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OBJECTIVE@#Anemoside B4 (AB4), the most abundant triterpenoidal saponin isolated from Pulsatilla chinensis, inhibited influenza virus FM1 or Klebsiella pneumoniae-induced pneumonia. However, the anti-SARS-CoV-2 effect of AB4 has not been unraveled. Therefore, this study aimed to determine the antiviral activity and potential mechanism of AB4 in inhibiting human coronavirus SARS-CoV-2 in vivo and in vitro.@*METHODS@#The cytotoxicity of AB4 was evaluated using the Cell Counting Kit-8 (CCK8) assay. SARS-CoV-2 infected HEK293T, HPAEpiC, and Vero E6 cells were used for in vitro assays. The antiviral effect of AB4 in vivo was evaluated by SARS-CoV-2-infected hACE2-IRES-luc transgenic mouse model. Furthermore, label-free quantitative proteomics and bioinformatic analysis were performed to explore the potential antiviral mechanism of action of AB4. Type I IFN signaling-associated proteins were assessed using Western blotting or immumohistochemical staining.@*RESULTS@#The data showed that AB4 reduced the propagation of SARS-CoV-2 along with the decreased Nucleocapsid protein (N), Spike protein (S), and 3C-like protease (3CLpro) in HEK293T cells. In vivo antiviral activity data revealed that AB4 inhibited viral replication and relieved pneumonia in a SARS-CoV-2 infected mouse model. We further disclosed that the antiviral activity of AB4 was associated with the enhanced interferon (IFN)-β response via the activation of retinoic acid-inducible gene I (RIG-1) like receptor (RLP) pathways. Additionally, label-free quantitative proteomic analyses discovered that 17 proteins were significantly altered by AB4 in the SARS-CoV-2 coronavirus infections cells. These proteins mainly clustered in RNA metabolism.@*CONCLUSION@#Our results indicated that AB4 inhibited SARS-CoV-2 replication through the RLR pathways and moderated the RNA metabolism, suggesting that it would be a potential lead compound for the development of anti-SARS-CoV-2 drugs.
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Objective:To summarize the clinical characteristics of pancreatic solid pseudopapillary neoplasm (pSPN).Methods:From March 2007 to August 2022, at the First Affiliated Hospital of Zhejiang University School of Medicine, the clinical data and follow-up results of 253 patients with pSPN and underwent surgical treatment were retrospectively analyzed. The diagnostic accuracy of preoperative ultrasound, magnetic resonance imaging (MRI), computed tomography (CT) and puncture biopsy of aspiration were compared and analyzed. Kaplan-Meier method was performed to calculate disease-free survival rate. The gender differences in the clinical and pathological features of pSPN were compared. According to whether recurrence and metastasis occurred after the primary operation, the patients with pSPN were divided into recurrence and metastasis group ( n=3) and disease-free survival group ( n=250). Univariate analysis was used to analyze whether gender, age, married status, with clinical symptoms, history of alcohol drinking or smoking, puncture biopsy of aspiration before operation, location and maximum diameter of tumor, surgical method (open surgery, minimally invasive surgery), type of surgery (parenchymal preservation surgery, conventional surgery), and pathological features (cellular atypia, invasion of adjacent organs, lymphovascular invasion, peripancreatic fat invasion, perineural invasion, capsular invasion and pancreatic parenchyma invasion) were predictive factors of recurrence and metastasis of pSPN. Two independent sample t test, chi-square test and Fisher′s exact test were used for statistical analysis. Results:Among 253 patients with pSPN, 49 (19.4%) were males and 204 (80.6%) were females. The diagnostic accuracy of MRI and CT were both higher than that of ultrasound (73.4%(124/169), 64.0%(146/228) vs. 33.3%(78/234)), and the diagnostic accuracy of MRI was higher than that of CT, and the differences were statistically significant ( χ2=62.93, 43.58 and 3.89, P<0.001, <0.001 and =0.049). The diagnostic accuracy of puncture biopsy of aspiration combined with immunochemistry was higher than that without combined immunochemistry (100.0%(23/23) vs. 8/13), and the difference was statistically significant (Fisher′s exact test, P=0.003). Eight cases missed during follow-up, and all the other 245 patients survived with the 5- year and 10-year disease-free survival rates of 99.2% and 97.2%, respectively. Male pSPN patients were older than female patients at diagnosis ((40.0±13.2) years old vs. (32.6±11.9) years old) and the maximum diameter of tumor was smaller than that of female patients ((3.88±2.05) cm vs. (4.87±3.05) cm), and the differences were statistically significant ( t=3.83 and -2.15, P<0.001 and =0.032). The results of univariate analysis showed that lymphovascular invasion was significantly correlated with pSPN recurrence and metastasis (Fisher′s exact test, P=0.012), and the other factors were not correlated with pSPN recurrence and metastasis (all P>0.05). Conclusions:CT, MRI and puncture biopsy of aspiration are comparatively reliable diagnostic method for pSPN before operation. There are significant differences in pSPN tumor growth and age of onset between genders. The postoperative recurrent rate of pSPN is low and the prognosis is good. Even if recurrence and metastasis occur, the patients can still survive for a long time after surgical treatment.
