Effect of repeated vitrification of human embryos on pregnancy and neonatal outcomes.

BACKGROUND: Repeated cryopreservation of embryos should occasionally be considered when embryos were not suitable for transfer. The effect of re-cryopreservation on embryos remains contentious. METHODS: This retrospective cohort study aimed to evaluate the pregnancy and neonatal outcomes of twice vitrificated blastocyst derived from once vitrified embryos. Total 410 vitrified-warmed blastocyst transfer cycles were divided into two groups according to the times of embryo vitrification: (1) vitrified blastocysts derived from fresh blastocysts (control group, n = 337); (2) twice vitrified blastocysts derived from once vitrified embryos (n = 73). The primary outcome was live birth rate. Multivariable logistic or linear regression analysis model was performed to describe the association between embryo cryopreservation times and clinical outcomes. RESULTS: No difference was observed in female age at retrieval and transfer, infertility period, body mass index (BMI), infertility type, endometrial thickness, and embryo transfer numbers between the two groups. The pregnancy outcomes of embryos in repeated cryopreservation group were comparable to those of embryos in control group, including implantation rate, clinical pregnancy rate, and live birth rate. In multivariate logistic regression analysis, the cryopreservation times did not affect the outcomes of biochemical pregnancy, clinical pregnancy, and live birth. Moreover, there was no difference in gestational age, birthweight and sex ratio of singleton newborns between groups. After correcting several possible confounding variables, no significant association was observed between cryopreservation times and neonatal birthweight. CONCLUSION: In conclusion, pregnancy and neonatal outcomes achieved with twice vitrified blastocyst transfer were comparable to those achieved with vitrified blastocyst transfer in control group.

Developing Ganoderma lucidum as a next-generation cell factory for food and nutraceuticals.

Ganoderma lucidum holds a colossal reservoir of hydrolytic enzymes and therapeutic compounds and can be a sustainable source of proteins and bioactive compounds. Its metabolic versatility, propelled by its rich genome content, provides excellent biosynthetic machinery for innovation-driven pathway engineering. However, robust regulatory networks and low frequency of homologous recombination are critical bottlenecks that limit the development of molecular tools and precise genetic markers for biomanufacturing innovations in this organism. Modern synthetic biology provides tools that could help to accelerate precise multiple gene targeting and editing and untangling the biosynthetic machinery of G. lucidum. This review provides insight into molecular strategies to unwind the regulatory bottlenecks and transform G. lucidum into efficient cell factories for food and nutraceuticals.

Oxidative stress and inflammatory markers in ovarian follicular fluid of women with diminished ovarian reserve during in vitro fertilization.

BACKGROUND: Follicular microenvironment has been proposed as an important factor for oocyte grown and maturation. We sought to evaluate the oxidative stress and inflammatory levels in follicular fluid (FF) and association with embryo quality in patients with diminished ovarian reserve (DOR). METHODS: The current research included 46 DOR cases and 56 normal ovarian reserve (NOR) cases. Twelve representative oxidative stress markers and eight representative inflammatory factors were measured in the FF. RESULTS: Oxidative stress markers total GSH (T-GSH) was decreased in the FF from women with DOR compared with that in NOR group (P = 0.041). More modest differences were observed for reduced GSH (rGSH) and rGSH/GSSG. Women with DOR compared to controls had higher level of TNF-α (P = 0.000) and lower level of IL-18 (P = 0.013). Correlation analysis revealed that GSSG was negatively correlated with normal fertilization rate in NOR group (r = -0.358, P = 0.008), and reduced GSH was negatively correlated with normal fertilization rate in DOR group (r = -0.299, P = 0.049). Moreover, as the regression analysis data showed, the GSSG level was significantly associated with embryo quality indicator. CONCLUSIONS: The FF in DOR patients was accompanied by increased oxidative stress and inflammatory levels. Follicular development of women with DOR might be influenced by unusual IL-18 and TNF-α levels in FF. And oxidative stress marker GSSG in NOR group was a negative predictor for embryo quality.

Advanced genome-editing technologies enable rapid and large-scale generation of genetic variants for strain engineering and synthetic biology.

Targeted genome editing not only improves our understanding of fundamental rules in life sciences but also affords us versatile toolkits to improve industrially relevant phenotypes in various host cells. In this review, we summarize the recent endeavor to develop efficient genome-editing tools, and emphasize the utility of these tools to generate massive scale of genetic variants. We categorize these tools into traditional recombination-based tools, and more advanced CRISPR as well as RNA-based genome-editing tools. This diverse panel of sophisticated tools has been applied to accelerate strain engineering, upgrade biomanufacturing, and customize biosensing. In parallel with high-throughput phenotyping and AI-based optimization algorithms, we envision that genome-editing technologies will become a driving force to automate and streamline biological engineering, and empower us to address critical challenges in health, environment, energy, and sustainability.

