancing effect on the fluorescence intensity, whereas thymine has a quenching effect. The transformation from a non-fluorescent species to fluorescent nanoclusters is responsible for the fluorescence enhancement with duplex formation of the elongated moiety. We hope that this design will aid future diversification of experimental designs to facilitate more applications that are currently limited by the aforementioned problems.

Effect of different artificial shrinkage methods, when applied before blastocyst vitrification, on perinatal outcomes.

BACKGROUND: In recent years, single blastocyst transfer combined with vitrification has been applied widely, which can maximize the cumulative pregnancy rate in per oocyte retrieval cycles and minimize the multiple pregnancy rate. Thus, the guarantee for these is the effectiveness of vitrified blastocyst. Studies has shown that AS of the blastocoel cavity prior to vitrification can reduce injuries, increase the thawed blastocyst survival rate and implantation rate. Several AS methods have been established. However, only a few studies have compared the effectiveness and safety of these AS methods. In this study, we aimed to compare the clinical outcomes and neonatal outcomes in FET cycles with single blastocyst that were artificially shrunk before vitrification by either LAS or MNAS method. METHODS: A retrospective comparative study of FET cycles in infertile patients which were at our clinic between January 2013 and December 2014. These FET cycles were divided into two groups by the shrinking methods used before vitrification and the clinical and neonatal outcomes were assessed. RESULTS: There were no statistically differences in blastocyst survival rates (95.40% vs 94.05%, P > 0.05) between the LAS and MNAS groups. However, compared with MNAS, LAS improved the warmed blastocyst implantation/clinical pregnancy rate (60.82% vs 54.37%, P < 0.05), live birth rate (50.43% vs 45.22%, P < 0.05) and also increased the monozygotic twin rate (4.07% vs 1.73%, P < 0.05). There were no differences in the average gestational weeks (38.83 ± 1.57 vs 38.74 ± 1.75), premature birth rate (0.30% vs 0.49%), average birth weight (3217.89 ± 489.98 g vs 3150.88 ± 524.03 g), low birth weight rate (5.60% vs 8.63%) and malformation rate (0.59% vs 0.48%) (P > 0.05). CONCLUSIONS: No significant differences in neonatal outcomes were observed, while in clinical outcomes, LAS improved the warmed blastocyst implantation/clinical pregnancy rate and live birth rate markedly, there was also an increased risk of monozygotic twin pregnancies.

A new insight into male fertility preservation for patients with completely immotile spermatozoa.

BACKGROUND: Sperm cryopreservation is the most effective method to preserve male fertility but this is normally used for motile spermatozoa. Thus, only motile spermatozoa are used for cryopreservation in most reproductive medicine centers worldwide. The immotile spermatozoa from some problematic patients are usually discarded, resulting in a missed opportunity of sterility cryopreservation for future assisted reproductive treatments. Many studies have shown that successful fertilization can be obtained after selection of viable sperm from the completely immotile spermatozoa before ICSI. Whether the completely immotile spermatozoa are worth of freezing has not been realized The aim of this study is to explore the clinical value of cryopreservation of immotile spermatozoa. METHODS: Completely immotile spermatozoa were collected and frozen, and subsequently viable but immotile frozen-thawed spermatozoa were selected by laser plus for ICSI. Main outcomes included spermatozoa survival index, fertilization rate and good quality embryo rate. RESULTS: After identification by laser, the fresh samples of spermatozoa presented with a mean survival rate of 54.86% and 26.05%, and this was reduced to 44.13% and 18.13% in frozen-thawed spermatozoa samples, which showed a frozen-thawed spermatozoa survival index of 0.80 and 0.70 in the testicular and ejaculate sperm, respectively. There were no statistically differences in fertilization rate (80% vs80.51%, 75.00% vs 81.48%), cleavage rate (95.45% vs 98.95%, 100.00% vs 95.45%) and good quality embryo rate (40.48% vs 52.13%, 33.33%vs38.10%) between the frozen-thawed immotile spermatozoa group and the routine fresh immotile spermatozoa ICSI group in both testicular and ejaculate sperm, respectively. CONCLUSIONS: The results of the study show that completely immotile spermatozoa can be frozen in order to preserve male fertility as long as viable spermatozoa are present. This procedure provides a further possibility for fertility preservation for patients with completely immotile spermatozoa.

A universal and enzyme-free immunoassay platform for biomarker detection based on gold nanoparticle enumeration with a dark-field microscope.

