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Yuan Huang,1,* Hong Yang,2,* Rui Ding,1,* Li Wang,3 Ji Li,2 Wenbo Li,4 Xuzhen Qin,1 Yingchun Xu,1 Jiaming Qian2 1Department of Laboratory Medicine, Peking Union Medical College Hospital & Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 2Department of Gastroenterology, Peking Union Medical College Hospital & Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 3Department of Epidemiology and Health Statistics, Institute of Basic Medicine, Peking Union Medical College, Beijing, People’s Republic of China; 4Department of Ultrasound, Peking Union Medical College Hospital & Chinese Academy of Medical Sciences, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Xuzhen Qin; Jiaming Qian, Email qxz_01@163.com; qianjiaming1957@126.comObjective: Serum pepsinogen (PG) is a good indicator of atrophic changes in the gastric mucosa. Gastric mucosal atrophy is a high-risk factor for gastric cancer. Serological testing for PG combined with endoscopy can help to improve gastric cancer screening. In this study, we established the reference ranges of serum PG-I, PG-II, and the PG-I/II ratio (PGR) in the Chinese population by chemiluminescence immunoassay (CLIA) and enzyme-linked immunosorbent assay (ELISA). Besides, in the real world, doctors are often confused by the results of different testing platforms. Thus, a comparison of methods CLIA and ELISA was performed.Methods: 2904 individuals were enrolled from six regions in China as part of the Chinese Adult Digestive Diseases Surveillance (2016) program. The individuals completed questionnaires and volunteered to undergo examinations, including gastroscopy, urea breath test, abdominal ultrasound examination and routine serologic tests. Serum was collected to measure PGs (including PG-I, PG-II and PGR) by CLIA and ELISA. Participants who were found obvious abnormalities or absent from the examinations were excluded. Ultimately, 747 healthy individuals were enrolled in this study. The Kolmogorov–Smirnov test was used to assess the distribution of variables. The Kruskal–Wallis H or Mann–Whitney U-tests were used to compare different sex, age, and geographical groups. The 95% reference ranges of PGs obtained by the two methods were established according to document CLSI-EP28-A3, with covariates of sex, age, and region. Spearman correlation analysis, linear regression analysis and allowable total error (ATE) zone analysis were utilized for comparing the two methods.Results: On overall, the 95% reference ranges of PG-I, PG-II, and PGR measured by CLIA were 23.00– 110.64 ng/mL, 2.50– 19.13 ng/mL, and 3.87– 13.30, respectively. Meanwhile, the reference ranges of PG-I, PG-II, and PGR measured by ELISA were 36.93– 205.06 ng/mL, 1.65– 17.96 ng/mL, and 7.50– 33.60, respectively. Both PG-I and PG-II levels measured by the two platforms were found to be influenced by sex and age. PGR measured by CLIA was influenced by age but not by sex, while PGR measured by ELISA was not affected by either age or sex. Regional factors did not significantly impact the PG results, except for PG-I detected by ELISA. Ultimately, reference ranges for PGs were established based on age and sex stratification. Additionally, the Spearman correlation analysis revealed that the correlation coefficients for PG-I, PG-II, and PGR detected by the two methods were 0.899, 0.887, and 0.777, respectively, indicating a strong correlation between the two methods. The regression equation for the PG levels detected by two methods was obtained through linear regression analysis. The ATE analysis provided a visual depiction of the consistency between the two methods, clearly indicating the poor agreement between them.Conclusion: This study established the reference ranges of PGs by strict and intact enrollment standard. In addition, the results indicated a strong linear relationship between the two methods, yet with a clear bias, which was valuable for laboratory interpretation.Keywords: pepsinogen, reference range, gastric cancer

