i-CRISPR: a personalized cancer therapy strategy through cutting cancer-specific mutations.

Developing a strategy to specifically kill cancer cells without inducing obvious damage to normal cells may be of great clinical significance for cancer treatment. In the present study, we developed a new precise personalized strategy named "i-CRISPR" for cancer treatment through adding DNA damage repair inhibitors(i) and inducing cancer cell-specific DNA double strand breaks by CRISPR. Through in vitro and in vivo experiments, we confirmed the efficacy of this strategy in multiple cancer models and revealed the mechanism of cell death. Our strategy might provide a novel concept for precise cancer therapy.

Endoplasmic reticulum stress triggered autophagy and regulated the phenotype transformation of rheumatoid arthritis synovial fibroblasts via the IRE1/JNK pathway.

Background: Previous studies have indicated that endoplasmic reticulum (ER) stress may actively promote the pathogenesis of rheumatoid arthritis (RA) by evoking autophagy. However, the underlying mechanism remains largely unknown. This study aimed to explore the mechanism of the ER stress-autophagy pathway in regulating the phenotype transformation of rheumatoid arthritis synovial fibroblasts (RASFs). Methods: Synovial tissue was obtained from RA and osteoarthritis (OA) patients during joint replacement surgery. ER stress/autophagy signature markers were examined in synovial tissue by real-time quantitative polymerase chain reaction (RT-PCR), western blot, and immunohistochemistry. Phenotype transformation of RASFs, including increased cell proliferation and invasion capability, was measured by CCK-8 assay and transwell invasion assay. Signaling pathways were further investigated and inositol requiring enzyme 1 (IRE1) was down-regulated in RASFs by transfecting specific short hairpin RNA-ERN1 (shRNA-ERN1) carried by lentiviral vectors. Results: The expression of ER stress/autophagy pathway-associated proteins, including GRP78, IRE1, protein kinase R-like endoplasmic reticulum kinase (PERK), and LC3, was significantly increased in RA synovium compared with OA synovium. After stimulation with tumor necrosis factor alpha (TNF-α) in vitro, the proliferation and invasion ability of RASFs were upregulated, while this phenomenon could be inhibited by 4-PBA (ER stress inhibitor) or 3-MA (autophagy inhibitor). The expression of IRE1 and p-JNK in particular, occurred in an obviously time-dependent manner after stimulation with TNF-α. Moreover, the proliferation and invasion of RASFs were inhibited after transfection with sh-RNA-ERN1 to downregulate IRE1 expression. Conclusions: ER stress triggered autophagy via the IRE1/JNK pathway to regulate the phenotype transformation of RASFs, indicating an important role of the ER stress-autophagy pathway in the pathological process of synovitis in RA.

A Novel α4/7-Conotoxin QuIA Selectively Inhibits α3ß2 and α6/α3ß4 Nicotinic Acetylcholine Receptor Subtypes with High Efficacy.

α6ß4 nAChR is expressed in the peripheral and central nervous systems and is associated with pain, addiction, and movement disorders. Natural α-conotoxins (α-CTxs) can effectively block different nAChR subtypes with higher efficacy and selectivity. However, the research on α6ß4 nAChR is relatively poor, partly because of the lack of available target-specific α-CTxs. In this study, we synthesized a novel α-4/7 conotoxin QuIA that was found from Conus quercinus. We investigated the efficacy of this peptide to different nAChR subtypes using a two-electrode voltage-clamp technique. Remarkably, we found α-QuIA inhibited the neuronal α3ß2 and α6/α3ß4 nAChR subtypes with significantly high affinity (IC50 was 55.7 nM and 90.68 nM, respectively), and did not block other nAChR subtypes even at a high concentration of 10 µM. In contrast, most α-CTxs have been determined so far to effectively block the α6/α3ß4 nAChR subtype while also maintaining a similar higher efficacy against the closely related α6ß2ß3 and/or α3ß4 subtypes, which are different from QuIA. In conclusion, α-QuIA is a novel α4/7-CTx, which has the potential to develop as an effective neuropharmacology tool to detect the function of α6ß4 nAChR.

Frequent injections of high-dose human umbilical cord mesenchymal stem cells slightly aggravate arthritis and skeletal muscle cachexia in collagen-induced arthritic mice.

