Performance comparison of multifarious deep networks on caries detection with tooth X-ray images.

OBJECTIVES: Deep networks have been preliminarily studied in caries diagnosis based on clinical X-ray images. However, the performance of different deep networks on caries detection is still unclear. This study aims to comprehensively compare the caries detection performances of recent multifarious deep networks with clinical dentist level as a bridge. METHODS: Based on the self-collected periapical radiograph dataset in clinic, four most popular deep networks in two types, namely YOLOv5 and DETR object detection networks, and UNet and Trans-UNet segmentation networks, were included in the comparison study. Five dentists carried out the caries detection on the same testing dataset for reference. Key tooth-level metrics, including precision, sensitivity, specificity, F1-score and Youden index, were obtained, based on which statistical analysis was conducted. RESULTS: The F1-score order of deep networks is YOLOv5 (0.87), Trans-UNet (0.86), DETR (0.82) and UNet (0.80) in caries detection. A same ranking order is found using the Youden index combining sensitivity and specificity, which are 0.76, 0.73, 0.69 and 0.64 respectively. A moderate level of concordance was observed between all networks and the gold standard. No significant difference (p > 0.05) was found between deep networks and between the well-trained network and dentists in caries detection. CONCLUSIONS: Among investigated deep networks, YOLOv5 is recommended to be priority for caries detection in terms of its high metrics. The well-trained deep network could be used as a good assistance for dentists to detect and diagnose caries. CLINICAL SIGNIFICANCE: The well-trained deep network shows a promising potential clinical application prospect. It can provide valuable support to healthcare professionals in facilitating detection and diagnosis of dental caries.

Co-exposure to 55 endocrine-disrupting chemicals linking diminished sperm quality: Mixture effect, and the role of seminal plasma docosapentaenoic acid.

Isolated effects of single endocrine-disrupting chemicals (EDCs) on male reproductive health have been studied extensively, but their mixture effect remains unelucidated. Previous research has suggested that consuming diet enriched in omega-3 polyunsaturated fatty acids (PUFA) might be beneficial for reproductive health, whether omega-3 PUFA could moderate the effect of EDCs mixture on semen quality remains to be explored. In this study of 155 male recruited from a reproductive health center in China, we used targeted-exposomics to simultaneously measure 55 EDCs in the urine for exposure burden. Regression analyses were restricted to highly detected EDCs (≥55%, n = 34), and those with consistently elevated risk were further screened and brought into mixture effect models (Bisphenol A, ethyl paraben, methyl paraben [MeP], benzophenone-1 [BP1], benzophenone-3, mono(3-carboxypropyl) phthalate [MCPP]). Bayesian Kernel Machine Regression (BKMR) and quantile-based g-computation (QGC) models demonstrated that co-exposure to top-ranked EDCs was related to reduced sperm total (ß = -0.18, 95%CI: -0.29 - -0.07, P = 0.002) and progressive motility (ß = -0.27, 95%CI: -0.43 - -0.10, P = 0.002), but not to lower semen volume. BP1, MeP and MCPP were identified as the main effect driver for deteriorated sperm motion parameters using mixture model analyses. Seminal plasma fatty acid profiling showed that high omega-3 PUFA status, notably elevated docosapentaenoic acid (DPA, C22:5n-3) status, moderated the association between MCPP and sperm motion parameters (total motility: ß = 0.26, 95%CI: 0.01 - -0.51, Pinteraction = 0.047; progressive motility: ß = 0.64, 95%CI: 0.23 - 1.05, Pinteraction = 0.003). Co-exposure to a range of EDCs is mainly associated with deteriorated sperm quality, but to a lesser extent on sperm quantity, high seminal plasma DPA status might be protective against the effect. Our work emphasizes the importance of exposomic approach to assess chemical exposures and highlighted a new possible intervention target for mitigating the potential adverse effect of EDCs on semen quality.