Zhaorui Wang,1 Jingjing Wang,1 Jing Pei,2 Yubo Pan,2 Rui Ding1 1Department of Emergency Surgery, Lu ‘an People’s Hospital, Lu ‘an Hospital Affiliated to Anhui Medical University, Lu ‘an, Anhui, 237000, People’s Republic of China; 2Department of General Surgery, First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230022, People’s Republic of ChinaCorrespondence: Rui Ding, Department of Emergency Surgery, Lu ‘an People’s Hospital, Lu ‘an Hospital Affiliated to Anhui Medical University, No. 21, Wanxi West Road, Lu ‘an, Anhui, 237000, People’s Republic of China, Email Dr20210908@163.comBackground: Commonly, the thyroid gland is regarded as an organ with fewer metastatic diseases, and colorectal metastasis to the thyroid (CMT) is rarely reported, especially, with that the clinical sign of thyroid metastasis nidus is the chief complaint. The CMT occurs in advanced colorectal cancer and is associated with poor prognosis and short survival.Case Report: In this case, we reported a patient with the sign of neck mass as the first manifestation of CMT. The patient underwent a partial thyroidectomy in June 2019, immunohistochemical findings of thyroid carcinoma suggested the possibility of adenocarcinoma of gastrointestinal tract. The patient underwent a colonoscopy in July 2019 and a colonic mass was found. Pathological examination diagnosed rectal adenocarcinoma. The patient underwent neoadjuvant chemotherapy, surgical treatment, postoperative adjuvant chemotherapy and targeted therapy. The patient died in June 2022.Conclusion: The metastasis disease would not be ignored at all, when a patient complains at signs of neck mass. Further, the possibility of metastasis cancer should be considered once thyroid nodules occur in patients with colorectal cancer. Even though the biological characteristics and stage of the primary tumor have an important impact on the prognosis, positive standardized treatments can also be helpful.Keywords: colorectum cancer metastases, secondary thyroid cancer, neck mass, case report, therapy, prognosis

Zhe Yu,1– 3 Ruoqi Ding,1– 3 Qiuju Yan,1– 3 Menghan Cheng,1– 3 Teng Li,1– 4 Fei Zheng,5 Lin Zhu,1– 4 Yang Wang,1– 4 Tao Tang,1– 4 En Hu1– 4 1Institute of Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 2NATCM Key Laboratory of TCM Gan, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 3Center for Interdisciplinary Research in Traditional Chinese Medicine, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China; 4Xiangya Hospital, Central South University, Nanchang, Jiangxi, 330004, People’s Republic of China; 5The College of Integrated Traditional Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha, 410008, People’s Republic of ChinaCorrespondence: En Hu; Ruoqi Ding, Integrative Medicine, Department of Integrated Traditional Chinese and Western Medicine, Xiangya Hospital, Central South University, Changsha, 410008, People’s Republic of China, Email znxyhe@csu.edu.cn; ruoqi_ding@outlook.comPurpose: Many herbs can promote neurological recovery following traumatic brain injury (TBI). There must lie a shared mechanism behind the common effectiveness. We aimed to explore the key therapeutic targets for TBI based on the common effectiveness of the medicinal plants.Material and methods: The TBI-effective herbs were retrieved from the literature as imputes of network pharmacology. Then, the active ingredients in at least two herbs were screened out as common components. The hub targets of all active compounds were identified through Cytohubba. Next, AutoDock vina was used to rank the common compound-hub target interactions by molecular docking. A highly scored compound-target pair was selected for in vivo validation.Results: We enrolled sixteen TBI-effective medicinal herbs and screened out twenty-one common compounds, such as luteolin. Ten hub targets were recognized according to the topology of the protein-protein interaction network of targets, including epidermal growth factor receptor (EGFR). Molecular docking analysis suggested that luteolin could bind strongly to the active pocket of EGFR. Administration of luteolin or the selective EGFR inhibitor AZD3759 to TBI mice promoted the recovery of body weight and neurological function, reduced astrocyte activation and EGFR expression, decreased chondroitin sulfate proteoglycans deposition, and upregulated GAP43 levels in the cortex. The effects were similar to those when treated with the selective EGFR inhibitor.Conclusion: The common effectiveness-based, common target screening strategy suggests that inhibition of EGFR can be an effective therapy for TBI. This strategy can be applied to discover core targets and therapeutic compounds in other diseases.Keywords: traditional Chinese medicine, medicinal plants, luteolin, AZD3759, epidermal growth factor receptor, astrocyte

