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Jie Liu, Lijuan Lu, Yu Wei, Yu Li, Qiong Wang, Lei Yu, Langen Zhuang, Guoxi Jin, Xiaoyan Pei Department of Endocrinology, the First Affiliated Hospital of Bengbu Medical University, Bengbu, Anhui Province, People’s Republic of ChinaCorrespondence: Guoxi Jin; Xiaoyan Pei, Department of Endocrinology, the First Affiliated Hospital of Bengbu Medical University, No. 287 Changhuai Road, Bengbu, Anhui Province, 233000, People’s Republic of China, Email jyzjyz1999@163.com; 245899985@qq.comAbstract: Pseudohypoparathyroidism (PHP) is a rare genetic disease characterized by hypocalcemia, hyperphosphatemia, and elevated parathyroid hormone (PTH) in serum. Here, we report a case of a patient with pseudohypoparathyroidism type IB (PHPIB) and subclinical hypothyroidism, analyze the clinical and genetic data of his family members, review the relevant literature, and classify and discuss the pathogenesis and clinical characteristics of each subtype. Finally, we discuss the treatment approach to improve clinicians’ understanding of the disease.Keywords: hypocalcemia, parathyroid hormone resistance, GNAS, STX16, molecular genetic mechanism

Le Zhuang,1 Weiyuan Ma,2 Jing Jiao1 1Department of Dermatology, Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong Province, People’s Republic of China; 2Department of Dermatology, Affiliated Hospital of Weifang Medical University, Weifang, Shandong Province, People’s Republic of ChinaCorrespondence: Jing Jiao, Department of Dermatology, Central Hospital Affiliated to Shandong First Medical University, No. 105, Jiefang Road, Lixia District, Jinan, Shandong Province, 250013, People’s Republic of China, Tel +8618100389680, Fax +8653185695114, Email 24588407@qq.com Weiyuan Ma, Department of Dermatology, Affiliated Hospital of Weifang Medical University, No. 2428, Yuhe Road, Kuiwen District, Weifang, Shandong Province, 261035, People’s Republic of China, Tel +861865362629, Fax +865363081201, Email fymaweiyuan@wfmc.edu.cnPurpose: Epidermal keratinocytes with an abnormal glucose metabolism have been identified in psoriasis. Hexokinase 2 (HK2) is a crucial enzyme involved in glycolytic metabolic pathways. However, the expression of HK2 and its potential therapeutic effects in psoriasis remains unclear. This study aimed to investigate the expression pattern of HK2 and evaluate its therapeutic effects in psoriasis.Patients and Methods: A gene expression dataset (GSE121212) downloaded from the Gene Expression Omnibus (GEO) database was used to examine the expression of HK2 in psoriasis. HK2 RNA and protein expression were investigated in psoriasis vulgaris (n=5) and healthy (n=5) samples. Immunohistochemistry for HK2 was performed on psoriasis vulgaris (n=22) and healthy skin (n=10) samples. Additionally, HaCaT cells were treated with M5 (interleukin [IL]-17A, tumor necrosis factor-α, IL-1α, IL-22, and Oncostatin-M) to induce a psoriatic inflammation cell model. A mouse model of psoriatic inflammation was established using topical 5% imiquimod cream. Psoriasis-like cells and mouse models were treated with the HK2 inhibitor 3-bromopyruvate (3-BrPA). Cell proliferation, glucose consumption, and lactate production were assessed. Furthermore, the activation of nuclear factor-kappa B (NF-Kb) and NOD-like receptor thermal protein domain associated protein 3 (NLRP3) was investigated using Western blot analysis.Results: According to the GEO dataset, HK2 expression was significantly elevated in psoriasis. Upregulation of HK2 in psoriatic tissues was confirmed by quantitative real-time polymerase chain reaction and Western blotting. The immunohistochemistry score for HK2 was higher in psoriatic lesions than in healthy skin. 3-BrPA inhibited the proliferation and glycolysis of M5-stimulated HaCaT cells. Topical 3-BrPA ameliorated imiquimod-induced psoriasis-like dermatitis. Activation of NF-kB and NLRP3 was downregulated by 3-BrPA treatment.Conclusion: Our study revealed that the glycolytic enzyme HK2 was upregulated in psoriasis and that the HK2 inhibitor 3-BrPA exhibited therapeutic effects in psoriasis cell and mouse models.Keywords: psoriasis, glucose metabolism, glycolysis, proliferation, 3-bromopyruvate