A single injection of low-dose human umbilical cord-derived mesenchymal stem cells (UC-MSCs) has been previously demonstrated to relieve synovitis and bone erosion in animal models of arthritis, but whether frequent injections of high-dose UC-MSCs relieve arthritis and inhibit loss of muscle mass has remained elusive. In the present study, DBA/1 mice were randomly divided into three groups: Normal (wild-type mice; n=11), collagen-induced arthritis (CIA; n=12) and CIA treated with UC-MSCs (n=11; 5x106 UC-MSCs per week for 3 weeks). Arthritis and skeletal muscle cachexia were evaluated until the end of the experiment on day 84. It was indicated that both the CIA and UC-MSC groups had lower body weights compared with the normal mice. Clinical arthritis scores, hind ankle diameters, synovitis and bone erosion progressively increased and were similar between the CIA and UC-MSC groups. Although there was no difference in food intake among the three groups, the normalized food intake of normal group was significantly higher than CIA group and UC-MSC group from day 42 onwards; there was no significance on day 77 but this could be neglected. Furthermore, gastrocnemius muscle weight in the UC-MSC group was significantly reduced compared with that in the CIA and normal groups. The UC-MSC group had higher levels of proinflammatory cytokines, such as TNF-α, IL-6 and IL-1ß than those in the CIA group. However, the other cytokines assessed and the fibrosis indices in the CIA and UC-MSC groups were not different from those in the control group and there was no inflammatory cell infiltration. Thus, frequent injections of high-dose UC-MSCs slightly aggravated synovitis and muscle cachexia in the murine CIA model and should therefore be avoided in the treatment of arthritis.

All optical metropolitan quantum key distribution network with post-quantum cryptography authentication.

Quantum key distribution (QKD) provides information theoretically secure key exchange requiring authentication of the classic data processing channel via pre-sharing of symmetric private keys to kick-start the process. In previous studies, the lattice-based post-quantum digital signature algorithm Aigis-Sig, combined with public-key infrastructure (PKI), was used to achieve high-efficiency quantum security authentication of QKD, and we have demonstrated its advantages in simplifying the MAN network structure and new user entry. This experiment further integrates the PQC algorithm into the commercial QKD system, the Jinan field metropolitan QKD network comprised of 14 user nodes and 5 optical switching nodes, and verifies the feasibility, effectiveness and stability of the post-quantum cryptography (PQC) algorithm and advantages of replacing trusted relays with optical switching brought by PQC authentication large-scale metropolitan area QKD network. QKD with PQC authentication has potential in quantum-secure communications, specifically in metropolitan QKD networks.

Treat-to-target strategies aiming at additional ultrasound remission is associated with better control of disease activity and less flare in rheumatoid arthritis.

INTRODUCTION: Treat-to-target (T2T) strategy has greatly improved the prognosis of rheumatoid arthritis (RA). However, the additional benefit of targeting ultrasound (US) remission in addition to clinical remission has been debated. METHODS: RA patients in clinical remission or low disease activity were enrolled. They were assorted into two groups according to the principle of T2T strategy adopted. In clinical group, treatment decision was made with the aim of maintaining DAS28(ESR) ≤ 3.2 only, while in clinical US group, the aim was to attain total power Doppler (PD) US score = 0 in addition to DAS28(ESR) ≤ 3.2. The time-averaged DAS28, flare, and changes of treatment strategy were compared. RESULTS: One hundred ninety-four patients completed 1-year follow-up, with 100 in clinical US and 94 in clinical group. Compared to clinical group, time-averaged DAS28 in clinical US group was significantly lower (1.89 ± 0.51 vs. 2.33 ± 0.71, P < 0.01) with less flare (20.0% vs. 36.2%, P < 0.05). Furthermore, at the end of 1 year, significantly more patients successfully achieved step-down therapy (66.0% vs. 44.7%, P < 0.01) and dramatically fewer patients with step-up therapy in the clinical US group (13.0% vs. 25.5%, P < 0.05) compared to clinical group. In clinical US group, baseline DAS28(ESR) > 2.29, presence of subclinical synovitis, and step-down strategy were independent risk factors for relapse after clinical remission or low disease activity was achieved. CONCLUSIONS: An US-driven T2T in addition to current clinical remission strategy is associated with better control of the disease activity, reduction of relapse, as well as long-term step-down therapy. Step-down strategy should be carefully applied to the patients with baseline DAS28(ESR) over 2.29 and presence of subclinical synovitis even after they have achieved clinical remission or low disease activity. Key Points • Targeting ultrasound remission in addition to current T2T strategy is associated with a better control of RA. • Step-down strategy should be cautiously considered in those with DAS28(ESR) > 2.29 and baseline subclinical synovitis after they have achieved clinical remission or low disease activity.