Huiying Zhang,1,* Daojun Chen,1,2,* Qianqian Ji,1 Meiyan Yang,1 Rui Ding1 1Department of Occupational Health and Environmental Health, School of Public Health, Anhui Medical University, Hefei, Anhui, 230032, People’s Republic of China; 2School of Medical Technology, Anhui Medical College, Hefei, Anhui, 230601, People’s Republic of China*These authors contributed equally to this workCorrespondence: Daojun Chen, Email daodaodao1978@163.comBackground: Microcystin-leucine-arginine (MC-LR) is the most abundant and most toxic variant of microcystin isomers. Various experiments have clearly shown that MC-LR has hepatotoxicity and carcinogenicity, but there are relatively few studies on its immune damage effect. In addition, numerous studies have shown that microRNAs (miRNAs) are involved in a wide range of biological processes. Do miRNAs also play a role in inflammatory response caused by microcystin exposure? This is the question to be answered in this study. Moreover, this study can also provides experimental evidence for the significance of miRNA applications.Objective: To investigate the effect of MC-LR on the expressions of miR-146a and pro/anti-inflammatory cytokines in human peripheral blood mononuclear cells (PBMCs) and to further explore the role of miR-146a in the inflammatory responses caused by MC-LR.Methods: Serum samples from 1789 medical examiners were collected and detect the concentrations of MCs, and 30 serum samples with concentrations of MCs around P25, P50, and p75 were randomly selected for the detection of inflammatory factors. PBMCs from fresh peripheral blood extracted from these 90 medical examiners were subsequently tested for relative miR-146a expression. In vitro, the MC-LR were exposed to the PBMCs to detect the levels of inflammatory factors as well as the relative expression of miR-146a-5p. Then, a miRNA transfection assay was performed to verify the regulation of inflammatory factors by miR-146a-5p.Results: In population samples, the expression of inflammatory factors and miR-146a-5p increased with increasing MCs concentration. In vitro experiments showed that the expression of inflammatory factors and miR-146a-5p in PBMCs increased with MC-LR exposure time or exposure dose too. In addition, inhibiting the expression of miR-146a-5p in PBMCs reduced inflammatory factor levels.Conclusion: miR-146a-5p exerts a promoting effect on the MC-LR-induced inflammatory response by positively regulating inflammatory factor levels.Keywords: microcystin-leucine-arginine, MC-LR, human peripheral blood, mononuclear cells, PBMCs, miR-146a, inflammatory cytokines

Xiubin Zhang,1 Rong Ding,2 Zhaoxin Zhang,3 Mengyun Chen,4 Yueheng Yin,5 Jennifer K Quint1 1National Heart and Lung Institute, Imperial College London, London, W12 0BZ, UK; 2School of Health in Social Science, The University of Edinburgh, Edinburgh, Scotland, EH8 9AG, UK; 3Medical college, Xijing University, Xi’an, 710123, People’s Republic of China; 4School of Nursing, Lanzhou University, Lanzhou, 730000, People’s Republic of China; 5School of Nursing, Nanjing Medical University, Nanjing, 210000, People’s Republic of ChinaCorrespondence: Jennifer K Quint, National Heart and Lung Institute, Imperial College London, White City Campus, London, W12 0BZ, UK, Email j.quint@imperial.ac.ukBackground: Increased medication adherence leads to better asthma control and health outcomes. However, many studies have found that patient adherence to maintenance medication is poor.Aim: We undertook a meta-synthesis of qualitative studies, to investigate asthma patient and healthcare professionals’ perspectives of medication adherence.Methods: This systematic review was reported by following the PRISMA guidelines. The Joanna Briggs Institute (JBI) meta-aggregative approach was used for the qualitative synthesis. The protocol was registered in PROSPERO (CRD42022346831).Results: In total, 12 articles were included in the review. These articles reported findings from 433 participants in total, which included 315 patients and 118 healthcare professionals. Four synthesised findings with sub-themes were identified from the reviewed studies. These synthesised findings were described as: 1) The role of relationship and communication with/between Healthcare Professionals in medication adherence; 2) Insufficient information from Healthcare Professionals acting as a barrier for adherence; 