Amplification editing enables efficient and precise duplication of DNA from short sequence to megabase and chromosomal scale.

Duplication is a foundation of molecular evolution and a driver of genomic and complex diseases. Here, we develop a genome editing tool named Amplification Editing (AE) that enables programmable DNA duplication with precision at chromosomal scale. AE can duplicate human genomes ranging from 20 bp to 100 Mb, a size comparable to human chromosomes. AE exhibits activity across various cell types, encompassing diploid, haploid, and primary cells. AE exhibited up to 73.0% efficiency for 1 Mb and 3.4% for 100 Mb duplications, respectively. Whole-genome sequencing and deep sequencing of the junctions of edited sequences confirm the precision of duplication. AE can create chromosomal microduplications within disease-relevant regions in embryonic stem cells, indicating its potential for generating cellular and animal models. AE is a precise and efficient tool for chromosomal engineering and DNA duplication, broadening the landscape of precision genome editing from an individual genetic locus to the chromosomal scale.

DNA topology regulates PAM-Cas9 interaction and DNA unwinding to enable near-PAMless cleavage by thermophilic Cas9.

CRISPR-Cas9-mediated genome editing depends on PAM recognition to initiate DNA unwinding. PAM mutations can abolish Cas9 binding and prohibit editing. Here, we identified a Cas9 from the thermophile Alicyclobacillus tengchongensis for which the PAM interaction can be robustly regulated by DNA topology. AtCas9 has a relaxed PAM of N4CNNN and N4RNNA (R = A/G) and is able to bind but not cleave targets with mutated PAMs. When PAM-mutated DNA was in underwound topology, AtCas9 exhibited enhanced binding affinity and high cleavage activity. Mechanistically, AtCas9 has a unique loop motif, which docked into the DNA major groove, and this interaction can be regulated by DNA topology. More importantly, AtCas9 showed near-PAMless editing of supercoiled plasmid in E. coli. In mammalian cells, AtCas9 exhibited broad PAM preference to edit plasmid with up to 72% efficiency and effective base editing at four endogenous loci, representing a potentially powerful tool for near-PAMless editing.

Efficient targeted insertion of large DNA fragments without DNA donors.

Targeted insertion of large DNA fragments holds great potential for treating genetic diseases. Prime editors can effectively insert short fragments (~44 bp) but not large ones. Here we developed GRAND editing to precisely insert large DNA fragments without DNA donors. In contrast to prime editors, which require reverse transcription templates hybridizing with the target sequence, GRAND editing employs a pair of prime editing guide RNAs, with reverse transcription templates nonhomologous to the target site but complementary to each other. This strategy exhibited an efficiency of up to 63.0% of a 150-bp insertion with minor by-products and 28.4% of a 250-bp insertion. It allowed insertions up to ~1 kb, although the efficiency remains low for fragments larger than 400 bp. We confirmed efficient insertion in multiple genomic loci of several cell lines and non-dividing cells, which expands the scope of genome editing to enable donor-free insertion of large DNA sequences.

Efficacy of Voluntary Medical Male Circumcision to Prevent HIV Infection Among Men Who Have Sex With Men : A Randomized Controlled Trial.

BACKGROUND: Observational studies suggest that voluntary medical male circumcision (VMMC) may lower HIV risk among men who have sex with men (MSM). A randomized controlled trial (RCT) is needed to confirm this. OBJECTIVE: To assess the efficacy of VMMC in preventing incident HIV infection among MSM. DESIGN: An RCT with up to 12 months of follow-up. (Chinese Clinical Trial Registry: ChiCTR2000039436). SETTING: 8 cities in China. PARTICIPANTS: Uncircumcised, HIV-seronegative men aged 18 to 49 years who self-reported predominantly practicing insertive anal intercourse and had 2 or more male sex partners in the past 6 months. INTERVENTION: VMMC. MEASUREMENTS: Rapid testing for HIV was done at baseline and at 3, 6, 9, and 12 months. Behavioral questionnaires and other tests for sexually transmitted infections were done at baseline, 6 months, and 12 months. The primary outcome was HIV seroconversion using an intention-to-treat analysis. RESULTS: The study enrolled 124 men in the intervention group and 123 in the control group, who contributed 120.7 and 123.1 person-years of observation, respectively. There were 0 seroconversions in the intervention group (0 infections [95% CI, 0.0 to 3.1 infections] per 100 person-years) and 5 seroconversions in the control group (4.1 infections [CI, 1.3 to 9.5 infections] per 100 person-years). The HIV hazard ratio was 0.09 (CI, 0.00 to 0.81; P = 0.029), and the HIV incidence was lower in the intervention group (log-rank P = 0.025). The incidence rates of syphilis, herpes simplex virus type 2, and penile human papillomavirus were not statistically significantly different between the 2 groups. There was no evidence of HIV risk compensation. LIMITATION: Few HIV seroconversions and limited follow-up period. CONCLUSION: Among MSM who predominantly practice insertive anal intercourse, VMMC is efficacious in preventing incident HIV infection; MSM should be included in VMMC guidelines. PRIMARY FUNDING SOURCE: The National Science and Technology Major Project of China.