Weiyuan Ma,1 Xiuwen Zhang,2 Le Zhuang3 1Department of Dermatology, Affiliated Hospital of Weifang Medical University, Weifang, Shandong Province, People’s Republic of China; 2Department of Dermatology, Weihai Municipal Hospital, Weihai, Shandong Province, People’s Republic of China; 3Department of Dermatology, Central Hospital Affiliated to Shandong First Medical University, Jinan, Shandong Province, People’s Republic of ChinaCorrespondence: Le Zhuang, Department of Dermatology, Central Hospital Affiliated to Shandong First Medical University, No.105, Jiefang Road, Lixia District, Jinan, Shandong Province, 250013, People’s Republic of China, Tel +8615966301378, Fax +86053185695114, Email zhuangle@aliyun.comPurpose: Melanomas are highly malignant and rapidly develop drug resistance due to dysregulated apoptosis. Therefore, pro-apoptotic agents could be effective for the management of melanoma. Hydrogen sulfide is ubiquitous in the body, and exogenous hydrogen sulfide has been reported to show inhibitory and pro-apoptotic effects on cancer cells. However, whether high concentrations of exogenous hydrogen sulfide have pro-apoptotic effects on melanoma and its mechanisms remain unknown. Hence, this study aimed to explore the pro-apoptotic effects and mechanisms of exogenous hydrogen sulfide on the A375 melanoma cell line treated with a hydrogen sulfide donor (NaHS).Methods: The cell proliferation test, flow cytometric analysis, Hoechst 33258 staining, and Western blotting of B-cell lymphoma 2 and cleaved caspase-3 were used to explore the pro-apoptotic effects of hydrogen sulfide on A375 cells. The transcriptional profile of NaHS-treated A375 cells was further explored via high-throughput sequencing. Western blotting of phosphorylated inositol-requiring enzyme 1α (p‐IRE1α), phosphorylated protein kinase R-like ER kinase (p-PERK), phosphorylated eukaryotic translation initiation factor 2α (p-eIF2α), C/EBP homologous protein, glucose-regulating protein 78, IRE1α, PERK, and eIF2α was performed to verify the changes in the transcriptional profile.Results: NaHS inhibited A375 melanoma cell proliferation and induced apoptosis. The endoplasmic reticulum stress unfolded protein response and apoptosis-associated gene expression was upregulated in NaHS-treated A375 melanoma cells. The overactivation of the unfolded protein response and increase in endoplasmic reticulum stress was verified at the protein level.Conclusion: Treatment with NaHS increased endoplasmic reticulum stress, which triggered the overactivation of the unfolded protein response and ultimately lead to melanoma cell apoptosis. The pro-apoptotic effect of NaHS suggests that it can be explored as a potential therapeutic agent in melanoma.Keywords: hydrogen sulfide, melanoma, apoptosis, unfolded proteins response, endoplasmic reticulum stress

Le Zhuang,1– 6,* Jie Tian,1– 4,7,* Binbin Lai,1– 4,7 Guohong Zhang,1– 4 Hang Li1– 4 1Department of Dermatology, Peking University First Hospital, Beijing, People’s Republic of China; 2National Clinical Research Center for Skin and Immune Diseases, Peking University First Hospital, Beijing, People’s Republic of China; 3Beijing Key Laboratory of Molecular Diagnosis on Dermatoses, Peking University First Hospital, Beijing, People’s Republic of China; 4NMPA Key Laboratory for Quality Control and Evaluation of Cosmetics, Peking University First Hospital, Beijing, People’s Republic of China; 5Dermatology Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 6Central Hospital Affiliated to Shandong First Medical University, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong Province, People’s Republic of China; 7Institute of Medical Technology, Peking University Health Science Center, Beijing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Hang Li, Peking University First Hospital, No. 8, Xishiku Street, Xicheng District, Beijing, 100034, People’s Republic of China, Tel +8613693058190, Fax +861083572350, Email drlihang@126.comBackground: Anti-programmed cell death ligand-1 (anti-PD-L1) immunotherapy is often used for advanced urothelial carcinoma and melanoma, including amelanotic melanoma, a relatively rare subtype with little to no pigment in the tumor cells. However, cellular heterogeneity of amelanotic melanoma during or after anti-PD-L1 immunotherapy treatments has not been described.Purpose: To investigate cellular heterogeneity in acral amelanotic melanoma after