Live-cell imaging reveals redox metabolic reprogramming during zygotic genome activation.

Metabolic programming is deeply intertwined with early embryonic development including zygotic genome activation (ZGA), the polarization of zygotic cells, and cell fate commitment. It is crucial to establish a noninvasive imaging technology that spatiotemporally illuminates the cellular metabolism pathways in embryos to track developmental metabolism in situ. In this study, we used two high-quality genetically encoded fluorescent biosensors, SoNar for NADH/NAD+ and iNap1 for NADPH, to characterize the dynamic regulation of energy metabolism and redox homeostasis during early zygotic cleavage. Our imaging results showed that NADH/NAD+ levels decreased from the early to the late two-cell stage, whereas the levels of the reducing equivalent NADPH increased. Mechanistically, transcriptome profiling suggested that during the two-cell stage, zygotic cells downregulated the expression of genes involved in glucose uptake and glycolysis, and upregulated the expression of genes for pyruvate metabolism in mitochondria and oxidative phosphorylation, with a decline in the expression of two peroxiredoxin genes, Prdx1 and Prdx2. Collectively, with the establishment of in situ metabolic monitoring technology, our study revealed the programming of redox metabolism during ZGA.

Partial reuse of circadian clock genes along parallel clines of diapause in two moth species.

Understanding the molecular basis of repeated evolution improves our ability to predict evolution across the tree of life. Only since the last decade has high-throughput sequencing enabled comparative genome scans to thoroughly examine the repeatability of genetic changes driving repeated phenotypic evolution. The Asian corn borer (ACB), Ostrinia furnacalis (Guenée), and the European corn borer (ECB), Ostrinia nubilalis (Hübner), are two closely related moths displaying repeatable phenological adaptation to a wide range of climates on two separate continents, largely manifesting as changes in the timing of diapause induction and termination across latitude. Candidate genes underlying diapause variation in North American ECB have been previously identified. Here, we sampled seven ACB populations across 23 degrees of latitude in China to elucidate the genetic basis of diapause variation and evolutionary mechanisms driving parallel clinal responses in the two species. Using pooled whole-genome sequencing (Pool-seq) data, population genomic analyses revealed hundreds of single nucleotide polymorphisms (SNP) whose allele frequencies covaried with mean diapause phenotypes along the cline. Genes involved in circadian rhythm were over-represented among candidate genes with strong signatures of spatially varying selection. Only one of two circadian clock genes associated with diapause evolution in ECB showed evidence of reuse in ACB (period [per]), but per alleles were not shared between species nor with their outgroup, implicating independent mutational paths. Nonetheless, evidence of adaptive introgression was discovered at putative diapause loci located elsewhere in the genome, suggesting that de novo mutations and introgression might both underlie the repeated phenological evolution.

Delamination of rotator cuff tears impairs healing after repair: a systematic review and meta-analysis.

PURPOSE: To compare the clinical outcomes and retear rates after rotator cuff repair (RCR) between delaminated and non-delaminated tears. METHODS: This systematic review was conducted according to the preferred reporting items for systematic reviews and meta-analyses guidelines using the PubMed, Cochrane Library, the Web of Science and Embase databases. Only articles on arthroscopic RCR with clinical outcome scores and data on the number of rotator cuff retears and complete healing were included. This study's relevant data were extracted and statistically analyzed. The methodological index for nonrandomized studies was used to assess the risk of bias in the included studies. After conducting a heterogeneity test and sensitivity analysis to determine whether the samples were heterogeneous, the study also detected publication bias. A sub-group test was used to evaluate the influences of the imaging follow-up period on retear rates. RESULTS: Ten eligible articles were identified with 2,061 patients (925 in the delaminated group and 1,136 in the non-delaminated group). The meta-analysis demonstrated that delamination was significantly associated with higher retear rates (P = 0.026; odds ratio = 1.873, 95% confidence interval 1.079-3.252; I2 = 51.6%) with an imaging follow-up period of > 1 year and lower rates of complete healing (P = 0.036; odds ratio = 0.659, 95% confidence interval 0.446-0.973; I2 = 9.0%) in patients after rotator cuff repair. However, no significant differences were observed between the two groups based on American Shoulder and Elbow Surgeons score, Constant score, visual analog scale score, external rotation, internal rotation, or forward elevation. CONCLUSIONS: This meta-analysis found that delamination was significantly associated with higher retear rates with imaging follow-up period of > 1 year, and lower rates of complete healing. In addition, the preoperative and postoperative clinical scores and shoulder joint range of motion were similar between patients with delaminated and non-delaminated tears. LEVEL OF EVIDENCE: Level IV.