Low power consumption grating magneto-optical trap based on planar elements.

The grating-based magneto-optical trap (GMOT) is a promising approach for miniaturizing cold-atom systems. However, the power consumption of a GMOT system dominates its feasibility in practical applications. In this study, we demonstrated a GMOT system based on planar elements that can operate with low power consumption. A high-diffraction-efficiency grating chip was used to cool atoms with a single incident beam. A planar coil chip was designed and fabricated with a low power consumption nested architecture. The grating and coil chips were adapted to a passive pump vacuum chamber, and up to 106 87Rb atoms were trapped. These elements effectively reduce the power consumption of the GMOT and have great potential for applications in practical cold-atom-based devices.

Robust enhancement of high-harmonic generation from all-dielectric metasurfaces enabled by polarization-insensitive bound states in the continuum.

The emerging all-dielectric platform exhibits high-quality (Q) resonances governed by the physics of bound states in the continuum (BIC) that drives highly efficient nonlinear optical processes. Here we demonstrate the robust enhancement of third-(THG) and fifth-harmonic generation (FHG) from all-dielectric metasurfaces composed of four silicon nanodisks. Through the symmetry breaking, the genuine BIC transforms into the high-Q quasi-BIC resonance with tight field confinement for record high THG efficiency of 3.9 × 10-4 W-2 and FHG efficiency of 4.8 × 10-10 W-4 using a moderate pump intensity of 1 GW/cm2. Moreover, the quasi-BIC and the resonantly enhanced harmonics exhibit polarization-insensitive characteristics due to the special C4 arrangement of meta-atoms. Our results suggest the way for smart design of efficient and robust nonlinear nanophotonic devices.

Acupoint Catgut Embedding as Adjunctive Therapy for Patients With Gallstones.

AIMS OF THIS STUDY: A randomized clinical trial was undertaken to investigate the efficacy of acupoint catgut embedding (ACE) as adjunctive therapy to tauroursodeoxycholic acid (TUDCA) therapy on gallbladder emptying and clinical symptoms in patients with gallstone disease. MATERIALS AND METHODS: Between August 2018 and January 2019, 70 patients with gallstones in our hospital were enrolled in this prospective clinical trial. All the patients were randomly divided into the ACE group (ACE+TUDCA treatment for 8 wk) and the Sham group (Sham ACE+TUDCA treatment for 8 wk). In the ACE group, all the patients were nightly given ACE every 2 weeks, and in 2 groups, every patient took TUDCA 500 mg at bedtime. The parameters about gallbladder emptying were detected by ultrasound before and after the treatment, and the clinical symptom scores were recorded at the same time points. RESULTS: A total of 63 patients with gallstone disease were included in our study, with 33 patients in the ACE group and 30 patients in the Sham group. In the ACE group, the empty volume (EV) and gallbladder ejection fraction (GBEF) were improved after treatment (P<0.05). Almost every symptom score (except symptom 7, P=0.15) and total score were decreased (P<0.05). In the Sham group, the symptom 1, 2, 4, 5 scores, and total score were significantly decreased (P<0.05). Moreover, the residual volume in the ACE group was significantly lower than in the Sham group (P=0.008). The EV and GBEF in the ACE group were higher than that in the Sham group (P<0.05). The score of symptom 6 in the ACE group was lower than that in the Sham group (P=0.008). CONCLUSION: ACE therapy could more effectively improve the gallbladder emptying with a shorter treatment course. Therefore, ACE+TUDCA therapy might be a time-saving treatment for gallstones.

Enhancing Goos-Hänchen shift based on magnetic dipole quasi-bound states in the continuum in all-dielectric metasurfaces.