immunotherapy exposure.Methods: We evaluated subtle visual changes of the melanoma by dermoscopy and performed a pathological examination to analyze the heterogeneity of microscopic morphological and immunohistochemistry changes. The cellular transcriptional heterogeneity and corresponding biological function profiles of the melanoma were determined by single-cell RNA sequencing (scRNA-seq).Results: The dermoscopic examination revealed black globules and scar-like depigmentation areas against a homogeneous red background. Pigmented and amelanotic melanoma cells were observed microscopically. The pigmented cells were large and contained melanin granules expressing Melan-A and HMB45; the amelanotic cells were small and did not express HMB45. Ki-67 immunohistochemical staining revealed that the pigmented melanoma cells had a higher proliferative ability than the amelanotic cells. scRNA-seq identified three cell clusters: amelanotic cell cluster 1, amelanotic cell cluster 2, and pigmented cell cluster. Furthermore, a pseudo-time trajectory analysis showed that amelanotic cell cluster 2 originated from amelanotic cell cluster 1 and transformed into the pigmented melanoma cell cluster. The expression pattern of melanin synthesis-related and lysosome-endosome-related genes in different cell clusters supported the cell cluster transformation results. Also, upregulated expression of cell cycle genes indicated that the pigmented melanoma cells had a high proliferative ability.Conclusion: Coexisting amelanotic and pigmented melanoma cells indicated cellular heterogeneity in an acral amelanotic melanoma from a patient who underwent immunotherapy treatment. Additionally, the pigmented melanoma cells acquired a higher proliferative ability than the amelanotic melanoma cells.Keywords: amelanotic, acral melanoma, cellular heterogeneity, immunotherapy, drug resistance

Lijuan Zhuang,&ast; Xiaoyan Xie,&ast; Lihua Wang, Xiulan Weng, Yingling Xiu, Dabin Liu, Liying Zhong Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou, 350001, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Liying Zhong; Dabin Liu, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, 18 Daoshan Road, Fuzhou, 350001, People’s Republic of China, Tel +86-13860610354 ; +86-13489997701, Fax +86-591-87551247, Email zhongliying@fjmu.edu.cn; 15968250@qq.comBackground: The characteristics of high-risk human papillomavirus (HR-HPV) infection in different pathological types of cervical cancer in China are unclear. The aim of this study was to evaluate HR-HPV genotypes and age stratification with cervical squamous cell carcinoma (SCC) and adenocarcinoma (ADC) in China.Materials and Methods: Patients diagnosed with cervical cancer by histopathology in Fujian Maternity and Child Health Hospital from January 1, 2014, to December 31, 2017, were included in this study. The HR-HPV genotype was analyzed in cervical specimens. Logistic regression was used to calculate odds ratios (ORs). All tests of statistical significance were two-sided, and the P value< 0.05.Results: A total of 1,590,476 women were screened for cervical cancer, and 688 cervical cancers were detected, including 554 SCC and 93 ADC. The overall HR-HPV infection rate in SCC was higher than that in ADC (91.2% vs 81.7%, P=0.005). HPV-16 was the most prevalent genotype in SCC (70.0%) but was only 31.2% in ADC (P< 0.001). However, the prevalence of HPV-18 in ADC was significantly higher than that in SCC (45.2% vs 7.0%; P< 0.001). In SCC, the prevalence of HPV-16 was consistently much higher than that of HPV-18 regardless of age group. Among ADC, the prevalence of HPV-18 was higher than that of HPV-16 in women aged ≥ 45 years. Interestingly, in those aged < 35 years, the highest prevalence was observed for HPV-16. HPV-18 infection has the highest risk of ADC (OR= 12.109; P< 0.001), and HPV-45 and HPV-51 were also found to be associated with the occurrence of ADC. However, HPV-16 infection greatly increased the risk of having histological SCC.Conclusion: HPV-16 and HPV-18 infections are key risk factors for SCC and ADC. The use of HR-HPV genotyping tests in cervical cancer screening and vaccination against major HPV genotypes could reduce the incidence of cervical cancer.Keywords: cervical squamous cell carcinoma, cervical adenocarcinoma, high-risk human papillomavirus