Association of obesity with high retears and complication rates, and low functional scores after rotator cuff repair: a systematic review and meta-analysis.

BACKGROUND: Obesity influences the outcomes of orthopedic surgeries such as total knee arthroplasty and spinal surgery. However, the effect of obesity on the outcomes of rotator cuff repair is unknown. This systematic review and meta-analysis aimed to examine the effect of obesity on rotator cuff repair outcomes. METHODS: PubMed, EMBASE, Web of Science, and Cochrane Library databases were searched to identify relevant studies published from their inception till July 2022. Two reviewers independently screened titles and abstracts using the specified criteria. Articles were included if they indicated the effect of obesity on rotator cuff repair and the related outcomes after surgery. Review Manager 5.4.1 software was used to perform statistical analysis. RESULTS: Thirteen articles involving 85,497 patients were included. Obese patients had higher retear rates than nonobese patients (odds ratio [OR] 2.58, 95% confidence interval [CI] 1.23-5.41, P = .01), lower American Shoulder and Elbow Surgeons scores (mean difference [MD]: -3.59, 95% CI: -5.45 to [-1.74]; P = .0001), higher visual analog scale for pain (mean difference: 0.73, 95% CI: 0.29-1.17; P = .001), higher reoperation rates (OR 1.31, 95% CI 1.21-1.42, P < .00001), and higher rates of complications (OR 1.57, 95% CI 1.31-1.87, P = .000). Obesity did not affect the duration of surgery (MD: 6.03, 95% CI: -7.63 to 19.69; P = .39) or external rotation of the shoulder (MD: -1.79, 95% CI: -5.30 to 1.72; P = .32). CONCLUSION: Obesity is a significant risk factor for retear and reoperation after rotator cuff repair. Furthermore, obesity increases the risk of postoperative complications and leads to lower postoperative American Shoulder and Elbow Surgeons scores and higher shoulder visual analog scale for pain.

Life history responses of the small brown planthopper Laodelphax striatellus to temperature change.

Temperature is a key environmental factor in ectotherms and influences many life history traits. In the present study, the nymphal development time, sex ratio and wing dimorphism of the small brown planthopper Laodelphax striatellus were examined under the conditions of constant temperatures, naturally varying temperatures (or different generations), and different temperatures combined with different photoperiod. The results showed that from 18 to 28 °C, the developmental time of nymphs was gradually shortened with the increase of temperature, whereas the high temperatures of 30 and 32 °C in the third to fifth instar nymphal stages and high summer temperature of 28.8 and 29.7 °C significantly delayed developmental time and resulted in higher mortality of nymphs. In all treatments, the developmental time was longer in females than males. The nymphs took significantly longer time to develop in the short daylength of 12 h than in longer daylengths of 13, 14, 15 and 16. Differences in developmental time were also found between wing morph, with long-winged individuals being significantly longer than the short-winged individuals at lower temperatures and significantly shorter than the short-winged individuals at higher temperatures. In all treatments, the sex ratio was stable, approaching 1:1, without being affected by temperature, generations and photoperiod. Photoperiod and temperature had significantly influence on the wing dimorphism. Long daylength combined with different temperatures resulted in significantly higher proportions of long-winged morph, whereas the low temperatures combined with the short daylengths in autumn and winter resulted in significantly high proportion of short-winged morph. This study broadens our understanding of the life-history traits of this planthopper and provides basic data for analyzing the effects of climate change on the planthopper reproduction.

Weak value amplification for angular velocity measurements.

