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BACKGROUND Splenic hamartoma (SH) is a rare, benign vascular proliferation that is often found incidentally. It may be misdiagnosed as a splenic aneurysm or splenic malignancy. CASE SUMMARY A 21-year-old male patient was admitted to our hospital with a complaint of an incidentally discovered asymptomatic splenic space-occupying lesion for 2 wk. Abdominal computed tomography (CT) scan showed a circular low-density shadow in the hilum of the spleen. Contrast-enhanced CT revealed an aneurysm located in the hilum of the spleen before operation. Laparoscopic splenectomy was performed and postoperative pathology revealed the presence of SH. CONCLUSION Imaging studies are insufficient for the differential diagnosis of SH from other diseases, and laparoscopic splenectomy is a less invasive procedure and useful for the diagnostic purpose as well.

Background Increasing numbers of studies have demonstrated that circulating tumor cells (CTCs) undergo a phenotypic change termed epithelial‐mesenchymal transition (EMT), and researchers have proposed that EMT might provide CTCs with increased potential to survive in the different microenvironments encountered during metastasis through various ways, such as by increasing cell survival and early colonization. However, the exact role of EMT in CTCs remains unclear. Methods In this study, we identified CTCs of 41 patients with gastric cancer using Cyttel‐CTC and im‐FISH (immune‐fluorescence in situ hybridization) methods, and tested the expression of EMT markers and ULBP1 (a major member of the NKG2D—natural killer [NK] group 2 member D—ligand family) on CTCs. Moreover, we investigated the relationship between the expression of EMT markers and ULBP1 on CTCs and gastric cancer cell lines. Results Our results showed that the CTCs of gastric cancer patients exhibited three EMT marker subtypes, and that the expression of ULBP1 was significantly lower on mesenchymal phenotypic CTCs (M+CTCs) than on epithelial phenotypic CTCs (E+CTCs). EMT induced by TGF‐β in vitro produced a similar phenomenon, and we therefore proposed that EMT might be involved in the immune evasion of CTCs from NK cells by altering the expression of ULBP1. Conclusions Our study indicated that EMT might play a vital role in the immune invasion of CTCs by regulating the expression of ULBP1 on CTCs. These findings could provide potential strategies for targeting the immune evasion capacity of CTCs., In this proof‐of‐principle study, we demonstrated that gastric CTCs in peripheral blood exhibited obvious EMT characteristics, as well as a downregulation of NKG2D ligands, with TGF‐β1‐induced EMT accompanied by this downregulation of NKG2D ligands in vitro. Considering these results, we conclude that EMT of gastric CTCs might be involved in the immune evasion of CTCs through shedding or restricting the expression of NKG2D ligands during the process of metastasis.

Background METTL3 is an RNA methyltransferase that mediates m6A modification and is implicated in mRNA biogenesis, decay, and translation. However, the biomechanism through which METTL3 regulates MALAT1-miR-1914-3p-YAP axis activity to induce NSCLC drug resistance and metastasis is not very clear. Methods The expression of mRNA was analyzed by qPCR assays. Protein levels were analyzed by western blotting and immunofluorescent staining. Cellular proliferation was detected by CCK8 assays. Cell migration and invasion were analyzed by wound healing and transwell assays, respectively. Promoter activities and gene transcription were analyzed by luciferase reporter assays. Finally, m6A modification was analyzed by MeRIP. Results METTL3 increased the m6A modification of YAP. METTL3, YTHDF3, YTHDF1, and eIF3b directly promoted YAP translation through an interaction with the translation initiation machinery. Moreover, the RNA level of MALAT1 was increased due to a higher level of m6A modification mediated by METTL3. Meanwhile, the stability of MALAT1 was increased by METTL3/YTHDF3 complex. Additionally, MALAT1 functions as a competing endogenous RNA that sponges miR-1914-3p to promote the invasion and metastasis of NSCLC via YAP. Furthermore, the reduction of YAP m6A modification by METTL3 knockdown inhibits tumor growth and enhances sensitivity to DDP in vivo. Conclusion Results indicated that the m6A mRNA methylation initiated by METTL3 promotes YAP mRNA translation via recruiting YTHDF1/3 and eIF3b to the translation initiation complex and increases YAP mRNA stability through regulating the MALAT1-miR-1914-3p-YAP axis. The increased YAP expression and activity induce NSCLC drug resistance and metastasis.