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Objective:To investigate the consistency between the iodine-unstained area and the pathological size of endoscopic submucosal dissection (ESD) specimens of superficial esophageal cancer.Methods:A retrospective study was performed on data of 32 patients with superficial esophageal cancer who accepted ESD from May 2019 to April 2020 in the First Affiliated Hospital, Zhejiang University School of Medicine. The maximum transverse diameter and maximum longitudinal diameter of the iodine-unstained area were compared with the tumor pathological area. A size difference no more than 0.5 cm was considered as conformity, any difference between 0.5 and 1.0 cm was considered as non-conformity, and any difference no less than 1.0 cm was considered as serious non-conformity. At the same time, pink sign after spraying Lugo solution and the consistency of pink sign area with the iodine free area were observed.Results:A total of 32 patients with 33 lesions were enrolled in this study, including 23 males and 9 females and the age of the patients was 59.5±7.3 years. There were 19 (57.6%) lesions whose size of iodine-unstained area was consistent with the tumor pathological area. These 19 lesions were all positive for the pink sign, and the pink sign area overlapped with the iodine-unstained area. In addition, 4 (12.1%) iodine-unstained areas of the lesions did not match the size of the pathological area, and 10 (30.3%) iodine-unstained areas of the lesions were seriously inconsistent with the size of the pathological area. These 14 (42.4%) lesions were all positive for pink sign, and the pink sign area was significantly smaller than the iodine-unstained area. Among the 14 discordant lesions, 2 lesions underwent ESD according to the iodine-unstained area, which resulted in excessive resection and postoperative stenosis.Conclusion:Determining the extent of superficial esophageal cancer by iodine-unstained areas before ESD may lead to excessive resection of the lesions, which is related to the fact that the iodine-unstained areas of the lesions are sometimes significantly larger than the pink sign areas. Therefore, in order to achieve precise treatment, endoscopists can choose the iodine-unstained area with positive pink sign as the first choice for resection.
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Objective:To screen the key exosomal long non-coding RNAs (lncRNAs) molecules that cause nasopharyngeal carcinoma cells to develop chemoradiotherapy resistance.Methods:In vitro, a model of concurrent chemoradiotherapy for human nasopharyngeal carcinoma cells was constructed, and the continuous shock method of high-dose concurrent chemoradiotherapy was used to induce the establishment of chemoradiotherapy-resistant nasopharyngeal carcinoma cell lines, and its resistance formation was verified. Exosomes produced by chemoradiotherapy-resistant cell lines and respective mother cell lines for nasopharyngeal carcinoma were extracted and identified. Finally, biochip technology was used to detect the differential expression levels of exosomal lncRNAs. Results:After 10 repeated treatments of concurrent chemoradiotherapy, CNE-1 CRR and CNE-2 CRR were successfully obtained. Compared with the mother cell lines, CNE-1 CRR and CNE-2 CRR had a tendency to transform from epithelial to interstitial morphology, and the number of cell clones was higher, and the values of average lethal dose (D 0), quasi-threshould dose (D q), survival fraction after 2 Gy irradiation (SF 2) and cell survival rate were higher. Nasopharyngeal carcinoma cells were detected by PCR chip of exosomal lncRNAs. Compared with their respective mother cell lines, 18 lncRNAs in CNE-1 CRR exosomes were significantly up-regulated and 31 lncRNAs were significantly down-regulated, and 15 lncRNAs were significantly up-regulated and 38 lncRNAs were significantly down-regulated in CNE-2 CRR exosomes. CNE-1 CRR also had similar expression profiles to CNE-2 CRR. Conclusion:There are significantly up-regulated and down-regulated lncRNAs in the exosomes of CNE-1 CRR and CNE-2 CRR.
