Comparison of frozen-thaw blastocyst transfer strategies in women aged 35-40 years: a retrospective study.

Objective: To compare the effects of five different frozen-thaw embryo transfer (FET) strategies in women aged 35-40 years. Methods: Data from 1,060 patients were divided into five groups according to the number and quality of transferred blastocysts: a high-quality single blastocyst group (group A, n= 303), a high-quality double blastocysts group (group B, n= 176), a high-quality plus poor-quality double blastocysts group (group C, n= 273), a poor-quality double blastocysts group (group D, n= 189), and a poor-quality single blastocyst group (group E, n= 119). Comparative analyses were then performed between groups with regard to primary conditions, pregnancy, and neonatal outcomes. Results: Group A had the lowest twin pregnancy rate (1.97%) and incidence of low-birth-weight infants (3.45%), which were significantly different from groups B, C, and D. In addition, the preterm birth rate (7.89%), neonatal birth weight (3300 g [3000, 3637.5]), and neonatal birth age (39.14 weeks [38.43, 39.61]) in group A were different from those in groups B and C. Double blastocyst transfer (DBT) was associated with a 20.558-fold (Risk Ratio [RR]=20.558, 95% confidence interval [CI], 6.628-63.763) and 3.091-fold (RR=3.091, 95% CI, 1.69-5.653) increased risk of twin pregnancy and preterm delivery in unadjusted analysis, respectively, when compared with single blastocyst transfer (SBT). In the adjusted analysis, we observed similar risk estimates (adjusted RR=26.501, 95% CI, 8.503-82.592; adjusted RR=3.586, 95% CI, 1.899-6.769). Conclusion: Although, high-quality SBT resulted in a lower live birth rate than high-quality DBT, it also significantly reduced the risk of adverse pregnancies, thus resulting in more benefits for both the mother and baby. Collectively, our data indicate that high-quality SBT remains the optimal FET strategy for women aged 35-40 years and warrants further clinical application.

Vaginal Epithelial Cell Membrane-Based Phototherapeutic Decoy Confers a "Three-in-One" Strategy to Treat against Intravaginal Infection of Candida albicans.

Phototherapy is an effective strategy to control Candida albicans (C. albicans) infection without raising the concern of drug resistance. Despite its effectiveness, a higher dose of phototherapeutic power is required for C. albicans elimination compared to bacteria that have to be used, which is readily accompanied by off-target heat and toxic singlet oxygen to damage normal cells, thus limiting its usefulness for antifungal applications. Here to overcome this, we develop a "three-in-one" biomimetic nanoplatform consisting of an oxygen-dissolved perfluorocarbon camouflaged by a photosensitizer-loaded vaginal epithelial cell membrane. With a cell membrane coating, the nanoplatform is capable of specifically binding with C. albicans at the superficial or deep vaginal epithelium, thereby centering the phototherapeutic agents on C. albicans. Meanwhile, the cell membrane coating endows the nanoplatform to competitively protect healthy cells from candidalysin-medicated cytotoxicity. Upon candidalysin sequestration, pore-forming on the surface of the nanoplatform accelerates release of the preloaded photosensitizer and oxygen, resulting in enhanced phototherapeutic power for improved anti-C. albicans efficacy under near-infrared irradiation. In an intravaginal C. albicans-infected murine model, treatment with the nanoplatform leads to a significantly decreased C. albicans burden, particularly when leveraging candidalysin for further elevated phototherapy and C. albicans inhibition. Also, the same trends hold true when using the nanoplatform to treat the clinical C. albicans isolates. Overall, this biomimetic nanoplatform can target and bind with C. albicans and simultaneously neutralize the candidalysin and then transform such toxins that are always considered a positive part in driving C. albicans infection with the power of enhancing phototherapy for improved anti-C. albicans efficacy.

Doxorubicin Detoxification in Healthy Organs Improves Tolerability to High Drug Doses for Enhanced Antitumor Therapy.