Asialoglycoprotein receptor 1 promotes SARS-CoV-2 infection of human normal hepatocytes.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes multi-organ damage, which includes hepatic dysfunction, as observed in over 50% of COVID-19 patients. Angiotensin I converting enzyme (peptidyl-dipeptidase A) 2 (ACE2) is the primary receptor for SARS-CoV-2 entry into host cells, and studies have shown the presence of intracellular virus particles in human hepatocytes that express ACE2, but at extremely low levels. Consequently, we asked if hepatocytes might express receptors other than ACE2 capable of promoting the entry of SARS-CoV-2 into cells. To address this question, we performed a genome-wide CRISPR-Cas9 activation library screening and found that Asialoglycoprotein receptor 1 (ASGR1) promoted SARS-CoV-2 pseudovirus infection of HeLa cells. In Huh-7 cells, simultaneous knockout of ACE2 and ASGR1 prevented SARS-CoV-2 pseudovirus infection. In the immortalized THLE-2 hepatocyte cell line and primary hepatic parenchymal cells, both of which barely expressed ACE2, SARS-CoV-2 pseudovirus could successfully establish an infection. However, after treatment with ASGR1 antibody or siRNA targeting ASGR1, the infection rate significantly dropped, suggesting that SARS-CoV-2 pseudovirus infects hepatic parenchymal cells mainly through an ASGR1-dependent mechanism. We confirmed that ASGR1 could interact with Spike protein, which depends on receptor binding domain (RBD) and N-terminal domain (NTD). Finally, we also used Immunohistochemistry and electron microscopy to verify that SARS-CoV-2 could infect primary hepatic parenchymal cells. After inhibiting ASGR1 in primary hepatic parenchymal cells by siRNA, the infection efficiency of the live virus decreased significantly. Collectively, these findings indicate that ASGR1 is a candidate receptor for SARS-CoV-2 that promotes infection of hepatic parenchymal cells.

"Cocktail" of environmental chemicals and early reproductive outcomes of IVF: The insight from paternal and maternal exposure.

BACKGROUND: Humans are exposed to various chemicals, including organophosphate esters (OPEs), phthalates (PAEs), and phenols. The effects on early reproductive outcomes of in vitro fertilization (IVF) remain unclear. METHODS: We recruited 192 women and 157 men who underwent IVF treatment. A total of forty-nine urinary chemicals were detected, including six OPEs, fifteen PAEs, six parabens, two chlorophenols, nine bisphenols, five benzophenones, and six synthetic phenolic antioxidants. We examined the individual and joint effects of parental chemical exposure on early reproductive outcomes. RESULTS: We found that certain chemicals were associated with early reproductive outcomes in Poisson regression models. For example, urinary diphenyl phosphate was negatively associated with high-quality embryos in both female (ß: -0.12, 95%CI: -0.17, -0.07) and male partners (ß: -0.09, 95%CI: -0.15, -0.03). A negative association was found between mixed chemicals and high-quality embryos in Bayesian kernel machine regression, weighted quantile sum regression (ß: -0.34, 95%CI: -0.60, -0.07), and quantile-based g-computation model (ß: -0.69, 95%CI: -1.34, -0.05) among female partners. Paternal mixture exposure was not associated with early reproductive outcomes. CONCLUSIONS: Our results indicated that increased exposure to environmental chemicals was associated with adverse early reproductive outcomes of IVF, especially female partners.

Spermatogenic cell-specific SPACA4 is essential for efficient sperm-zona pellucida binding in vitro.

Fertilization is a complex and highly regulated process that involves a series of molecular interactions between sperm and oocytes. However, the mechanisms of proteins involved in human fertilization, such as that of testis-specific SPACA4, remain poorly understood. Here we demonstrated that SPACA4 is a spermatogenic cell-specific protein. SPACA4 is expressed during spermatogenesis, upregulated in early-stage spermatids, and downregulated in elongating spermatids. SPACA4 is an intracellular protein that locates in the acrosome and is lost during the acrosome reaction. Incubation with antibodies against SPACA4 inhibited the binding of spermatozoa to zona pellucida. SPACA4 protein expression levels across different semen parameters were similar but varied significantly among patients. A prospective clinical study found no association between SPACA4 protein levels and fertilization or cleavage rates. Thus, the study suggests a novel function for SPACA4 in human fertilization in a non-dose-dependent manner. However, a larger clinical trial is required to evaluate the potential use of sperm SPACA4 protein levels to predict fertilization potential.