Purpose: It has been reported that miRNA-124 inhibits hepatocellular carcinoma (HCC) progression, while lncRNA-UCA1 promotes HCC. The aim of this study is to find whether miRNA-124, as a tumor suppressor in HCC can inhibit lncRNA-UCA1 in HCC cell. Methods: Tumor tissues and adjacent healthy tissues were obtained from 66 patients diagnosed with HCC in Binzhou Medical University Hospital from January 2011 to January 2013. Cell proliferation assay, in vitro cell migration and invasion assay were applied for the research. Results: In the present study we found that miRNA-124 was downregulated, while lncRNA-UCA1 was upregulated in tumor tissues comparing to adjacent healthy tissues of HCC patients. Expression of miRNA-124 and lncRNA-UCA1 in tumor tissues were not affected by HBV or HCV infection. Analysis of followed-up data revealed that low miRNA-124 level and high lncRNA-UCA1 level were closely correlated with poor survival. Overexpression of miRNA-124 led to inhibited lncRNA-UCA1 expression in cells of HCC cell lines, while overexpression of lncRNA-UCA1 failed to significantly affect miRNA-124 expression. Expression levels of miRNA-124 and lncRNA-UCA1 were inversely and significantly correlated in tumor tissues but not in adjacent healthy tissues. Overexpression of miRNA-124 led to inhibited, while overexpression of lncRNA-UCA1 led to increased proliferation, migration and invasion rates of HCC cell lines. In addition, lncRNA-UCA1 overexpression attenuated the inhibitory effects of miRNA-124 overexpression on cancer cell proliferation, migration and invasion. Conclusion: Therefore, miRNA-124 may inhibit the proliferation, migration and invasion of cancer cell in hepatocellular carcinoma by downregulating lncRNA-UCA1.

Background: Recent evidence indicates that UBE2C participates in carcinogenesis by regulating the cell cycle, apoptosis, metastasis, and transcriptional processes. Additionally, miR-548e-5p dysregulation plays a vital role in tumor progression. However, the molecular mechanism via which UBE2C is directly targeted by miR-548-5p, resulting in increase in cellular growth and invasiveness of cancer cells, and its interactions with the epithelial-mesenchymal transition (EMT) marker protein ZEB1/2 in non-small cell lung cancer (NSCLC) is not understood. Methods: Expression of UBE2C and miR-548e-5p was analyzed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The protein level of UBE2C and ZEB1/2 was analyzed using western blotting and immunofluorescence staining. Cellular proliferation was detected using the cell counting kit 8 (CCK8) and 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Cell migration, invasion, and growth were analyzed using the wound healing and transwell assay. Promoter activity and transcription was analyzed using the luciferase reporter assay. Chromatin immunoprecipitation was used to detect binding of UBE2C to 5′UTR-ZEB1/2. Results: We observed that 4,5-ubiquitin-conjugating enzyme E2C (UBE2C) expression was higher in NSCLC tissue than in the adjacent normal tissue and was associated with increased cell proliferation and invasion. UBE2C enhanced NSCLC progression and metastasis by affecting the cell cycle and inhibiting apoptosis. We also observed that miR-548e-5p was significantly downregulated in lung cancer tissue specimens, which decreased the expression of its direct substrate, UBE2C. Moreover, miR-548e-5p overexpression and UBE2C under-expression significantly suppressed lung cancer cell proliferation, migration, and invasion. Luciferase reporter and chromatin immunoprecipitation assays indicated that miR-548e-5p directly binds to the 3′-UTR of UBE2C and decreases UBE2C mRNA expression. Furthermore, UBE2C knockdown downregulated the mesenchymal marker vimentin and upregulated the epithelial marker E-cadherin. Bioinformatics assays, coupled with western blotting and luciferase assays, revealed that UBE2C directly binds to the 5′-untranslated region (UTR) of the transcript of the E-cadherin repressor ZEB1/2 and promotes EMT in lung cancer cells. Conclusion: miR-548e-5p directly binds to the 3′-UTR of UBE2C and decreases UBE2C mRNA expression. UBE2C is an oncogene that promotes EMT in lung cancer cells by directly targeting the 5′-UTR of the transcript encoding the E-cadherin repressor ZEB1/2. miR-548e-5p, UBE2C, and ZEB1/2 constitute the miR-548e-5p-UBE2C-ZEB1/2 signal axis, which enhances cancer cell invasiveness by directly interacting with e EMT marker proteins. We believe that the miR-548e-5p-UBE2C-ZEB1/2 signal axis may be a suitable diagnostic marker and a potential target for lung cancer therapy.