With its well-documented toxicity, the use of doxorubicin (Dox) for cancer treatment requires trade-offs between safety and effectiveness. This limited use of Dox also hinders its functionality as an immunogenic cell death inducer, thus impeding its usefulness for immunotherapeutic applications. Here, we develop a biomimetic pseudonucleus nanoparticle (BPN-KP) by enclosing GC-rich DNA within erythrocyte membrane modified with a peptide to selectively target healthy tissue. By localizing treatment to organs susceptible to Dox-mediated toxicity, BPN-KP acts as a decoy that prevents the drug from intercalating into the nuclei of healthy cells. This results in significantly increased tolerance to Dox, thereby enabling the delivery of high drug doses into tumor tissue without detectable toxicity. By lessening the leukodepletive effects normally associated with chemotherapy, dramatic immune activation within the tumor microenvironment was also observed after treatment. In three different murine tumor models, high-dose Dox with BPN-KP pretreatment resulted in significantly prolonged survival, particularly when combined with immune checkpoint blockade therapy. Overall, this study demonstrates how targeted detoxification using biomimetic nanotechnology can help to unlock the full potential of traditional chemotherapeutics.

Biocompatible Assessment of Erythrocyte Membrane-Camouflaged Polymeric PLGA Nanoparticles in Pregnant Mice: Both on Maternal and Fetal/Juvenile Mice.

Purpose: Poly(lactic-co-glycolic) acid (PLGA) nanoparticles coated with the membrane of red blood cells (RBC-NP) have been applied in various biomedical fields. Despite the well-documented great biocompatibility, the potential toxicity of RBC-NP on maternal mice or their developing fetuses during pregnancy, or juvenile mice post-birth, remains unclear, which warrants a systematic evaluation. Methods: We fabricate an RBC-NP with approximately 50 nm in diameter (RBC-NP-50). Upon RBC-NP-50, pregnant mice are intravenously injected with this nanoparticle either at a single high dose of 400 mg/kg (1HD) or a low dose of 200 mg/kg for 3 times (3LD). Afterwards, the biocompatible assessments are performed at 48 h after the final injection or 21 d post-birth/partum both on maternal and fetal/juvenile mice. Results: RBC-NP-50 is capable of accumulating in the placenta and then passing through the blood-fetal barrier (BFB) into the fetus. On 48 h after RBC-NP-50 exposure, no significant dose-dependent toxicity is observed in maternal mice including blood biochemistry, inflammatory factors, progesterone level, histological analysis, etc, whereas fetal brains reveal remarkable differentially expressed genes analyzed by transcriptome sequencing. On 21 d post-birth, those genes' expression in juvenile mice is alleviated, along with negligible differences in behavioral evaluations including surface righting test, negative geotaxis test, cliff avoidance test, and olfactory orientation test. Conclusion: These results indicate that RBC-NP is considered to be generally safe and biocompatible both for maternal mice and fetus during pregnancy, and for the subsequent juvenile mice post-birth, although future studies will need to examine higher dosage or longer-term measurements.

Comparison of frozen-thawed embryo transfer strategies for the treatment of infertility in young women: a retrospective study.

Objective: To investigate transfer strategies in the frozen-thawed embryo transfer (FET) cycle. Methods: The clinical data of 1,652 FET patients were divided into five groups according to the number and quality of the transferred blastocyst: high-quality single blastocyst group (group A, n = 558), high-quality plus poor-quality double blastocyst group (group B, n = 435), poor-quality double blastocyst group (group C, n = 241), high-quality double blastocyst group (group D, n = 298), and poor-quality single blastocyst group (group E, n = 120). Inter-group comparison analyses of primary conditions, pregnancy outcomes and neonatal outcomes were then performed. Results: Group A had the highest embryo implantation rate (67.38%), significantly different from the implantation rates of the other four groups. The gemellary pregnancy rate (1.60%), preterm birth rate (5.58%), neonatal birth weight (3,350g [3,000g, 3,650g]), neonatal birth age (39.57 weeks [38.71, 40.34]), and incidence of low birth weight (7.02%) in group A were different from those in groups B, C, and D, but did not significantly differ from those in group E. Moreover, the proportions of male infants born in groups A (56.86%) and D (59.41%) were significantly higher than those in the other three groups. Double blastocyst transfer (0.528, 95% CI [0.410-0.680], P < 0.001) and high-quality blastocyst transfer (0.609, 95% CI [0.453-0.820], P = 0.001) were found to be protective factors for live birth. In addition, double blastocyst transfer was also the largest risk factor for pregnancy complications (3.120, 95% CI [2.323-4.190], P < 0.001) and neonatal complications (2.230, 95% CI [1.515-3.280], P < 0.001), especially for gemellary pregnancy (59.933, 95% CI [27.298-131.58], P < 0.001) and preterm birth (3.840, 95% CI [2.272-6.489], P < 0.001). Based on the ROC curves, a double blastocyst transfer could predict gemellary pregnancy reliably with a high area under the curve (AUC = 78.53%). Additionally, a double blastocyst transfer could effectively predict a high risk of pregnancy complications (AUC = 65.90%), neonatal complications (AUC = 64.80%) and preterm birth (AUC = 66.20%). Conclusion: The live birth rate of frozen-thawed high-quality single blastocyst transfer is lower than that of double high-quality blastocyst transfer, which can significantly increase the embryo implantation rate. High-quality single blastocyst transfer also significantly lowers the risk of gemellary pregnancy, preterm birth, and low birth weight, and can significantly improve maternal and infant outcomes. After weighing the pros and cons of live birth with pregnancy and neonatal complications, the authors believe that high-quality single blastocyst transfer is the optimal FET strategy for young women and is worthy of further clinical application. Despite this recommendation, high-quality single blastocyst transfer can increase the risk of monozygotic twins, as well as significantly increase the proportion of male infants born.

