Objective:To investigate the expression of nuclear matrix protein 4 (NMP4) in hepatocellular carcinoma (HCC), and its relationship with clinicopathological features and survival prognosis of patients.Methods:The clinical data of 100 HCC patients who were treated with radical resection of liver cancer in the Department of Hepatobiliary Surgery of the Third Affiliated Hospital of Wenzhou Medical University from July 1, 2014 to July 1, 2019 were retrospectively analyzed. There were 63 males and 37 females, aged (58.5±10.4) years old. Immunohistochemical method was used to detect the expression of NMP4 protein in HCC cancer tissue and the corresponding adjacent normal tissue. According to the expression of NMP4 in HCC tissues, 100 patients were divided into two groups: the NMP4-positive expression group ( n=62) and the NMP4-negative expression group ( n=32). Univariate analysis was performed on the relationship between NMP4 expression and clinical pathological features as well as overall survival of HCC patients. Cox multivariate analysis was performed on the factors influencing postoperative prognosis of HCC patients. Results:Immunohistochemistry results showed that NMP4 was primarily expressed in the nucleus, the positive expression rate of NMP4 in HCC tissues was higher than that in adjacent non-cancerous tissues [62.0% (62/100) vs. 8.0%(8/100)], and the difference was statistically significant ( χ2=2.12, P=0.003). Univariate analysis revealed that the overall survival of HCC patients was correlated with the degree of tumor differentiation, tumor length, BCLC stage, number of tumor foci, vascular tumor thrombus and expression of NMP4 (all P<0.05). Cox multivariate analysis revealed that low differentiation, high BCLC stage (stage C), number of tumor foci (≥3), and positive expression of NMP4 were independent risk factors affecting postoperative survival and recurrence-free survival of HCC patients. The median overall survival and median recurrence-free survival of HCC patients in the NMP4-positive expression group were 22.3 months and 11.5 months, respectively. In contrast, that in the NMP4-negative expression group were 40.6 months and 19.4 months, respectively. The cumulative survival rate and recurrence-free survival rate of HCC patients in the NMP4-positive expression group were lower than those in the NMP4-negative expression group, and the differences were statistically significant (both P<0.05). Conclusion:Positive NMP4 expression was closely correlated with malignant biological progression and poor prognosis of HCC patients.

Objective:To investigate the expression characteristics of mucin 5B (MUC5B) protein and programmed cell death factor 4 (PDCD4) protein in patients with intrahepatic cholangiocarcinoma (ICC), and to construct a nomogram model for prognosis prediction.Methods:Clinical data of 100 patients who underwent radical surgical resection and were diagnosed as ICC by postoperative pathology from September 2009 to September 2020 in the Third Affiliated Hospital of Wenzhou Medical University were retrospectively selected, including 46 males and 54 females, aged (56.9±12.2) years old. Immunohistochemistry was used to detect the expression of MUC5B and PDCD4 protein in 100 cases of ICC and corresponding adjacent tissues respectively, and the relationship between them and clinicopathological factors of ICC patients was analyzed. Univariate and multivariate Cox regression analysis were performed to analyze the influencing factors on postoperative prognosis of ICC patients. The nomogram model was constructed using rms package and performed internal verification.Results:The positive expression rate of MUC5B protein in ICC was 76.0% (76/100), which was higher than that in para-cancer tissues 27.0%(27/100), and the difference was statistically significant ( χ2=11.33, P=0.015). While the positive expression rate of PDCD4 protein in ICC was 21.0%(21/100), which was lower than that in normal tissues 73.0% (73/100), and the difference was statistically significant ( χ2=15.57, P=0.007). Multivariate Cox regression analysis showed that ICC patients with carbohydrate antigen 19-9>37 kU/L, tumor length>5 cm, tumor TNM stage Ⅱ/Ⅲ, tumor medium/low differentiation, MUC5B positive expression, and PDCD4 negative expression had a high risk of short survival after resection (all P<0.05). The nomogram model was constructed based on the above indicators, and the C-index was 0.801. The postoperative survival calibration curve showed that the high predictive survival fit of the nomogram model, and the area under the receiver operating characteristic curve was 0.862. Conclusions:Positive expression of MUC5B protein and negative expression of PDCD4 protein in ICC tissue suggest poor prognosis of ICC patients. The nomogram model constructed on the basis of MUC5B and PDCD4 protein is well distinguished and has ideal predictive efficacy.