Lijuan Zhuang &ast;, Xiulan Weng &ast;, Lihua Wang, Xiaoyan Xie, Liying Zhong, Dabin Liu, Yingling Xiu Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, Fuzhou, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yingling Xiu; Dabin Liu, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Fujian Medical University, 18 Daoshan Road, Fuzhou, People’s Republic of China, Tel +86-13860610354 ; +86-13489997701, Fax +86-591-87551247, Email xiuyingling@fjmu.edu.cn; xiuyingling1985@163.com; 15968250@qq.comPurpose: A high-risk human papillomavirus E6/E7 mRNA (HR-HPV mRNA) assay is widely used in cervical cancer screening in China. However, it is still unclear whether stand-alone HR-HPV mRNA testing is sufficient for primary screening. The purpose of this study was to investigate the feasibility of a stand-alone HR-HPV mRNA assay for primary screening of cervical cancer.Methods: Women aged 21 and older were recruited in Fujian Province, China, from January 2020 to January 2022. Cervical exfoliated cells were collected for cervical cytology and HR-HPV mRNA assays, and women with positive results on either assay were referred for colposcopy. The screening effectiveness of the assay was calculated based on the cervical histology. When comparing the efficacy of the different screening strategies, only women aged 25 and older were included.Results: A total of 9927 women were recruited. This study identified 217 cases of high-grade squamous intraepithelial disease or worse (HSIL+). The overall age-specific HR-HPV infection rate showed a U-shaped distribution. The sensitivity of the HR-HPV mRNA assay to identify CIN2+ and CIN3+ was 97.2% and 97.9%, respectively, which was significantly higher than that of cytology (82.9% and 88.6%, P< 0.001 and 0.002). The sensitivity of the HR-HPV mRNA primary screening strategy to identify CIN2+ and CIN3+ was 92.2% and 94.3%, respectively, which was similar to the co-testing strategy (P=0.336 and 0.394) and higher than the cytology primary screening (P=0.002 and 0.048). In addition, the HR-HPV primary screening strategy had a lower referral rate for colposcopy than cytology primary screening (5.4% vs 6.6%, P< 0.001), and the screening cost was lower than co-testing (&dollar;29,594.3 per 1000 screened women vs &dollar;55,140 per 1000 screened women, P< 0.001).Conclusion: In conclusion, the detection of CIN2+/CIN3+ by HR-HPV mRNA is both specific and sensitive. It may be suitable for primary screening of cervical cancer in China.Keywords: cervical cancer, HR-HPV, E6/E7 mRNA, primary screening, cervical intraepithelial neoplasia

Yao Tong,&ast; Chengrong Bao,&ast; Yi-Qiong Xu,&ast; Lei Tao,&ast; Yao Zhou, Lei Zhuang, Ying Meng, Hui Zhang, Jingjing Xue, Weijun Wang, Lele Zhang, Qingbo Pan, Zhenzhen Shao, Tianran Hu, Qian Guo, Qingsheng Xue, Han Lu, Yan Luo Department of Anesthesiology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Yan Luo; Han Lu Email ly11087@rjh.com.cn; luhan0301@163.comBackground: Acute lung injury (ALI) is a severe respiratory disease with high rates of morbidity and mortality. Many mediators regarding endogenous or exogenous are involved in the pathophysiology of ALI. Here, we have uncovered the involvement of integrins and matrix metalloproteinases, as critical determinants of excessive inflammation and endothelial permeability, in the regulation of ALI.Methods: Inflammatory cytokines were measured by quantitative real-time PCR for mRNA levels and ELISA for secretion levels. Endothelial permeability assay was detected by the passage of rhodamine B isothiocyanate-dextran. Mice lung permeability was assayed by Evans blue albumin (EBA). Western blot was used for protein level measurements. The intracellular reactive oxygen species (ROS) were evaluated using a cell-permeable probe, DCFH-DA. Intratracheal injection of lipopolysaccharide (LPS) into mice was conducted to establish the lung injury model.Results: Exogenous MMP-9 significantly aggravated the inflammatory response and permeability in mouse pulmonary microvascular endothelial cells (PMVECs) treated by LPS, whereas knockdown of MMP-9 exhibited the opposite phenotypes. Knockdown of integrin β 3 or β 5 in LPS-treated PMVECs significantly downregulated MMP-9 expression and decreased inflammatory response and permeability in the presence or absence of exogenous