Methylene Blue Combined with Ropivacaine for Intercostal Nerve Block After Autologous Costal Cartilage Removal in Juvenile Patients.

OBJECTIVES: Autologous costal cartilage is commonly used as a graft material in plastic surgery. However, after autologous costal cartilage removal, the pain at the surgical site is particularly strong. We conducted this controlled clinical study to verify the efficacy of methylene blue (MB) in intercostal nerve block after autologous costal cartilage removal and to provide a reference for the application of MB in postoperative analgesia after autologous costal cartilage removal. METHODS: In this study, 90 adolescent patients with congenital microtia who underwent autologous rib cartilage graft for auricular reconstruction were randomly allocated to one of three groups (Group A: intercostal nerve block was performed with 0.75% ropivacaine; Group B: intercostal nerve block was performed with 1% MB; and Group C: intercostal nerve block was performed with 1% MB and 0.75% ropivacaine mixture). Two trained researchers observed and recorded the pain status of the children at 6 hours (T1), 24 hours (T2), 48 hours (T3), and 72 hours (T4) after surgery, respectively. Numerical rating pain scale (NRS) was used for scoring. And adverse reactions such as nausea, vomiting, and skin itching were recorded. RESULTS: In this study, there was no statistical difference in age and gender of patients in Groups A, B, and C (P >0.05). In terms of NRS comparison, 6 hours after operation (T1), Group B > Group A > Group C (P< 0.05); 24 hours after operation (T2), Group B > Group A > Group C (P< 0.05); 48 hours after operation (T3), Group B > Group A > Group C (P< 0.05); 72 hours after operation (T4), Group A > Group B > Group C (P< 0.05). There were no statistically significant differences in postoperative nausea, vomiting, and skin itching among the three groups (P>0.05). CONCLUSION: The analgesic effect of IV self-controlled analgesia combined with ropivacaine is quick, but the maintenance time is short. The analgesic effect of IV self-controlled analgesia combined with MB is slow to onset but long to maintain. The analgesic effect of IV self-controlled analgesia combined with MB and ropivacaine mixture is quick and maintained for a long time. Therefore, in patients after removal of costal cartilage, we recommend the analgesic treatment method of IV self-controlled analgesia combined with MB and ropivacaine mixture. LEVEL OF EVIDENCE I: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 . Special Topic.

Research Progress on the Microregulatory Mechanisms of Fertilization: A Review.

The process of fertilization includes sperm capacitation, hyperactivation, an acrosome reaction and the release of acrosome enzymes, membrane fusion and channel formation, the release of the sperm nucleus, and gamete fusion. This process is closely related to the shape and vitality of the sperm, acrosome enzyme release, and the zona pellucida structure of the egg, as well as the opening and closing of various ion (e.g., calcium) channels, the regulation of signaling pathways such as cyclic adenosine monophosphate-protein kinase A, the release of progesterone, and the coupling of G-proteins. The interaction among multiple factors and their precise regulation give rise to multiple cascading regulatory processes. Problems with any factor will affect the success rate of fertilization. Recent studies have shown that with rapid societal development, the incidence of male infertility is increasing and occurs at younger ages. According to World Health Organization statistics, 15% of couples of childbearing ages have infertility problems, of which 50% are caused by male factors. Additionally, the cause of infertility cannot be identified in as many as 60% to 75% of male infertility patients. In this article, we review the research progress on the microregulation of fertilization and mechanisms underlying this process to identify causes and develop novel prevention and treatment strategies for male infertility.

Can SpRY recognize any PAM in human cells?