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Cereblon (CRBN), a substrate receptor of cullin 4-RING E3 ligase (CRL4) regulates the ubiquitination and degradation of c-Jun, mediating the lipopolysaccharide-induced cellular response. However, the upstream signaling pathway that regulates this process is unknown. In this study, we describe how endoplasmic reticulum (ER) stress reversely regulates sequestosome-1 (p62)and c-Jun protein levels. Furthermore, our study reveals that expression of p62 attenuates c-Jun protein levels through the ubiquitinproteasome system. Conversely, siRNA knockdown of p62 elevates c-Jun protein levels. Immunoprecipitation and immunoblotting experiments demonstrate that p62 interacts with c-Jun and CRBN to form a ternary protein complex. Moreover, we find that CRBN knockdown completely abolishes the inhibitory effect of p62 on c-Jun. Using brefeldin A as an inducer of ER stress, we demonstrate that the p62/c-Jun axis participates in the regulation of ER stress-induced apoptosis, and that CRBN is required for this regulation. In summary, we have identified an upstream signaling pathway, which regulates p62-mediated c-Jun degradation. Our findings elucidate the underlying molecular mechanism by which p62/c-Jun axis regulates the ER stress-induced apoptosis, and provide a new molecular connection between ER stress and apoptosis.
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Autophagy is a highly conservative cellular self-protective behavior dependent on lysosomes, and can be used as an important factor in promoting or preventing cancer, and its effect is related to the type and development of tumors. A full understanding of autophagy pathway is helpful to improve the diagnosis and treatment of tumors. Studies have shown that autophagy is closely related to the occurrence and development of oral tumors. Autophagy-related genes and signal pathways play a dual regulatory role on oral tumors. This article reviews the latest progress in the regulatory mechanism and therapeutic effect of autophagy on oral tumors.
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Objective:To explore the diagnostic value of endoscopic ultrasonography (EUS) for duodenal accessory papilla.Methods:Data of 122 cases of duodenal accessory papilla diagnosed by EUS at the endoscopy center of the First Affiliated Hospital of Zhejiang University School of Medicine from February 28, 2006 to February 28, 2018 were analyzed and summarized.Results:Of the 122 duodenal accessory papilla cases, the age was 52.1±12.9, with more males than females. The most common site of duodenal accessory papillae was the descending part above the papilla (88/122, 72.13%), followed by the junction of duodenal bulb and descending part (29/122, 23.77%), and a small proportion of lesions located in the duodenal bulb (5/122, 4.10%). Duodenal accessory papillae were all solitary, whose diameter mostly ranged 0.5-1.0 cm (88/122, 72.13%), a smaller proportion of diameter larger than 1.0 cm (23/122, 18.85%), and only a few with diameter less than 0.5 cm (11/122, 9.02%). Most duodenal accessory papillae were hypoechoic (71/122, 58.20%) or moderate to low echogenic (35/122, 28.68%), and the echoes were mostly homogeneous. The mucosa layer was smooth, with a sphincteroid structure in the submucosa and below. The boundary of the duodenal accessory papillae was mostly clear (121/122, 99.18%) and characteristic lacunar cavity structures were often seen in the center (83/122, 68.03%). The surrounding intestinal wall was normal and no associated enlarged lymph nodes were found around the intestine.Conclusion:EUS can clearly show the structure of duodenal accessory papilla and adjacent organs, and is of high value for the diagnosis of duodenal accessory papilla.
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Objective@#To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).@*Methods@#From January 2018 to January 2019, at the First Affiliated Hospital of School of Medicine of Zhejiang University, patients with UC or IC, and health controls, each 10 cases, were enrolled into UC group, IC group and normal control (NC) group, respectively. Fasting serum samples of all the subjects were collected. After removal of high-abundance protein, followed by proteolysis, peptide labeling and fractionating, the samples were then processed by mass spectrometry. The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed. Heat map of protein was constructed. The differential protein was defined as the protein fold change over 1.5 or less than 0.67. The Reactome database was used to cluster the pathways of differential proteins among groups. Statistical methods included t test, hypergeometry test and corrected by BH multiple test.@*Results@#A total of 357 serum proteins were identified by proteomic profiling. There were 27 differential proteins between the IC group and the NC group, including six up-regulated proteins and 21 down-regulated proteins. There were 228 differential proteins between the UC group and the NC group, including 75 up-regulated proteins and 153 down-regulated proteins. There were 49 differential proteins between UC group and IC group, including 22 up-regulated proteins and 27 down-regulated proteins. In the comparison of differential proteins between the NC group, IC group and UC group, only the expression of fibrin 3 was statistically significant (the fold change between UC and NC, between UC and IC, between IC and NC were 0.24, 0.46 and 0.53, respectively; t=-5.089, -7.298 and -3.919, all P<0.01). The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group, only the platelet degranulation pathway was enriched, and 10 proteins were involved in this pathway (P<0.01). In the comparison of differential proteins between UC group and NC group, there were 58 pathways enriched, of which 38 proteins were involved in the platelet degranulation pathway, 16 proteins were involved in the initial complement trigger pathway, 13 proteins were involved in the complement cascade pathway, and 11 proteins were involved in antibody-mediated complement activation pathway (all P<0.01). In the comparison of differential proteins between UC group and IC group, three different pathways were obtained. Among them, nine proteins were involved in the platelet degranulation pathway, seven proteins were involved in the initial complement trigger pathway, and five proteins were involved in the complement cascade pathway (all P<0.01).@*Conclusions@#The difference in serum proteome between IC patients and UC patients was significant, and the differential proteins are mainly involved in platelet activation and complement activation. The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.