Objective:To study the roles of hepatoma-derived growth factor (HDGF), vascular endothelial growth factor (VEGF) and mucin 5B (MUC5B) in hepatolithiasis associated with intrahepatic cholangiocarcinoma (HICC).Methods:The clinical data and tissue specimens of 116 patients who underwent hepatectomy at the Third Affiliated Hospital of Wenzhou Medical University from October 1999 to October 2019 were retrospectively analyzed. There were 41 patients with HICC (the HICC group), 38 patients with intrahepatic cholangiolithiasis (the intrahepatic cholangiolithiasis group), and 37 patients with benign liver tumor who underwent hepatectomy (the control group). There were 47 males and 69 females, with age of (66.1±3.2) years old. The positive expressions of HDGF, VEGF and MUC5B in the three groups were compared. In 41 patients with HICC, the correlation between positive expressions of HDGF, VEGF and MUC5B with patients’ clinical characteristics were studied.Results:Compared with the control group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group and the intrahepatic cholangiolithiasis group were significantly increased, ( P<0.05). Compared with the intrahepatic cholangiolithiasis group, the positive expression rates of HDGF, VEGF and MUC5B in the HICC group were significantly increased ( P<0.05). Positive expression of VEGF in HICC patients was correlated with tumor differentiation, tumor local invasion, tumor length, tumor stage, tumor carbohydrate antigen CA19-9 level and lymph node metastasis ( P<0.05). Spearman correlation analysis showed that HDGF was positively correlated with VEGF expression in HICC tissues specimens ( r=0.935, P<0.01). Survival analysis showed that the cumulative survival rates (36.7%, 17.1%, 7.3%) of patients with positive expression of VEGF were significantly lower than that of patients with negative expression (51.2%, 26.8%, 19.5%) at 1, 3, 5 years after surgery ( P<0.01). The cumulative survival rate (34.1%, 17.1%, 4.9%) of patients with MUC5B positive expression were significantly lower than that of patients with negative expression (53.7%, 31.7%, 17.1%) at 1, 3, 5 years after surgery ( P<0.01). Conclusion:HDGF can be used as a reference indicator for early assessment of HICC. Overexpressions of VEGF and MUC5B can be used as important indicators for HICC in evaluating disease progression and prognosis.

[摘 要] 目的：分析FOXD1在食管鳞状细胞癌（ESCC）组织中的表达及其与临床病理特征和患者预后的关系，探讨其对ESCC TE1细胞增殖、侵袭能力的影响及其对TGF-β1诱导TE1细胞EMT进程的影响。方法：采用qPCR和IHC法检测ESCC组织和细胞中FOXD1的表达，并分析其与临床病理特征和患者预后的关系；构建FOXD1敲减质粒并转染TE1细胞，检测其对TE1细胞增殖、侵袭能力的影响；用qPCR和WB法检测TGF-β1处理前后FOXD1及EMT相关基因和蛋白的表达变化及敲减FOXD1对EMT相关基因和蛋白表达的影响。结果：ESCC组织和细胞中FOXD1均呈高表达（均P<0.01），并与患者OS呈负相关；FOXD1表达水平、肿瘤TNM分期以及淋巴结转移均是影响ESCC患者预后的独立危险因素（均P<0.01）。TGF-β1可促进TE1细胞FOXD1的表达，并诱发其EMT进程（均P<0.05）；敲减FOXD1可抑制TE1细胞的增殖和侵袭能力，并可部分逆转由TGF-β1诱发的TE1细胞EMT进程。结论：FOXD1在ESCC组织及TE1细胞中呈高表达且是影响ESCC患者预后的独立危险因素，敲低FOXD1可显著抑制TE1细胞的增殖、侵袭及TGF-β1介导的EMT进程。