Developing an enzyme-free, non-amplification strategy for biomarker detection with universality and easy implementation is of central importance in clinical diagnosis and therapeutic monitoring. Herein, we report for the first time a universal and enzyme-free magnetic bead-based sandwich-format immunoassay platform for biomarker detection by combining secondary antibody functionalized AuNPs and automatic AuNP counting readout. For the prostate specific antigen (PSA), the detection limit is found to be 1 ng mL-1, and the spike recoveries (n = 3) with 10% fetal bovine serum are 113.5% for 2 ng mL-1 and 107.7% for 10 ng mL-1. The assay also presents reasonable repeatability as indicated by the coefficient of variance of 13.1% with 5 measurements in 60 days. This strategy has been successfully applied to the determination of carcinoembryonic antigen (CEA) and alpha-fetoprotein (AFP), demonstrating the universality of this strategy. Our proposed non-amplification platform presents sensitivity comparable to that of the enzyme-linked immunosorbent assay (ELISA) with better repeatability; and more importantly, our method has better simplicity than most of the amplification-based methods, and thus is more suitable for routine analysis. The highlights of our work suggest that it is a promising method and would be potentially an alternative for ELISA in laboratories where routine analyses are intensively performed.

Blastocoele re-expansion time in vitrified-warmed cycles is a strong predictor of clinical pregnancy outcome.

AIM: To assess the predictive value of blastocoele re-expansion time in clinical pregnancy outcome in vitrified-warmed cycles. METHODS: Data on 468 single vitrified-thawed blastocyst transfer cycles (in patients aged <38 years) carried out from January 2012 through December 2012, at the Reproductive Medicine Center, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, were analyzed. Vitrified-warmed blastocysts were divided into three groups according to blastocoele re-expansion time: group A, <1 h; group B, 1-2 h; and group C, >2 h, and the clinical pregnancy outcomes (i.e. live birth rate, miscarriage rate and occurrence of singleton pregnancies) compared between the groups. RESULTS: Significant differences were observed in the implantation/clinical pregnancy rate between groups A, B and C (70.10%, 51.76% and 28.74%, respectively, P < 0.01). There was a significant linear decline in this rate with increasing blastocyst re-expansion time. The rate of miscarriage also tended to increase with increasing blastocyst re-expansion time, but the difference was not statistically significant (P > 0.05). Of the pregnant patients, no significant difference was observed in the rates of monozygotic twins and ectopic pregnancy between the three groups. For the newborns, similar live birth, low-birthweight and premature delivery rates were observed between the groups. CONCLUSIONS: Timing of blastocoele re-expansion in vitrified-warmed cycles is a strong predictor of clinical pregnancy outcome. The faster the re-expansion of the blastocoele, the higher the developmental potential of the blastocysts.

[Fluorescence Anisotropy and Applications Based on Functional Nucleic Acid Recognition].

Based on the difference of the depolarization of the emitted light before and after the interaction of molecules, fluorescence anisotropy, also known as fluorescence polarization, can be beneficial for the study of interactions and the detection of the targets. In 1950s, Gregorio Weber first studied the interactions between dansyl chloride and bovine serum albumin or ovalbumin with fluorescence anisotropy method, which paved the way for fluorescence anisotropy in biochemical applications. Since the early 1990s, functional nucleic acids (FNAs, including aptamers, and nucleic acid enzymes) were discovered and synthesized, which have been widely used in functional nucleic acid-based sensing. Aptamers are oligonucleotides that can recognize and bind specifically to various molecular targets. The fluorescence anisotropy methods which are based on aptamer recognition have the advantages of high selectivity, sensitivity and through-put. They play an important role in the study of interaction with protein, nucleic acids and small molecules. However, the way of enhancing the fluorescence anisotropy change of small molecules associated with the binding events is challenging. This paper reviews the basic principles and designs of fluorescence anisotropy methods bases on functional nucleic acid recognition for study and detection of proteins, nucleic acids and small molecules that play an important role in the life activity.

Nonamplification Sandwich Assay Platform for Sensitive Nucleic Acid Detection Based on AuNPs Enumeration with the Dark-Field Microscope.

A simple and efficient assay platform with high sensitivity, convenient implementation, and moderate cost in reagents and instrumentation is most appropriate for routine applications. On the basis of the gold nanoparticle (AuNP) enumeration signal readout mode established in our laboratory, we have developed a nonamplification sandwich assay for nucleic acid detection with the 3 fM limit of detection for a sequence related to Alzheimer's disease. This AuNP counting based method takes advantages of the distinctive and strong localized surface plasmon resonance light scattering with the dark-field microscope and magnetic separation. It is shown that the presence of 20 nM random DNA sequence or calf thymus DNA with a mass up to 10(6)-fold of the targets do not significantly interfere with the assay signal. The spike recoveries of Hela cell lysate sample at 109.3% for 20 pM target and 110.5% for 100 pM target indicate the potential of this proposed method in practical sample applications. This nonamplification sandwich assay platform in principle is applicable to other assays such as the immunoassay and thus would be expected to find a breadth of applications that can make the best use of the simplicity and sensitivity.