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Objective To screen and identify serum protein biomarkers for the differential diagnosis between ischemic colitis (IC) and ulcerative colitis (UC) by tandem mass tag (TMT) combined with liquid chromatography/tandem mass spectrometry (LC-MS/MS).Methods From January 2018 to January 2019,at the First Affiliated Hospital of School of Medicine of Zhejiang University,patients with UC or IC,and health controls,each l0 cases,were enrolled into UC group,IC group and normal control (NC) group,respectively.Fasting serum samples of all the subjects were collected.After removal of high-abundance protein,followed by proteolysis,peptide labeling and fractionating,the samples were then processed by mass spectrometry.The protein with TMT data of three groups was obtained and protein with TMT value 0 were removed.Heat map of protein was constructed.The differential protein was defined as the protein fold change over 1.5 or less than 0.67.The Reactome database was used to cluster the pathways of differential proteins among groups.Statistical methods included t test,hypergeometry test and corrected by BH multiple test.Results A total of 357 serum proteins were identified by proteomic profiling.There were 27 differential proteins between the IC group and the NC group,including six up-regulated proteins and 21 down-regulated proteins.There were 228 differential proteins between the UC group and the NC group,including 75 up-regulated proteins and 153 down-regulated proteins.There were 49 differential proteins between UC group and IC group,including 22 up-regulated proteins and 27 down-regulated proteins.In the comparison of differential proteins between the NC group,IC group and UC group,only the expression of fibrin 3 was statistically significant (the fold change between UC and NC,between UC and IC,between IC and NC were 0.24,0.46 and 0.53,respectively;t =-5.089,-7.298 and -3.919,all P ＜ 0.01).The results of pathway cluster analysis showed that in the comparison of differential proteins between IC group and NC group,only the platelet degranulation pathway was enriched,and 10 proteins were involved in this pathway (P ＜ 0.01).In the comparison of differential proteins between UC group and NC group,there were 58 pathways enriched,of which 38 proteins were involved in the platelet degranulation pathway,16 proteins were involved in the initial complement trigger pathway,13 proteins were involved in the complement cascade pathway,and 11 proteins were involved in antibody-mediated complement activation pathway (all P ＜ 0.01).In the comparison of differential proteins between UC group and IC group,three different pathways were obtained.Among them,nine proteins were involved in the platelet degranulation pathway,seven proteins were involved in the initial complement trigger pathway,and five proteins were involved in the complement cascade pathway (all P ＜ 0.01).Conclusions The difference in serum proteome between IC patients and UC patients was significant,and the differential proteins are mainly involved in platelet activation and complement activation.The candidate proteins identified in this study may be used as biomarkers for the differential diagnosis of UC and IC in the future.