The weak-value-amplification technique has shown great importance in the measurement of tiny physical effects. Here we introduce a polarization-dependent angular velocity measurement system consisting of two Glan prisms and a true zero-order half-wave plate, where a non-Fourier-limited Gaussian pulse acts as the meter. The angular velocities measurements results agree well with theoretical predictions, and its uncertainties are bounded by the Cramér-Rao bound. We also investigate uncertainties of angular velocities for different numbers of detected photons and the smallest reliable postselection probability, which can reach ${3.42*10^{- 6}}$.

Inheritance of key life-history traits in crosses between northern and southern populations of the cabbage beetle Colaphellus bowringi (Coleoptera: Chrysomelidae).

A southern population (S) from Xiushui County (29°1'N, 114°4'E) and a northern population (N) from Shenyang city (41°48'N, 123°23'E) of the cabbage beetle, Colaphellus bowringi vary greatly in their life-history traits, and may serve as an excellent model with which to study the inheritance of life-history traits. In the present study, we performed intraspecific hybridization using the two populations, comparing the key life-history traits (fecundity, development time, body weight, growth rate, and sexual size dimorphism (SDD)) between the two populations (S♀ × S♂ and N♀ × N♂) and their two hybrid populations (S♀ × N♂ and N♀ × S♂ populations) at 19, 22, 25, and 28°C. Our results showed that there were significant differences in life-history traits between the two parental populations, with the S population having a significantly higher fecundity, shorter larval development time, larger body weight, higher growth rate, and greater weight loss during metamorphosis than the N population at almost all temperatures. However, these life-history traits in the two hybrid populations were intermediate between those of their parents. The life-history traits in the S × N and N × S populations more closely resembled those of the maternal S population and N population, respectively, showing maternal effects. Weight loss for both sexes was highest in the S population, followed by the S × N, N × S, and N populations at all temperatures, suggesting that larger pupae lost more weight during metamorphosis. The changes in SSD with temperature were similar between the S and the S × N populations and between the N and the N × S populations, also suggesting a maternal effect. Overall, our results showed no drastic effect of hybridization on C. bowringi, being neither negative (hybrid inferiority) nor positive (heterosis). Rather, the phenotypes of hybrids were intermediate between the phenotypes of their parents.

Risk HLA class II alleles and amino acid residues in myeloperoxidase-ANCA-associated vasculitis.

A genome-wide association study (GWAS) indicated that myeloperoxidase-ANCA associated vasculitis (AAV) is associated with HLA-DQ. However, susceptibility alleles in these loci have been under-investigated. Here we genotyped 258 Chinese patients with myeloperoxidase-AAV and 597 healthy control individuals at HLA DRB1, DQA1, DQB1 and DPB1, and extracted the encoded amino acid sequences from the IMGT/HLA database. The replication cohort included 97 cases and 107 controls. T cell epitopes of myeloperoxidase were predicted and docked to the HLA molecules. We found DQA1∗0302 (odds ratio 2.34 (95% confidence interval 1.75-3.14)) and DQB1∗0303 (odds ratio 1.89 (1.45-2.48)) were risk alleles for myeloperoxidase-AAV. They are in overt linkage disequilibrium (r2 0.69) and the haplotype DQA1∗0302-DQB1∗0303 presents a significant risk (haplotype score 6.39) as well. Aspartate160 on the DQ α chain (odds ratio 2.06 (1.60-2.67)), encoded by DQA1∗0302, and isoleucine185 on the DQ ß chain (odds ratio 1.73 (1.38-2.18)), encoded by DQB1∗0303, both located in the α2ß2 domains, conferred significant risk for myeloperoxidase-AAV. Homologous modeling showed that DQα∗160D may confer susceptibility to myeloperoxidase-AAV by altering dimerization of the HLA molecules. Thus, more attention should be paid to the roles of amino acids in the α2ß2 domains in addition to the α1ß1 binding groove of HLA class II molecules.

The analysis of clinical and laboratory data: a large outbreak of dengue fever in Chaozhou, Guangdong province, China.