3) How patient’s attitude/beliefs effect their adherence to medication; and 4) Patients’ personal behaviour and other relevant barriers.Conclusion: The synthesized findings provide a strong evidence-base of patient and health professionals’ perspectives and behaviours toward medication adherence, which helps to identify and address non-adherence. Healthcare providers can use these findings to support patients’ adherence to asthma medications. The findings suggest that empowering people to make informed decisions around medication adherence rather than “adherence controlling” by health professionals is very important. Effective dialogue and appropriate education are critical approaches to increase medication adherence.Keywords: asthma patients, healthcare professional, medication adherence, perspectives, systematic review

Xianbin Ding,1 Liling Chen,1 Wenge Tang,1 Ting Chen,1 Jingru Xu,1 Xianxian Yang,1 Rui Ding,2 Xiaojun Tang3 1Chongqing Municipal Center for Disease Control and Prevention, Chongqing, People’s Republic of China; 2First Medical College, Chongqing Medical University, Chongqing, People’s Republic of China; 3School of Public Health and Management, Medical and Social Development Research Center, Chongqing Medical University, Chongqing, People’s Republic of ChinaCorrespondence: Liling Chen, Chongqing Municipal Center for Disease Control and Prevention, Chongqing, People’s Republic of China, Email mbcllgz@163.comObjective: The prevalence of hyperuricemia appears to be high worldwide. We aimed to explore the interaction between harmful alcohol use and tea consumption on hyperuricemia.Methods: This study recruited 22,449 Han residents based on the data from the China Multi-Ethnic Cohort (CMEC) study, Chongqing province, to have a face-to-face electronic questionnaire, physical examination, and clinical laboratory tests. The difference in hyperuricemia between the different populations was compared by the Chi-square test. The interaction between harmful alcohol use and tea consumption was analyzed by the multivariate logistic regression model.Results: Amongst 22,449 participants, the mean age was 51.5± 11.8 years, and 46.83% of them were males. The proportion of harmful alcohol use, tea consumption, and harmful alcohol use and tea consumption were 14.01%, 21.01%, and 6.54%, respectively. Multivariate logistic regression showed that the odds ratio (OR) of harmful alcohol use and tea consumption (OR=2.21, 95% CI: 1.58– 3.10) was greater than that of harmful alcohol use (OR=1.63, 95% CI:1.17– 2.27) and tea consumption (OR=1.34, 95% CI:1.10– 1.63). Among males, the results were similar (harmful alcohol use and tea consumption: OR=2.02, 95% CI: 1.43– 2.84; harmful alcohol use: OR=1.61, 95% CI: 1.14– 2.27; tea consumption: OR=1.28, 95% CI: 1.05– 1.57). However, among females, the odds ratio of harmful alcohol use and tea consumption (OR=15.50, 95% CI: 1.36– 176.50) was more than 10 times than that of only harmful alcohol use (OR=1.55, 95% CI: 0.42– 5.69) or tea consumption (OR=1.22, 95% CI: 0.52– 2.82).Conclusion: The interaction of harmful alcohol use and tea consumption was a positive risk for hyperuricemia in Han residents aged 30– 79 years in China.Keywords: hyperuricemia, tea consumption, harmful alcohol use, interaction, Han residents

Hongguan Jiao,1 Ran Ding,2 Pukai Jin,3 Junwen Wang,4 Jun Xu,5 Weiyi Tian6 1School of Information Engineering, Guizhou University of Traditional Chinese Medicine, Guiyang, People’s Republic of China; 2School of Health Preservation of Traditional Chinese Medicine, Guizhou University of Traditional Chinese Medicine, Guiyang, People’s Republic of China; 3Department of Radiology, Inova Mount Vernon Hospital, Alexandria, VA, USA; 4Institute of Basic Theory of Traditional Chinese Medicine, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 5Information Institute of Traditional Chinese Medicine, China Academy of Chinese Medical Sciences, Beijing, People’s Republic of China; 6School of Basic Medicine, Guizhou University of Traditional Chinese Medicine, Guiyang, People’s Republic of ChinaCorrespondence: Hongguan Jiao, School of Information Engineering, Guizhou University of Traditional Chinese Medicine, Guiyang, People’s Republic of China, Tel +86-18886094606, Fax +86-851-88233048, Email joe7741@163.comIntroduction: With the wide acceptance of acupuncture, many papers and guidelines recommend that acupuncture is