In this study, an innovative transdermal peptide, #PKU12, was developed based on transdermal peptide TD-1, and the anti-tumor effect of PKU12-based siRNA against HPV was investigated in vivo. Furthermore, transcriptome differences between PKU12 + siRNA treatment and control groups were compared to assess treatment effects. The top five upregulated and downregulated genes identified by RNA sequencing were further subjected to survival analysis. The present study, for the first time, showed that this novel peptide could enhance the transdermal delivery of the siRNA targeting HPV16 L1, E6, and E7. PKU12-based siRNA delivery significantly repressed the mRNA expression levels of HPV16 L1, E6, and E7 in the SiHa xenograft tumors and attenuated tumor growth as well. The RNA-sequencing results showed that a total of 586 DEGs were detected in the PKU12 + siRNA-treated tumor tissues compared to the control tumor tissues. The GSEA analysis revealed that DEGs were inversely associated with the HIF-1 signaling pathway, the TNF signaling pathway, the AGE-RAGE signaling pathway, the NF-kappa B signaling pathway, ferroptosis, the IL-17 signaling pathway, ovarian steroidogenesis, and rheumatoid arthritis. Further functional enrichment analysis revealed that DEGs were significantly enriched in several key pathways, including cytokine-cytokine receptor interaction, the TNF signaling pathway, and the IL-17 signaling pathway. High expression of MYH1, MYH4, FGG, DEPP1, and ZBTB16 was associated with shorter overall survival of patients with cervical cancer; high expression of SULT1E1, RAB3C, CXCR3, and PROX2 was associated with longer overall survival of patients with cervical cancer. In conclusion, the transdermal peptide PKU12 is potentially a good candidate for a siRNA delivery vehicle for the treatment of cervical cancer.

目的 筛选晒后修复乳剂的最佳处方,并对其进行质量评价.方法 采用单因素、Box-Behnken中心组合设计原理和响应面分析法对晒后修复乳剂的处方进行优化,通过考察乳剂外观性状及稳定性,确定最佳的处方配比,最后对乳剂保湿性、抗氧化性和安全性进行评价.结果 晒后修复乳剂最佳处方为单硬脂酸甘油酯4.0％、白凡士林3.0％、二甲基硅油5.0％、硬脂酸3.0％、三乙醇胺0.4％、十二烷基硫酸钠0.2％、透明质酸钠0.1％、尼泊金乙酯0.1％、青刺果油3.0％、金缕梅提取液8.0％.该处方所得基质均匀细腻,离心及加速试验未见分层,易于涂抹、安全温和、质量稳定;当青刺果油加入量为3.0％时,乳剂具有很好的稳定性、保湿性和抗氧化性.结论 所制晒后修复乳剂质量稳定、安全温和,有望开发成一种新型晒后修复乳剂.

A randomized, double-blind, placebo-controlled, parallel phase II study was conducted for generalized myasthenia gravis (gMG) in Chinese population to evaluate the efficacy and safety of batoclimab as a novel anti-FcRn therapy. Eligible patients were randomized in a 1:1:1 ratio to placebo, batoclimab 340 mg, 680 mg groups, respectively. In double-blinded period, patients received batoclimab or placebo on days 1, 8, 15, 22, 29, and 36. In the open-label period, all patients received batoclimab 340 mgon days 50, 64, and 78. In follow-up period, patients were examined on days 92, 106, and 120. The primary endpoint was myasthenia gravis activities of daily living (MG-ADL) score change on day 43 from baseline. Secondary endpoints included assessment on Quantitative Myasthenia Gravis (QMG), Myasthenia Gravis Composite (MGC), Myasthenia Gravis Quality of Life (MG-QoL), pharmacokinetics, pharmacodynamics and safety events. Totally 30 subjects were randomized, with 9,10 and 11 in the placebo, batoclimab 340mg and 680mg groups respectively.MG-ADL score changes from baseline to day 43 were -2.2±0.9, -4.7±0.6, and -4.4±1.0 in the placebo, batoclimab 340 mg, and 680 mg groups, respectively. Similar changes were observed in QMG, MGC, and MG-QoL scores. The proportion of patients with clinically significant improvement on day 43 was higher in batoclimab groups. In open-label period, symptom recurrence and IgG levels rise again except the former placebo group. On day 120 the placebo group had more improvement compared to batoclimab groups, with IgG levels reaching a plateau. Treatment-emergent adverse events (TEAEs) were comparable among the three groups. No death or TEAEs leading to study discontinuation was observed. The batoclimab groups experienced mild-to-moderate albumin reductions and total cholesterol elevation, all returned to baseline in follow-up without clinical interference. In conclusion, batoclimab is effective and safe in Chinese patients with gMG.

