Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9.

The type II CRISPR/Cas9 system has been used widely for genome editing in zebrafish. However, the requirement for the 5'-NGG-3' protospacer-adjacent motif (PAM) of Cas9 from Streptococcus pyogenes (SpCas9) limits its targeting sequences. Here, we report that a Cas9 ortholog from Staphylococcus aureus (SaCas9), and its KKH variant, successfully induced targeted mutagenesis with high frequency in zebrafish. Confirming previous findings, the SpCas9 variant, VQR, can also induce targeted mutations in zebrafish. Bioinformatics analysis of these new Cas targets suggests that the number of available target sites in the zebrafish genome can be greatly expanded. Collectively, the expanded target repertoire of Cas9 in zebrafish should further facilitate the utility of this organism for genetic studies of vertebrate biology

Enhanced excitability of small dorsal root ganglion neurons in rats with bone cancer pain

<p>Abstract</p> <p>Background</p> <p>Primary and metastatic cancers that affect bone are frequently associated with severe and intractable pain. The mechanisms underlying the development of bone cancer pain are largely unknown. The aim of this study was to determine whether enhanced excitability of primary sensory neurons contributed to peripheral sensitization and tumor-induced hyperalgesia during cancer condition. In this study, using techniques of whole-cell patch-clamp recording associated with immunofluorescent staining, single-cell reverse-transcriptase PCR and behavioral test, we investigated whether the intrinsic membrane properties and the excitability of small-sized dorsal root ganglion (DRG) neurons altered in a rat model of bone cancer pain, and whether suppression of DRG neurons activity inhibited the bone cancer-induced pain.</p> <p>Results</p> <p>Our present study showed that implantation of MRMT-1 tumor cells into the tibial canal in rats produced significant mechanical and thermal hyperalgesia in the ipsilateral hind paw. Moreover, implantation of tumor cells provoked spontaneous discharges and tonic excitatory discharges evoked by a depolarizing current pulse in small-sized DRG neurons. In line with these findings, alterations in intrinsic membrane properties that reflect the enhanced neuronal excitability were observed in small DRG neurons in bone cancer rats, of which including: 1) depolarized resting membrane potential (RMP); 2) decreased input resistance (R_in); 3) a marked reduction in current threshold (CT) and voltage threshold (TP) of action potential (AP); 4) a dramatic decrease in amplitude, overshot, and duration of evoked action potentials as well as in amplitude and duration of afterhyperpolarization (AHP); and 5) a significant increase in the firing frequency of evoked action potentials. Here, the decreased AP threshold and increased firing frequency of evoked action potentials implicate the occurrence of hyperexcitability in small-sized DRG neurons in bone cancer rats. In addiotion, immunofluorescent staining and single-cell reverse-transcriptase PCR revealed that in isolated small DRG neurons, most neurons were IB4-positive, or expressed TRPV1 or CGRP, indicating that most recorded small DRG neurons were nociceptive neurons. Finally, using in vivo behavioral test, we found that blockade of DRG neurons activity by TTX inhibited the tumor-evoked mechanical allodynia and thermal hyperalgesia in bone cancer rats, implicating that the enhanced excitability of primary sensory neurons underlied the development of bone cancer pain.</p> <p>Conclusions</p> <p>Our present results suggest that implantation of tumor cells into the tibial canal in rats induces an enhanced excitability of small-sized DRG neurons that is probably as results of alterations in intrinsic electrogenic properties of these neurons. Therefore, alterations in intrinsic membrane properties associated with the hyperexcitability of primary sensory neurons likely contribute to the peripheral sensitization and tumor-induced hyperalgesia under cancer condition.</p

On leptonic width of X(4260)X(4260)

New measurements on cross sections in $e^+e^-\to J/\psi\pi^+\pi^-$, $h_c\pi^+\pi^-$, $D^0D^{*-}\pi^++c.c.$, $\psi(2S)\pi^+\pi^-$, $\omega\chi_{c0}$ and $J/\psi\eta$ channels have been carried out by BESIII, Belle and BABAR collaborations, and also in the $D_s^*\bar D_s^*$ channel. We perform extensive numerical analyses by combining all these data available, together with those in $D\bar D^*$ and $D^*\bar D^*$ channels. Though the latter show no evident peak around $\sqrt{s}=4.230$ GeV, the missing $X(4260)$ is explained as that it is concealed by the interference effects of the well established charmonia $\psi(4040)$, $\psi(4160)$ and $\psi(4415)$. Our analyses reveal that the leptonic decay width of $X(4260)$ ranges from $O(10^2)$ eV to $O(1)$ keV, and hence may be explained in the conventional quark model picture. That is, the $X(4260)$ may well be interpreted as a mixture of $4^3S_1$ and $3^3D_1$ states.Comment: two small mistakes are fixed, figures redrawn, major physical outputs remain unchanged. Version published in EPJ

The diagnostic value of elastography and ultrasound contrast in papillary thyroid microcarcinoma