The application of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated proteins (Cas) can be limited due to a lack of compatible protospacer adjacent motif (PAM) sequences in the DNA regions of interest. Recently, SpRY, a variant of Streptococcus pyogenes Cas9 (SpCas9), was reported, which nearly completely fulfils the PAM requirement. Meanwhile, PAMs for SpRY have not been well addressed. In our previous study, we developed the PAM Definition by Observable Sequence Excision (PAM-DOSE) and green fluorescent protein (GFP)|-reporter systems to study PAMs in human cells. Herein, we endeavored to identify the PAMs of SpRY with these two methods. The results indicated that 5'-NRN-3', 5'-NTA-3', and 5'-NCK-3' could be considered as canonical PAMs. 5'-NCA-3' and 5'-NTK-3' may serve as non-priority PAMs. At the same time, PAM of 5'-NYC-3' is not recommended for human cells. These findings provide further insights into the application of SpRY for human genome editing.

Deficiency of C1QL1 Reduced Murine Ovarian Follicle Reserve Through Intraovarian and Endocrine Control.

Ovarian aging is associated with depletion of the ovarian follicle reserve, which is the key determinant of fertility potential in females. In this study, we found that the small, secreted protein complement 1Q-like (C1QL1) is involved in the regulation of follicle depletion through intraovarian and endocrine control in a multidimensional collaborative manner. C1ql1 was detected to be conserved in the ovary and showed high transcript levels during folliculogenesis. Blockade of C1QL1 by IP and ovarian intrabursal injection of C1QL1 antiserum into prepubertal mice impaired folliculogenesis accompanied by reductions in body weight, fat mass, and intraovarian lipid accumulation. An elevation of circulating estradiol levels, reduction of hypothalamic KISS1 and GnRH expression, and a decrease in serum FSH levels were found in C1QL1-deficient mice. In C1QL1-deficient ovaries, many primordial follicles were recruited and developed into medium follicles but underwent atresia at the large follicle stages, which resulted in depletion of follicle reserve. Depletion of C1QL1 alleviated the inhibitory effect of C1QL1 on granulosa cell apoptosis and the stimulatory effect of C1QL1 on granulosa cell autophagy, which resulted in accumulation in the preantral and early antral follicles and an increase in the atretic follicles. The abnormal profile of endocrine hormones accelerated the intraovarian effect of C1QL1 deficiency and further led to depletion of ovarian reserve. Altogether, this study revealed the expression patterns and the mechanism of action of C1QL1 during folliculogenesis and demonstrated that deficiency of C1QL1 caused ovarian follicular depletion.

The feasibility of switching from IVF to IVM combined with all-blastocyst-culture and transfer for patients with ovarian hyperstimulation syndrome tendency.

OBJECTIVE: To investigate the feasibility of switching from in vitro fertilization (IVF) to in vitro maturation (IVM) combined with all-blastocyst-culture and transfer as a supplementary infertility treatment in patients with ovarian hyperstimulation syndrome (OHSS) tendency METHODS: Retrospective cohort study including 184 patients who switched from IVF and underwent 192 IVM cycles between January 2016 and December 2020. The outcomes were compared between cleavage-stage embryo transfer (group A, n = 74) and blastocyst-stage transfer (group B, n = 52) groups. RESULTS: The OHSS rate is 0%. 66 cycles were canceled for transfer. Among the 126 transfer cycles, number of retrieved oocytes, proportion of metaphase II oocytes, cleavage rate, and proportion of high-quality embryos on day 3 post-fertilization are significantly lower in group A than that in group B. On the contrary, number of transferred embryos is significantly lower in group B than that in group A, whereas the rates of implantation, clinical pregnancy, and live births are significantly higher in group B than that in group A. CONCLUSION: Timely switching to IVM combined with all-blastocyst-culture and transfer for patients undergoing controlled ovarian hyperstimulation and exhibiting characteristics of OHSS tendency is feasible as a supplementary infertility treatment.

A patient with severe congenital neutropenia harbors a missense ELANE mutation due to paternal germline mosaicism.