MMP-9. Additionally, the interaction of MMP-9 and integrin β 5 was impaired by a ROS scavenger, which further decreased the pro-inflammatory cytokines production and endothelial leakage in PMVECs subjected to co-treatment (LPS with exogenous MMP-9). In vivo studies, exogenous MMP-9 treatment or knockdown β 3 integrin significantly decreased survival in ALI mice. Notably, knockdown of β 5 integrin alone had no remarkable effect on survival, but which combined with anti-MMP-9 treatment significantly improved the survival by ameliorating excessive lung inflammation and permeability in ALI mice.Conclusion: These findings support the β 3/5 integrin-MMP-9 axis as an endogenous signal that could play a pivotal role in regulating inflammatory response and alveolar-capillary permeability in ALI.Keywords: acute lung injury, endothelial cells, integrin, MMP-9

Zetao Ma,1,2,&ast; Xizhi Wang,1,&ast; Qingbo Lv,1 Yingchao Gong,1 Minghong Xia,1 Lenan Zhuang,1 Xue Lu,1 Ying Yang,1 Wenbin Zhang,1 Guosheng Fu,1 Yang Ye,1 Dongwu Lai1 1Key Laboratory of Cardiovascular Intervention and Regenerative Medicine of Zhejiang Province, Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang Province, 310016, People’s Republic of China; 2Department of Cardiology, Zhongshan People’s Hospital, Zhongshan, Guangdong Province, 528403, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Dongwu Lai; Yang YeKey Laboratory of Cardiovascular Intervention and Regenerative Medicine of Zhejiang Province, Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 3 East Qingchun Road, Hangzhou, Zhejiang Province, 310016, People’s Republic of ChinaTel +86-571-86006241Fax +86-571-86006246Email laidw@zju.edu.cn; yeyang1222@zju.edu.cnBackground: Considered as one of the major reasons of sudden cardiac death, hypertrophic cardiomyopathy (HCM) is a common inherited cardiovascular disease. However, effective treatment for HCM is still lacking. Identification of hub gene may be a powerful tool for discovering potential therapeutic targets and candidate biomarkers.Methods: We analysed three gene expression datasets for HCM from the Gene Expression Omnibus. Two of them were merged by “sva” package. The merged dataset was used for analysis while the other dataset was used for validation. Following this, a weighted gene coexpression network analysis (WGCNA) was performed, and the key module most related to HCM was identified. Based on the intramodular connectivity, we identified the potential hub genes. Then, a receiver operating characteristic curve analysis was performed to verify the diagnostic values of hub genes. Finally, we validated changes of hub genes, for genetic transcription and protein expression levels, in datasets of HCM patients and myocardium of transverse aortic constriction (TAC) mice.Results: In the merged dataset, a total of 455 differentially expressed genes (DEGs) were identified from normal and hypertrophic myocardium. In WGCNA, the blue module was identified as the key module and the genes in this module showed a high positive correlation with HCM. Functional enrichment analysis of DEGs and key module revealed that the extracellular matrix, fibrosis, and neurohormone pathways played important roles in HCM. FRZB, COL14A1, CRISPLD1, LUM, and sFRP4 were identified as hub genes in the key module. These genes showed a good predictive value for HCM and were significantly up-regulated in HCM patients and TAC mice. We also found protein expression of LUM and sFRP4 increased in myocardium of TAC mice.Conclusion: This study revealed that five hub genes are involved in the occurrence and development of HCM, and they are potentially to be used as therapeutic targets and biomarkers for HCM.Keywords: hypertrophic cardiomyopathy, HCM, weighted gene coexpression network analysis, WGCNA, hub gene, biomarkers, bioinformatics analysis

