Precise and efficient genome editing is very important for gene functional characterization. In recent years, sequence-specific DNA nucleases have been developed to increase the efficiency of gene targeting or genome editing in animals and plants. Among them, Zinc-Finger Nucleases (ZFNs) and Transcription Activator-Like Effector Nucleases (TALENs) are two most commonly used sequencespecific chimeric proteins (Gaj et al., 2013). Recently, a breakthrough gene-targeting tool based on RNA-guided Cas9 nuclease from type II prokaryotic Cluster Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated system has been developed (Jinek et al., 2012). CRISPR/Cas system is an adaptive defense system in prokaryotes to fight against alien nucleic acids (Horvath and Barrangou, 2010). The CRISPR loci are variable short spacers separated by short repeats, which are transcribed into synthetic single-guide RNA (sgRNA). The sgRNA forms a functional complex with CRISPR-associated nuclease (Cas9) and guide the nuclease to genomic loci matching a 20-bp complementary invading DNA, cleaving it immediately upstream of a required 5'-NGG Protospacer Adjacent Motif (PAM). A chimeric sgRNA that mimics the natural synthetic sgRNA can be used to target Cas9 for genome editing in eukaryotic cells.Until now, the CRISPR/Cas system has been successfully applied to efficient genome editing in bacteria, animals and plants (Jiang et al., 2013; Mali et al., 2013; Xie and Yang, 2013). To inform the selection of target sites and avoid off-target effects, Hsu and colleagues characterized Cas9 targeting specificity in human cells by a volume of experiments, and built a computational tool for optimized sgRNA selection in human and animal genomes (Hsu et al., 2013). However, with the wide application of CRISPR-system in plant genome editing, tools for optimized sgRNA selection in plants are highly needed. In the current analysis, we present a web application tool-CRISPR-P, for CRISPR sgRNA design in more than 20 plant species. CRISPR-P allows users to search for highly specific Cas9 target sites within DNA sequences of interest, which also provides off-target loci prediction for further analysis and marks restriction enzyme cutting sites for convenience. CRISPR-P is freely available at http://cbi.hzau.edu.cn/crispr/.Genomic data and annotation of the analyzed plant genomes are obtained from public databases. The genome and annotation of Arabidopsis lyrata (v.1.0), Arabidopsis thaliana (TAIR10), Brachypodium distachyon (v1.0), Brassica

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High tibial osteotomy (HTO) is a promising surgery that can treat osteoarthritis of the medial septum of the knee. However, the extensive release of soft tissue and the osteotomy gap may produce intraoperative and postoperative bone bleeding. Tranexamic acid (TXA) is an effective blood management strategy, as it competitively inhibits the activation process of plasminogen and prevents fibrinolytic enzymes from degrading fibrin. Therefore, we compared the operative bone bleeding of patients who underwent HTO who received either intravenous (IV) or topical TXA in this research. The medical records of a total of 191 patients (including 72 who received IV TXA, 64 who received topical TXA and 55 control patients) who received open-wedge HTO were retrospectively reviewed from January 2016 to August 2019. There were no obvious demographic differences between the groups. Here, we used independent parameters to assess the efficacy of topical and IV TXA in reducing blood loss. Compared with the IV TXA group, patients receiving topical TXA therapy had greater blood loss (622 ± 231 ml versus 451 ± 231 ml, mean difference 171 mL [95% CI, 87–254]; p < 0.001). The hemoglobin concentration of the IV TXA group was obviously higher than that of the topical medication group. No patients had thromboembolic complications during the entire study period. In our study, it seemed that either IV or topical use of TXA might reduce blood loss after open-wedge HTO, and the blood loss and amount of drainage in the IV TXA group showed huge decreases compared to those in the topical group.

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Our study first established a connection between MMP9 and OPCAB procedure, suggesting that MMP9 could be a novel biomarker for identifying perioperative myocardial injury in patients undergoing OPCAB.

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Varicocele is a common abnormality, but the conventional microsurgical subinguinal varicocelectomy (CMSV) has some disadvantages. We invented Modified Inguinal Microscope-Assisted Varicocelectomy (MIMV) under local anesthesia. This study aims to evaluate MIMV by comparing it to CMSV in operating duration, time to return to normal activity, postoperative complications, achievement of natural pregnancy and improvement of semen quality for patients with infertility, pain score for those with scrotal pain, and so on. We enrolled 3089 patients who underwent MIMV and 476 who underwent CMSV in our hospital. Both the operating duration and the time to return to normal activity of MIMV was shorter than that of CMSV (P < 0.001). The recurrence rate (P < 0.001) and injury rate of vas deferens (P = 0.011) after MIMV were lower than that after CMSV. Moreover, patients with MIMV showed higher degree of satisfaction with the surgery experience and outcome than those with CMSV (P < 0.001). However, no statistical difference was found between the two groups in scores of pain due to surgery, postoperative varicose veins diameters, reflux duration, and the postoperative complications of wound infection, hydrocele, atrophy of testis, epididymitis, and scrotal hematoma. In summary, MIMV is a promising varicocelectomy and could be applied more in clinical practice.

Purpose This study aimed to explore the feasibility and safety of the tunnel approach in laparoscopic radical right hemicolectomy for colon cancer. Methods From July 2016 to October 2018, a total of 106 consecutive patients with colon cancer who underwent laparoscopic radical right hemicolectomy at the Affiliated Cancer Hospital of Zhengzhou University were enrolled. The patients were stratified into either a tunnel approach (TA) (n = 56) group or traditional medial approach (MA) (n = 50) group according to the surgical technique performed. The baseline demographics, perioperative outcomes and oncologic outcomes were compared between the two groups. Results The baseline characteristics did not differ between groups. The TA group had significantly less blood loss [20.0 (10.0–40.0) vs. 100 (100.0–150.0) ml, p < 0.001] and a shorter operation time [128.4 ± 16.7 vs. 145.6 ± 20.3 min, p < 0.001] than the MA group. The time to first flatus and postoperative hospital stay were similar [3.0 (2.0–4.0) vs. 3.0 (3–4.0) days, p = 0.329; 10.4 ± 2.6 vs. 10.7 ± 3.0 days, p = 0.506] between the two groups. The conversion to laparotomy and complication rates were similar between groups (0 vs. 6.0%, p = 0.203; 14.3% vs. 18.0%, p = 0.603, respectively). No treatment-related deaths occurred in either group. The TA group did not have significantly better survival outcomes than the MA group (p = 0.372). Conclusions The TA seems to allow for more favourable results in terms of blood loss and operative time than the MA, with similar results regarding time to first flatus, hospital stay, postoperative complication rate, conversion rate and oncologic outcomes; moreover, the TA is easier for beginners to master.