Clinical application of next generation sequencing-based haplotype linkage analysis in the preimplantation genetic testing for germline mosaicisms.

BACKGROUND: Preimplantation genetic testing (PGT) for monogenic disorders (PGT-M) for germline mosaicism was previously highly dependent on polymerase chain reaction (PCR)-based directed mutation detection combined with linkage analysis of short tandem repeats (STRs). However, the number of STRs is usually limited. In addition, designing suitable probes and optimizing the reaction conditions for multiplex PCR are time-consuming and laborious. Here, we evaluated the effectiveness of next generation sequencing (NGS)-based haplotype linkage analysis in PGT of germline mosaicism. METHODS: PGT-M with NGS-based haplotype linkage analysis was performed for two families with maternal germline mosaicism for an X-linked Duchenne muscular dystrophy (DMD) mutation (del exon 45-50) or an autosomal TSC1 mutation (c.2074C > T). Trophectoderm biopsy and multiple displacement amplification (MDA) were performed for a total of nine blastocysts. NGS and Sanger sequencing were performed in genomic DNA of family members and embryonic MDA products to detect DMD deletion and TSC1 mutation, respectively. Single nucleotide polymorphism (SNP) sites closely linked to pathogenic mutations were detected with NGS and served in haplotype linkage analysis. NGS-based aneuploidy screening was performed for all embryos to reduce the risk of pregnancy loss. RESULTS: All nine blastocytes showed conclusive PGT results. Each family underwent one or two frozen-thawed embryo transfer cycles to obtain a clinical pregnancy, and the prenatal diagnosis showed that the fetus was genotypically normal and euploid for both families. CONCLUSIONS: NGS-SNP could effectively realize PGT for germline mosaicism. Compared with PCR-based methods, the NGS-SNP method with increased polymorphic informative markers can achieve a greater diagnostic accuracy. Further studies are warranted to verify the effectiveness of NGS-based PGT of germline mosaicism cases in the absence of surviving offsprings.

Radiographic evaluation of the tenting screw technique in horizontal alveolar bone augmentation: A retrospective study.

OBJECTIVES: To radiographically analyze the effects of tenting screw technique (TS) and onlay bone grafts (OG) in horizontal bone augmentation. MATERIALS AND METHODS: Patients receiving horizontal bone augmentation by TS or OG were selected. The clinical outcomes and cone beam computed tomography (CBCT) data were documented pre-grafting, immediately post-grafting, before and after implantation. The survival rates, clinical complications, alveolar bone width, and volumetric bone augmentation were evaluated and statistically analyzed. RESULTS: A total of 25 patients and 41 implants were involved in this study, with no grafting failures observed in either the TS group (n = 20) or the onlay group (n = 21). Volumetric bone resorption rate in the TS group (21.34%) was significantly lower than that of the OG group (29.38%). In addition, significant horizontal bone gain was achieved in both groups (TS: 6.15 ± 2.12 mm; OG: 4.86 ± 1.40 mm) during the recovery period, with higher gain in the TS group. No apparent statistical difference in terms of volumetric bone gain was observed between the TS (748.53 mm3 , 607.47 mm3 ) and OG group (811.77 mm3 , 508.49 mm3 ) immediately post-grafting or after the recovery period. CONCLUSION: Both TS and OG achieved satisfactory bone augmentation effects, yet TS resulted in more bone augmentation and better stability than OG, with a reduced use of autogenous bone. Overall, the tenting screw technique can serve as an effective alternative to autogenous bone grafts.

Risk factors associated with loss of variable-thread tapered implants: A retrospective observational study of 1-5 years.