A large-scale dengue fever (DF) outbreak occurred in Chaozhou, Guangdong province, China 2015. In our study, 528 dengue-positive patient samples were collected for clinical and laboratory data analysis. 491 cases (93.0%) were primary dengue fever (PDF), 22 cases (4.2%) were dengue hemorrhagic fever (DHF) and 15 cases (2.8%) were diagnosed with severe dengue fever (SDF). All cases were infected by dengue virus serotype 2 (DENV-2), and the isolated strains belonged to cosmopolitan genotype, which were grouped closely with Malaysia strains from 2010 to 2014. Moreover, the study showed that laboratory indices have significantly difference in PDF, DHF and SDF patients. A comprehensive analysis of these data could assist and guide the clinical diagnosis for DF, which has an important significance for the control of dengue virus infection.

Experimental Demonstration of Higher Precision Weak-Value-Based Metrology Using Power Recycling.

The weak-value-based metrology is very promising and has attracted a lot of attention in recent years because of its remarkable ability in signal amplification. However, it is suggested that the upper limit of the precision of this metrology cannot exceed that of classical metrology because of the low sample size caused by the probe loss during postselection. Nevertheless, a recent proposal shows that this probe loss can be reduced by the power-recycling technique, and thus enhance the precision of weak-value-based metrology. Here we experimentally realize the power-recycled interferometric weak-value-based beam-deflection measurement and obtain the amplitude of the detected signal and white noise by discrete Fourier transform. Our results show that the detected signal can be strengthened by power recycling, and the power-recycled weak-value-based signal-to-noise ratio can surpass the upper limit of the classical scheme, corresponding to the shot-noise limit. This work sheds light on higher precision metrology and explores the real advantage of the weak-value-based metrology over classical metrology.

A novel Brassica-rhizotron system to unravel the dynamic changes in root system architecture of oilseed rape under phosphorus deficiency.

BACKGROUND AND AIMS: An important adaptation of plants to phosphorus (P) deficiency is to alter root system architecture (RSA) to increase P acquisition from the soil, but soil-based observations of RSA are technically challenging, especially in mature plants. The aim of this study was to investigate the root development and RSA of oilseed rape (Brassica napus L.) under low and high soil P conditions during an entire growth cycle. METHODS: A new large Brassica-rhizotron system (approx. 118-litre volume) was developed to study the RSA dynamics of B. napus 'Zhongshuang11' in soils, using top-soils supplemented with low P (LP) or high P (HP) for a full plant growth period. Total root length (TRL), root tip number (RTN), root length density (RLD), biomass and seed yield traits were measured. KEY RESULTS: TRL and RTN increased more rapidly in HP than LP plants from seedling to flowering stages. Both traits declined from flowering to silique stages, and then increased slightly in HP plants; in contrast, root senescence was observed in LP plants. RSA parameters measured from the polycarbonate plates were empirically consistent with analyses of excavated roots. Seed yield and shoot dry weights were closely associated positively with root dry weights, TRL, RLD and RTN at both HP and LP. CONCLUSIONS: The Brassica-rhizotron system is an effective method for soil-based root phenotyping across an entire growth cycle. Given that root senescence is likely to occur earlier under low P conditions, crop P deficiency is likely to affect late water and nitrogen uptake, which is critical for efficient resource use and optimal crop yields.

Life-history responses of the rice stem borer Chilo suppressalis to temperature change: Breaking the temperature-size rule.

Temperature is a key environmental factor for ectotherms and affects a large number of life history traits. In the present study, development time from hatching to pupation and adult eclosion, pupal and adult weights of the rice stem borer, Chilo suppressalis were examined at 22, 25, 28 and 31°C under L18:D 6. Larval and pupal times were significantly decreased with increasing rearing temperature and growth rate was positively correlated with temperature. Larval and pupal developmental times were not significantly different between females and males. The relationship between body weight and rearing temperature in C. suppressalis did not follow the temperature-size rule (TSR), both males and females gained the highest body weight at 31°C. Females were significantly larger than males at all temperatures, showing a female biased sex size dimorphism (SSD). Contrary to Rensch's rule, SSD and body weight in C. suppressalis tended to increase with rising temperature. Male pupae lost significantly more weight at metamorphosis compared to females. We discuss the adaptive significance of the reverse-TSR in the moth's life history.