目的 评估弹性成像与超声造影（CEUS）两种检查技术对鉴别诊断甲状腺微小乳头状癌（TMC）的价值。方法  对常规超声检测出且定性困难的73例80个甲状腺微小结节进行弹性成像及CEUS检查，所有结节均经手术病理证实。比较两种检查方法的准确性。结果 80个结节中CEUS诊断正确率为85.0%（68/80）,其中6例TMC误诊为良性病变，6例良性结节误诊为TMC；弹性成像5分法诊断正确率为92.5%（74/80）,其中3例TMC误诊为良性结节，3例良性结节误诊为TMC。性成像诊断甲状腺微小癌的敏感性94.0%，特异性90.0%，准确性92.5%；CEUS诊断甲状腺微小癌的敏感性88.0%，特异性80.0%，准确性85.0%。结论 CEUS和弹性成像对于诊断TMC方面均有价值，但弹性评分≥3作为诊断TMC的敏感性、特异性及准确性均高于CEUS。Objective: To assess the value of elastic imaging and CEUS two inspection techniques for differential diagnosis of thyroid papillary carcinoma (TMC). Method: To do elastic imaging and CEUS checks to 73 cases of 80 thyroid nodules which was tested by conventional ultrasonic and difficult to quantify. All nodules were confirmed by surgery and pathologic examination. Comparing the accuracy of both detection methods. Result: Of the 80 nodules, the accuracy of CEUS diagnosis was 85.0%（68/80）,  of which 6 cases were misdiagnosed as benign lesions, and 6cases of benign nodules were misdiagnosed as TMC: the accuracy of 5-point scale criteria of elastography was 92.5%（74/80）, of which 3 TMC were misdiagnosed as benign nodules: and 3 benign nodules were misdiagnosed as TMC. The application of elastography in the diagnosis of thyroid microcarcinoma displayed a sensitivity of 94.0%, a specificity of 90.0% and an accuracy of 92.5%. Elastography detection was more advantagerous than CEUS in the diagnosis of thyroid microcarcinoma, and compared to CEUS , the differences were statistically significant（P &lt;0.05）.Conclusion: Elastography and Ultrasound Contrast have highly practical value to diagnosis of TMC.  The sensitivity specificity and accuracy of using elastic score ≥3 as criteria of diagnosis of TMC was higher than that of CEUS

Relationship between Angiotensin-Converting Enzyme Insertion/Deletion Gene Polymorphism and Susceptibility of Minimal Change Nephrotic Syndrome: A Meta-Analysis

Aim. This meta-analysis was performed to evaluate the association between ACE I/D gene polymorphism and MCNS susceptibility. Method. A predefined literature search and selection of eligible relevant studies were performed to collect the data from electronic databases. Results. Six articles were identified for the analysis of association between ACE I/D gene polymorphism and MCNS risk, including 4 for Asians, one in Caucasian population and one for Africans. There was a markedly positive association between D allele or DD genotype and MCNS susceptibility in Asians (D: P = .01, DD: P = .02), but not for Caucasians and Africans (Caucasians: D: P = .16, DD: P = .98; Africans: D: P = .81, DD: P = .49). Furthermore, the II genotype seemed not to play a protective role against MCNS risk for Asians, Caucasians and Africans (P = .12, P = .09, P = .76, resp.). Interestingly, there was also significant association between ACE I/D gene polymorphism and MCNS susceptibility in overall populations (D: P = .007, DD: P = .04, II: P = .03). Conclusion. D allele or DD genotype might be a significant genetic molecular marker for MCNS susceptibility in Asians and overall populations, but not for Caucasians and Africans. More larger and rigorous genetic epidemiological investigations are required to further explore this association

An optical fibre sensor for remotely detecting water traces in organic solvents

First published online 26 Aug 2016Tetraphenylpyrazine-triphenylamine (TPP-TPA) was used to detect water traces in organic solvents by monitoring the shift of the fluorescence peak wavelength. This wavelength based method avoids the intrinsic problems of fluorescence intensity change based methods. The use of optical fibres for the detection provides a remote and field-deployable sensing ability.Mengke Han, Ming Chen, Heike Ebendorff-Heidepriem, Cheng Fang, Anjun Qin, Hu Zhang, Ben Zhong Tang, Youhong Tang and Yinlan Rua

Lipid alternations in the plasma of COVID-19 patients with various clinical presentations.

BACKGROUND: COVID-19 is a highly infectious respiratory disease that can manifest in various clinical presentations. Although many studies have reported the lipidomic signature of COVID-19, the molecular changes in asymptomatic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-infected individuals remain elusive. METHODS: This study combined a comprehensive lipidomic analysis of 220 plasma samples from 166 subjects: 62 healthy controls, 16 asymptomatic infections, and 88 COVID-19 patients. We quantified 732 lipids separately in this cohort. We performed a difference analysis, validated with machine learning models, and also performed GO and KEGG pathway enrichment analysis using differential lipids from different control groups. RESULTS: We found 175 differentially expressed lipids associated with SASR-CoV-2 infection, disease severity, and viral persistence in patients with COVID-19. PC (O-20:1/20:1), PC (O-20:1/20:0), and PC (O-18:0/18:1) better distinguished asymptomatic infected individuals from normal individuals. Furthermore, some patients tested positive for SARS-CoV-2 nucleic acid by RT-PCR but did not become negative for a longer period of time (≥60 days, designated here as long-term nucleic acid test positive, LTNP), whereas other patients became negative for viral nucleic acid in a shorter period of time (≤45 days, designated as short-term nucleic acid test positive, STNP). We have found that TG (14:1/14:1/18:2) and FFA (4:0) were differentially expressed in LTNP and STNP. CONCLUSION: In summary, the integration of lipid information can help us discover novel biomarkers to identify asymptomatic individuals and further deepen our understanding of the molecular pathogenesis of COVID-19