BACKGROUND: Clinical and genetic characteristics of ELANE mutation of a 3-year-old male who had a severe congenital neutropenia (SCN) were examined. We then investigated whether CRISPR/Cas9-mediated gene editing could correct the mutation. PROCEDURE: The proband underwent extensive clinical assessments, such as exome sequencing and bioinformatics analysis, so that pathogenic genes could be identified. Sanger sequencing was also utilized for confirmation. The cell line, 293-ELANE, harboring ELANE mutation was generated, and the mutation was then corrected by CRISPR/Cas9-mediated homology-directed repair (HDR). RESULTS: The ELANE gene test in the proband unveiled a heterozygous de novo missense mutation: c. 248T > A (p.V83D), which was not detected in his asymptomatic parents who had provided peripheral blood samples. We found that 46.01% of his father's sperm cells had the same mutation. These results demonstrate that the proband inherited the ELANE mutation from his father, who had an average neutrophil count but had a germline mosaicism. The highest repair efficiency of CRISPR/Cas9-mediated HDR for 293-ELANE is 4.43%. CONCLUSIONS: We identified a missense mutation (p.V83D) in ELANE that causes SCN. This is the first report on paternal semen mosaicism of an ELANE mutation. Our study paves the way for preimplantation genetic diagnosis (PGD) based on ELANE mutation prevention and clinical treatment of congenital disabilities.

Clinical and perinatal outcomes of fresh single-blastocyst-transfer cycles under an early follicular phase prolonged protocol according to day of trigger estradiol levels.

BACKGROUD: This study's objectives were to compare the clinical, perinatal, and obstetrical outcomes of patients with different estradiol (E2) levels in fresh single-blastocyst-transfer (SBT) cycles under an early follicular phase prolonged regimen on the day of trigger. METHODS: We recruited patients in fresh SBT cycles (n = 771) undergoing early follicular phase prolonged protocols with ß-hCG values above 10 IU/L between June 2016 and December 2018. Patients who met the inclusion and exclusion criteria were divided into four groups according to their serum E2 level percentages on the day of trigger: <25th, 25th-50th, 51st-75th, and >75th percentile groups. RESULTS: Although the rates of clinical pregnancy (85.57% (166/194)), embryo implantation 86.60% (168/194), ongoing pregnancy (71.13% (138/194)), and live birth (71.13% (138/194)) were lowest in the >75th percentile group, we did not observe any significant differences (all P > 0.05). We used this information to predict the rate of severe ovarian hyperstimulation syndrome (OHSS) area under the curve (AUC) = 72.39%, P = 0.029, cut off value of E2 = 2,893 pg/ml with the 75% sensitivity and 70% specificity. The 51st-75th percentile group had the highest rates of low birth weight infants (11.73% (19/162), P = 0.0408), premature delivery (11.43% (20/175), P = 0.0269), admission to the neonatal intensive care unit (NICU) (10.49% (17/162), P = 0.0029), twin pregnancies (8.57% (15/175), P = 0.0047), and monochorionic diamniotic pregnancies (8.57% (15/175); P = 0.001). We did not observe statistical differences in obstetrics complications, including gestational diabetes mellitus (GDM), gestational hypertension, placenta previa, premature rupture of membranes (PROM), and preterm premature rupture of membranes (PPROM). CONCLUSION: We concluded that serum E2 levels on the day of trigger were not good predictors of live birth rate or perinatal and obstetrical outcomes. However, we found that high E2 levels may not be conducive to persistent pregnancies. The E2 level on the day of trigger can still be used to predict the incidence of early onset severe OHSS in the fresh SBT cycle.

Screening a novel signature and predicting the immune landscape of metastatic osteosarcoma in children via immune-related lncRNAs.