Inflammation-Targeted Celastrol Nanodrug Attenuates Collagen-Induced Arthritis through NF-κB and Notch1 Pathways.

Rheumatoid arthritis (RA) is a systemic inflammatory disorder which can cause bone and cartilage damage leading to disability, yet the treatment remains unsatisfactory nowadays. Celastrol (Cel) has shown antirheumatic activity against RA. However, the frequent parenteral delivery and poor water solubility of Cel restrict its further therapeutic applications. Here, aiming at effectively overcoming the poor water solubility and short half-life of Cel to boost its beneficial effects for treating RA, we developed a polymeric micelle for Cel delivery based on a reactive oxygen species (ROS) sensitive polymer. Our results demonstrated that Cel may inhibit the repolarization of macrophages toward the pro-inflammatory M1 pheno-type via regulating the NF-κB and Notch1 pathways, which resulted in significantly decreased secretion of multiple pro-inflammatory cytokines to suppress the RA progression. Consequently, the Cel-loaded micelle effectively alleviated the major RA-associated symptoms including articular scores, ankle thickness, synovial inflammation, bone erosion, and cartilage degradation.

Heterogeneous Responses to Mechanical Force of Prostate Cancer Cells Inducing Different Metastasis Patterns.

The physical cues in the extracellular environment play important roles in cancer cell metastasis. However, how metastatic cancer cells respond to the diverse mechanical environments of metastatic sites is not fully understood. Here, substrates with different mechanical properties are prepared to simulate the extracellular mechanical environment of various human tissues. The prostate cancer (PC) cells derived from different cancer metastasis sites show heterogeneity in mechanical response. This heterogeneity mediates two distinct metastasis patterns. High stiffness promotes individual cell migration and proliferation by inducing Yes-associated protein and tafazzin (YAP/TAZ) nuclear localization in bone metastasis-derived cells, whereas low stiffness promotes cell migration and proliferation by inducing lymphatic metastasis-derived cells to form clusters characterized by high expression of CD44. The different metastasis patterns induced by the mechanical properties of the extracellular environment are crucial in the development of PC.

BACE1-AS prevents BACE1 mRNA degradation through the sequestration of BACE1-targeting miRNAs.

Abnormal long noncoding RNAs (lncRNAs) and microRNAs (miRNAs) participate in the pathophysiology of Alzheimer's disease (AD). However, it remains unclear whether these two types of noncoding RNAs functionally interact and which factors mediate these interactions in AD. ß-secretase 1 (BACE1) is the enzyme responsible for amyloid plaque formation, which is a central pathological feature of AD. The lncRNA BACE1-AS and some miRNAs have been implicated in the regulation of BACE1. In this study, we reveal that BACE1-AS shares many miRNA-response elements with BACE1. The overexpression of BACE1-AS results in the repression of miRNAs that target BACE1, thus preventing BACE1 mRNA from being degraded. The knockdown of BACE1-AS increases the levels of these miRNAs, thereby reducing the expression of BACE1. Thus, BACE1-AS functions as a competing endogenous RNA (ceRNA). Our results also deepen the understanding of the regulation of BACE1 by BACE1-AS. In addition to increasing the stability of BACE1 mRNA through the formation of RNA duplexes, BACE1-AS can regulate BACE1 indirectly by acting as a ceRNA. Therefore, we propose that BACE1 functions as a ceRNA and forms a network through its associations with protein-coding genes, lncRNAs and miRNAs in the pathophysiology of AD.

The agreement between ultrasound-determined joint inflammation and clinical signs in patients with rheumatoid arthritis.