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Objective To set up a computer-assisted polyp detection system under colonoscopy,and to preliminarily verify its effectiveness.Methods Based on Faster R-CNN algorithm and the open source implementation of the open source framework tensorflow and Faster R-CNN,a computer-assisted polyp detection system under colonoscopy was constructed.According to the size and difficulty of the training set,five test groups were set up:test group one,two,three and four contained 1 000,2 000,4 000 and 6 000 training samples,respectively.Test group five increased the probability of selecting the difficult samples based on 6 000 training samples.In different training sets,the sensitivity,specificity,other classification evaluation parameters,and the evaluation parameters of target detection such as recall and precision of this polyps detection system were calculated.Results Classification evaluation parameters showed that the sensitivities of test group one,two,three,four and five were 90.1％,93.3％,93.3％,93.3 ％ and 93.5 ％,respectively,and the difference was statistically significant (x2 =25.324,P＜0.01).The sensitivities of test group two,three,four and five were all higher than that of test group one,and the differences were statistically significant (x2 =13.964,13.508,13.508 and 13.386,all P＜ 0.006 25).There were no significant differences in specificity and positive predictive value among test groups (both P＞0.05).The negative predictive values of test group one,two,three,four and five were 90.4％,93.3％,93.3％,93.3％ and 93.5％,respectively,and the differences were statistically significant (x2 =21.862,P＜0.01).The negative predictive values of test group two,three,four and five were higher than that of test group one,and the differences were statistically significant (x2=11.447,11.564,11.755,13.760;all P＜0.006 25).As the training sample size increased from 1 000 to 2 000,the area under curve (AUC) increased by 2％,and further increased the sample size to 6 000,AUC increased by less than 1 ％.At this point maintaining the same sample size while increasing the proportion of difficult samples,AUC increased by 0.4％.The results of evaluation parameters of target detection showed that the recall rate of each test group was 73.6％,79.8％,79.5％,79.8％ and 83.3％,respectively,and the differences were statistically significant (x2 =71.936,P＜0.01).Among them,the recall rates of test group two,three and four were higher than that of test group one,and the differences were statistically significant (x2 =25.960,23.492 and 25.960,all P＜0.006 25),and the recall rate of test group five was higher than those of test group one,two,three and four,and the differences were statistically significant (x2=67.361,9.899,11.527 and 9.899;all P＜0.006 25).In addition,the precision rates of test group one,two,three,four and five were 87.9％,85.3％,90.2％,91.4％ and 89.2％,respectively,and the difference was statistically significant (x2=48.194,P＜0.01).The precision rates of test group three and five were higher than that of test group two,and the differences were statistically significant (x2 =24.508 and 15.223,both P＜0.006 25),and the precision rate of test group four was higher than those of test group one and two,and the differences were statistically significant (x2=13.524 and 39.120,both P＜0.006 25).As samples size and training difficulty increased,the values of F1-score and mean average precision increased steadily.Conclusions This study initially constructed a computer-assisted polyp detection system under colonoscopy.Currently the maximum sensitivity reached 93.5％,and the maximum recall rate reached 83.3％.Increasing the training set size may improve the polyp detection result to a certain degree,however it will reach a bottleneck.At this time,increasing the training difficulty can further improve the detection scores,especially the recall rate.
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Oral cancer is not only a serious threat to human health and life but also its incidence rate has been increasing year by year. Oral cancer animal model is indispensable for the research of clinical diagnosis and treatment of oral cancer. Therefore, it is of great importance to establish an animal model of disease which is similar to the natural occurrence of human oral cancer and to study its pathogenesis, prevention and treatment. At present, animal models of oral cancer mainly include spontaneous animal model, induced animal model and genetically modified animal model. The research progress, current problems and the future development perspectives of animal models of oral cancer in recent years are summarized in this paper.
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Objective To analyze the correlation between serum leptin and osteoprotegerin with IL-6 and TNF-α in the pa-tients with gout arthritis osteoporosis .Methods Forty-eight cases of gout arthritis were selected as the research subjects (GA group) ,including 19 cases of complicating osteoporosis(OP group) and 29 cases of non-complicating osteoporosis(non-OP group) . Other 45 individuals undergoing healthy physical examination were selected as the control group .The bone mineral density (BMD) was measured by dual energy X-ray BMD absorptionmetry instrument and the levels of serum LEP ,OPG ,IL-6 and TNF-αwere de-tected by ELISA .Results The incidence rate of osteoporosis in the GA group was significantly higher than that in the control group (39 .6% vs .13 .3% ,P=0 .004) ,and BMD in various sites was significantly lower than that of the control group (P<0 .05) . The serum LEP level in the OP group was significantly increased compared with that in the non-OP group and control group(P<0 .05) ,and the OPG level was significantly decreased ,the difference was statistically significant (P<0 .05) .The serum LEP level in the non-OP group was significantly higher than that in the control group (P<0 .05) ,but the OPG level had no statistical difference between non-OP group and control group(P>0 .05) .The serum IL-6 and TNF-αlevels in the OP group were significantly higher than those in the non-OP group and control group(P<0 .05) ,and the non-OP group was significantly higher than the control group (P<0 .05) .The serum LEP level in the OP group was positively correlated with IL-6 and TNF-α(P<0 .05) ,and the serum OPG level was negatively correlated with IL-6 and TNF-α(P<0 .05) .Conclusion The serum LEP and OPG levels have abnormal change in the patients with gout arthritis osteoporosis ,which has a certain correlation with IL-6 and TNF-α.