BACKGROUND: The immune microenvironment plays an essential role in osteosarcoma (OSs); however, differences in immune-related long non-coding ribonucleic acids (irlncRNAs) in children with localized OSs and metastatic OSs have not yet been investigated. METHODS: The clinical data and the transcriptome of OSs were obtained from the Therapeutically Applicable Research to Generate Effective Treatments (TARGET) database, and the immune-related genes were derived from the imported dataset. The correlations between immune-related genes and lncRNAs were examined. Next, the differential expressions of the irlncRNA pairs (IRLPs) in localized OSs and distant metastatic OSs were analyzed, and a prognostic model was constructed based on the significant differentially expressed IRLPs. We also analyzed the association between the IRLPs' signature risk score and the infiltration of the immune cells. Finally, we investigated the correlation between risk score and drug resistance. RESULTS: Thirty upregulated and 22 downregulated lncRNAs were identified in the localized and metastatic OSs samples. Univariate and multivariate cox regression analyses were undertaken to select 6 lncRNA pairs to establish the prognostic signature, the model was valuable in predicting OSs prognosis. Further, the expression of the finally selected irlncRNAs indicated that VPS9D1-AS1 (P=0.031), AP003086.2 (P=0.041), AL031847.1 (P=0.008), AL020997.3 (P=0.020), AC011444.1 (P=0.025), and AC006449.2 (P=0.003) were significantly upregulated in metastasis patients, but USP27X-AS1 (P=0.046), AL008721.2 (P=0.005), AC002091.1 (P=0.033), and AL118558.4 (P=0.049) were significantly overexpressed in localized patients. The overexpression of AC002091.1 (P=0.038) and AL118558.4 (P=0.004) resulted in better overall survival, but the upregulation of AC011444.1 (P=0.045), AL031847.1 (P=0.020), VPS9D1-AS1 (P=0.039), and AC006449.2 (0.006) led to a poor outcome. Differences in immune cell infiltration indicated that metastatic patients and localized have significant difference of 4 (CD4) T cells (P=0.006), monocytes (P=0.029), activated mast cells (P=0.018), and neutrophils (P=0.026), and a high abundance of activated dendritic cells (P=0.010) and activated mast cells (P=0.049) resulted in poor prognosis. Patients in the high-risk-score group were resistant to axitinib, but sensitive to dasatinib, bortezomib, and cisplatin. CONCLUSIONS: In the present study, IRLPs were used to construct a novel and practical model for predicting the prognosis of localized and metastatic OSs in children.

[Genetic Diagnosis of Thalassemia in Baise, Guangxi Zhuang Autonomous Region].

OBJECTIVE: To understand the genotypes and distribution characteristics of thalassemia in Baise, Guangxi Zhuang Autonomous Region, to provide references for the prevention and diagnosis of thalassemia in the region and improve the quality of eugenics. METHODS: 3 482 pregnant women and their spouses from January 2019 to August 2019 in Baise Maternal and Child Health Hospital for prenatal genetic diagnosis were selected, α, ß- thalassemia genes were detected by Gap-PCR, PCR and DNA reverse dot hybridization, cases carrying thalassemia gene were confirmed and statistical analyzed. RESULTS: 2 260 samples (64.90%) carrying thalassemia gene were found, among which 1 459 cases (64.56%) were diagnosed as α- thalassemia, 617 cases (27.30%) as ß- thalassemia, 184 cases (8.14%) as α complex ß- thalassemia. Among 1 459 α- thalassemia genes, --SEA /αα(637 cases, 43.66%), -α3.7 /αα (306 cases, 20.97%), -αCS /αα(143 cases, 9.80%), -α4.2 /αα(124 cases, 8.50%) and -αWS /αα(77 cases, 5.27%) were the most common, while among 617 ß- thalassemia genes, CD17 (229 cases, 37.12%), CD41-42 (213 cases, 34.52%), IVS-I-1 (41 cases, 6.65%), ßE (38 cases, 6.16%) and CD71-72 (34 cases, 5.51%) were the most common. And --SEA /αα/ CD17 (24 cases, 13.04%), -α4.2 /αα/ CD17 (13 cases, 7.07%), -α3.7 /αα/ CD41-42 (12 cases, 6.52% ) and --SEA /αα/ CD41-42 (12 cases, 6.52%) were mainly found in 184 cases of α complex ß - thalassemia. CONCLUSION: Genotyes of thalassemia in Baise, Guangxi Zhuang Autonomous Region are complex and diverse. The prenatal screening and diagnosis of thalassemia in the region should be strengthened in accordance with the characteristics of genetypes in the region, in order to reduce birth defects and improve eugenics quality.

Finite Element Analysis of an Improved Correction System for Spinal Deformity.

BACKGROUND/AIM: Surgical treatment for spinal deformity aims to correct malformation, release the nerves, and reconstruct spinal stability. To explore and develop a new improved spinal correction system (ISCS) for clinical application, we studied the stability and biomechanical characteristics of the ISCS through finite element analysis and comparison of the ISCS with the pedicle screw and rod system (PSRS). PATIENTS AND METHODS: Using L1-L3 CT image data of a normal adult male lumbar spine for establishment of L1-L3 finite element model, we established posterior internal fixation models for a comparative finite element analysis of PSRS and ISCS. An axial load of 500 N and a moment of 10 N•m were applied to L1 to simulate flexion, extension, lateral bending, and axial rotation. Stress distribution characteristics, load sharing, strain bending stiffness and strain angle change of the models were measured. RESULTS: In flection and extension directions, the maximum stress of the L2 vertebral body and the L1/2 and L2/3 discs in PSRS was less than that of ISCS. In lateral bending and axial rotation directions, the maximum stress between PSRS and ISCS was similar. However, the stress shielding rate of L2, L1/2, and L2/3 intervertebral discs in ISCS was significantly lower than that of PSRS. We also found that both models had similar angular displacement and maximum displacement in lateral bending direction, but PSRS had a lower angular displacement and maximum displacement in flection and extension directions. Finally, we showed that PSRS had similar angular displacement and a lower maximum displacement compared with ISCS in axial rotation, whereas ISCS had lower bending stiffness than PSRS in different directions. CONCLUSION: ISCS can effectively fix spinal deformities compared to PSRS. ISCS provides a new option for orthopedic surgery treatment of scoliosis and, therefore, warrants further clinical studies in patients with other spinal deformities.