Metasurface-mediated bound states in the continuum (BIC) provides a versatile platform for light manipulation at the subwavelength dimension with diverging radiative quality factor and extreme optical localization. In this work, we theoretically propose the magnetic dipole quasi-BIC resonance in asymmetric silicon nanobar metasurfaces to realize giant Goos-Hänchen (GH) shift enhancement by more than three orders of wavelength. In sharp contrast to GH shift based on the Brewster dip or transmission-type resonance, the maximum GH shift here is located at the reflection peak with unity reflectance, which can be conveniently detected in the experiment. By adjusting the asymmetric parameter of metasurfaces, the Q-factor and GH shift can be modulated accordingly. More interestingly, it is found that GH shift exhibits an inverse quadratic dependence on the asymmetric parameter. Furthermore, we theoretically design an ultrasensitive environmental refractive index sensor based on the quasi-BIC enhanced GH shift, with a maximum sensitivity of 1.5×107 µm/RIU. Our work not only reveals the essential role of BIC in engineering the basic optical phenomena but also suggests the way for pushing the performance limits of optical communication devices, information storage, wavelength division de/multiplexers, and ultrasensitive sensors.

Rabi resonance in coherent population trapping: microwave mixing scheme.

Coherent population trapping (CPT) resonance signals have promise in a wide range of applications involving precision sensing. Generally, the CPT phenomenon occurs in a three-level Λ system with a bichromatic phase-coherent light fields. We theoretically and experimentally studied an Rb vapor-cell-based atomic system involving bichromatic CPT optical fields and an external microwave (MW) field simultaneously. In such a mixing scheme, the coherence of the ground states could be controlled either by the Rabi frequency of the microwave field or by the relative phase between the optical fields and the MW field. Moreover, we investigated the Rabi resonance in this mixing scheme. The Rabi frequency of the MW field can be measured SI (International System of Units)-traceably based on the Rabi resonance lineshape, and thus holds the potential to realize intensity stabilization of the optical field in this system. Simple theoretical models and numerical calculations are also presented to explain the experimental results. There is scope to use the proposed technique in future development of SI-traceable optical field strength standards.

Salivary microbial diversity at different stages of human immunodeficiency virus infection.

Human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) disrupts the host microbial balance. During disease progression, the oral microbial environment is altered in untreated people living with HIV/AIDS (PLWHA); however, no studies have reported changes in salivary microbial diversity during different stages of HIV infection. Therefore, in this study, we aimed to assess the relationships between immune dysfunction and changes in saliva microbiota. To this end, we collected saliva samples from 11 HIV-negative individuals and 44 PLWHA during different stages based on the Centers for Disease Control and Prevention criteria (stage 0, early stage during the first 6 months after infection; stages 1, 2, and 3 associated with CD4+ T-lymphocyte counts of ≥500, 200-499, and ≤200 or opportunistic infection, respectively). We analyzed salivary microbial community diversity using polymerase chain reaction amplification and Illumina MiSeq sequencing. We found that HIV-positive individuals had significantly greater alpha-diversity in the microbial community composition compared with HIV-negative controls (P < 0.05) except for AIDS (stage 3); however, the predominant salivary microbiota in the five groups remained similar. Porphyromonas in the four positive groups was the only genus that was significantly less abundant in the HIV-positive groups than in the control group (P < 0.05). There were some consistencies between the general abundance of salivary microbiota and AIDS disease progression. Lots of bacterial abundances in the saliva increased dramatically during the acute HIV infection (stage 0), and some of the negligible and abnormally proliferating bacteria in the asymptomatic stage showed a downward trend. Additionally, in the AIDS stage, partial inhibition was observed. Notably, Porphyromonas was closely related to the immune activation of HIV, showing a decline in abundance once infected with HIV. Solobacterium, which induces inflammation, was negatively correlated with CD4 counts. Overall, our findings provided important insights into changes in salivary microbial diversity in PLWHA.

Novel computer-generated hologram encoding method based on partially temporal coherent light.

Partially temporal coherent light (PTCL) has been applied to holographic reconstruction to reduce speckle noise in display systems, while the encoding methods of computer-generated hologram (CGH), based on PTCL, have not been reported. We propose a novel method to encoding CGH, in which a PTCL with a broadband continuous spectrum is used to illuminate the object image. The continuous spectrum is discretized into different wavelengths and a weight value associated with PTCL power spectrum is assigned to each wavelength. The diffractive transmission is based on Fresnel diffraction theory. The phase distribution of the encoded CGH is obtained using the sum of multiplying the different CGH phase distributions of corresponding discrete wavelengths by the corresponding weight values. The modulation results without iteration are performed to verify the feasibility of the proposed method and the iterative algorithm is introduced to improve the quality of the modulation. The reconstructed images from the proposed encoding method exhibit high quality as compared with that obtained from the encoding method based on ideal temporal coherent light. Numerical simulations and optical experiments are good consistent with each other. The proposed method can provide a reference for various wave-front modulations.