Salmonella is a crucial food‐borne pathogen causing food poisoning, leading to severe public health events. Here, we developed a technique by integrating recombinase polymerase amplification with CRISPR‐LbCas12a and employing two targets with engineered crRNA for detection of Salmonella (RPA‐LbCas12a‐TTECDS). Our findings revealed that this novel method rapidly detects trace Salmonella in food through fluorescence intensity and provides a template for other food‐borne pathogen detection methods. Further, crRNA was optimized to increase detection sensitivity. Double targets were used to enhance the detection accuracy, reaching the level of qPCR, which was superior to fluorescent RPA. The RPA‐LbCas12a‐TTECDS system specifically detected Salmonella levels as low as 50 CFU per ml at 37°C in 1 h. In summary, a simple, rapid, sensitive and high accuracy detection technique based on CRISPR‐Cas12a was created for Salmonella detection without complicated equipment.

WangShiBaoChiWan(WSBCW)is a traditional Chinese medicine with a recorded administration history of more than 180 years.In the present study,the preclinical safety of WSBCW was evaluated the preclinical safety of WSBCW using a toxicity test,which consisted of an administration period of 28 d and a recovery period of 15 d.During the test,male and female SD rats were administered the medicine once a day by oral gavage,at a dose of 60 mg/kg/day,600 mg/kg/day,or 1500 mg/kg/day.As a reference medicine,mosapride citrate was administered at a dose of 37.5 mg/kg/day,which was clinically equivalent to the high-dosage treatment of WSBCW.With all the dosage groups,statistically,no adverse effect was observed in terms of clinical observation,food intake,body weights,organ coefficient,blood biochemistry,and histopathology examination.No intestinal melanosis was observed in the rats.When the data were examined animal by animal,test substance-related adverse effects were found with the high-dosage rats in hematology assay.The deranged,however,reversible changes suggested a compromised intestinal barrier,which was also observed with in mosapride citrate-treated rats.In addition to the histopathology assay,molecular toxicology was explored using high-throughput gene sequencing.No evident toxicity was revealed.In summary,administration of WSBCW was well tolerated within a treatment of 28 d.

In contrast to the well-established genomic 5-methylcytosine (5mC), the existence of N6-methyladenine (6 mA) in eukaryotic genomes was discovered only recently. Initial studies found that it was actively regulated in cancer cells, suggesting its involvement in the process of carcinogenesis. However, the contribution of 6 mA in tongue squamous cell carcinoma (TSCC) still remains uncharacterized. In this study, a pan-cancer type analysis was first performed, which revealed enhanced 6 mA metabolism in diverse cancer types. The study was then focused on the regulation of 6 mA metabolism, as well as its effects on TSCC cells. To these aspects, genome 6 mA level was found greatly increased in TSCC tissues and cultured cells. By knocking down 6 mA methylases N6AMT1 and METTL4, the level of genomic 6 mA was decreased in TSCC cells. This led to suppressed colony formation and cell migration. By contrast, knockdown of 6 mA demethylase ALKBH1 resulted in an increased 6 mA level, enhanced colony formation, and cell migration. Further study suggested that regulation of the NF-κB pathway might contribute to the enhanced migration of TSCC cells. Therefore, in the case of TSCC, we have shown that genomic 6 mA modification is involved in the proliferation and migration of cancer cells.

