bluuebunny/crossref_metadata_2025_split huggingface dataset:
A multiplex guide RNA expression system and its efficacy for plant genome engineering
Authors: Oh Youngbin , Lee Bora , Kim Hyeonjin , Kim Sang-Gyu 
Published: February 2020

Abstract
Background
The Streptococcus pyogenes CRISPR system is composed of a Cas9 endonuclease (SpCas9) and a single-stranded guide RNA (gRNA) harboring a target-specific sequence. Theoretically, SpCas9 proteins could cleave as many targeted loci as gRNAs bind in a genome.
Results
We introduce a PCR-free multiple gRNA cloning system for editing plant genomes. This method consists of two steps: (1) cloning the annealed products of two single-stranded oligonucleotide fragments harboring a complimentary target-binding sequence on each strand between tRNA and gRNA scaffold sequences in a pGRNA vector; and (2) assembling tRNA-gRNA units from several pGRNA vectors with a plant binary vector containing a SpCas9 expression cassette using the Golden Gate assembly method. We validated the editing efficiency and patterns of the multiplex gRNA expression system in wild tobacco (Nicotiana attenuata) protoplasts and in transformed plants by performing targeted deep sequencing. Two proximal cleavages by SpCas9-gRNA largely increased the editing efficiency and induced large deletions between two cleavage sites.
Conclusions
This multiplex gRNA expression system enables high-throughput production of a single binary vector and increases the efficiency of plant genome editing.
Optical coherence tomography angiography evaluation of the effects of phacoemulsification cataract surgery on macular hemodynamics in Chinese normal eyes
Authors: Jia Xinyu , Wei Yinjuan , Song Hui 
Published: February 2020

Abstract
Background: To quantitatively evaluate the possible effects of phacoemulsification cataract surgery on macular hemodynamics using optical coherence tomography angiography (OCTA). Methods ： Prospective observational study. Superficial and deep macular vascular densities as well as parameters of foveal avascular zone (FAZ) were measured preoperatively (baseline) and at 1 day, 1 week and 4 weeks postoperation in normal eyes (≥22 mm and≤24 mm) of patients scheduled for phacoemulsification cataract surgery with intraocular lens implantation. Correlations between the rate of change of pre- and post-operative vascular densities and surgical parameters were analyzed. Results : 123 eyes of 107 patients were recruited. Compared with baseline measurements, no statistically significant variation was found in macular vascular densities of day one after surgery (P>0.05). While both superficial and deep macular vascular densities were significantly increased postoperatively on week 1 and week 4 (P<0.05; P<0.05). There were no statistically significant differences in any of the FAZ parameters between the baseline measurements and the entire follow-up period (P>0.05 for all). There were no statistically significant correlations between main surgical parameters and the macular vascular densities. Conclusions : In normal eyes, macular blood perfusion gradually increased after phacoemulsification cataract surgery and was stabilized in one week. Foveal avascular zone was basically stable before and after surgery. Main parameters and intraoperative perfusion of phacoemulsification surgery may not be the key factors affecting macular hemodynamics.
Management of Retinitis Pigmentosa via Platelet Rich Plasma or Combination with Electromagnetic Stimulation: Retrospective Analysis of One-year Results
Authors: Arslan Umut , Özmert Emin 
Published: February 2020