Formulas of partially spatial coherent light and design algorithm for computer-generated holograms.

Formulas of partially spatial coherent light are derived and its corresponding design algorithm is proposed for generating computer-generated holograms based on partially spatial coherent light. The partially coherent light is divided into two terms: spatially absolute coherent part and incoherent part. The former is propagated by angular spectrum method, while the latter is based on the optical transfer function. The related formulas are derived where the coherent function (degree of coherence) is related. A modified iterative algorithm is further developed for optimizing the phase distributions. Numerical simulations and optical experiments are both performed to verify the proposed algorithm. The results obtained by the proposed method and the traditional method are compared, and it is clear that the speckle contrasts in optical experiments are improved more than 46%, and the image quality is obviously improved. This method could also provide new applications for three-dimensional holographic display, beam shaping, and other wave-front modulation techniques.

Multi-region phase calibration of liquid crystal SLM for holographic display.

The liquid crystal spatial light modulator (SLM) is able to provide flexible wavefront control, whereas the initial phase and its response distortions will heavily influence the modulation accuracy. The currently existing calibration methods are tedious and time consuming. A novel multi-region calibration method for minimizing those distortions is proposed. The entire panel is divided into several local regions based on the similarity of phase response characteristic. The nonlinear phase response and static phase distortion of each local region are calibrated in the iterative division procedure. The calibration method is theoretically analyzed and experimentally verified. For the Jasper 4 K SLM panel, when five local regions are built, the root mean error of linear phase shifts is reduced to 0.1 rad and the compensation accuracy of the static phase distortion reaches 0.24 wavelength. The calibrated SLM is applied for the color holographic display and the results show that the reconstructed image quality is improved significantly. The proposed method is simpler and faster because of the reasonable regional division and lower calibration complexity. It could be used for the calibration of various phase only or complex modulators with high space bandwidth product in the future.

Minimizing the ratio of ionizable lipid in lipid nanoparticles for in vivo base editing.

Lipid nanoparticles (LNPs) have gained clinical approval as carriers for both siRNA and mRNA. Among the crucial components of LNPs, ionizable lipids play a pivotal role in determining the efficiency of RNA delivery. In this study, we synthesized a series of ionizable lipids, denoted as HTO, with a higher count of hydroxyl groups compared to SM-102. Remarkably, LNPs based on HTO12 lipid demonstrated comparable mRNA delivery efficiency and biosafety to those based on SM-102. However, the former reduced the ratio of ionizable lipid/total lipids to mRNA in LNPs by 2.5 times compared to SM-102. The HTO12 LNP efficiently encapsulated adenine base editor mRNA and sgRNA targeting Pcsk9, leading to substantial gene editing within the liver of mice and effective reduction of the target protein. Our study underscores that ionizable lipids with multiple hydroxyl groups may facilitate an improved lipid-to-mRNA ratio to minimize the dosage of ionizable lipids for in vivo delivery.

SARS-CoV-2-specific T-cell responses are induced in people living with human immunodeficiency virus after booster vaccination.