Whole-exome sequencing analysis in 10 families of sporadic microtia with thoracic deformities.

BACKGROUND: Microtia is a congenital malformation of the external ear and may occur as an isolated deformity or as part of a syndrome. Our previous study found a high correlation between microtia and thoracic deformities, thus, we propose that external ear and thorax development may be regulated by certain genes in common. METHODS: We performed exome sequencing on 10 families of sporadic microtia with thoracic abnormalities. We identified mutated genes under different models of inheritance, and checked them through Mouse Genome Informatics and association analysis. RESULTS: We identified 45 rare mutations, including 9 de novo mutations, 20 heterozygous mutations, 3 homozygous mutations, and 13 hemizygous mutations, of which 2 are likely to be causative. They are de novo missense variant in PHF5A and compound heterozygous mutations in CYP26B1, of which CYP26B1 mutation is highly likely pathogenic. CONCLUSION: The results indicate that certain genes may affect both external ear and thorax development, and demonstrate the benefits of whole-exome sequencing in identifying candidate genes of microtia. This study provides a new way for genetic exploration in microtia.

Comparison of Pregnancy Outcomes Between Single-Morula Embryo Transfer and Single-Blastocyst Transfer in Fresh IVF/ICSI Cycles.

BACKGROUND This study investigated the effectiveness and feasibility of day 4 (D4) morula embryo transfer (ET) in comparison with day 5 (D5) blastocyst ET, with regards to their clinical data, laboratory test results, and pregnancy outcomes. MATERIAL AND METHODS This retrospective cohort study enrolled 1070 patients, including 178 cases in group D4 and 892 cases in group D5. The endpoint was live birth rate after fresh embryo transfer. Furthermore, the clinical outcomes of D4 embryos with different morphology were compared and assigned to 3 groups: in group 1 (n=66) the embryos were compacted but not expanded, in group 2 (n=102) the embryos were compacted and expanded (early blastocyst), and in group 3 (n=10) the embryos were not compacted. RESULTS Groups D4 and D5 had comparable clinical pregnancy rates (53.37% vs. 59.97%) and live birth rates (43.25% vs 50.89%), and there were no significant differences between the 2 groups. In group 3, there was only 1 clinical pregnancy and no live birth. In comparison between group 1 and group 2, the clinical pregnancy rate of group 2 showed an upward trend (48.48% vs 60.78%), but there was no significant difference. There was also no statistically significant difference in the live birth rate between the 2 groups (42.42% vs 49.01%). CONCLUSIONS Transferring of compacted embryos or early blastocysts can result in high clinical pregnancy rates and live birth rates. In addition to the cleavage and blastocyst ET, morula ET may serve as an alternative option for the clinician.

Targeting of keloid with TRAIL and TRAIL-R2/DR5.

Keloid is a common and frequently-occurring disease in plastic surgery, and its ugly appearance and itching symptoms bring mental and life pain to patients. However, the clinical treatment of keloid, such as drug injection treatment, surgical resection, cryotherapy, laser treatment and other therapeutic effects are poor. Since the discovery of tumor necrosis factor related apoptosis inducing ligand (TRAIL) in 1995, its selective apoptosis on tumor cells makes it have a great prospect in the targeted treatment of tumor. In recent years, it has been found that the formation of keloid is related to the imbalance of apoptosis of fibroblasts in scar and the binding of TRAI to its receptor mediates the apoptosis of fibroblasts. Therefore, the use of TRAIL and TRAIL-R2/death receptor 5 (DR5) in the treatment of keloid has become a hot research topic. In this paper, the present situation, mechanism and development prospect of TRAIL and TRAIL-R2/DR5 targeted treatment of keloid were reviewed, which provided a reference for promoting the development of keloid treatment.