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Objective To analyze the clinical manifestations, computed tomography scan (CT), gastroscope, endo-scopic ultrasonography (EUS), and therapy method of gastritis cystica profunda. Methods Retrospectively analyzed clinical manifestations, CT, gastroscope, EUS, and pathological results of 6 cases of gastritis cystica profunda. Results In these 6 cases, 3 of them were doubted gastric carcinoma, 3 cases were considered stomach mass by CT. Gastroscope hinted apophysis lesions, but all cases were suggested gastritis cystica profunda by EUS. And all cases were removed through endoscopic submucosal dissection (ESD). Pathology were confirmed the diagnosis. Conclusion EUS combined with endoscopic mucosal resection (EMR) or ESD technique can improve the diagnostic rate. For gas-tritis cystica profunda which are not associated with malignant tumor can be treated through ESD.
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Proteolytic cleavage is one of the post-translational modifications and plays important roles in many biological processes, such as apoptosis and tumor cell metastasis. The identification of the cleavage events can improve our understanding of their biological functions in these processes. Although proteomic approaches using N-terminal labeling have resulted in the discovery of many proteolytic cleavages, this strategy has its own inherent drawbacks. Labeling of protein C-termini is an alternative approach. Here, we optimized the labeling procedure in the profiling protein C-termini by enzymatic labeling (ProC-TEL) and improved the labeling efficiency for the positive isolation of protein C-terminal peptides and mass spectrometric identification. We applied this approach to a complex protein mixture from Escherichia coli and identified many C-terminal peptides and internal cleaved peptides from more than 120 proteins. From the identified cleavages, we found several previously known internal proteolytic cleavage sites and many novel ones which may play roles in regulating normal biological processes. This work provides a potential new way, complementary to the N-terminomics, for the identification of proteolytic cleavages in complex biological systems.
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Cathepsin A , Chemistry , Protein C , Chemistry , Protein Processing, Post-Translational , Proteolysis , ProteomicsABSTRACT
Objective To explore the diagnostic model and clinical application value of serum proteomic fingerprint in inflammatory bowel disease (IBD) .Methods Serum proteome profiles of 72 IBD patients (54 Crohn′s disease (CD) and 18 ulcerative colitis (UC) and 44 healthy controls were analyzed by the weak cation exchange (WCX) beads combined matrix‐assisted laser desorption/ionization time of flight mass spectrometry (MALDI‐TOF‐MS ) technique . Among three groups , every two groups were compared .Wilcoxon rank sum test was used to screen out the peaks of difference expressed protein (P<0 .05) .Genetic algorithm combining with support vector machine (SVM ) was utilized to select the best diagnostic model .The predictive effects of this model was evaluated by leave one out method (LOO ) . Results The 10 most discriminating protein peaks were screened out between CD group and healthy control group , between UC group and healthy control group , between CD group and UC group . A diagnostic model established with four protein peaks ,the mass‐to‐charge ratio (M /Z ) of them was 3 275 .29 ,4 963 .91 ,4 980 .53 and 5 336 .90 ,could better distinguish CD and healthy controls .The specificity was 97 .7% ,and the sensitivity was 92 .6% in CD diagnosis .A diagnostic model established with four protein peaks ,the M /Z of them was 2 272 .41 ,2 660 .42 ,3 029 .77 and 5 002 .78 ,could better distinguish UC and healthy controls .The specificity was 100 .0% ,and the sensitivity was 94 .4% .A specificity was 50 .0% and sensitivity was 88 .9% in CD diagnosis with the diagnostic model of six protein peaks and the M /Z of them was 2 082 .63 ,2 210 .64 ,4 039 .02 ,4 298 .30 ,4 978 .03 ,5 002 .22 .Conclusion The diagnostic model of serum difference expressed protein in CD and UC is established by MALDI‐TOF‐MS technique and genetic algorithm combining with SVM ,which has high diagnostic value in IBD .