Abstract
Purpose To investigate whether the natural progression rate of retinitis pigmentosa can be decreased with subtenon autologous platelet rich plasma application alone or combination with retinal electromagnetic stimulation.Material and methods The study includes retrospective analysis of 60 retinitis pigmentosa patients. Patients constitute 3 groups with similar demographic characteristics. The combined management group consists of 20 retinitis pigmentosa patients (40 eyes) who received combined retinal electromagnetic stimulation and subtenon platelet rich plasma as Group1; The subtenon platelet rich plasma-only group consisted of 20 retinitis pigmentosa patients (40 eyes) as Group2; The natural course (control) group consists of 20 retinitis pigmentosa patients (40 eyes) who did not receive any treatment were classified as Group3. Horizontal and vertical ellipsoid zone width, fundus perimetry deviation index and best corrected visual acuity changes were compared within and between groups after one year follow up period.Results Horizontal ellipsoid zone percentage changes were detected +1 % in Group1, -2.85% in Group2, -9.36% in Group3 (Δp 1>2>3). Vertical ellipsoid zone percentage changes were detected +0.34 % in Group1, -3.05% in Group2, -9.09% in Group3 (Δp 1>2>3). Fundus perimetry deviation index percentage changes were detected +0.05% in Group1, -2.68% in Group2 and -8.78% in Group3 (Δp 1>2>3). Conclusion Platelet-rich plasma is a good source of growth factors, but its half-life is 4-6 months. Subtenon autologous platelet rich plasma might more effectively slow down photoreceptor loss when repeated as booster injections and combined with retinal electromagnetic stimulation.
Combined and interaction effect of Chlamydia pneumoniae infection and smoking on lung cancer: a case-control study in southeast China
Authors: Xu Xin , Liu Zhiqiang , Xiong Weimin , Qiu Minglian , Kang Shuling , Xu Qiuping , Cai Lin , He Fei 
Published: February 2020

Abstract
Background: This case-control study investigated the role of Chlamydia pneumoniae (Cpn) infection in the pathogenesis of lung cancer and the combined and interaction effect of Chlamydia pneumoniae infection and smoking or other environmental factors. Methods: The study was comprised of 449 lung cancer patients and 512 age- and gender-matched healthy controls. All participants provided a 5-ml fasting peripheral venous blood sample for testing Cpn-specific IgG and IgA by using micro-immunofluorescence. Besides analyzing the associations between Cpn and lung cancer, combined effect analysis, logistic regression, and the excel table made by Andersson were used to analyze the combined and interaction effects of Cpn and environmental factors on lung cancer. Results: Compared to those with no evidence of serum Cpn IgA or Cpn IgG, those with both Cpn IgG+ and IgA+ had 2.00 times the risk (95% CI: 1.34 - 3.00) of developing lung cancer. Smokers with Cpn IgG+ or IgA+ were associated with a significantly increased risk of lung cancer, the adjusted OR was 1.79 (95% CI: 1.10-2.91) or 2.27(95% CI:1.38-3.72), respectively. Those exposed to passive smoking with Cpn IgG+ or IgA+ also increased the risk of lung cancer, the adjusted OR was 1.82 (95% CI: 1.20-2.77) or 1.87(95% CI:1.22-2.87), respectively. The similar results were also observed among alcohol drinking people. Multiplicative and additive interactions were not observed between Cpn infection and environmental factors. The combined effects of Cpn IgG+ or IgA+ and smoking, passive smoking, family history of cancer were found on lung cancer. Conclusion: The Cpn infection was potentially associated with primary lung cancer in the Chinese Han population and had combined effects with smoking, passive smoking, and the family history of cancer. Keywords: Chlamydia pneumoniae infection, case-control study, environmental factors, lung cancer
The exploration of use of an online knowledge base on patient experiences and health care quality: An observational cohort study in primary health care
Authors: Gerdeskold Christian , Toth-Pal Eva , Wårdh Inger , Nilsson Gunnar H. , Nager Anna 
Published: February 2020

Abstract
Background Evidence-based information available at the point of care improves patient care outcomes. Online knowledge bases can increase the application of evidence-based medicine and influence patient outcome data which may be captured in quality registries. The aim of this study was to explore the effect of use of an online knowledge base on patient experiences and health care quality. Methods The study was conducted as a retrospective, observational study of 24 primary health care centers in Sweden exploring their use of an online knowledge base. Frequency of use was compared to patient outcomes in two national quality registries. A socio-economic care need index was applied to assess whether the burden of care influenced the results from those quality registries. Non-parametric statistical methods and linear regression were used. Results Frequency of knowledge base use showed two groups: frequent and non-frequent users, with a significant use difference between the groups (p<0.001). Outcome data showed significant higher values for all seven National Primary Care Patient Survey dimensions in the frequent compared to the non-frequent knowledge base users (p<0.001), whereas 10 out of 11 parameters in the National Diabetes Register showed no differences between the groups (p>0.05). Adjusting for care need index had almost no effect on the outcomes for the groups. Conclusions Frequent users of a national online knowledge base received higher ratings on patient experiences, but figures on health care quality in diabetes showed near to no correlation. The findings indicate that some effects may be attributed to the use of knowledge bases and requires a controlled evaluation.
Inhibition of HIF1A-AS1 promoted starvation-induced hepatocellular carcinoma cells apoptosis by reducing HIF-1α/mTOR mediated autophagy
Authors: Hong Fenfen , Li Yang , Gao Yu , Zheng Linfeng , Li Xianpeng , Xu Feng 
Published: February 2020