Effect of photoperiod and temperature on the intensity of pupal diapause in the cotton bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae).

The intensity of pupal diapause in the cotton bollworm, Helicoverpa armigera (Hübner) was investigated under both laboratory and natural conditions. By transferring diapausing pupae induced under LD 11:13, LD 12:12 and LD 13:11 at 20, 22 and 25 °C to 25 °C combined with LD 15:9 to terminate diapause the rearing day length of 11 h evoked greater intensity of diapause than did 12 and 13 h at 25 °C; whereas the rearing temperature of 25 °C evoked more intense diapause than did 20 and 22 °C under LD 11:13. By transferring diapausing pupae induced under LD 12:12 at 20 and 22 °C to six temperatures of 18, 20, 22, 25, 28 and 31 °C combined with LD 15:9 to terminate diapause, the duration of diapause was significantly shortened from 146 days at 18 °C to 24 days at 31 °C, showing that high temperatures significantly accelerate diapause development. Furthermore, the duration of diapause was significantly longer at the rearing temperature of 22 °C than that at 20 °C when the diapause-terminating temperatures were 20 and 22 °C. Chilling at 5 °C did not shorten the duration of diapause but lengthened it when chilling period was included. However, chilling plays an important role in synchronizing adult emergence. Rearing temperature of 22 °C also evoked more intense diapause than did 20 °C in most chilling treatments. When the overwintering pupae were transferred at different times from natural temperatures to 25 °C, it was found that the earlier the transfer took place, the earlier the adults emerged when the time spent under natural conditions was included. However, cool temperatures before March showed an enhanced effect on diapause development at 20 °C, suggesting that the high diapause-terminating temperature can offset the effect of chilling on diapause development. The result of diapause termination under natural conditions suggests that the developmental threshold for post-diapause development in H. armigera should be around 17.5 °C.

A comparison of the life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera.

In order to understand the differences of life-history traits between diapause and direct development individuals in the cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae), the development time, body size, growth rate, and adult longevity were investigated between the two populations, which were induced under 12:12 L:D and 16:8 L:D photoperiods, respectively, at 20, 22, and 25°C. The results indicated that the larval development time, pupal weight, adult weight, and growth rate were significantly different between diapause and direct developing individuals. The diapause developing individuals had a significantly higher pupal and adult weight and a longer larval time compared with direct developing individuals. However, the growth rate in diapause developing individuals was lower than that in the direct developing individuals. Analysis by GLM showed that larval time, pupal and adult weight, and growth rate were significantly influenced by both temperature and developmental pathway. The pupal and adult weights were greater in males than females in both developmental pathways, exhibiting sexual size dimorphism. The dimorphism in adult weight was more pronounced than in pupal weight because female pupae lost more weight at metamorphosis compared to male pupae. Protogyny was observed in both developmental pathways. However, the protogyny phenomenon was more pronounced at lower temperatures in direct developing individuals, whereas it was more pronounced in diapause developing individuals when they experienced higher temperatures in their larval stage and partial pupal period. The adult longevity of diapause developing individuals was significantly longer than that of direct developing individuals. The results reveal that the life-history strategy was different between diapause and direct developing individuals.

Development and Validation of a Predictive Model for Early Identification of Cognitive Impairment Risk in Community-Based Hypertensive Patients.