effective for chronic pain (CP). In this study, we applied bibliometric methods to analyze the current research situation of acupuncture intervention in CP, to gain insight into the current situation and future development trend of this field.Material and Methods: Science Citation Index Expanded was searched for publications related to acupuncture for CP between 1900 and 2022. VOSviewer, CiteSpace and Thomson Data Analyzer were used to analyze the annual publication, authors and cited authors and their countries (regions) and institutions, journals and cited journals, cited references, co-occurrence keywords, burst keywords, and the relevant centrality.Results: A total of 1968 papers were retrieved, the annual publications have shown a rapid growth trend in the recent 20 years. The USA (708) and the Kyung Hee University (31) were the most productive country and institution, respectively, while the USA (0.37) and University of Maryland (0.13) had the highest centrality. MacPherson, Hugh published the most papers in this field (29), and Vickers, A J were the most influential author (289 times cited). Journal of Alternative and Complementary Medicine was the most productive journal (92), whereas PAIN was the most influential one (4743 times cited). Breivik, H’s (2006) paper had the most citation count (3025), while Furlan’s (2005) paper had the highest centrality (0.23). The research focuses in this field mainly include pain, electroacupuncture, Low back pain, Systematic review, Randomized controlled trial etc. Researchers are currently paying more attention to the psychological problems caused by CP.Conclusion: The research of acupuncture for CP will be further expanded. International cooperation of this research field needs to be further strengthened. More high-quality designed trials need to be conducted.Keywords: chronic pain, acupuncture, bibliometric analysis, VOSviewer, CiteSpace

Menghan Li,1,2,* Ran Ding,3,* Xinming Yang,1,2 Dawei Ran1,2 1Acupuncture-Moxibustion Clinical Department, First Teaching Hospital of Tianjin University of Traditional Chinese Medicine, National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin, 300381, People’s Republic of China; 2Graduate School, Tianjin University of Traditional Chinese Medicine, Tianjin, People’s Republic of China; 3School of Medical Informatics Engineering, Anhui University of Chinese Medicine, Hefei, 230012, People’s Republic of China*These authors contributed equally to this workCorrespondence: Menghan Li, Email limenghan01@126.comPurpose: PSD is a syndrome that occurs after a stroke, which manifests as a series of depressive symptoms and corresponding physiological symptoms. Relevant studies have shown that the drug therapy is often accompanied by drug side effects and patient resistance. Acupuncture has attracted attention as a treatment method without adverse reactions of patients. The purpose of this study was to investigate the possible mechanism of action of acupuncture in PSD.Patients and Methods: Download depression and stroke datasets from public databases. Bioinformatics methods were used to analyze the key gene targets related to stroke and depression. Functional enrichment analysis assesses important pathways. Further screen PSD-related biological pathways and genes. After the experimental model was established, the expression differences of key genes and related pathways were compared between the model group and the control group through acupuncture treatment and qPCR verification.Results: Depression and stroke-related genes were obtained by bioinformatics methods, and then important biological processes and biological pathways related to depression and stroke were analyzed by GO and KEGG. And further screen out the disease targets closely related to PSD. In the follow-up animal experiments, we confirmed that acupuncture can intervene on these key pathways and targets, and then play a role in the targeted therapy of diseases.Conclusion: The results of this study show that five genes (“NRBP1”, “SIRT1”, “BDNF”, “MAPK3”, “CREB1”.) and key biological pathways such as NFkB, PI3K/AKT activation, and MAPK are the keys to the occurrence and development of PSD biomarkers, which can also be therapeutically intervened by acupuncture.Keywords: PSD, bioinformation, biomarkers, acupuncture