Automatic enumeration of gold nanomaterials at the single-particle level.

In this study, we developed a highly sensitive automatic counting method for gold nanomaterials at the single particle level, which can serve as a general sensing platform based on counting of gold nanomaterials. This method substantially improved the sensitivity and accuracy for AuNP counting by adopting the color image processing based on the distinctive localized plasmonic light-scattering of gold nanomaterials. The 60-nm AuNPs, with concentrations down to 4 fM, can be detected with our method. As a universal counting approach for gold nanomaterials, such as gold nanospheres, nanorods, and aggregates from particles under detectable size, this quantification method should be versatile to a breadth of applications.

[Successful pregnancy following laser-assisted selection of viable but immotile spermatozoa for intracytoplasmic sperm injection: A report of 2 cases].

OBJECTIVE: To investigate the feasibility and clinical application value of selecting viable spermatozoa by noncontact diode laser. METHODS: We obtained immotile spermatozoa from 2 infertile men with obstructive azoospermia or severe asthenospermia and selected viable spermatozoa using a single laser shot at the sperm tail. Those that responded to the laser shot by a curling reaction of the tail were regarded as presumably viable and used for intracytoplasmic sperm injection (ICSI). RESULTS: The mean fertilization rate was 88.89% after ICSI with the laser-selected viable spermatozoa. Both of the embryo transfers resulted in a single pregnancy. CONCLUSION: Noncontact diode laser is a useful alternative for the assessment of sperm viability, which may help to achieve successful pregnancy.

[Effects of IVF versus ICSI on the outcomes of elective blastocyst culture].

OBJECTIVE: To explore the effects of different fertilization methods on the outcomes of elective blastocyst culture. METHODS: We retrospectively analyzed the outcomes of elective blastocyst culture for 1 153 cycles of IVF and 205 cycles of ICSI performed between january 2009 and December 2012. RESULTS: A total number of 14 748 embryos in the IVF group and 2 655 embryos in the ICSI group underwent sequential blastocyst culture, with 7 871 blastocysts formed in the former and 1 210 in the latter. No cycles were canceled for no blastocyst formation in either of the two groups. The rates of quality embryos, blastocyst formation and embryo utilization were significantly higher in the IVF than in the ICSI group (64.77 vs 58.72%, 53.37 vs 45.57%, and 60.06 vs 52.17%, all P < 0.05), but the rates of implantation, clinical pregnancy and abortion showed no significant differences between the two groups (48.94 vs 51.43%, 49.03 vs 52.02%, and 11.69% vs 15.56, all P > 0.05). CONCLUSION: With the same inclusion criteria of selective blastocyst culture, IVF has a lower risk of cycle cancellation due to no blastocyst formation and therefore may effect higher rates of blastocyst formation and embryo utilization than ICSI. Our study suggested that appropriate inclusion criteria of selective blastocyst culture should be laid down according to different fertilization methods.

Identification of Differentially Expressed mRNAs and miRNAs and Related Regulatory Networks in Cumulus Oophorus Complexes Associated with Fertilization.

Cumulus oophorus complexes (COCs) are the first extracellular barriers that sperm must pass through to fuse with oocytes, which have an important role in oocyte maturation and fertilization. However, little is known about the molecular mechanisms of COCs involved in fertilization. In this study, COCs were collected and then randomly divided into a test group that interacted with sperm and a control group that did not interact with sperm. Then, the total RNA was extracted; RNA transcriptome and small RNA libraries were prepared, sequenced, and analyzed. The results showed that 1283 differentially expressed genes (DEGs), including 560 upregulated and 723 downregulated genes. In addition, 57 differentially expressed miRNAs (DEMIs) with 35 upregulated and 22 downregulated were also detected. After the RNA-seq results were verified by RT-qPCR, 86 effective DEGs and 40 DEMIs were finally screened and a DEMI-DEG regulatory network was constructed. From this, the top ten hub target genes were HNF4A, SPN, WSCD1, TMEM239, SLC2A4, E2F2, SIAH3, ADORA3, PIK3R2, and GDNF, and they were all downregulated. The top ten hub DEMIs were miR-6876-5p, miR-877-3p, miR-6818-5p, miR-4690-3p, miR-6789-3p, miR-6837-5p, miR-6861-5p, miR-4421, miR-6501-5p, and miR-6875-3p, all of which were upregulated. The KEGG signaling pathway enrichment analysis showed that the effective DEGs were significantly enriched in the calcium, AMPK, and phospholipase D signaling pathways. Our study identified several DEGs and DEMIs and potential miRNA-mRNA regulatory pathways in COCs and these may contribute to fertilization. This study may provide novel insights into potential biomarkers for fertilization failure.