OBJECTIVES: This study aimed to evaluate the survival rate of variable-thread tapered implants (VTTIs) and identify risk factors for early/late implant loss. MATERIALS AND METHODS: From January 2016 to December 2019, patients who received VTTIs were included in this study. The cumulative survival rates (CSRs) at implant/patient levels were calculated by the life table method and presented via Kaplan-Meier survival curves. The relation between investigated variables and early/late implant loss was analyzed by the multivariate generalized estimating equation (GEE) regression model on the implant level. RESULTS: A total of 1528 patients with 2998 VTTIs were included. 95 implants from 76 patients were lost at the end of observation. At the implant level, the CSRs at 1, 3, and 5 years were 98.77%, 96.97%, and 95.39%, respectively, whereas they were 97.84%, 95.31%, and 92.96% at the patient level, respectively. The multivariate analysis revealed that non-submerged implant healing (OR = 4.63, p = .037) was associated with the early loss of VTTIs. Besides, male gender (OR = 2.48, p = .002), periodontitis (OR = 3.25, p = .007), implant length <10 mm (OR = 2.63, p = .028), and overdenture (OR = 9.30, p = .004) could significantly increase the risk of late implant loss. CONCLUSION: Variable-thread tapered implants could reach an acceptable survival rate in clinical practice. Non-submerged implant healing was associated with early implant loss; male gender, periodontitis, implant length <10 mm, and overdenture would significantly increase the risk of late implant loss.

Performance of indirect technique in peri-implant soft tissue contour duplication in the anterior maxilla: A clinical pilot study.

OBJECTIVE: To evaluate the performance of the indirect technique in peri-implant soft tissue contour duplication after the delivery procedure in the anterior maxilla. MATERIALS AND METHODS: Patients with single implant-supported fixed restorations in the anterior maxilla were recruited. For the impression procedure, an intraoral scan was acquired by both the direct and the indirect techniques. For the delivery procedure, implants were randomly allocated into one of the two groups according to the approaches of digital impression preceding definite crown fabrication (A-direct technique; B-indirect technique) and were scanned again after the definite crown delivery. The stereolithography files were superimposed to analyze changes in peri-implant soft tissue contour after the delivery procedure. The main outcomes were dimensional deviations of peri-implant mucosa, and the secondary outcome was differences in the pink esthetic score (PES). RESULTS: A total of 20 implants that underwent the complete workflow were included. After the delivery procedure, significant deviations in palatal tissue thickness between the provisional and definite crowns were observed in Group A but these were absent in Group B. Additionally, deviations in labial thickness (0.27 ± 0.12 mm vs. 0.08 ± 0.09 mm) and palatal thickness (0.17 ± 0.15 mm vs. 0.03 ± 0.08 mm), and labial volume of soft tissue (1.87 ± 0.94 mm3 vs. 0.75 ± 0.74 mm3 ) in Group A were significantly higher than those in Group B. No significant differences in PES were found. CONCLUSION: The indirect technique of scanning the provisional crown can more accurately duplicate the peri-implant soft tissue contour than the direct technique, resulting in a smaller deviation of the soft tissue in the delivery procedure.

Chemokine Receptors CCR6 and PD1 Blocking scFv E27 Enhances Anti-EGFR CAR-T Therapeutic Efficacy in a Preclinical Model of Human Non-Small Cell Lung Carcinoma.

Chimeric antigen receptor (CAR)-T cells, a therapeutic agent for solid tumors, are not completely effective due to a lack of infiltration of T cells into the tumor site and immunity caused by Programmed Death Receptor 1(PD1). Here, an epidermal growth factor receptor (EGFR) CAR-T cell was engineered to express the chemokine receptor CCR6 and secrete PD1 blocking Single-chain antibody fragment (scFv) E27 to enhance their anti-tumor effects. The findings showed that CCR6 enhanced the migration of EGFR CAR-E27-CCR6 T cells in vitro by the Transwell migration assay. When incubated with tumor cells, EGFR CAR-E27-CCR6 T cells specifically exerted potent cytotoxicity and produced high levels of pro-inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin-2 (IL-2), and interferon-γ (IFN-γ). A non-small cell lung carcinoma (NSCLC) cell line-derived xenograft model was constructed by implanting modified A549 cell lines into immunodeficient NOD.PrkdcscidIl2rgem1/Smoc (NSG) mice. In comparison with traditional EGFR CAR-T cells, live imaging indicated that EGFR CAR-E27-CCR6 T cells displayed superior anti-tumor function. In addition, the histopathological examination of mouse organs showed no obvious organic damage. Our findings confirmed that PD1 blocking and CCR6 can enhance the anti-tumor function of EGFR CAR-T cells in an NSCLC xenograft model, providing an effective treatment strategy to improve the efficacy of CAR-T in NSCLC.