BACKGROUND: T-cell-mediated immunity is crucial for the effective clearance of viral infection, but the T-cell-mediated immune responses that are induced by booster doses of inactivated severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccines in people living with human immunodeficiency virus (PLWH) remain unclear. METHODS: Forty-five PLWH who had received antiretroviral therapy (ART) for more than two years and 29 healthy controls (HCs) at Beijing Youan Hospital were enrolled to assess the dynamic changes in T-cell responses between the day before the third vaccine dose (week 0) and 4 or 12 weeks (week 4 or week 12) after receiving the third dose of inactivated SARS-CoV-2 vaccine. Flow cytometry, enzyme-linked immunospot (ELISpot), and multiplex cytokines profiling were used to assess T-cell responses at the three timepoints in this study. RESULTS: The results of the ELISpot and activation-induced marker (AIM) assays showed that SARS-CoV-2-specific T-cell responses were increased in both PLWH and HCs after the third dose of the inactivated SARS-CoV-2 vaccine, and a similar magnitude of immune response was induced against the Omicron (B.1.1.529) variant compared to the wild-type strain. In detail, spike-specific T-cell responses (measured by the ELISpot assay for interferon γ [IFN-γ] release) in both PLWH and HCs significantly increased in week 4, and the spike-specific T-cell responses in HCs were significantly stronger than those in PLWH 4 weeks after the third vaccination. In the AIM assay, spike-specific CD4+ T-cell responses peaked in both PLWH and HCs in week 12. Additionally, significantly higher spike-specific CD8+ T-cell responses were induced in PLWH than in HCs in week 12. In PLWH, the release of the cytokines interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-α), and IL-22 by peripheral blood mononuclear cells (PBMCs) that were stimulated with spike peptides increased in week 12. In addition, the levels of IL-4 and IL-5 were higher in PLWH than in HCs in week 12. Interestingly, the magnitude of SARS-CoV-2-specific T-cell responses in PLWH was negatively associated with the extent of CD8+ T-cell activation and exhaustion. In addition, positive correlations were observed between the magnitude of spike-specific T-cell responses (determined by measuring IFN-γ release by ELISpot) and the amounts of IL-4, IL-5, IL-2 and IL-17F. CONCLUSIONS: Our findings suggested that SARS-CoV-2-specific T-cell responses could be enhanced by the booster dose of inactivated COVID-19 vaccines and further illustrate the importance of additional vaccination for PLWH.

The deubiquitinating enzyme USP4 regulates BRCA1 stability and function.

BRCA1 plays a suppressive role in breast tumorigenesis. Ubiquitin-dependent degradation is a common mechanism that regulates BRCA1 protein stability, and several ubiquitin ligases involved have been identified. However, the deubiquitinating enzyme for BRCA1 remains less defined. Here, we report that the deubiquitinase USP4 interacts with, deubiquitinates and stabilizes BRCA1, maintaining the protein level of BRCA1. USP4 knockdown results in a decreased BRCA1 protein level, impairment in homologous recombination mediated double-stranded break repair, and increased genome instability, and confers resistance to DNA damage-inducing agents and PARP inhibitors. Ectopic expression of USP4 stabilizes BRCA1 and reverse the effects caused by USP4 knockdown. Moreover, USP4 is low expressed in human breast cancer tissues and its low expression correlates with poorer survival of patients. Furthermore, we identified several loss-of-function mutations of USP4 in human gynecological cancers, the catalytic activity of which or their interaction with BRCA1 is disrupted. Together, we reveal that USP4 is a deubiquitinase for BRCA1. USP4 positively regulates the stability and function of BRCA1 through de-ubiquitination, and plays important role in the suppression of breast cancer.

Crosstalk between human immunodeficiency virus infection and salivary bacterial function in men who have sex with men.

Background: Engaging in anal sexual intercourse markedly increases the risk of developing HIV among men who have sex with men (MSM); oral sexual activities tend to uniquely introduce gut-derived microbes to salivary microbiota, which, combined with an individual's positive HIV status, may greatly perturb oral microecology. However, till date, only a few published studies have addressed this aspect. Methods: Based on 16S rRNA sequencing data of bacterial taxa, MicroPITA picks representative samples for metagenomic analysis, effectively revealing how the development and progression of the HIV disease influences oral microbiota in MSM. Therefore, we collected samples from 11 HIV-negative and 44 HIV-positive MSM subjects (stage 0 was defined by HIV RNA positivity, but negative or indeterminate antibody status; stages 1, 2, and 3 were defined by CD4+ T lymphocyte counts ≥ 500, 200-499, and ≤ 200 or opportunistic infection) and selected 25 representative saliva samples (5 cases/stage) using MicroPITA. Metagenomic sequencing analysis were performed to explore whether positive HIV status changes salivary bacterial KEGG function and metabolic pathway in MSM. Results: The core functions of oral microbiota were maintained across each of the five groups, including metabolism, genetic and environmental information processing. All HIV-positive groups displayed KEGG functions of abnormal proliferation, most prominently at stage 0, and others related to metabolism. Clustering relationship analysis tentatively identified functional relationships between groups, with bacterial function being more similar between stage 0-control groups and stage 1-2 groups, whereas the stage 3 group exhibited large functional changes. Although we identified most metabolic pathways as being common to all five groups, several unique pathways formed clusters for certain groups; the stage 0 group had several, while the stage 2 and 3 groups had few, such clusters. The abundance of K03046 was positively correlated with CD4 counts. Conclusion: As HIV progresses, salivary bacterial function and metabolic pathways in MSM progressively changes, which may be related to HIV promoting abnormal energy metabolism and exacerbate pathogen virulence. Further, infection and drug resistance of acute stage and immune cell destruction of AIDS stage were abnormally increased, predicting an increased risk for MSM individuals to develop systemic and oral diseases.