BACKGROUND: Ultrasound (US) is sensitive for detecting joint and tendon inflammation in patients with rheumatoid arthritis (RA). So far, which grade of abnormalities on US corresponds to clinical manifestations is unclear. This study aimed to investigate the agreement between US-detected joint inflammation and clinical signs (joint swelling and tenderness). METHODS: In this cross-sectional study, 22 joints of the wrists and hands were, respectively, evaluated by physical examination (PE) and ultrasound in RA patients. Gray scale (GS) and power Doppler (PD) of synovitis, detected by ultrasound, were graded by semi-quantitative scoring systems (0-3). Tenosynovitis and peritendinitis were assessed qualitatively (0/1). RESULTS: A total of 258 consecutive RA patients were included, with median disease duration of 57 months and mean Disease Activity Score based on 28 joints (DAS28)-ESR/DAS28-CRP of 4.47/3.99. In a total of 5676 joints assessed, the overall concordance rate between positive clinical signs and ultrasound-determined joint inflammation was fair (κ = 0.365, p < 0.01). In wrists, joint tenderness showed higher κ coefficient (κ = 0.329, p < 0.01) with ultrasound-determined joint inflammation than swelling (κ = 0.263, p < 0.01); however, swelling showed higher κ coefficient (κ = 0.156-0.536, p < 0.01) with ultrasound-determined joint inflammation than tenderness (κ = 0.061-0.355, p < 0.01) in metacarpophalangeal (MCP) and proximal interphalangeal (PIP) joints. Synovitis had consistently higher agreement with tenderness and swelling than tenosynovitis/peritendinitis. Tenderness and swelling had the highest κ coefficient with GS ≥ 1 synovial hyperplasia in most MCP and PIP joints, while with GS ≥ 2 synovial hyperplasia in wrists. For all 22 joints, PD ≥ 1 synovitis had the highest κ coefficient with clinical tenderness and swelling. CONCLUSIONS: Synovitis had better agreement with clinical signs than tenosynovitis/peritendinitis. Joint swelling showed better agreement with US-determined inflammation than tenderness for MCP and PIP joints, while the opposite for wrists. Both tenderness and swelling are more likely to correspond to GS ≥ 2 for wrists, GS ≥ 1 for MCP and PIP joints, and PD ≥ 1 for any joint.

A combination model to predict relapse and successful conventional DMARDs de-escalation in rheumatoid arthritis patients with sustained clinical remission.

OBJECTIVES: To determine the long-term outcomes of RA patients in sustained clinical remission under different therapeutic strategies and explore the risk factors to relapse. METHODS: RA patients in sustained clinical remission (DAS28(CRP) ≤2.6 for at least 6 months) were enrolled. Their baseline clinical features, ultrasonography and x-ray of hands were collected. The usage of conventional synthetic disease-modified anti-rheumatic drugs (csDMARDs) at baseline and every follow-up visits were recorded. Patients were divided into maintain-therapy group or de-escalate-therapy group according to their treatment during follow-up. The time-point of follow-up visits reaching 2 years or flare (DAS28(CRP)>2.6) was defined as the endpoint of the study. The risk factors to predict flare was analysed by logistic regression model. RESULTS: 94 patients were enrolled in the study, with 59 in de-escalate-therapy group and 35 in maintain-therapy group. During an average of 20.8 months of follow-up, 40 (42.6%) patients relapsed, with 31 (52.5%) from de-escalate-therapy group and 9 (25.7%) from maintain-therapy group. De-escalate-therapy increased the risk of flare by 2.3 times (OR=3.38, p=0.044). Baseline DAS28(CRP) (OR=6.97, p=0.038), presence of subclinical synovitis (OR=3.67, p=0.024), combination of 2 csDMARDs (OR=3.72, p=0.030) were the risk factors for relapse, and the best cut-off value of DAS28(CRP) for relapse prediction through ROC curve was 1.82. Taking the three parameters into the model for a combined prediction probability, the area under the ROC curve was 0.722 (95% CI 0.61, 0.82, p=0.000). CONCLUSIONS: De-escalation therapy was associated with higher risk of relapse in RA patients with sustained clinical remission. A combination model of DAS28(CRP)<1.82 and no subclinical synovitis may help to predict successful csDMARDs reduction in RA patients with sustained clinical remission receiving csDMARDs monotherapy.