Allogeneic gene-edited HIV-specific CAR-T cells secreting PD-1 blocking scFv enhance specific cytotoxic activity against HIV Env+ cells invivo.

HIV-specific chimeric antigen receptor (CAR) T-cells have been developed to target HIV-1 infected CD4+ T-cells that express HIV Env proteins. However, T cell exhaustion and the patient-specific autologous paradigm of CAR-T cell hurdled clinical applications. Here, we created HIV-specific CAR-T cells using human peripheral blood mononuclear cells and a 3BNC117-E27 (3BE) CAR construct that enabled the expression of programmed cell death protein (PD-1) -blocking scFv E27 and the single-chain variable fragment of the HIV-1-specific broadly neutralizing antibody 3BNC117 to target native HIV Env. Compared with T cells expressing 3BNC117-CAR alone, 3BE CAR-T cells showed greater cytotoxic activity against HIV Env+ cells with stronger proliferation capability, higher killing efficiency, and enhanced cytokine secretion in the presence of HIV Env-expressing cells. Furthermore, we manufactured TCR-deficient 3BE CAR-T cells through gene editing and demonstrated that these CAR-T cells could effectively kill HIV Env â€‹+ â€‹cells in vivo without the occurrence of severe graft-versus-host disease (GvHD) in NSG mice. These data suggest that we have provided a feasible approach to the generation of "off-the-shelf" anti-HIV CAR-T cells in combination with PD-1 checkpoint blockade immunotherapy, which can be a powerful therapeutic candidate for the functional cure of HIV.

Marginal bone loss of tissue- or bone-level implants after simultaneous guided bone regeneration in the posterior mandibular region: A retrospective cohort study.

PURPOSE: To analyze the marginal bone loss (ΔMBL) of tissue- or bone-level implants after placed with simultaneous guided bone regeneration (GBR). MATERIALS AND METHODS: A total of 151 patients who received 104 tissue-level or 128 bone-level implants placement with simultaneous GBR in the mandibular posterior region between January 2011 and December 2016 were included in this study. The marginal bone level (MBL) was recorded using the radiographic data obtained at implant placement, second-stage surgery, and the follow-up visit. Generalized estimating equation (GEE) was used to compare the ΔMBL of tissue- and bone-level implants, and the influencing factors of ΔMBL were further analyzed. RESULTS: At the last follow-up visit, the MBL of tissue-level implants was 0.73 ± 0.86 mm, above the rough-smooth interface, while that of bone-level implants was 0.82 ± 1.05 mm, above the implant platform. The ΔMBL of tissue-level implants was 1.03 mm, which was slightly higher than 0.81 mm of bone-level implants, but there was no significant difference (p > 0.05). No contributing factor associated with ΔMBL was identified by multivariate regression analysis in this study. CONCLUSION: Within the limits of this retrospective analysis, the ΔMBL of tissue-level implants is similar to that of bone-level implants after placed with simultaneous GBR, and both types of implants can achieve desirable marginal bone stability.

Luteinizing hormone stimulates the expression of amphiregulin in human theca cells.