Background: Paraganglioma (PGL) is a rare neuroendocrine tumour derived from the extra- adrenal gland that is also known as ectopic pheochromocytoma (PHEO). The typical clinical symptoms are paroxysmal hypertension with headache, sweating and palpitation. However, PGL with abnormal glucose metabolism as the main manifestation is rare.Case Presentation: In this case, the patient had a history of diabetes for 13 years and showed persistently increased blood glucose. The application of multiple hypoglycaemic drugs to control the patient’s blood glucose was ineffective, and a limited effect of high-dose insulin was observed. This patient underwent a contrast-enhanced computed tomography scan(CT),showing a soft tissue density shadow (3.2×2.4cm) with clear edges and slight enhancement in the medial side of inferior vena cava. Exploratory laparotomy was performed and significant blood pressure changes were observed when the tumourwas removed during the operation. The patient’s blood glucose decreased significantly and hypoglycaemiawas noted when the patient’s original dose of insulin was maintained. The patient recovered uneventfully and was discharged after the operation.Conclusions: Hyperglycaemia caused by PGL is rare in the clinic; therefore, it is necessary to pay attention to the relevant blood glucose indexes during the preoperative examination and diagnosis process to improve the examination process and further assist in the preoperative diagnosis of PGL. It is necessary to pay attention to changes in blood pressure during and after the operation to avoid hypoglycaemia. Lifetime follow up is necessary after the operation.

Background. Intraoperative fluid (IOF) administration plays an important role during major abdominal surgery although increased fluid intake can adversely influence postoperative outcomes. However, the effect of the IOF rate on the outcomes of pancreatoduodenectomy (PD) is unclear. Methods. 151 patients, who underwent PD at Binzhou Medical University Hospital between January 2010 and May 2017, were categorized into three groups according to IOF rates (ml/kg/hr): restricted (n=47), standard (10–15, n=76), and liberal (>15, n=28). Results. The overall postoperative morbidity was 56.95%. The incidence of postoperative pancreatic fistula (POPF) was 11.26%. The in-hospital mortality rate was 7.28% with the most common cause being grade C POPF and secondary intra-abdominal infections. The patients in the liberal group had significantly higher incidences of POPF (25%) and respiratory complications (21.43%). The other outcome parameters such as recovery of bowel function, hospital stay, and postoperative daily drainage were similar among the groups. Multivariable analysis confirmed the IOF rate to be most strongly associated with POPF (odds ratio: 5.195, confidence interval: 1.142–23.823, P=0.023) and respiratory complications (odds ratio: 7.302, confidence interval: 0.676–58.231, P=0.025). Conclusions. The IOF rate significantly affects the incidence of POPF and respiratory complications after PD. Careful patient-oriented fluid therapy may help to prevent these complications.

METTL3 is an RNA methyltransferase that mediates mThe expression of mRNA was analyzed by qPCR assays. Protein levels were analyzed by western blotting and immunofluorescent staining. Cellular proliferation was detected by CCK8 assays. Cell migration and invasion were analyzed by wound healing and transwell assays, respectively. Promoter activities and gene transcription were analyzed by luciferase reporter assays. Finally, mMETTL3 increased the mResults indicated that the m

Additional file 1: Figure S1. METTL3 increases the m6A modification and protein level of YAP. Figure S2. m6A mRNA methylation promotes cellular growth and invasion in H1299 cells. Figure S3. The cellular growth, invasion, migration and EMT were increased in the A549/DDP and 95-D cells. Figure S4. The expressions of YTHDF1 and YTHDF2 in the NSCLC tumors. Figure S5. miR-1914-3p together with MALAT1 modified by m6A modification regulates YAP activity in H1299 cells. Figure S6. miR-1914-3p down regulates YAP activity. Figure S7. miR-1914-3p sponged by MALAT1 regulates cellular growth in NSCLC cells. Figure S8. METTL3 induced NSCLC drug resistance and metastasis via increasing stability of MALAT1.