One-Pot Synthesis of Monofunctionalized Oligoethylene Glycols through Ring-Opening and Heterogeneous Hydrolysis of Macrocyclic Sulfates.

The one-pot nucleophilic ring-opening reaction of oligoethylene glycol macrocyclic sulfates provides an efficient strategy for the monofunctionalization of oligoethylene glycols without protecting or activating group manipulation. In this strategy, the hydrolysis process is generally promoted by sulfuric acid, which is hazardous, difficult to handle, environmentally unfriendly, and unfit for industrial operation. Here, we explored a convenient handling solid acid, Amberlyst-15, as a replacement for sulfuric acid to accomplish the hydrolysis of sulfate salt intermediates. With this method, 18 valuable oligoethylene glycol derivatives were prepared with high efficiency, and gram-scale applicability of this method has been successfully demonstrated to afford a clickable oligoethylene glycol derivative 1b and a valuable building block 1g for F-19 magnetic resonance imaging traceable biomaterial construction.

All-in-One Heptamethine Cyanine Amphiphiles for Dual Imaging-Guided Chemo-Photodynamic-Photothermal Therapy of Breast Cancer.

Developing a theranostic system that integrates multimodal imaging, synergistic therapeutic, and formulation entities is a promising strategy for efficient cancer treatment. However, the complexity and safety concerns of multiple functional entities hinder their clinical translation. Herein, versatile "all-in-one" heptamethine cyanine amphiphiles (PEG-Cy-Fs) with multiple favorable capabilities, including fluorine-19 magnetic resonance imaging (19 F MRI), near-infrared fluorescence imaging (NIR FLI), photodynamic therapy (PDT), photothermal therapy (PTT), polyethylene glycolation (PEGylation) and high biocompatibility, are developed for the convenient construction of theranostic platforms. Amphiphiles PEG-Cy-Fs are synthesized on a multi-hundred-milligram scale with high efficacy, which self-assembled with a chemotherapy drug tamoxifen (TAM) into monodisperse and stable nanoparticles (SoFoTm/PEG-Cy-F18 ) with "turned on" FLI, sensitive 19 F MRI, mitochondria-targeting ability, high PDT and PTT efficacy, and PEGylation-optimized pharmacokinetics. The selective accumulation of SoFoTm/PEG-Cy-F18 in xenograft MCF-7 tumor with a long retention time (>10 days) enabled 19 F MRI-NIR FLI-guided chemo-photodynamic-photothermal therapy (chemo-PDT-PTT) of breast cancer with high therapeutical index in mice. The "all-in-one" heptamethine cyanine amphiphile may facilitate the convenient and standardized preparation of high-performance theranostics systems for clinical translation.

A point of confusion for embryologists in the identification of viable spermatozoa by the eosin-nigrosin test.

A viable spermatozoon is a prerequisite for fertilization in intracytoplasmic sperm injection (ICSI). Thus, it is crucial to select viable but immotile spermatozoa on the day of ICSI. We report conflicting results in the identification of viable but immotile spermatozoa between the eosin-nigrosin staining and the laser test, which resulted in confusion for embryologists during assisted reproductive technology (ART). Three patients' semen samples that showed no motile spermatozoa are described in this report. To identify viable spermatozoa, we used both the eosin-nigrosin test and the laser test for each sample, and repeated the semen analysis twice in each patient. Viable but immotile spermatozoa selected by the laser test were used for ICSI. Viable spermatozoa were detected by both the eosin-nigrosin and laser tests in two patients (case 1, 95.00% vs. 24.21% and 92.68% vs. 22.22%; case 2, 41.18% vs. 23.48% and 39.81% vs. 22.52%), indicating consistent results between the two methods. In the third patient, the eosin-nigrosin test yielded viability rates of 20.75% and 19.14%, while the result of the laser test was 0%. Thus, testicular aspiration was performed to collect viable sperm from this patient. Normal fertilization was achieved after the injection of viable but immotile spermatozoa selected from these patients by the laser test, resulting in the birth of two healthy babies. Our study documents a case where the eosin-nigrosin test showed a limitation in identifying viable but immotile spermatozoa for ART, while the laser test may overcome this limitation. Larger samples may be required to corroborate the clinical value of the laser test.