Monolithic metallic glasses often exhibit work softening induced by high atom mobility in the supercooled liquid region in tension. In this work, we report an unusual viscous flow of the Pd42.5Ni42.5P15 metallic glass in its supercooled liquid region, which is characterized by a remarkable hardening behavior with decent plasticity during tension. To unravel the causes of this unusual hardening, we conducted structural and thermodynamic analyses by employing several experiments, including dynamic mechanical analysis, differential scanning calorimetry, high-resolution transmission electron microscopy, and in situ heating synchrotron high-energy x-ray diffraction. It was found that the unusual hardening behavior was attributed to structural ordering during deformation, rather than nanocrystallization or structural relaxation observed, sometimes, in metallic glasses deformed in supercooled liquid region. We also incorporated an additional work hardening region in the modified deformation map for metallic glasses.

This study was conducted to collect and review clinical data in patients who had ever been on teriflunomide 14 mg/day in the West China hospital, and to assess effectiveness and safety of teriflunomide in a cohort of MS patients. In this single-center, retrospective, observational, real world study, we collected clinical data of patients treated with teriflunomide 14 mg/day for at least 3 months. Primary endpoint was the annualized relapse rate (ARR). Secondary endpoints were the mean change from baseline in expanded disability status scale (EDSS), the proportion of relapse-free and disability progression-free patients at last follow-up visit and the overall safety of teriflunomide. Of 90 patients identified, 74 were eligible and included in the analysis. Women represented 68.9% of the cohort. Median age and median disease duration were 32.81 and 5.92 years, respectively. A total of 35.0% of the patients received prior disease-modifying therapies. Median duration of teriflunomide exposure was 9 months. The mean ARR was significant lower at last follow-up visit (0.20 ± 0.47) compared to the prior to teriflunomide treatment (1.02 ± 0.81) (p = 0.005), with a 78.0% reduction. A total of 75.0% of teriflunomide-treated patients remained relapse-free during the data collection period. The mean EDSS score showed a reduction from a baseline score of 1.90 ± 1.71 to a mean post-treatment score of 1.88 ± 1.85. Proportion of patients free from disability progression was 82.0%. The most common adverse events (AEs) were hair thinning (22.9%), leukopenia (5.4%), and cheilitis (2.7%), followed by abnormal liver function and abnormal menstruation (1.3% each). Although no serious AEs were reported, 18.9% of patients discontinued teriflunomide. The real-world teriflunomide benefits observed in this study were similar than those observed in previous clinical trials. Teriflunomide was well tolerated and no new safety findings were observed in this study.

As one of the most abundant DNA methylation form in prokaryotes, N 6 -methyladenine nucleotide (6 mA) was however only recently identified in eukaryotic genomes. To explore the implications of N 6 -adenine methylation in adipogenesis, genomic N 6 -adenine methylation was examined across adipocyte differentiation stages of 3T3-L1 cells. When the N 6 -adenine methylation profiles were analyzed and compared with the levels of gene expression, a positive correlation between the density of promoter 6 mA and gene expression level was uncovered. By means of in vitro methylation and gene knockdown assay, METTL4, a homologue of Drosophila methylase CG14906 and C. elegans methylase DAMT-1, was demonstrated to be a mammalian N 6 -adenine methylase that functions in adipogenesis. Knockdown of Mettl4 led to altered adipocyte differentiation, shown by defective gene regulation and impaired lipid production. We also found that the effects of N 6 -adenine methylation on lipid production involved the regulation of INSR signaling pathway, which promotes glucose up-taking and lipid production in the terminal differentiation stage.