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Objective To investigate the role and possible mechanism of caveolin-1 (CAV1) in the forming of cholesterol gallstone in mice fed with lithogenic diet.Methods Cholesterol gallstone susceptible C57BL/6 mice were study objects.The mice of control group (n=6) and experiment group (n=6) were fed with normal diet and lithogenic diet for four weeks respectively.The condition of cholesterol gallstone forming,changes of serum lipid and bile composition were measured,and the expressions of CAV1 and scavenger receptor classB member Ⅰ (SR-BⅠ) at mRNA and protein level in the liver and gallbladder were detected by realtime-polymerase chain reaction and Western blot,respectively.The t test was performed for mean comparsion between the two groups.Results The incidence rate of gallstone in experimental group was 100％ after fed with lithogenic diet for four weeks,the lipid level significantly increased,and the proportion of cholesterol in bile raised and bile salt decreased.Compared with those of control group,the expressions of CAV1 at mRNA and protein level in the liver and gallbladder tissues siginificantly decreased (in liver tissue,mRNA 0.53 ± 0.13 vs 1.00 ± 0.32,t =3.330,protein level 0.39 ± 0.07vs 0.92±0.06,t=10.280; in gallbladder tissue,mRNA 0.40±0.22 vs 1.00±0.22,t=3.823,protein level 1.04±0.07 vs 1.34 ± 0.04,t =6.367,all P＜0.01).There was no significant difference in the relative expression of SR-BⅠ at mRNA and protein level in the liver and gallbladder tissues between the mice of experiment group and control group.Conclusion The changes of CAV 1 expression at mRNA and protein level in liver and gallbladder tissues may affect lipids metabolism and cholesterol transportation in liver and gallbladder tissues of experiment mice,which might play an important role in the formation of cholesterol gallstone.
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BACKGROUND:In order to optimize the biological activity of hydroxyapatite, previous experiments have used plasma spraying technique to prepare a piezoelectric ceramic coating on the surface of hydroxyapatite, but the cytotoxicity of this new material is not clear. OBJECTIVE: To evaluate the cytotoxicity of hydroxyapatite/barium titanate biological piezoelectric ceramic coatingin vitro. METHODS: The 3rd generation beagle bone marrow mesenchymal stem cels were seeded on hydroxyapatite/barium titanate piezoelectric ceramic specimens and hydroxyapatite specimens, respectively. After 5 days, the celladhesion was detected by scanning electron microscopy. The 3rd generation bone marrow mesenchymal stem cels were also co-cultured with hydroxyapatite/barium titanate piezoelectric ceramic specimen extract, hydroxyapatite specimen extract, low-glucose Dulbecco’s modified Eagle’s medium containing 5% dimethylsulfoxide and 15% fetal bovine serum, and low-glucose Dulbecco’s modified Eagle’s medium containing 15% fetal bovine serum, respectively. The cytotoxicity was tested by cellCounting Kit-8 assay at days 1, 3, 5 after co-culture. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cels on the surface of hydroxyapatite/barium titanate piezoelectric ceramic specimens and hydroxyapatite specimens grew proliferatively and presented with multi-layer growth. The connection between cels and pseudopodia was very close, which indicates that the two kinds of materials both have good cytocompatibility. cellCounting Kit-8 assay showed that the cels cultured in the extracts of hydroxyapatite/barium titanate biological piezoelectric ceramic and hydroxyapatite specimens proliferated more than 80%, and the toxicity was grade 1 that meant no cytotoxicity.
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Objective To investigate the clinical significance of serum anti-Saccharomyces cerevisias antibody (ASCA),anti-outer membrane porin C (anti-OmpC),antibody to Pseudomonas fluorescens-associated sequence I2 (anti-I2 )and antibody to bacterial flagellin (anti-CBirl )in the diagnosis and treatment of inflammatory bowel disease (IBD).Methods From 2011 to 2013,87 patients with IBD were enrolled and divided into Crohn′s disease (CD)group (66 cases)and ulcerative colitis (UC)group (21 cases).A total of 62 age and gender matched healthy individuals were enrolled as the control group. Fasting blood samples (2 mL)of the subjects were collected.The expression of ASCA,anti-OmpC,anti-I2 and anti-Cbirl antibodies was detected with enzyme-linked immunosorbent assay (ELISA)kits.The diagnosis value of each antibody in IBD and the differential diagnostic value of in UC and CD were compared by receiver operating characteristic (ROC)curve.Results The area under the curve (AUC)of ASCA between IBD and the healthy control group,between CD group and UC group was 0.580 and 0.512, respectively;the accuracy in diagnosis was low.The AUC of anti-CBirl between IBD and the healthy control group was 0.617.There was no differential diagnosis significance of the other antibodies.The positive rate of ASCA in IBD group was 62.1 % (54/87),which was significantly higher than that in the control group (38.7%,24/62).The positive rates of anti-OmpC and anti-I2 in IBD group was significantly lower than those in the control group and the differences were statistically significant (both P 0.05).The specificity,sensitivity,positive predictive value (PPV)and negative predictive value (NPV)of ASCA in differential diagnosis of CD and UC was 52.4%,66.7%,81 .48% and 33.33%,respectively.The specificity and sensitivity of anti-OmpC,anti-I2 and anti-CBirl in differential diagnosis of CD and UC was 81 .0% to 100.0% and 9.1 % to 37.9%,respectively.The specificity,sensitivity,PPV and NPV of double-positive ASCA and anti-I2 in the diagnosis of CD was 57.1 %,86.4%,82.6% and 50.0%, respectively.The positive rate of ASCA and anti-I2 in CD group was significantly higher than that in UC group (84.8%(56/66)vs 57.1 % (12/21 );χ2 =5 .633,P =0.018 ).Conclusions Positive ASCA has some significance in the diagnosis of patients with IBD in our country.The detection of anti-I2 can help to diagnose ASCA negative CD.Because of low sensitivity and positive rate,anti-OmpC and anti-CBirl have limited value in the diagnosis of IBD and the differential diagnosis of UC and CD in our country.