Xin Liu,1 Ziqiao Xu,1 Yuting Wang,1 Huiling Luo,1 Donglei Zou,1 Ziyuan Zhou,2 Lixing Zhuang3 1Clinical Medical College of Acupuncture Moxibustion and Rehabilitation, Guangzhou University of Chinese Medicine, Guangzhou, 510000, People’s Republic of China; 2Faculty of Science, The University of Hong Kong, Hong Kong, 999077, People’s Republic of China; 3Department of Rehabilitation, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510000, People’s Republic of ChinaCorrespondence: Lixing ZhuangDepartment of Rehabilitation, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, No. 16 Airport Road, Baiyun District, Guangzhou, People’s Republic of ChinaTel +86 13822287775Email zhuanglixing@163.comObjective: This study aims to improve the reporting quality of randomized controlled trials (RCTs) by evaluating RCTs of acupuncture for low back pain (LBP) based on the CONSORT and STRICTA statements.Methods: Literature from the Cochrane Library, Medline, Embase, Ovid, China National Knowledge Infrastructure (CNKI), WanFang database, and Chongqing Weipu (VIP) was systematically searched from 2010 to 2020. The general characteristics and the overall quality score (OQS) of the literature were evaluated by two investigators. The agreement between investigators was calculated using Cohen’s kappa statistics.Results: A total of 31 RCTs were extracted in the final analysis. Based on the CONSORT statement, the items “title and abstract”, “background and objectives”, “intervention”, “outcomes”, “statistical methods”, “baseline data”, “outcomes and estimation” and “interpretation” have a positive rate of greater than 80%. The items “implementation”, “generalizability” and “protocol” have a positive rate of less than 30%. Based on the STRICTA statement, the items “style of acupuncture”, “needle retention time”, “number of treatment sessions”, “frequency and duration of treatment” and “precise description of the control or comparator” have a positive rate of greater than 80%. The item “extent to which the treatment was varied” has a positive rate of less than 30%. The agreements among most items are determined to be moderate or good.Conclusion: The reporting quality of RCTs of acupuncture for LBP is moderate. Researchers should rigidly follow the CONSORT and STRICTA statements to enhance the quality of their studies.Keywords: acupuncture, quality of reporting, low back pain, CONSORT, STRICTA

Langen Zhuang, Ziwei Wang, Xiaolei Hu, Qingqing Yang, Xiaoyan Pei, Guoxi Jin Department of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233004 Anhui, People’s Republic of ChinaCorrespondence: Guoxi JinDepartment of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, 233004, People’s Republic of ChinaTel +86-18096530238Email guoxijinnfm@163.comBackground: The intervention of circular RNA HIPK3 (circHIPK3) in diabetes has drawn increasing attention in recent years. However, the underlying mechanism of circHIPK3 in diabetic nephropathy (DN) has not been fully elucidated. Thus, the current study aims to investigate the role of circHIPK3 in high glucose (HG)-induced toxicity to human renal tubular epithelial HK-2 cells.Methods: The expression of circHIPK3 in HK-2 cells induced by HG was determined by qRT-PCR and Western blot. The regulatory effects of circHIPK3 and miR-326/miR-487a-3p on cells proliferative and apoptosis were evaluated by CCK-8 and flow cytometry. Dual-luciferase reporter assay was applied to predict the target genes of miR-326 or miR-487a-3p.Results: Expression level of circHIPK3 in HK-2 cells was remarkably decreased after the treatment of HG. The overexpression of circHIPK3 effectively reversed the HG-induced HK-2 cell proliferation inhibition and apoptosis. Furthermore, SIRT1 was confirmed to be the target gene of miR-326 and miR-487a-3p, which were showed to be the downstream genes of circHIPK3. The silencing of miR-326 or miR-487a-3p was also proved to induce proliferation and reduce apoptosis in HG-induced HK-2 cells.Conclusion: Our data suggest that overexpression of circHIPK3 can attenuate the proliferation inhibition of HK-2 induced by HG and inhibit apoptosis through sponging miR-326 or miR-487a-3p to regulate SIRT1.Keywords: diabetic nephropathy, high glucose, circHIPK3, miR-326, miR-487a-3p, SIRT1