During the development of primary Sjogren's syndrome (pSS), aberrant expression of autoantigen is a hallmark event. To explore the regulation of autoantigen tripartite motif containing 21 (Ro/SSA, TRIM21), microRNA profiling was performed in our previous study. In which, twoTRIM21-targeting microRNAs were identified, namely miR-1207-5p and miR-4695-3p. To further pursue their roles in the development of pSS, assays were performed with cultured human submandibular gland (HSG) cells, and salivary gland tissues. Results showed that transfection of miR-1207-5p or miR-4695-3p mimics down-regulated not only the expression ofTRIM21, but also the levels of pro-apoptotic genes B cell lymphoma 2 associated X (BAX), Caspase 9 (CASP-9) and Caspase 8 (CASP-8). This finally led to antiapoptotic phenotypes in HSG cells. Consistent with the antiapoptotic activity, transfection of microRNA inhibitors up-regulated the expression ofTRIM21and led to a pro-apoptotic phenotype. These therefore propose miR-1207-5p and miR-4695-3p as two antiapoptotic microRNAs functioning through apoptosis pathway. Supporting this speculation, assays performed with salivary gland tissues revealed down-regulation of miR-1207-5p and miR-4695-3p, as well as up-regulation ofTRIM21and pro-apoptoticCASP-8gene in pSS samples. Significance of the study For pSS patients, apoptosis of acinar and ductal epithelial cells has been proposed to be a potential mechanism that impairs the secretion of salivary glands. In our study, two autoantigen-targeting microRNAs were characterized as antiapoptotic microRNAs functioning through apoptosis pathway, which may be potential targets for the treatment of pSS.

Change history: In the HTML version of this Letter, Extended Data Fig. 4 incorrectly corresponded to Fig. 4 (the PDF version of the figure was correct). This has been corrected online.

To explore the editing specificity of CRISPR/Cpf1 system, effects of target mutation were systematically examined using a reporter activation assay, with a set of single-nucleotide mutated target site. Consistent with our previous study performed with CRISPR/Cas9, a “core” sequence region that is highly sensitive to target mutation was characterized. The region is of 4-nucleotide long, located from +4 to +7 position of the target site, and positioned within a positively charged central channel when assembled into Cpf1 endonuclease. Single-nucleotide mutation at the core sequence could abolish gene editing mediated by a however active sgRNA. With a great majority of the target sites, a kind of ‘super’ off-target gene editing was observed with both CRISPR/Cpf1 and CRISPR/Cas9. For a given target site, mutation at certain positions led to greatly enhanced off-target gene editing efficacy, even up to 10-fold of that of the fully-matched target. Study further found that these effects were determined by the identity of target nucleotide, rather than the nucleotide of crRNA. This likely suggests that the interactions between target nucleotide and the endonuclease are involved in this process.

We report results on the searches of weakly interacting massive particles (WIMPs) with sub-GeV masses (m_{χ}) via WIMP-nucleus spin-independent scattering with Migdal effect incorporated. Analysis on time-integrated (TI) and annual modulation (AM) effects on CDEX-1B data are performed, with 737.1 kg day exposure and 160 eVee threshold for TI analysis, and 1107.5 kg day exposure and 250 eVee threshold for AM analysis. The sensitive windows in m_{χ} are expanded by an order of magnitude to lower DM masses with Migdal effect incorporated. New limits on σ_{χN}^{SI} at 90% confidence level are derived as 2×10^{-32}∼7×10^{-35}  cm^{2} for TI analysis at m_{χ}∼50-180  MeV/c^{2}, and 3×10^{-32}∼9×10^{-38}  cm^{2} for AM analysis at m_{χ}∼75  MeV/c^{2}-3.0  GeV/c^{2}.