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Objective To explore the role of caveolin-1 in nonalcoholic fatty liver disease (NAFLD) caused by high-fat diet.Methods A total of 12 ten-week-old male C57BL/6 mice were fed with high-fat and high-cholesterol diet for 14 weeks to establish the NAFLD animal model.And six syngeneic mice fed with normal diet at the same time were taken as control.All the mice were sacrificed by the end of 14th week,body weight,liver weight and the changes of serum lipids of the two groups were compared.The changes of caveolin-1 at mRNA and protein levels in the liver of mice with NAFLD were detected by quantitative polymerase chain reaction (qPCR) and Western blot.The liver steatosis of the mice was observed under light microscopy after stained by hematoxylin and eosin.The changes of distribution of caveolin-1 in liver were examined by immunohistochemistry.The differences of caveolin-1 at mRNA and protein level in livers between the two groups were compared by t test.The differences of immunohistochemical scores of caveolin-1 expression in the livers of mice with different degree of fatty liver were analyzed by ordinal variables of two independent samples ranksum test analysis.Results After 14 weeks high-fat and high-cholesterol diet,all the mice of experiment group developed NAFLD.Nine of which were severe and three were moderate.Compared with the control group,serum total cholesterol,high density lipoprotein cholesterol and low density lipoprotein cholesterol of experiment group significantly increased ((1.940 ± 0.300) mmol/L vs (3.771±0.800) mmol/L,(1.120±0.066) mmol/L vs (2.224±0.420) mmol/L,(0.510±0.191) mmol/L vs (1.241±0.660) mmol/L,t=-3.760,-5.474,-3.332,all P＜0.01),however there was no significant difference in triglyceride (P＞0.05).The caveolin-1 of experiment group significantly increased at mRNA (1.536 ±0.226 vs 0.980± 0.272,t=3.371,P＜0.05) and protein levels (0.643±0.240 vs 0.100±0.130,t=4.847,P＜0.01).The immunohistochemical results indicated that the increased caveolin-1 expression mainly distributed in the membrane of hepatocytes,cytoplasm and membrane of lipid droplets.Conclusion The up-regulated caveolin-1 expression in the livers of NAFLD mice induced by high-fat and high-cholesterol may be involved in the mechanism of NAFLD.
ABSTRACT
Objective To analyze endoscopic ultrasonography (EUS) image of esophageal granular cell tumor (GCT) by computer image analysis software,and to explore its diagnostic and differential diagnostic value.Methods Eight esophageal GCT and eight leiomyomas confirmed by pathological and immunohistochemical examination were collected,mean gray value and gray value standard deviation of EUS image of the leisons were analyzed by computer image analysis software.The comparison of two groups was analyzed by t test.Results The endoscopic and ultrasound images of esophageal GCT and leiomyomas were similar.The mean gray value of EUS image of esophageal GCT and esophageal leiomyomas was 67.51 and 51.76,the difference was statistically significant (t=2.225,P=0.043).However,there was no significant difference in the standard deviation of gray value (13.54 vs 13.12,t=0.225,P=0.825).Conclusion The accuracy of GCT diagnosed by EUS could be increased by comparing the gray values of EUS images of esophageal GCT and leiomyomas.