Rationale: Duodenal neuroendocrine tumor (d-NET) is a rare tumor originating in the neuroendocrine system. The clinical manifestations of d-NET are similar to those of other digestive tract tumors, resulting in a lack of specificity and complex clinical symptoms. Patient concerns: A 55-year-old female patient was admitted to our hospital with a chief complaint of an abdominal mass that had been present for more than 4 months. Diagnoses: The upper abdomen enhanced computed tomography scan showed an uneven density mass across the upper abdomen, and the tumor size was approximately 6.2 × 5.8 cm with obvious visible enhancement present in 1 area and a cystic nonenhanced area. The postoperative pathology showed the tumor cells to be positive for chromogranin, synaptophysin, cytokeratin, CD56 (partial weak), negative for vimentin, CD117, DOG-1, CD34, S-100, SMA, desmin, and Ki-67 approximately 2%, which confirmed the diagnosis of d-NETs. Interventions: We preferred laparoscopic surgical exploration, but the tumor started at the ascending part of the duodenum and involved the mesenteric artery. As the branches of the superior mesenteric artery were intertwined with the tumor, it was difficult to operate with the endoscope, so we converted to open laparotomy. The postoperative pathology revealed the presence of d-NET. Outcomes: The patient recovered uneventfully and was discharged after the operation. One-month and 3-month follow-up after surgery, showed no evidence of recurrence. Lessons: Radiological imaging studies are insufficient for the differential diagnosis of abdominal mass from other diseases, whereas surgery is the only radical treatment method, and the preferred surgical method is still active radical resection of the tumor.

Pancreatic cystic lesion (PCL) is a relatively low incidence of pancreatic disease. No clinical symptoms of PCL prevalence rate is 2.4% ~ 13.5% and has a tendency to increase with the increase of age. With the development of modern imaging technology and the improvement of people health consciousness, there has been a dramatic increase in the prevalence of PCL. PCLs involving a wide range of pathology can range from obviously benign to borderline malignant potential lesions to overt malignancy. Pancreatic cystadenocarcinoma belongs to one of the malignant tumor of PCLs, may be malignant from pancreatic cystadenoma, and it is clinically rare, accounting for only 1% of the malignant tumors of the pancreas. Preoperative diagnosis is difficult of the disease, it mostly need to postoperative pathological diagnosis. The main symptoms of pancreatic cystadenocarcinoma are dull pain or low back pain in the upper and middle abdomen, and mass in the upper abdomen. Abdominal pain is not intense, some patients only for the feeling of fullness discomfort. Other symptoms may include loss of appetite, nausea, indigestion, weight loss, jaundice, and in a few patients, gastrointestinal bleeding. In this paper, we report a rare case of pancreatic cystadenocarcinoma with a huge cystic lesion that grows rapidly in a short period of time in pregnant women, which preoperative diagnosis is considered pancreatic cystadenoma in the preoperative. Complete resection was performed and histological examination confirmed the diagnosis of pancreatic cystadenocarcinoma.

Lei Zhou, Yanyan Du, Lingqun Kong, Xingyuan Zhang, Qiangpu Chen Department of Hepatobiliary Surgery, The Affiliated Hospital of Binzhou Medical University, Binzhou, China Background and aim: Hepatocellular carcinoma (HCC) is a major cause of cancer mortality and is increasing incidence worldwide. The aim of this study was to identify the key genes and microRNAs in HCC and explore their potential mechanisms. Methods: The gene expression profiles of GSE76427, GSE64041, GSE57957, and the microRNA dataset GSE67882 were downloaded from the Gene Expression Omnibus database. The online tool GEO2R was used to obtain differentially expressed genes (DEGs) and miRNAs (DEMs). The gene ontology and the Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed for DEGs using the Database for Annotation, Visualization, and Integrated Discovery. A protein–protein interaction (PPI) network of the DEGs was constructed by Search Tool for the Retrieval of Interacting Genes and visualized by Cytoscape. Moreover, miRecords was used to predict the target genes of DEMs. Results: In total, 106 DEGs were screened out in HCC, consisting of 89 upregulated genes and 17 downregulated genes, which were mainly enriched in biological processes associated with oxidation–reduction process. Besides, the Kyoto Encyclopedia of Genes and Genomes pathways including chemical carcinogenesis, drug metabolism-cytochrome P450, tryptophan metabolism, and retinol metabolism were involved. A PPI network was constructed consisting of 105 nodes and 66 edges. A significant module including nine hub genes, ASPM, AURKA, CCNB2, CDKN3, MELK, NCAPG, NUSAP1, PRC1, and TOP2A, was detected from the PPI network by Molecular Complex Detection. The enriched functions were mainly associated with the mitotic cell cycle process, cell division, and mitotic cell cycle. In addition, a total of 21 DEMs were identified, including 9 upregulated and 12 downregulated miRNAs. Interestingly, ZBTB41 was the potential target of seven miRNAs. Finally, the nine hub genes and three miRNA-target genes expression levels were validated by reverse transcription-polymerase chain reaction. The relative expression levels of nine genes (ASPM, AURKA, CDKN3, MELK, NCAPG, PRC1, TOP2A, ZBTB41, and ZNF148) were significantly upregulated in cancer tissues. Conclusion: This study identified the key genes and potential molecular mechanisms underlying the development of HCC, which could provide new insight for HCC interventional strategies. Keywords: hepatocellular carcinoma, bioinformatic analysis, differentially expressed genes, differentially expressed microRNAs