The Long Noncoding RNA TTTY15, Which Is Located on the Y Chromosome, Promotes Prostate Cancer Progression by Sponging let-7.

BACKGROUND: The link between prostate cancer (PCa) development and aberrant expression of genes located on the Y chromosome remains unclear. OBJECTIVE: To identify Y-chromosomal long noncoding RNAs (lncRNAs) with critical roles in PCa and to clarify the corresponding mechanisms. DESIGN, SETTING, AND PARTICIPANTS: Aberrantly expressed lncRNAs on the Y chromosome were identified using transcriptome analysis of PCa clinical samples and cell lines. Biological functions and molecular mechanisms of the lncRNAs were revealed using in vitro and in vivo experimental methods. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Experiments and outcome measurements were performed in duplicate or triplicate. Wilcoxon signed-rank test was employed for comparison of RNA levels in clinical cohorts. Analysis of variance was employed for comparisons among multiple groups. RESULTS AND LIMITATIONS: In most patients with PCa, TTTY15 was the most elevated lncRNA located on the Y chromosome. Knockout of this lncRNA by two different CRISPR-Cas9 strategies suppressed PCa cell growth both in vitro and in vivo. TTTY15 promoted PCa by sponging the microRNA let-7, consequently increasing CDK6 and FN1 expression. FOXA1 is an upstream regulatory factor of TTTY15 transcription. CONCLUSIONS: The Y-chromosomal lncRNA TTTY15 was upregulated in most PCa tissues and could promote PCa progression by sponging let-7. PATIENT SUMMARY: We found that TTTY15 levels were frequently elevated in prostate cancer (PCa) tissues compared with those in paracancerous normal tissues in a large group of PCa patients, and we observed a tumour suppressive effect after TTTY15 knockout using CRISPR/Cas9. These results may have therapeutic implications for PCa patients.

Human Adipose-Derived Stem Cells Promote Seawater-Immersed Wound Healing by Activating Skin Stem Cells via the EGFR/MEK/ERK Pathway.

Seawater (SW) immersion can increase the damage of skin wounds and produce refractory wounds. However, few studies have been conducted to investigate the mechanisms of SW immersion on skin wounds. In our current study, we investigated the effect of human adipose-derived stem cells (hADSCs) on the repair of SW-treated full-thickness skin wounds and the underlying mechanisms. The results showed that SW immersion could reduce the expression of EGF and suppress the activation of the MEK/ERK signaling pathway. At the same time, the proliferation and migration of skin stem cells were inhibited by SW immersion, resulting in delayed wound healing. However, hADSCs significantly accelerated the healing of SW-immersed skin wounds by promoting cell proliferation and migration through the aforementioned mechanisms. Our results indicate a role for hADSCs in the repair of seawater-immersed skin wounds and suggest a potential novel treatment strategy for seawater-immersed wound healing.

Trends in the activity of rheumatoid arthritis as the consequence of treat-to-target strategy: eight-year data from 2009 to 2016.

OBJECTIVES: To investigate the trends in the activity of rheumatoid arthritis (RA) over the past 8 years and evaluate the value of treat-to target (T2T) strategy in daily practice. METHODS: All the medical records of RA patients from 2009 to 2016 were retrospectively reviewed. Disease activity scores at obtained visits were measured by DAS28-CRP, DAS28-ESR, SDAI and CDAI. To display trends over years, both mean and time-adjusted methods were applied in calculation of annual disease activity and remission rate. Disease activity and remission rate were also compared before and after the year 2011 when application of T2T strategy was initiated in our centre. Furthermore, a sub-cohort study including T2T and non-T2T period groups was conducted with outcome of cumulative percentage of remission and time to achieve first remission during the first year follow-up. RESULTS: In total, 1,001 patients with 6,944 clinical visits were included. Over an eight-year period, significant improvements were witnessed in disease activity and remission rate, measured by all four indices (p<0.0001). More patients achieved lower disease activity and higher remission rates after T2T adherence in 2011 compared to those in the years 2009 and 2010 (p<0.0001). Moreover, sub-cohort study revealed that more patients (49.3-73.2% vs. 19.1-34.5%, OR=2.4-3.0) achieved remission with a shorter median time compared with the non-T2T period group (p<0.0001), particularly in DAS28-CRP (21 vs. >52 weeks), DAS28-ESR (37 vs. >52 weeks). CONCLUSIONS: Over the past 8 years, the RA activity has substantially decreased and T2T strategy was directly attributable to the favourable changes in clinical practice.