Abstract
Background: Hepatocellular carcinoma (HCC) is still a major health burden in China considering its high incidence and mortality. Long non-coding RNAs (lncRNAs) were found playing vital roles in tumor progression, suggesting a new way of diagnosis and prognosis prediction, or treatment of HCC. This study was designed to investigate the role of HIF1A-AS1 during the progression of HCC, and to explore its related mechanisms. Methods: The expression of HIF1A-AS1 was detected in 50 paired carcinoma tissues and adjacent normal tissues by quantitative real-time PCR assay. HCC cells apoptosis was induced by nutrient-deficient culture medium, and detected by Cell Counting Kit‐8 and flow cytometer assays. HIF1A-AS1 inhibition in HCC cells was accomplished by small interfering RNA transfection. Results: HIF1A-AS1 was overexpressed in HCC tissues and was associated with tumor size, TNM stage and lymph node metastasis. Compared with low HIF1A-AS1 group, high HIF1A-AS1 group had a shorter overall survival and a worse disease-free survival. HIF1A-AS1 expression was significantly higher in HCC cell lines (7721 and Huh7) than that in normal hepatocyte cell line L02 under normal culture condition. However, under nutrient-deficient condition, HIF1A-AS1 expression was significantly increased in both HCC and normal hepatocyte cell lines, and was increased with the prolongation of nutrient-free culture. inhibition of HIF1A-AS1 promoted starvation-induced HCC cells apoptosis. Furthermore, inhibition of HIF1A-AS1 could also reduce starvation-induced HCC cells autophagy. The expression of HIF-1α and phosphorylated mTOR was significantly decreased in HCC cells after HIF1A-AS1 inhibition. Conclusions: HIF1A-AS1, overexpressed in HCC and associated with HCC prognosis, could regulate starvation-induced HCC cells apoptosis by reducing HIF-1α/mTOR mediated autophagy, promoting HCC cell progression. Trial registration: This research is retrospectively registered.
Clinical Application of the Reverse Puncture Technique for Total Laparoscopic Colorectal Resection with Natural Orifice Specimen Extraction
Authors: Kong Qi , Xia Yabin , Hao Hu , Jina Yan , Wu Longchao , Xu Li , Huang Xiaoxu 
Published: February 2020

Abstract
Objective To explore the feasibility, safety and short-term effects of the reverse puncture technique for total laparoscopic colorectal resection with natural orifice specimen extraction surgery (NOSES). Materials and Methods The clinical data of 28 patients undergoing total laparoscopic anterior resection with NOSE from July 2016 to July 2019 were retrospectively analysed. The operation was performed following the principle of total mesorectal excision. The reverse puncture technique was used to put the anvil into the proximal sigmoid colon. Then, intra-abdominal anastomosis of the proximal colon and distal rectum was carried out, and the short-term efficacy was evaluated. Results All 28 patients successfully underwent surgery, and the average operative time was 186 (160~220) min. The average time of anvil placement was 15 (12~18) min, the blood loss volume ranged from 60 to 150 ml, and the average length of hospital stay was 9.2 (7-11) d. All of the distal resection margins were confirmed to be negative by the pathological results. No surgery-associated complications, such as anastomosis stricture, leakage, or bleeding, were observed, and no local recurrence or metastasis occurred after 6 months of follow-up. Conclusion Reverse puncture for total laparoscopic colorectal resection with NOSE can avoid purse-string sutures under laparoscopy and avoid the auxiliary incision at the abdominal wall, which minimizes pain after operation. This approach is safe and feasible with a shortened hospital stay and rapid recovery, which results in good short-term clinical outcomes.
Efficacy of Hyperthermic Intraperitoneal Chemotherapy (HIPEC) in the Management of Malignant Ascites
Authors: Jiao Jie , Li Chengzhen , Yu Guanying , Zhang Lei , Shi Xiaoyan , Yan Jingdu , Zhang Houjun , Guo Peiming 
Published: February 2020