The Efficacy of Drug Injection in the Treatment of Pathological Scar: A Network Meta-analysis.

BACKGROUND: Pathological scars mainly include hyperplastic scars and keloids, and there is no uniform treatment standard for the treatment of pathological scar in clinic now. Drug injection in the treatment of pathological scar is widely used because of its advantages of less trauma and simple operation. Therefore, we used a network meta-analysis to compare the curative effect of four kinds of drugs which are commonly used in the treatment of pathological scar such as botulinum toxin type A, corticosteroids (including diprospan and triamcinolone acetonide (TAC)), verapamil and 5-fluorouracil (5-FU), systematically. It is hoped that our study will provide evidence for the choice of drugs in the treatment of pathological scar by injection. METHODS: Relevant articles from Wanfang, VIP, CNKI, PubMed, Cochrane Library and Embase databases were extracted by us. They were included into a network meta-analysis to compare the four kinds of drugs which are commonly used in the treatment of pathological scar. RESULTS: The network meta-analysis included a total of 1513 patients from 23 studies. Through meta-analysis, we found that the efficacy of botulinum toxin type A combined with corticosteroid drugs was best in the treatment of pathological scar by injection. There was no significant difference between botulinum toxin type A, corticosteroids combined with 5 Fu, verapamil and 5-FU. The efficacy of corticosteroids combined with 5-FU was better than that of corticosteroids alone and verapamil alone, but there was no significant difference between them and 5-FU. Further, the order of efficacy predicted by the SUCRA curve was as follows: botulinum toxin type A combined with corticosteroids > corticosteroids combined with 5-FU > botulinum toxin type A > corticosteroids > 5-FU > verapamil. Moreover, no publication bias was found in the funnel diagram. CONCLUSION: In the injection treatment of pathological scar, we recommend the combined injection of two drugs, especially botulinum toxin type A combined with corticosteroids. The effective treatment of botulinum toxin type A combined with corticosteroids in the treatment of pathological scar is as follows: Patients were treated once monthly with intralesional injection of TAC (0.1 ml/cm3) mixed with botulinum toxin type A (2.5 IU/cm3) for a total of 3 treatments. However, there are still limitations in this network meta-analysis, and its conclusion still needs to be further confirmed by more randomized controlled trials. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Candesartan modulates microglia activation and polarization via NF-κB signaling pathway.

Microglia are diverse cells that acquire different functional phenotypes in response to microenvironment in which they reside. Several transcriptional regulators have been identified that regulate different microglia phenotypes. They are mainly stimulated into two opposing phenotypes, classically (M1) and alternatively (M2) phenotype. Regulating microglia polarization from M1 to M2 state has been suggested as a potential therapeutic approach in treatment of CNS disorders. Candesartan, an angiotensin II type I receptors antagonist, exerts beneficial effects for antioxidant, anti-inflammation, neurotrophic, and anti-apoptotic function. However, the effect of candesartan on microglia polarization and underlying mechanisms remain unknown. In this study, the resting microglia were stimulated to M1 microglia with lipopolysaccharide (LPS) and interferon-γ (IFN-γ), and then treated with vehicle or candesartan for 24 h. RT-PCR was utilized to detect the mRNA expression of microglia phenotype markers and inflammatory cytokines. Microglia phenotype markers and toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) pathway were determined by western blot. A neuron-microglia co-culture system was used to determine whether candesartan could ameliorate the neurotoxic effect of M1 microglia to oxygen-glucose deprivation (OGD) neuron. Candesartan treatment reduced the expression of M1 markers, and increased M2 markers. Meanwhile, candesartan reduced fluorescence intensity and protein level of M1 marker and enhanced M2 marker. Candesartan also regulated the neuroinflammatory response via reducing the release of pro-inflammatory cytokines and increasing anti-inflammatory cytokines in LPS + IFN-γ stimulated BV2 cells. Candesartan markedly inhibited the protein level of TLR4, the phosphorylation of IKBα and p65, and suppressed nuclear translocation of NF-κB p65. BAY 11-7085, a NF-κB inhibitor, remarkably enlarged the inhibitory effect of candesartan on NF-κB pathway. In addition, M1 phenotype microglia exacerbated post-OGD N2a cells death and LDH release, whereas candesartan reversed such neurotoxic effect. Candesartan treatment may ameliorate stroke-induced neuronal damage through shifting microglia to M2 phenotype in a TLR4/NF-κB-dependent manner.