Objective: To investigate the risk factors for the development of mild cognitive dysfunction in hypertensive patients in the community and to develop a risk prediction model. Method: The data used in this study were obtained from two sources: the China Health and Retirement Longitudinal Study (CHARLS) and the Chinese Longitudinal Healthy Longevity Survey (CLHLS). A total of 1121 participants from CHARLS were randomly allocated into a training set and a validation set, following a 70:30 ratio. Meanwhile, an additional 4016 participants from CLHLS were employed for external validation of the model. The patients in this study were divided into two groups: those with mild cognitive impairment and those without. General information, employment status, pension, health insurance, and presence of depressive symptoms were compared between the two groups. LASSO regression analysis was employed to identify the most predictive variables for the model, utilizing 14-fold cross-validation. The risk prediction model for cognitive impairment in hypertensive populations was developed using generalized linear models. The model's discriminatory power was evaluated through the area under the receiver operating characteristic (ROC) curve and calibration curves. Results: In the modeling group, eight variables such as gender, age, residence, education, alcohol use, depression, employment status, and health insurance were ultimately selected from an initial pool of 21 potential predictors to construct the risk prediction model. The area under the curve (AUC) values for the training, internal, and external validation sets were 0.777, 0.785, and 0.782, respectively. All exceeded the threshold of 0.7, suggesting that the model effectively predicts the incidence of mild cognitive dysfunction in community-based hypertensive patients. A risk prediction model was developed using a generalized linear model in conjunction with Lasso regression. The model's performance was evaluated using the area under the receiver operating characteristic (ROC) curve. Hosmer-Lemeshow test values yielded p = .346 and p = .626, both of which exceeded the 0.05 threshold. Calibration curves demonstrated a significant agreement between the nomogram model and observed outcomes, serving as an effective tool for evaluating the model's predictive performance. Discussion: The predictive model developed in this study serves as a promising and efficient tool for evaluating cognitive impairment in hypertensive patients, aiding community healthcare workers in identifying at-risk populations.

Quantum spin driven Yu-Shiba-Rusinov multiplets and fermion-parity-preserving phase transition in K3C60.

Magnetic impurities in superconductors are of increasing interest due to emergent Yu-Shiba-Rusinov (YSR) states and Majorana zero modes for fault-tolerant quantum computation. However, a direct relationship between the YSR multiple states and magnetic anisotropy splitting of quantum impurity spins remains poorly characterized. By using scanning tunneling microscopy, we systematically resolve individual transition-metal (Fe, Cr, and Ni) impurities induced YSR multiplets as well as their Zeeman effects in the K3C60 superconductor. The YSR multiplets show identical d orbital-like wave functions that are symmetry-mismatched to the threefold K3C60(1 1 1) host surface, breaking point-group symmetries of the spatial distribution of YSR bound states in real space. Remarkably, we identify an unprecedented fermion-parity-preserving quantum phase transition between ground states with opposite signs of the uniaxial magnetic anisotropy that can be manipulated by an external magnetic field. These findings can be readily understood in terms of anisotropy splitting of quantum impurity spins, and thus elucidate the intricate interplay between the magnetic anisotropy and YSR multiplets.

Large-scale 2D heterostructures from hydrogen-bonded organic frameworks and graphene with distinct Dirac and flat bands.

The current strategies for building 2D organic-inorganic heterojunctions involve mostly wet-chemistry processes or exfoliation and transfer, leading to interface contaminations, poor crystallizing, or limited size. Here we show a bottom-up procedure to fabricate 2D large-scale heterostructure with clean interface and highly-crystalline sheets. As a prototypical example, a well-ordered hydrogen-bonded organic framework is self-assembled on the highly-oriented-pyrolytic-graphite substrate. The organic framework adopts a honeycomb lattice with faulted/unfaulted halves in a unit cell, resemble to molecular "graphene". Interestingly, the topmost layer of substrate is self-lifted by organic framework via strong interlayer coupling, to form effectively a floating organic framework/graphene heterostructure. The individual layer of heterostructure inherits its intrinsic property, exhibiting distinct Dirac bands of graphene and narrow bands of organic framework. Our results demonstrate a promising approach to fabricate 2D organic-inorganic heterostructure with large-scale uniformity and highly-crystalline via the self-lifting effect, which is generally applicable to most of van der Waals materials.

Realization of monolayer ZrTe5 topological insulators with wide band gaps.