BACKGROUND: Luteinizing hormone (LH) can stimulate mural granulosa cells to produce Amphiregulin (AREG), which can induce the resumption of meiosis in oocytes. Theca cells are present in the outer layer of follicles, providing communication with the pituitary axis through the established vascular system around the follicle. As LH target cells, it is unknown whether theca cells can produce AREG after LH stimulation. METHODS: Primary cultured human theca cells were treated with LH (with or without the inhibitor of PKA, H89), or agonists of adenylate cyclase (forskolin or db-cAMP). The mRNA and protein levels of AREG were evaluated by RT-qPCR, immunochemistry, immunofluorescence, western blotting, and ELISA. RESULTS: Immunohistochemistry of normal ovarian tissue obtained in the early-mid follicle phase showed that AREG expression was absent in both the theca layer and the granulosa cell layer of antral follicles. Double immunofluorescent staining revealed colocalization of AREG and CYP17A1 in human theca cells and colocalization of FSHR and AREG in human granulosa cells isolated from follicular fluid collected during IVF/ICSI after hCG trigger. LH significantly increased the mRNA and protein levels of AREG in human theca cells and the concentration of AREG in the culture medium. Forskolin and db-cAMP, activators of the cAMP/PKA signalling pathway, also significantly increased the mRNA and protein levels of AREG in human theca cells and the concentration of AREG in the culture medium. H89 antagonized the stimulating effect of LH on AREG expression in human theca cells. In addition, the concentration of AREG was lower in polycystic ovarian syndrome (PCOS) follicular fluid than in normal follicular fluid. The mRNA levels of AREG were significantly lower in PCOS granulosa cells and theca cells than in normal granulosa cells and theca cells. CONCLUSION: LH can stimulate the expression of AREG in human theca cells, and the adenylate cyclase/cAMP/PKA cascade may mediate this process. Expression of AREG is decreased in PCOS theca cells compared to normal theca cells, with or without LH stimulation.

NOP53 undergoes liquid-liquid phase separation and promotes tumor radio-resistance.

Aberrant DNA damage response (DDR) axis remains the major molecular mechanism for tumor radio-resistance. We recently characterized liquid-liquid phase separation (LLPS) as an essential mechanism of DDR, and identified several key DDR factors as potential LLPS proteins, including nucleolar protein NOP53. In this study, we found that NOP53 formed highly concentrated droplets in vivo and in vitro, which had liquid-like properties including the fusion of adjacent condensates, rapid fluorescence recovery after photobleaching and the sensitivity to 1,6-hexanediol. Moreover, the intrinsically disordered region 1 (IDR1) is required for NOP53 phase separation. In addition, multivalent-arginine-rich linear motifs (M-R motifs), which are enriched in NOP53, were essential for its nucleolar localization, but were dispensable for the LLPS of NOP53. Functionally, NOP53 silencing diminished tumor cell growth, and significantly sensitized colorectal cancer (CRC) cells to radiotherapy. Mechanically, NOP53 negatively regulated p53 pathway in CRC cells treated with or without radiation. Importantly, data from clinical samples confirmed a correlation between NOP53 expression and tumor radio-resistance. Together, these results indicate an important role of NOP53 in radio-resistance, and provide a potential target for tumor radio-sensitization.

Neutralization of five SARS-CoV-2 variants of concern by convalescent and BBIBP-CorV vaccinee serum.

The prevalence of SARS-CoV-2 variants of concern (VOCs) is still escalating throughout the world. However, the level of neutralization of the inactivated viral vaccine recipients' sera and convalescent sera against all VOCs, including B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma), B.1.617.2 (Delta), and B.1.1.529 (Omicron) remains to be lack of comparative analysis. Therefore, we constructed pseudoviruses of five VOCs using a lentiviral-based system and analyzed their viral infectivity and neutralization resistance to convalescent and BBIBP-CorV vaccinee serum at different times. Our results show that, compared with the wild-type strain (WT), five VOC pseudoviruses showed higher infection, of which B.1.617.2 and B.1.1.529 variant pseudoviruses exhibited higher infection rates than wild-type or other VOC strains, respectively. Sera from 10 vaccinated individuals at the 1, 3 and 5-month post second dose or from 10 convalescent at 14 and 200 days after discharge retained neutralizing activity against all strains but exhibited decreased neutralization activity significantly against the five VOC variant pseudoviruses over time compared to WT. Notably, 100% (30/30) of the vaccinee serum samples showed more than a 2.5-fold reduction in neutralizing activity against B.1.1.529, and 90% (18/20) of the convalescent serum samples showed more than 2.5-fold reduction in neutralization against B.1.1.529. These findings demonstrate the reduced protection against the VOCs in vaccinated and convalescent individuals over time, indicating that it is necessary to have a booster shot and develop new vaccines capable of eliciting broad neutralization antibodies.