The coupling of transport code SOLPS with the turbulence code BOUT++ was reported in Reference [D. R. Zhang et al., Phys. Plasmas 26, 012508 (2019)], while the grids of SOLPS and BOUT++ are not completely consistent with each other, especially in the divertor region. In the present work, a method of replacing the grids of BOUT++ with the grids of SOLPS is proposed to make the simulation region fully consistent with each other for the SOLPS/BOUT++ coupling. A SOLPS grid file is generated with an MHD equilibrium and used in BOUT++ code to simulate the profiles of plasma density, ion temperature, and electron temperature with the six‐field two‐fluid model. The profiles of the main plasma parameters simulated with the SOLPS grids are similar with the profiles simulated with the BOUT++ grids at the midplane, while the profiles are deformed compared with the profiles simulated with the BOUT++ grids at the outer divertor target because of the differences of the distributions of SOLPS grids and BOUT++ grids in the divertor region. The radial particle transport coefficient and heat transport coefficients are also calculated by using the BOUT++ code with the two grids, and the comparisons of the radial particle transport coefficient and heat transport coefficients simulated with the two grids at the midplane and outer divertor target plate are discussed. [ABSTRACT FROM AUTHOR]

Qian Yang,1,* Laichao Ni,1,* Saber Imani,1,* Zhangqiang Xiang,1 Rui Hai,1 Ruilin Ding,1 Shaozhi Fu,1 Jing bo Wu,1,2 Qinglian Wen1 1Department of Oncology, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People’s Republic of China; 2Nuclear Medicine and Molecular Imaging Key Laboratory of Sichuan Province, Department of Nuclear Medicine, The Affiliated Hospital of Southwest Medical University, Luzhou, Sichuan 646000, People’s Republic of China*These authors contributed equally to this workCorrespondence: Qinglian WenDepartment of Oncology, The Affiliated Hospital of Southwest Medical University, Southwest Medical University, 3-319 Zhongshan Road, Luzhou, Sichuan 646000, People’s Republic of ChinaTel/ Fax +86-830-3160283Email wql73115@hotmail.comBackground: Anlotinib is a highly potent multi-target tyrosine kinase inhibitor, with very good anti-tumor activity against a variety of solid tumors. However, its effect on colorectal cancer (CRC) is not yet clearly understood. The objective of this study was to investigate the anti-tumor effect and underlying mechanism of anlotinib in the pathogenesis of CRC.Materials and Methods: Effects of anlotinib on CT26 cells proliferation and microvessel formation in endothelial cells were determined by MTT assay and tube formation assay. Cell migration and invasion were analyzed by using the wound healing assay and transwell assay. Cell cycle and apoptosis were detected by flow cytometry. A CRC xenograft mouse model was used for conducting in-vivo studies to verify the effect of anlotinib. The expression of Ki-67 and CD31 in the tumor tissue was detected by immunohistochemistry and protein expression was measured by Western blot.Results: In-vitro studies revealed that anlotinib inhibited the proliferation, migration, and invasion of CT26 cells and the tube formation of HUVECs in a dose-dependent manner. Anlotinib also significantly induced cell apoptosis and G2/M arrest. It effectively inhibited tumor growth and prolonged survival time in the CRC xenograft mouse model. Immunohistochemical analysis of the tumor tissue revealed that anlotinib downregulated CD31 and Ki-67 which are the biomarkers of microvessel density and proliferation. Furthermore, anlotinib was able to inhibit the activation of VEGFR-2/AKT and FGFR, PDGFRβ and their downstream signaling ERK.Conclusion: The findings of the present study suggested that anlotinib suppressed cell proliferation and angiogenesis via inhibition of AKT/ERK signaling pathway in colorectal cancer and could be a novel therapeutic strategy for treatment of CRC.Keywords: anlotinib, colorectal cancer, angiogenesis, proliferation, tyrosine kinase inhibitor