// Baoguang Hu 1, * , Xiaokun Tian 1, 2, * , Yanbin Li 1 , Yuming Li 1 , Zhaodong Han 3 , Jiajia An 3 and Lingqun Kong 4 1 Department of Gastrointestinal Surgery, The Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong, China 2 Department of Burn and Plastic Surgery, The Sixth People’s Hospital of Zibo, Zibo, Shandong, China 3 Department of Clinical Laboratory, The Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong, China 4 Department of Hepatobiliary Surgery, The Affiliated Hospital of Binzhou Medical University, Binzhou, Shandong, China * These authors have contributed equally to this work Correspondence to: Yu-ming Li, email: PIliym@126.com Jiajia An, email: jiajiabest_007@163.com Keywords: gastric cancer; circulating tumor cells; immune evasion; epithelial-mesenchymal transition; transforming growth factor-beta 1 Received: July 11, 2017 Accepted: November 17, 2017 Published: January 02, 2018 ABSTRACT Increasing numbers of studies have demonstrated that circulating tumor cells (CTCs) undergo a phenotypic change termed epithelial-mesenchymal transition (EMT), and researchers have proposed that EMT might provide CTCs with increased potential to survive in the different microenvironments encountered during metastasis through various ways, such as by enhancing metastatic competence, and increasing cell survival and early colonization. However, the exact role of EMT in CTCs remains unclear. In this study, we identified CTCs in the peripheral blood of 41 patients with gastric cancer using Cyttel-CTC and im-FISH (immune-fluorescence in situ hybridization) methods, and tested the expression of EMT markers and NK-cell receptor D (NKG2D) ligands on CTCs. Moreover, we investigated the relationship between the expression of EMT markers and NKG2D ligands on CTCs and gastric cancer cell lines. The results showed that the CTCs in the peripheral blood of gastric cancer patients exhibited three EMT marker subtypes, and that the expression of NKG2D ligands was significantly lower on mesenchymal phenotypic CTCs (M + CTCs) than on epithelial phenotypic CTCs (E + CTCs). EMT induced by TGF-β in vitro produced a similar phenomenon, and we therefore proposed that EMT might be involved in the immune evasion of CTCs from NK cells by altering the expression of NKG2D ligands. In conclusion, our study indicated that EMT might play a key role in the immune invasion of gastric CTCs by regulating the expression of NKG2D ligands on the tumor cells. These findings could provide potential strategies for targeting the immune evasion capacity of CTCs.