Jingyi Dai,1,2,* Jun Zhao,3,* Yingrong Du,1 Lin Zhuang,1 Edward B McNeil,2 Virasakdi Chongsuvivatwong2 1Department of Liver Diseases, The Third People’s Hospital of Kunming City, Kunming, Yunnan, People’s Republic of China; 2Epidemiology Unit, Faculty of Medicine, Prince of Songkla University, Hat Yai, Songkhla, Thailand; 3Department of Preventive Medicine, School of Public Health and Management, Hubei University of Medicine, Shiyan, Hubei, People’s Republic of China*These authors contributed equally to this workCorrespondence: Virasakdi Chongsuvivatwong Email cvirasak@medicine.psu.ac.thBackground: Data concerning adherence to hepatocellular carcinoma (HCC) surveillance among chronic liver disease (CLD) patients at high risk of developing HCC in China are limited. We aimed to examine the relationship between HCC-related knowledge dimensions and adherence to HCC surveillance procedures among chronic liver disease patients at high risk of developing HCC and to identify potential barriers.Methods: A total of 380 patients with chronic liver disease at high risk of developing HCC were recruited between May and August 2018 to complete a survey during the first week of their first hospitalization at the Third People’s Hospital of Kunming in China. We followed up each patient up to 7 months by telephone to confirm whether the patient returned to complete investigations for HCC surveillance. Patient’s socio-demographic characteristics, HCC-related knowledge, and perceived barriers to HCC surveillance were measured using a structured questionnaire during their hospitalization. Factor analysis was performed on the knowledge questions to reduce the dimensions. Univariate and multivariate analyses were performed to examine the association between dimensions of HCC-related knowledge and patients’ adherence to HCC surveillance.Results: A total of 327 eligible patients had been successfully contacted in the follow-up phase. Only a quarter of patients completed HCC surveillance within 7 months after their first admission to hospital. High costs and perceived poor test efficacy were the two major barriers for HCC surveillance. Three common factors were derived from the factor analysis of HCC-related knowledge, namely, “Surveillance”, “Lifestyle”, and ‘Prognosis’. Knowledge of HCC surveillance and lifestyle but not prognosis had an influence on adherence to HCC surveillance. Patients with better surveillance and lifestyle knowledge domain had better adherence to HCC surveillance.Conclusion: Adherence to HCC surveillance procedures is low in the study area. Closing the gap in HCC-related knowledge, particularly regarding surveillance and lifestyle, may help to increase adherence rates.Keywords: HCC surveillance, adherence, knowledge, barriers

Xiaolei Hu, 1 Hengyan Zhang, 1 Langen Zhuang, 1 Guoxi Jin, 1 Qingqing Yang, 1 Min Li, 1 Weihua Sun, 1 Fengling Chen 2 1Department of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, People’s Republic of China; 2Department of Endocrinology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, People’s Republic of ChinaCorrespondence: Xiaolei HuDepartment of Endocrinology, The First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui 233004, People’s Republic of ChinaTel +86-0552-3086113Email caesar80@163.comFengling ChenDepartment of Endocrinology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, People’s Republic of ChinaTel +86-021-56691101Email Prof_flchen@163.comBackground: Ubiquitin-fold modifier-1 (Ufm1) is a recently identified ubiquitin-like protein. We previously confirmed that Ufm1 expression was increased in diabetic mice. However, its role in the development of diabetes remains undefined.Methods: Lentivirus-mediated gene knockdown and overexpression techniques were used to observe the effect of Ufm1 on the expression of inflammatory factors, adhesion molecules and chemokines, as well as the transcriptional activity of nuclear factor kappa-B (NF-κB) in macrophages. Western blot and immunofluorescence analyses were used to analyse the mechanism by which Ufm1 affects the transcriptional activity of NF-κB. Finally, the effects of Ufm1 on inflammation and pancreatic, renal and myocardial damage were observed in db/db mice.Results: Knockdown of Ufm1 by lentivirus shRNA targeting Ufm1 (Lv-shUfm1) led to decreased secretion of IL-6, IL-1β, ICAM-1, VCAM-1, MCP-1 and CXCL2 in RAW264.7 cells that were exposed to LPS and TNF-α, while lentiviral overexpression of Ufm1 (Lv-Ufm1) caused the opposite effect. Interestingly, further investigation indicated that Ufm1 induced NF-κB p65 nuclear translocation in RAW264.7 cells via increasing the ubiquitination and degradation of IκBα. In an in vivo experiment, pretreatment of db/db mice with Lv-shUfm1 reduced the mRNA levels of TNF-α, IL-6, IL-1β, ICAM-1, VCAM-1, MCP-1 and CXCL2 in resident peritoneal macrophages (RPMs) and decreased the