FBXO34 promotes latent HIV-1 activation by post-transcriptional modulation.

ABSTRACTAcquired immunodeficiency syndrome (AIDS) cannot be completely cured, mainly due to the existence of a latent HIV-1 reservoir. However, our current understanding of the molecular mechanisms underlying the establishment and maintenance of HIV-1 latent reservoir is not comprehensive. Here, using a genome-wide CRISPR-Cas9 activation library screening, we identified E3 ubiquitin ligase F-box protein 34 (FBXO34) and the substrate of FBXO34, heterogeneous nuclear ribonucleoprotein U (hnRNP U) was identified by affinity purification mass spectrometry, as new host factors related to HIV-1 latent maintenance. Overexpression of FBXO34 or knockout of hnRNP U can activate latent HIV-1 in multiple latent cell lines. FBXO34 mainly promotes hnRNP U ubiquitination, which leads to hnRNP U degradation and abolishment of the interaction between hnRNP U and HIV-1 mRNA. In a latently infected cell line, hnRNP U interacts with the ReV region of HIV-1 mRNA through amino acids 1-339 to hinder HIV-1 translation, thereby, promoting HIV-1 latency. Importantly, we confirmed the role of the FBXO34/hnRNP U axis in the primary CD4+ T lymphocyte model, and detected differences in hnRNP U expression levels in samples from patients treated with antiretroviral therapy (ART) and healthy people, which further suggests that the FBXO34/hnRNP U axis is a new pathway involved in HIV-1 latency. These results provide mechanistic insights into the critical role of ubiquitination and hnRNP U in HIV-1 latency. This novel FBXO34/hnRNP U axis in HIV transcription may be directly targeted to control HIV reservoirs in patients in the future.

The neutralization of B.1.617.1 and B.1.1.529 sera from convalescent patients and BBIBP-CorV vaccines.

The SARS-CoV-2 variants B.1.617.1 (Kappa) contain multiple mutations in the spike protein. However, the effect of B.1.617.1 lineage-related mutants on viral infectivity and inactivated-virus vaccine efficacy remains to be defined. We therefore constructed 12 B.1.617.1-related pseudoviruses and systematically studied the effects of mutations on virus infectivity and neutralization resistance to convalescent and inactivated virus vaccine sera. Our results show that the B.1.617.1 variant exhibited both higher infectivity and neutralization resistance in sera at 1 or 3 months after vaccination of 28 individuals and at 14 and 200 days after discharge of 15 convalescents. Notably, 89% of vaccines and 100% of the convalescent serum samples showed more than 2.5-fold reduction in neutralization against one single mutation: E484Q. Besides, we found a significant decrease in neutralizing activity in convalescent patients and BBIBP-CorV vaccines for B.1.1.529. These findings demonstrate that inactivated-virus vaccination or convalescent sera showed reduced, but still significant, neutralization against the B.1.617.1 variant.

Reduction of Pre-Existing Adaptive Immune Responses Against SaCas9 in Humans Using Epitope Mapping and Identification.

The CRISPR-Cas9 system is increasingly being used as a gene editing therapeutic technique in complex diseases but concerns remain regarding the clinical risks of Cas9 immunogenicity. In this study, we detected antibodies against Staphylococcus aureus Cas9 (SaCas9) and anti-SaCas9 T cells in 4.8% and 70% of Chinese donors, respectively. We predicted 135 SaCas9-derived B cell epitopes and 50 SaCas9-derived CD8+ T cell epitopes for HLA-A*24:02, HLA-A*11:01, and HLA-A*02:01. We identified R338 as an immunodominant SaCas9 B cell epitope and SaCas9_200-208 as an immunodominant CD8+ T cell epitope for the three human leukocyte antigen allotypes through immunological assays using sera positive for SaCas9-specific antibodies and peripheral blood mononuclear cells positive for SaCas9-reactive T cells, respectively. We also demonstrated that an SaCas9 variant bearing an R338G substitution reduces B cell immunogenicity and retains its gene-editing function. Our study highlights the immunological risks of the CRISPR-Cas9 system and provides a solution to mitigate pre-existing adaptive immune responses against Cas9 in humans.