Abstract
Objective: The purpose of this study is to compare the difference of clinical efficacy between conventional intraperitoneal chemotherapy and HIPEC, so as to explore the clinical application value and advantages of HIPEC. Design: A retrospective analysis was conducted on 80 patients with malignant ascites admitted to our hospital from June 2017 to June 2019. The general clinical data and qualitative data of the treatment results of 80 patients with malignant ascites were processed by SPSS19.0 using χ2 test and quantitative data were processed by t test. P <0.05, statistical data can be considered statistically significant. Results: 1. There was no significant change in vital signs and temperature in the observation group during the treatment, and the difference was not statistically significant 2. The short-term total effective rate of patients in the observation group was 91.11%, and the short-term total effective rate of the patients in the control group was 40%. 3. There was no significant difference in the incidence of adverse reactions between the two groups of patients. Conclusion: Intraperitoneal hyperthermic chemotherapy combined with intravenous chemotherapy can significantly control malignant ascites, and has small adverse reactions, which is worthy of clinical promotion and application. Key words: efficacy; malignant ascites; intraperitoneal hyperthermic chemotherapy.
Functional production of human antibody by the filamentous fungus Aspergillus oryzae
Authors: Huynh Hung Hiep , Morita Naoki , Sakamoto Toshihiro , Katayama Takuya , Miyakawa Takuya , Tanokura Masaru , Chiba Yasunori , Shinkura Reiko , MARUYAMA Jun-ichi 
Published: February 2020

Abstract
Background: Monoclonal antibodies (mAbs) as one of the biopharmaceuticals take a pivotal role in the current therapeutic application. Generally, mammalian cell lines such as Chinese hamster ovary (CHO) cell lines are used to produce the recombinant antibody. However, there are still concerns about the high cost and the risk of pathogenic contamination when using mammalian cells. Aspergillus oryzae, the filamentous fungus recognized as a GRAS (Generally Regarded As Safe) organism, has an ability to secrete a large amount of proteins into the culture supernatant, and thus the fungus has been used as one of the cost-effective microbial hosts for heterologous protein production. Pursuing this strategy, human anti-TNFα antibody adalimumab, one of the world’s best-selling antibodies for the treatment of immune-mediated inflammatory diseases including rheumatoid arthritis, was chosen to attempt for producing the full length of mAbs by A. oryzae. Generally, N-glycosylation in antibody affects the immune effector functions such as antibody-dependent cell-mediated cytotoxicity (ADCC) via binding to the Fc receptor (FcγR) on immune cells. The CRISPR/Cas9 system was used to firstly delete the Aooch1 encoding a key enzyme for the hyper-mannosylation process in fungi to investigate the binding ability of antibody with FcγRIIIa.Results: The adalimumab was expressed in A. oryzae by the fusion protein system with α-amylase AmyB. The full-length adalimumab consisting of two heavy and two light chains was successfully produced in the culture supernatants. Among the producing strains, the highest amount of antibody was obtained from the ten-protease deletion strain (39.7 mg/L). Two-step purifications by Protein A and size-exclusion chromatography were applied to obtain the high purity sample for further analysis. The antigen-binding and TNFα neutralizing activities of the adalimumab produced by A. oryzae were comparable with those of the commercial product – Humira. No apparent binding with the FcγRIIIa was detected with the recombinant adalimumab even by altering the N-glycan structure under the Aooch1 deletion, which suggests only a little additional activity of immune effector functions.Conclusion: These results demonstrated an alternative low-cost platform for human antibody production by using A. oryzae, possibly offering a reasonable expenditure for patient’s welfare.
Determinants of Obesity and Metabolic Health in the Afghan Population: Protocol, Methodology and Preliminary Results
Authors: Sahrai Mohammad Sediq , Huybrechts Inge , Biessy Carine , Rinaldi Sabina , Ferrari Pietro , Wasiq Abdul Wahed , Gunter Marc J. , Dossus Laure 
Published: February 2020