Rationale: Distant metastasis and chemoresistance are the major causes of short survival after initial chemotherapy in lung adenocarcinoma patients. However, the underlying mechanisms remain elusive. Our pilot study identified high expression of the homeodomain transcription factor HOXB13 in chemoresistant lung adenocarcinomas. We aimed to investigate the role of HOXB13 in mediating lung adenocarcinoma chemoresistance. Methods: Immunohistochemistry assays were employed to assess HOXB13 protein levels in 148 non-small cell lung cancer patients. The role of HOXB13 in lung adenocarcinoma progression and resistance to cisplatin therapy was analyzed in cells, xenografted mice, and patient-derived xenografts. Needle biopsies from 15 lung adenocarcinoma patients who were resistant to cisplatin and paclitaxel therapies were analyzed for HOXB13 and EZH2 protein levels using immunohistochemistry. Results: High expression of HOXB13 observed in 17.8% of the lung adenocarcinoma patients in this study promoted cancer progression and predicted poor prognosis. HOXB13 upregulated an array of metastasis- and drug-resistance-related genes, including ABCG1, EZH2, and Slug, by directly binding to their promoters. Cisplatin induced HOXB13 expression in lung adenocarcinoma cells, and patient-derived xenografts and depletion of ABCG1 enhanced the sensitivity of lung adenocarcinoma cells to cisplatin therapy. Our results suggest that determining the combined expression of HOXB13 and its target genes can predict patient outcomes. Conclusions: A cisplatin-HOXB13-ABCG1/EZH2/Slug network may account for a novel mechanism underlying cisplatin resistance and metastasis after chemotherapy. Determining the levels of HOXB13 and its target genes from needle biopsy specimens may help predict the sensitivity of lung adenocarcinoma patients to platinum-based chemotherapy and patient outcomes.

CRISPR/Cas has been coming to prosperity since its discovery and application.It becomes a standard solution for gene editing in the past few years.A guide RNA is used to lead the endonuclease,such as Cas9 and Cpf1,to specific sites and break the double strand.However,there is also possibility that the system will cut a non-specific position,which is called "off-target effect".The off-target cleavage may cause trouble to gene function research or clinic treatment.In order to reveal the target specificity of Cpf1,this study explored the single-nucleotide mismatches by a dual-luciferase system.Our results showed that the poly(T) structure was prohibitive in spacer for Cpf1 targeting.Moreover,rA mismatches seemed to be of the least tolerance for CRISPR/Cpf1,which was same as CRISPR/Cas9.The phenomenon might be attributed to the homology of the two enzymes.In summary,our research suggest that more attention should be paid to off-target effects when using CRISPR/Cpf1 or CRISPR/Cas9,as this is an intrinsic characteristic of the system.

The development of adipose tissue is a precisely coordinated cellular process, in which both protein-coding and non-coding genes are involved. To characterize the in vivo function of a novel long non-coding RNA (lncRNAs), loss-of-function assays were performed with slincRAD knockdown mice. Down-regulation of slincRAD expression was found to impair the development of adipose tissue, leading to a slim phenotype for both of the male and female mice. Compared to normal adipocytes, slincRAD knockdown cells had defective differentiation features, such as smaller sizes and decreased lipid production. For elder mice, slincRAD knockdown led to abnormal glucose and lipid metabolism. Therefore, a physiologically important lncRNA was characterized in the development of adipose tissue.

Accumulating experimental evidence has shown that the aberrant expression of microRNAs (miRNAs) is involved in the development and progression of human cervical cancer. Previously, we identified miR-182 as an oncomiRNA in cervical cancer. However, the mechanism by which miR-182 is regulated and the interaction between human papillomavirus (HPV) and miR-182 in cervical cancer development remains unknown. In the present study, we explored the link between HPV E7 and miR-182 and verified that high-risk HPV E7 upregulated miR-182 expression through TGF-β/Smad4 signaling pathway in cervical cancer. By contrast, low-risk HPV E7 did not affect the expression of TGF-β and miR-182. Mechanistically, as high-risk HPV E7 bound to pRb, E2F was released from the complex and bound to the TGF-β promoter region, resulting in TGF-β overexpression. Furthermore, the Smad4 signaling pathway was activated upon TGF-β overexpression, which led to an interaction between Smad4 and the miR-182 promoter region, subsequently inducing the upregulation of miR-182 in both cervical cancer cells and the surrounding normal cells. In conclusion, this newly identified high-risk HPV E7/TGF-β/miR-182 regulatory network might inform the development of specific therapeutic strategies for cervical cancer.