Yiqixue buganshen recipe regulates the expression of integrin ανß3 in the endometrium of controlled ovarian hyperstimulation mice.

OBJECTIVE: To observe the effect of Yiqixue Buganshen recipe(, YBR) on the expression of integrin ανß3 in the endometrium of controlled ovarian hyperstimulation mice. METHODS: A total of 180 mice were divided into three groups: model group, treatment group and control group. The treatment and model groups were intraperitoneally injected with gonadotropin-releasing hormone analogue for 7 days; pregnant mare serum gonadotropin was also injected on the 7th day. After 48 h, human chorionic gonadotropin was injected. The control group was injected with an equal volume of saline at the same time. From the start of the experiment, the treatment group was intragastrically administered Jinghouzengzhi Recipe() and Cuhuangti Recipe(). The model group and the control group were intragastrically administered an equal volume of saline. Real-time reverse transcription polymerase chain reaction and Western blotting were used to detect the mRNA and protein expression of integrin ανß3 in mouse endometrium. RESULTS: Integrin ανß3 was expressed in mouse endometrium in all groups. Integrin αßß3 expression increased gradually along with pregnancy, progressing from pregnant day (Pd) 1. Integrin ανß3 expression significantly increased on Pd 4, then began to decrease on Pd 6. Integrin ανß3 expression in the treatment group was higher than in the model group, and the difference was statistically significant (P <0.05). The difference between the treatment group and the control group was not statistically significant (P >0.05). CONCLUSION: YBR improves endometrial receptivity, and may play an important role in embryonic implantation.

[Fertilization method for primary infertility patients without definite cause undergoing in vitro fertilization and embryo transfer].

OBJECTIVE: To determine the optimal fertilization method for primary infertility patients without definite causes undergoing in vitro fertilization and embryos transfer (IVF-ET). METHODS: A total of 321 IVF-ET cycles for primary infertility without definite causes were divided into two groups, namely group A with infertility period ≥ 5 years (165 cycles) and group B with infertility period <5 years (156 cycles). Each group was further divided into IVF, ICSI, and partial ICSI subgroups. The fertilization rate, incidence of low fertilization rate and clinical pregnancy rate were analyzed. RESULTS: The fertilization rate of IVF in group A was 67.5%, significantly lower than that of ICSI and partial ICSI in the same group (82.0% and 77.7% respectively) and that in IVF control group (76.3%, P<0.05). The incidence of low fertilization rate of IVF in group A was 33.3%, significantly lower than that of ICSI and partial ICSI (8.3% and 15.8%, P<0.05); in group B, the incidence of low fertilization rate of IVF was 12.3%, significantly lower than that of IVF in group A but showed no significant differences from that of ICSI and partial ICSI in group B (P>0.05). In group A, IVF resulted in a significantly lower clinical pregnancy rate (21.1%) than ICSI (43.3%, P<0.05), half ICSI (40.0%, P<0.05), IVF in the control group (48%, P<0.05), and IVF in group B (50.0%, P<0.05). CONCLUSION: ICSI treatment can increase the fertilization rate in IVF-ET cycles in patients with primary infertility for unknown causes, and may improve the clinical outcome of patients with long infertility period.

[Fertilization rates following 109 half intracytoplasmic sperm injection cycles for in vitro fertilization-embryo transfer].

OBJECTIVE: To evaluate the clinical outcomes of half intracytoplasmic sperm injection (partial ICSI) treatment in infertile patients with potential fertilization failure. METHODS: A total of 109 partial ICSI cycles of in vitro fertilization-embryo transfer (IVF-ET) were classified into 5 groups, namely group A (infertile patients for unidentified causes, 17 cycles), group B (oligo-asthenozoospermia patients, 28 cycles), group C (teratozoospermia patients, 8 cycles), group D (primary infertile patients without definite causes, 31 cycles), and group E (secondary infertile patients without definite causes, 25 cycles). The fertilization rate and normal fertilization rate after IVF and ICSI were compared between the groups. RESULTS: Significant differences were found in the fertilization rate following conventional IVF and ICSI in group A (53.1-/+38.8% vs 72.2-/+34.1%) and group D (58.8-/+31.6% vs 82.7-/+21.4%) (P<0.05), but not in groups B, C and E (P>0.05). The normal fertilization rates following IVF and ICSI in groups A, B, D, E were statistically different (P<0.05), but similar in group C (P>0.05). CONCLUSION: ICSI treatment may increase the fertilization rate of IVF-ET in patients with unexplained infertility and primary infertility, but not in patients with oligo-asthenozoospermia, teratozoospermia or secondary infertility.