Abstract
Background: Non-communicable diseases (NCDs) cause more than 70% of deaths worldwide and share modifiable risk factors including obesity and metabolic abnormalities. Over the past 15 years many changes in lifestyle, dietary patterns, physical activity and socioeconomic status have been observed in the Afghan population. This study aims to investigate which specific lifestyle factors, dietary patterns and characteristics of Westernization are associated with an increased risk of being overweight or obese and with poor metabolic health in the Afghan population.Methods: A population-based cross-sectional study was conducted where a total of 729 male and female participants were recruited. Face to face interviews and anthropometric measurements were conducted by trained health staff using standardized questionnaires which included information on socio-demographic and housing characteristics, income, occupation, ethnicity, personal and family medical history, stress, anthropometry, diet and physical activity. Bioelectric impedance analysis (BIA) was used to estimate body composition, including overall body fatness. Physical activity was measured by using the short version of the International Physical Activity Questionnaire (IPAQ). For a comprehensive assessment of dietary intake, a food frequency questionnaire (FFQ) specific to the Afghan population was developed which included all local food items relevant to the population. Lipid profile and fasting glucose were measured in a local laboratory. Biospecimens were collected using dried blood spots (DBS) and faecal occult blood test (FOBT) cards to perform microbiome and biomarker-based research.Discussion: This is the first study which will assess dietary patterns, lifestyle factors and their association with obesity and metabolic health in Afghanistan. Such a study will aid the development of dietary and lifestyle guidelines in Afghanistan which will promote better health and educate people to make healthy food choices. The findings will also help in designing and implementing effective public health strategies to promote a healthy lifestyle and prevent the epidemic of overweight and obesity, and hence, reduce the burden of non-communicable diseases in the region.
Juvenile Spondyloarthritis: focus on uveitis.
Authors: Marino Achille , Weiss Pamela F. , Brandon Timothy G. , Lerman Melissa Ann 
Published: February 2020

Abstract
Background Juvenile spondyloarthritis (JSpA) represents a group of inflammatory arthritides with several distinctive features (enthesitis, involvement of spine and sacroiliac joint, HLA-B27 association and development of uveitis). There are limited data on the course of uveitis in children with JSpA. This study aims to estimate the prevalence of uveitis and to look at the presence of HLA-B27 in relation to uveitis occurrence and ocular symptoms in a cohort of JSpA patients. Findings This is a cross sectional/retrospective study involving patients with JSpA followed in a tertiary referral hospital. Two hundred twenty-three patients were enrolled in the study. The prevalent diagnosis was enthesitis-related arthritis (ERA) (62%) followed by juvenile psoriatic arthritis (PsA), undifferentiated arthritis (UA), and the arthropathies associated with inflammatory bowel disease (IBD-A) (18%, 13%, 12%, respectively). Uveitis was reported in twenty-four patients (11%) of the JSpA cohort (JSpA-U). ERA patients had the highest uveitis prevalence (ERA-U) (13%) with similar prevalences in UA, PsA (7%) and in IBD-A (7% each). The prevalence of HLA-B27 positivity was 45% amongst JSpA-U (N=22), with fewer than half of patients with ERA-U HLA-B27 positive (44%). The overall prevalence of symptomatic uveitis was 79%. Neither the likelihood of uveitis, nor of symptomatic uveitis, varied by HLA-B27 status either in the entire cohort nor in those with ERA. Conclusions About one-tenth of patients developed uveitis, the majority of which was symptomatic. Fewer than half of the patients with uveitis were HLA-B27 positive. HLA-B27 status was not statistically associated with either the development of uveitis or symptomaticity of uveitis.