plasma levels of TNF-α, IL-6, IL-1β, ICAM-1, VCAM-1, MCP-1 and CXCL2. Additionally, in Lv-Ufm1-treated mice, the inverse results were observed. Following treatment with Lv-shUfm1 and Lv-Ufm1, NF-κB p65 nuclear translocation in RPMs was decreased and increased, respectively. Importantly, we observed that Lv-shUfm1 injection led to a decrease in plasma glycaemia, a reduction in urinary albuminuria and cardiomyocyte hypertrophy and an improvement in the histopathological appearance of pancreatic, kidney and myocardial tissue. Pretreatment of the mice with Lv-shUfm1 inhibited macrophage infiltration in the pancreas, kidney and myocardial tissue.Conclusion: Our data elucidate a new biological function of Ufm1 that mediates inflammatory responses. Ufm1-mediated p65 nuclear translocation occurs by modulating the ubiquitination and degradation of IκBα. Moreover, downregulating Ufm1 is an effective strategy to prevent the development of type 2 diabetes and its complications.Keywords: ubiquitin-fold modifier-1, macrophage, NF-κB, IκBα, diabetes, diabetic complications

Weidong Shen,1,* Zhonghai Jin,2,* Xiuping Tong,2 Haiying Wang,2 Lilei Zhuang,2 Xiaofeng Lu,2 Shenbao Wu21Department of Gastroenterology, Jiangyin Hospital Affiliated to Nantong University, Jiangyin, People’s Republic of China; 2Department of Gastroenterology, Yiwu Hospital, Wenzhou Medical University, Yiwu, People’s Republic of China*These authors contributed equally to this work Background: Colorectal cancer (CRC) is among the most frequent and lethal malignancies worldwide. Although great advances have been made in the treatment of CRC, prognosis remains poor. Our previous study indicated that tripartite motif-containing 14 (TRIM14) was upregulated in CRC samples. Methods: In the current study, the association between TRIM14 and CRC was investigated. Protein expression was determined by Western blotting and immunohistochemistry. Further, the biological roles of TRIM14 in CRC cell proliferation and apoptosis were explored both in vitro and in vivo. Results: We observed that increased TRIM14 expression in CRC tissues was closely related with aggressive clinicopathological characteristics and poor prognosis. TRIM14 knockdown markedly reduced proliferation and increased apoptosis in HT-29 and SW620 cells, whereas TRIM14 overexpression in LoVo cells displayed opposite results. Xenograft experiments using HT-29 cells confirmed suppression of tumor growth and induction of apoptosis upon TRIM14 knockdown in vivo. Furthermore, downregulation of TRIM14 inhibited the AKT pathway, as indicated by reduced levels of phosphorylated AKT, Bcl-2 and Cyclin D1, and elevated levels of phosphatase andtensin homology (PTEN) and p27. In addition, TRIM14 colocalized with PTEN in the cytoplasm and induced PTEN ubiquitination. Moreover, PTEN overexpression significantly inhibited pro-proliferative effects of TRIM14, indicating an involvement of PTEN/AKT signaling in mediating TRIM14 functions. Conclusions: The present data demonstrate that TRIM14 overexpression promotes CRC cell proliferation, suggesting TRIM14 as an attractive therapeutic target for CRC.Keywords: colorectal cancer, TRIM14, PTEN, AKT

Liping Zhuang,1,2,* Xia Yan,1,2,* Zhiqiang Meng1,2 1Department of Integrative Oncology, Fudan University Shanghai Cancer Center, Shanghai, China; 2Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China *These authors contributed equally to this work Background: A population-based estimate of risk of second primary malignancy (SPM) in patients with cholangiocarcinoma (CCA) is still lacking.Objectives: To investigate the overall and site-specific risk of SPM in patients with CCA. To identify risk factors of SPM and further evaluate the impact of SPM on overall survival (OS) and disease specific survival (DSS) in patients with CCA.Methods: Patients with histologically diagnosed CCA between 1973 and 2015 were identified from the Surveillance, Epidemiology and End Results database. Standardized incidence ratio (SIR) was calculated. Risk factors for SPM and CCA survival were evaluated by logistic, Cox, and nomogram methods.Results: We found that the overall risk of SPM in patients with CCA was significantly higher than that in the general population (SIR =1.27, 95% CI =1.03–1.55, P

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