Exploitation of heterosis loci for yield and yield components in rice using chromosome segment substitution lines.

We constructed 128 chromosome segment substitution lines (CSSLs), derived from a cross between indica rice (Oryza sativa L.) 9311 and japonica rice Nipponbare, to investigate the genetic mechanism of heterosis. Three photo-thermo-sensitive-genic male sterile lines (Guangzhan63-4s, 036s, and Lian99s) were selected to cross with each CSSL to produce testcross populations (TCs). Field experiments were carried out in 2009, 2011, and 2015 to evaluate yield and yield-related traits in the CSSLs and TCs. Four traits (plant height, spikelet per panicle, thousand-grain weight, and grain yield per plant) were significantly related between CSSLs and TCs. In the TCs, plant height, panicle length, seed setting rate, thousand-grain weight, and grain yield per plant showed partial dominance, indicating that dominance largely contributes to heterosis of these five traits. While overdominance may be more important for heterosis of panicles per plant and spikelet per panicle. Based on the bin-maps of CSSLs and TCs, we detected 62 quantitative trait loci (QTLs) and 97 heterotic loci (HLs) using multiple linear regression analyses. Some of these loci were clustered together. The identification of QTLs and HLs for yield and yield-related traits provide useful information for hybrid rice breeding, and help to uncover the genetic basis of rice heterosis.

Design considerations of high-performance InGaAs/InP single-photon avalanche diodes for quantum key distribution.

InGaAs/InP single-photon avalanche diodes (SPADs) are widely used in practical applications requiring near-infrared photon counting such as quantum key distribution (QKD). Photon detection efficiency and dark count rate are the intrinsic parameters of InGaAs/InP SPADs, due to the fact that their performances cannot be improved using different quenching electronics given the same operation conditions. After modeling these parameters and developing a simulation platform for InGaAs/InP SPADs, we investigate the semiconductor structure design and optimization. The parameters of photon detection efficiency and dark count rate highly depend on the variables of absorption layer thickness, multiplication layer thickness, excess bias voltage, and temperature. By evaluating the decoy-state QKD performance, the variables for SPAD design and operation can be globally optimized. Such optimization from the perspective of specific applications can provide an effective approach to design high-performance InGaAs/InP SPADs.

Experimental measurement-device-independent quantum key distribution.

Quantum key distribution is proven to offer unconditional security in communication between two remote users with ideal source and detection. Unfortunately, ideal devices never exist in practice and device imperfections have become the targets of various attacks. By developing up-conversion single-photon detectors with high efficiency and low noise, we faithfully demonstrate the measurement-device-independent quantum-key-distribution protocol, which is immune to all hacking strategies on detection. Meanwhile, we employ the decoy-state method to defend attacks on a nonideal source. By assuming a trusted source scenario, our practical system, which generates more than a 25 kbit secure key over a 50 km fiber link, serves as a stepping stone in the quest for unconditionally secure communications with realistic devices.

Metropolitan all-pass and inter-city quantum communication network.

We have demonstrated a metropolitan all-pass quantum communication network in field fiber for four nodes. Any two nodes of them can be connected in the network to perform quantum key distribution (QKD). An optical switching module is presented that enables arbitrary 2-connectivity among output ports. Integrated QKD terminals are worked out, which can operate either as a transmitter, a receiver, or even both at the same time. Furthermore, an additional link in another city of 60 km fiber (up to 130 km) is seamless integrated into this network based on a trusted relay architecture. On all the links, we have implemented protocol of decoy state scheme. All of necessary electrical hardware, synchronization, feedback control, network software, execution of QKD protocols are made by tailored designing, which allow a completely automatical and stable running. Our system has been put into operation in Hefei in August 2009, and publicly demonstrated during an evaluation conference on quantum network organized by the Chinese Academy of Sciences on August 29, 2009. Real-time voice telephone with one-time pad encoding between any two of the five nodes (four all-pass nodes plus one additional node through relay) is successfully established in the network within 60 km.