Wei Chen, 1 Ruixian Ding, 1 Jinlu Tang, 1 Haodong Li, 2 Chonghua Chen, 2 Yaru Zhang, 1 Qinxian Zhang, 1 Xiaoyan Zhu 1 1Department of Histology and Embryology, School of Basic Medical Sciences, Zhengzhou University, Henan, People’s Republic of China; 2Department of Clinical Medicine, School of Basic Medical Sciences, Zhengzhou University, Henan, People’s Republic of ChinaCorrespondence: Xiaoyan ZhuDepartment of Histology and Embryology, School of Basic Medical Sciences, Zhengzhou University, No. 100 Science Road, Henan 450000, People’s Republic of ChinaTel +86 139 3829 0386Fax +86 371 6778 1953Email zhxy@zzu.edu.cnBackground: The impact and potential molecular mechanisms of SST in the occurrence and development of GC have not been determined.Materials and Methods: Two pairs of sgRNA and reporter were designed according to targeting sequence of SST gene for double-nicking. Plasmids were transfected into 293T for selecting sgRNA with higher cutting efficiency. The subline which has knocked-out SST gene were selected by FACS and verified by sequencing and expression level. Moreover, the migration and invasion ability was evaluated by wound healing and transwell after knocking out SST. Besides, the protein expression of SEMA5A and KLF2 were observed by Western blotting and LSCM. Last, we detected the expression levels of SST, SEMA5A, and KLF2 in GC tissues by Western blotting.Results: The results revealed that the new subline 1E9, which had knocked out SST gene, was established by CRISPR/Cas9. In addition, the knockout of SST in GC cells markedly increased migration and invasion ability. The results also demonstrated that the knockout of SST increased the expression of SEMA5A and KLF2. The expression level of SST was decreased in GC tissues, and its decrease was associated with overexpression of SEMA5A and KLF2.Conclusion: SST plays an inhibitory role in the migration and invasion of GC cell BGC823. The protein expression levels of SEMA5A and KLF2 were enhanced in GC cells and tissues lacking SST expression.Keywords: somatostatin, CRISPR/Cas9, gastric cancer, migration, invasion

Wen-Shen Gu,1,2,* Wei-Zhen Fang,1,2,* Chun-Yue Liu,1,2 Kun-Yi Pan,1,2 Rui Ding,1,2 Xiao-Hui Li,3 Chao-Hui Duan1,21Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-Sen Memorial Hospital, Sun Yat-Sen University, Guangzhou 510120, People’s Republic of China; 2Department of Laboratory Medicine, Sun Yat-Sen Memorial Hospital, Guangzhou 510120, People’s Republic of China; 3Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Department of Clinical Laboratory Medicine, Sun Yat-Sen University Cancer Center, Guangzhou 510060, People’s Republic of China*These authors contributed equally to this work Background: Body mass index (BMI) has been associated with a risk of esophageal cancer. However, the influence of BMI and BMI loss on people with esophageal cancer that were treated with different therapies has not been described in China.Methods: In total, 615 consecutive patients that underwent esophagectomy and/or chemotherapy/radiotherapy were classified according to the Asian-specific BMI (kg/m2,) cutoff values. The impact of BMI and BMI loss on long-term overall survival (OS) was estimated using the Kaplan–Meier method and Cox proportional hazard models.Results: Multivariate analysis showed that overweight and obese patients had a more favorable survival than normal weight and underweight patients (p=0.017). Patients with a low BMI and high BMI loss before therapy had worse OS than others (p=0.001). Subgroup analysis showed that patients with a high BMI were more likely to suffer hypertension (p

Gene expression profiling of transplant recipient blood and urine can potentially be used to monitor graft function, but the multitude of protocols in use make sharing data and comparing results from different laboratories difficult. The goal of this study was to evaluate the performance of current methods of RNA isolation, reverse transcription and quantitative polymerase chain reaction (qPCR) and to test whether multiple centers using a standardized protocol can obtain the same results. Samples, reagents and detailed instructions were distributed to six participating sites that performed RNA isolation, reverse transcription and qPCR for 18S, PRF, GZB, IL8, CXCL9 and CXCL10 as instructed. All data were analyzed at a single site. All sites demonstrated proficiency in RNA isolation and qPCR analysis. Gene expression measurements for all targets and samples had correlations >0.938. The coefficient of variation of fold-changes between pairs of samples was less than 40%. All sites were able to accurately quantify a control sample of known concentration within a factor of 1.5. Collectively, we have formulated and validated detailed methods for measuring gene expression in blood and urine that can yield consistent results in multiple laboratories.