Background: Recent evidence indicates that miR-548-5p dysregulation plays a vital role in tumor progression. Aberrant miR-5485p expression may be linked to tumorigenesis since miRNAs play important roles in various cellular processes and cancer metastasis. UBE2C is a type of ubiquitination enzyme catalyzing the degradation of proteins into smaller polypeptides and amino acids. UBE2C participates in carcinogenesis by regulating the cell cycle, apoptosis, metastasis and transcriptional processes. However, the molecular mechanism of UBE2C as a growth factor regulating the tumorigenesis and metastasis of lung cancer is not clearly. EMT progression is mediated by the EMT-inducing transcriptional factors ZEB1/2, which is an important characteristic for cancer metastasis. What molecules regulate the expression of ZEB1/2 and then regulate the EMT in the lung cancer is also not clearly. We found that miR-548-5p, UBE2C, ZEB1/2 have the similar functions in tumor progression and metastasis. However, the relationship between miR-548-5p, UBE2C and ZBE1/2 in the tumorigenesis and development of lung cancer is still unknown. Methods: the mRNA expression of miR-548-5p, UBE2C was analyzed by RT-PCR assay. The protein level of UBE2C and ZEB1/2 was analyzed by western blot and Immunofluorescent staining. The cellular proliferation was detected by CCK8 and MTT. The cell migration and invasion growth were analyzed by wound healing assay and transwell assay. The activities of promoter and gene transcription were analyzed by luciferase reporter assay. Chromatin immunoprecipitation detected the combining ability of UBE2C and 5'UTRZEB1/ 2. Results: In our study, we found that miR-548-5p was significantly downregulated in lung cancer tissue specimens. Our objective was to elucidate the functions, direct target genes, and molecular mechanisms of miR-548-5p in lung cancer tissue. We found that miR-548-5p downregulated the expression of its direct substrate, 4,5-ubiquitin-conjugating enzyme E2C (UBE2C). UBE2C is a proto-oncogene that is activated in lung cancer. However, its role in lung cancer metastasis is unclear. We found here that UBE2C expression was higher in non-small-cell lung cancer (NSCLC) than in the normal tissue adjacent to it and was associated with increased cell proliferation and invasion. UBE2C enhanced NSCLC progression and metastasis by arresting the cell cycle and inhibiting apoptosis. We used reverse transcription (RT)-PCR assay to determine miR548-5p and UBE2C expression in lung cancer patients. Overexpression of miR-548-5p and UBE2C were established in lung cancer cells. Methyl thiazolyl tetrazolium (MTT)- and colony formation assays were performed to evaluate their effects on cell proliferation. We also performed cell migration- and invasion assays on lung cancer cells overexpressing miR548-5p and under expressing UBE2C. The results indicated that the aforementioned treatments significantly suppressed lung cancer cell proliferation, migration, and invasion. Moreover, luciferase reporter and Chromatin immunoprecipitation assay indicated that miR-548-5p directly binds to the 3'-UTR of UBE2C and decreases UBE2C mRNA expression. Using RT-PCR and western blot (WB), we also assessed epithelial-mesenchymal transition (EMT). We found that UBE2C knockdown downregulated the expression of the mesenchymal marker vimentin. In contrast, the epithelial marker E-cadherin was upregulated. Therefore, UBE2C promoted EMT. Bioinformatics assays coupled with WB and luciferase assays revealed that UBE2C directly binds to the 5′-untranslated region of the E-cadherin repressor, ZEB1/2 and promotes EMT in lung cancer cells. Conclusion: the miR-548-5p directly binds to the 3'-UTR of UBE2C and decreases UBE2C mRNA expression. UBE2C is an oncogene promoting EMT in lung cancer cells by directly targeting the 5'-UTR of E-cadherin repressor ZEB1/2. The miR-548-5p, UBE2C, and ZEB1/2 constitute the miR548-5p-UBE2C-ZEB1/2 signal axis which enhances cancer cell invasiveness by directly interacting with the EMT maker protein. We believe that the miR-548-5p-UBE2C-ZEB1/2 signal axis may be a suitable diagnostic marker and a potential target in lung cancer therapy Funding: This work was supported by the Science and Technology Development Foundation of Yantai (2015ZH082), Natural Science Foundation of Shandong Province (ZR2018QH004, ZR2016HB55, ZR2017PH067, ZR2017MH125 and ZR2015PH004), and Research Foundation of Binzhou Medical University (BY2015KYQD25 and BY2015KJ14). Declaration of Interest: The authors declare no competing financial interests. Ethical Approval: The experimental protocol was approved by the Research Ethics Committee of Binzhou Medical University, China (No. 2017-016-01 for human lung cancer specimen and No. 2017-009-09 for mouse experiments in vivo) and the written informed consent was obtained from all subjects. Informed consent was obtained from all individual participants included in the study. All patients were staged based on the criteria of the 7th Edition of the AJCC Cancer Staging Manual: Stomach (2010)

The energy-related carbon emissions of China’s manufacturing increased rapidly, from 36988.97 × 104 tC in 1996 to 74923.45 × 104 tC in 2012. To explore the factors to the change of the energy-related carbon emissions from manufacturing sector and the decoupling relationship between energy-related carbon emissions and economic growth, the empirical research was carried out based on the LMDI method and Tapio decoupling model. We found that the production scale contributed the most to the increase of the total carbon emissions, while the energy intensity was the most inhibiting factor. And the effects of the intrastructure and fuel mix on the change of carbon emissions were relatively weak. At a disaggregative level within manufacturing sector, EI subsector had a greater impact on the change of the total carbon emissions, with much more potentiality of energy conservation and emission reduction. Weak decoupling of manufacturing sector carbon emissions from GDP could be observed in the manufacturing sector and EI subsector, while strong decoupling state appeared in NEI subsector. Several advices were put forward, such as adjusting the fuel structure and optimizing the intrastructure and continuing to improve the energy intensity to realize the manufacturing sustainable development in low carbon pattern.