Exploitation of a shared genetic signature between obesity and endometrioid endometrial cancer.

Aims: The findings in epidemiological studies suggest that endometrioid endometrial cancer (EEC) is associated with obesity. However, evidence from gene expression data for the relationship between the two is still lacking. The purpose of this study was to explore the merits of establishing an obesity-related genes (ORGs) signature in the treatment and the prognostic assessment of EEC. Methods: Microarray data from GSE112307 were utilized to identify ORGs by using weighted gene co-expression network analysis. Based on the sequencing data from TCGA, we established the prognostic ORGs signature, confirmed its value as an independent risk factor, and constructed a nomogram. We further investigated the association between grouping based on ORGs signature and clinicopathological characteristics, immune infiltration, tumor mutation burden and drug sensitivity. Results: A total of 10 ORGs were identified as key genes for the construction of the signature. According to the ORGs score computed from the signature, EEC patients were divided into high and low-scoring groups. Overall survival (OS) was shorter in EEC patients in the high-scoring group compared with the low-scoring group (P < 0.001). The results of the Cox regression analysis showed that ORGs score was an independent risk factor for OS in EEC patients (HR = 1.017, 95% confidence interval = 1.011-1.023; P < 0.001). We further revealed significant disparities between scoring groups in terms of clinical characteristics, tumor immune cell infiltration, and tumor mutation burden. Patients in the low-scoring group may be potential beneficiaries of immunotherapy and targeted therapies. Conclusions: The ORGs signature established in this study has promising prognostic predictive power and may be a useful tool for the selection of EEC patients who benefit from immunotherapy and targeted therapies.

Shared sex hormone metabolism-related gene prognostic index between breast and endometrial cancers.

Aims: As sex hormone-dependent tumors, it remains to be clarified whether there is a common genetic signature and its value between breast and endometrial cancers. The aim of this study was to establish the shared sex hormone metabolism-related gene prognostic index (SHMRGPI) between breast and endometrial cancers and to analyze its potential role in the therapeutic and prognostic assessment of endometrial cancers. Methods: Using transcriptome data from TCGA, tumor-associated gene modules were identified by weighted gene co-expression network analysis, and the intersection of module genes with female sex hormone synthesis and metabolism genes was defined as sex hormone metabolism-related gene. SHMRGPI was established by the least absolute shrinkage and selection operator and Cox regression. Its prognostic value of patients with endometrial cancer was validated, and a nomogram was constructed. We further investigated the relationship between SHMRGPI groups and clinicopathological features, immune infiltration, tumor mutation burden, and drug sensitivity. Results: A total of 8 sex hormone metabolism-related gene were identified as key genes for the construction of prognostic models. Based on SHMRGPI, endometrial cancer patients were divided into high and low SHMRGPI groups. Patients in the low SHMRGPI group had longer overall survival (OS) compared with the high group (P< 0.05). Furthermore, we revealed significant differences between SHMRGPI groups as regards tumor immune cell infiltration, somatic mutation, microsatellite instability and drug sensitivity. Patients with low SHMRGPI may be the beneficiaries of immunotherapy and targeted therapy. Conclusions: The SHMRGPI established in this study has prognostic power and may be used to screen patients with endometrial cancer who may benefit from immunotherapy or targeted therapy.

Combined Influences of Water Content and Coarse Grain Content on Shear Strength of Unsaturated Soil Mixture.

An interlayer existed between the ballast layer and subgrade in the conventional railway substructure. Considering that the shear strength τ of the interlayer soil was influenced by the changes in the ballast grain content and water content, this aspect was explored in the present study. Monotonic triaxial tests were fulfilled, which considered five coarse grain contents fv and three water contents of fine soil wf. The results showed that the growth in fv contributed to an increment in τ of the soil mixture under both saturation and unsaturation. Conversely, in previous studies, the growth of fv induced an increment in τ under saturation, but a decline in that under unsaturation. This was explained by the competing influences of fv and suction ψ: in previous studies, increasing fv induced a decline in the dry density of the fine soil fraction ρd-f, which contributed to a decline in ψ. When the negative influence of declining ψ outweighed the positive influence of the incrementing fv, the τ of the soil mixture decreased. Meanwhile, modelling of the τ-ψ relationship in the soil mixture with varying fv was performed. This proposed model was examined using the test results from both the present and previous studies, which shows its reasonably good performance.