Two-dimensional topological insulators hosting the quantum spin Hall effect have application potential in dissipationless electronics. To observe the quantum spin Hall effect at elevated temperatures, a wide band gap is indispensable to efficiently suppress bulk conduction. Yet, most candidate materials exhibit narrow or even negative band gaps. Here, via elegant control of van der Waals epitaxy, we have successfully grown monolayer ZrTe5 on a bilayer graphene/SiC substrate. The epitaxial ZrTe5 monolayer crystalizes in two allotrope isomers with different intralayer alignments of ZrTe3 prisms. Our scanning tunneling microscopy/spectroscopy characterization unveils an intrinsic full band gap as large as 254 meV and one-dimensional edge states localized along the periphery of the ZrTe5 monolayer. First-principles calculations further confirm that the large band gap originates from strong spin-orbit coupling, and the edge states are topologically nontrivial. These findings thus provide a highly desirable material platform for the exploration of the high-temperature quantum spin Hall effect.

High-efficiency targeted transgene integration via primed micro-homologues.

Due to the difficulties in precisely manipulating DNA repair pathways, high-fidelity targeted integration of large transgenes triggered by double-strand breaks is inherently inefficient. Here, we exploit prime editors to devise a robust knock-in (KI) strategy named primed micro-homologues-assisted integration (PAINT), which utilizes reverse-transcribed single-stranded micro-homologues to boost targeted KIs in different types of cells. The improved version of PAINT, designated PAINT 3.0, maximizes editing efficiency and minimizes off-target integration, especially in dealing with scarless in-frame KIs. Using PAINT 3.0, we target a reporter transgene into housekeeping genes with editing efficiencies up to 80%, more than 10-fold higher than the traditional homology-directed repair method. Moreover, the use of PAINT 3.0 to insert a 2.5-kb transgene achieves up to 85% KI frequency at several therapeutically relevant genomic loci, suggesting its potential for clinical applications. Finally, PAINT 3.0 enables high-efficiency non-viral genome targeting in primary T cells and produces functional CAR-T cells with specific tumor-killing ability. Thus, we establish that the PAINT method is a powerful gene editing tool for large transgene integrations and may open new avenues for cell and gene therapies and genome writing technologies.

[Meta-analysis of obesity on the outcome of rotator cuff repair].

OBJECTIVE: To systematically evaluate obesity on the outcome of rotator cuff repair. METHODS: Literatures on the relationship between obesity and outcomes after rotator cuff repair were searched from PubMed, Embase, Cochrane Library, Web of Science, China biology medicine(CBM), CNKI, Wanfang and VIP databases from building database to August 1, 2022, and were screened independently by two authors according to inclusion and exclusion criteria. Endnote X9 and Excel 2019 were used for literature extraction, management and data entry, and Newcastle-Ottawa Scale (NOS) was used to evaluate quality of the included literatures. STATA 16.0 and RevMan 5.4 softwares were used to evaluate postoperative retear rate, reoperation rate, complication rate, American Shoulder and Elbow Surgeons (ASES) score, visual analogue scale (VAS), operative time and external rotation angle of shoulder joint pain were analyzed. RESULTS: Totally 13 literatures were included, including 6 retrospective studies, 5 case-control studies, 1 prospective cohort study, and 1 abstract of a study for which the full text was not available, with 85 503 patients (31 973 in obese group and 53 530 in non-obese group). Meta-analysis showed there were statistical differences between two groups in retear rate [OR=2.58, 95%CI(1.23, 5.41), P=0.01], reoperation rate[OR=1.31, 95%CI(1.21, 1.42), P<0.00], complication rate [OR=1.57, 95%CI(1.31, 1.87), P=0.00], ASES score[MD=-3.59, 95%CI(-5.45, -1.74), P=0.00], and VAS[MD=0.24, 95%CI(0.00, 0.49), P=0.05]. While there were no differences between two groups in operative time[MD=6.03, 95%CI(-7.63, 19.69), P=0.39], external rotation angle of shoulder joint[MD=-1.79, 95%CI(-5.30, 1.71), P=0.32]. CONCLUSION: Obesity is associated with higher rates of retear, resurgery, complications, poorer shoulder function and pain after rotator cuff repair.