EK-16A liposomes enhance HIV replication in ACH2 or J-Lat 10.6 cell engrafted NSG mice.

Background: Numbers of HIV latency reversal agents (LRAs) have been tested in clinical trials, but with limited effect. EK-16A is an ingenol derivative that isolated from Euphorbia kansui. Our prior studies have suggested that it could reactivate latent HIV and meanwhile inhibit HIV infection in vitro. Here, we further advanced the research in vivo. Methods:In vitro, the activity of EK-16A liposomes was measured in HIV latently infected cells. In serum pharmacology test, BALB/c mice were orally administered with EK-16A liposomes, serum was separated and co-cultured with cells, HIV reactivation was measured. In vivo, NSG mice were transplanted with human cells for 3 weeks and then administered with EK-16A liposomes for 3 days. In ACH2 cell engrafted NSG mice, P24 in plasma and cell-associated HIV RNA in tissues was measured. In J-Lat 10.6 cell engrafted NSG mice, GFP expression of J-Lat 10.6 cells in diverse tissues was measured. Hematoxylin and eosin (HE) staining was carried out for histopathological examination in both mice. Results: EK-16A liposomes can reactivate latent HIV in ACH2 and J-Lat 10.6 cells. Serum pharmacological test showed that EK-16A retained activity after oral administration. Importantly, in ACH2 cell engrafted NSG mice, EK-16A liposomes increased the secretion of P24 in plasma and the expression of cell-associated HIV RNA in tissues. In J-Lat 10.6 cell engrafted NSG mice, EK-16A liposomes increased the GFP expression of J-Lat 10.6 cells in diverse tissues, including the bone marrow, spleen, liver, lung and peripheral blood. Furthermore, there was no obvious histopathological change associated with the use of EK-16A liposomes in both mice. Conclusions: Our results confirmed the enhancing HIV replication activity and preliminary security of EK-16A in human cell engrafted NSG mice, laying the foundation for research in clinical trials.

Analysis of implant loss risk factors after simultaneous guided bone regeneration: A retrospective study of 5404 dental implants.

PURPOSE: The purpose was to analyze the risk factors for implant loss after simultaneous guided bone regeneration (GBR). MATERIALS AND METHODS: Patients who underwent implant placement with simultaneous GBR between January 2011 and December 2018 were screened for this study. The cumulative survival rate (CSR) was calculated using the life table method. Log-rank test and Kaplan-Meier survival estimates were used to identify potential risk factors for implant loss. The association between the investigated variables and implant loss was determined using hazard ratios (HRs) obtained from a multivariate Cox regression analysis. RESULTS: A total of 3973 patients with 5404 implants were included in this study. The CSRs of the implants at 1, 5, and 10 years were 99.6%, 98.9%, and 98.7%, respectively. Male patient (HR = 2.94, 95% CI: 1.41-6.14), periodontitis (HR = 4.26, 95% CI: 2.05-9.86), tissue-level implants (HR = 3.02, 95% CI: 1.30-6.98), narrow implants (HR = 2.71, 95% CI: 1.12-6.57), and implant length ≤10 mm (HR = 2.91, 95% CI: 1.41-6.02) significantly increased the risk of implant loss (p < 0.05). The risk of implant loss was significantly higher in the maxillary posterior region (HR = 2.26, 95% CI: 1.04-4.90) than in the maxillary anterior region (p < 0.05). Compared to Straumann, Nobel (HR = 4.07, 95% CI: 1.75-9.44) and other implant systems (HR = 14.23, 95% CI: 4.32-46.85) showed a significantly higher risk of implant loss (p < 0.05). CONCLUSION: Male patient, periodontitis, maxillary posterior region, Nobel implant system, other implant systems, tissue-level implants, narrow implants, and implant length ≤10 mm were considered risk factors for implant loss after simultaneous GBR.