Objective To investigate the function of mucin 2,mucin 4 on formation of lithogenic bile in patients with calculus of intrahepatic duct.Methods Bile duct mucosa,bile duct wall,bile and plasma were collected from 56 patients with calculus of intrahepatic duct (CID group) and 17 individuals without calculus of intrahepatic duct (control group).The bile duct wall was stained with mucin 2 (MUC2) and mucin 4 (MUC4).Reverse-transcription polymerase chain reaction (RT-PCR) was applied to study the mRNA expressions of MUC2 and MUC4 in the bile duct mucosa.The correlation of the bile duct and serum lipid index was analyzed.Results Serum lipid index in the CID group was significantly higher than control group (all P＜0.05).Biliary total bile acids and bile acids mol percentage were lower in the CID group than control group (both P＜0.05).The expressions of MUC2 was not increased significantly in CID group than the control group (all P＞0.05).The expressions of MUC4 were more significantly increased in CID group than the control group (P＜0.05).The mRNA of MUC4 in the CID group was also more significantly increased than in control group (P＜0.01).There were no correlations between MUC4 expression and the level of biliary total bile acid in the CID group (r=-0.374,P＜0.05).Conclusion The expression of MUC4 in patients with CID was enhanced,which promoted the absorption of bile acid by the mucosal epithelium of the bile duct,and caused a large amount of mucin to be secreted into bile,which may be related to the formation of stony bile.

Objective: To assess the clinical characteristics and identify the risk factors in the acute myocardial infarction (AMI) patients complicating with ventricular septal rupture (VSR). Methods: A retrospective study was performed on 96 AMI patients complicating with VSR, who were hospitalized in the Second Xiangya Hospital of Central South University, Hunan Provincial Peoples′ Hospital, the First Affiliated Hospital of University of South China, the Second Affiliated hospital of University of south China, Xiangtan Central Hospital from December 2007 to May 2017. There were 46 females and the age was (66.2±10.7) years (from 43 to 90 years). Patients were divided into in-hospital survival group (n=64) and in-hospital death group (n=32). The 96 patients were also divided into the early death group (survived ≤2 weeks after admission, n=50) and non-early death group (survived>2 weeks after admission, n=46). Multivariate logistic regression was used to analyze the independent risk factors of the early death. Results: Location of VSR was available in 71 patients, VSR was located at the apical or anterior septum near the apical region in 64.0% (32/50) patients with the anterior AMI, VSR was located at the posterior wall and basal inferior segment in 57.1% (12/21) patients with non-anterior AMI. Compared to the in-hospital survival group, patients in the in-hospital death group were older ((69.6±11.3) years vs. (64.6±10.1) years, P=0.031), incidence of non-ventricular aneurysm (71.9% (23/32) vs. 37.5% (24/64), P=0.001) and anterior AMI (84.4%(27/32) vs. 62.5%(40/64), P=0.028) was significantly higher in the in-hospital death group than in the in-hospital survival group. The comparison between the early death group and non-early death group showed that older age, female, no history of angina or myocardial infarction, Killip grade>Ⅲ, and non-ventricular aneurysm were related to increased risk of the early mortality in this patient cohort. Logistic regression analysis revealed that female (OR=5.109,95%CI 1.19-22.00, P=0.012), no history of angina or myocardial infarction (OR=23.34, 95%CI 3.44-158.37, P=0.001), Killip grade>Ⅲ(OR=5.35, 95%CI 1.26-22.66, P=0.019) and non-ventricular aneurysm (OR=6.30,95%CI 1.67-23.73, P=0.005) were independent risk factors for early death in this patient cohort. Conclusion The risk factors of in-hospital death include older age, non-ventricular aneurysm and anterior AMI. Female, no history of angina or myocardial infarction, Killip grade>Ⅲ and non-ventricular aneurysm are independent risk factors for the early death of AMI patients complicating VSR.

Objective To investigate the expression of early growth response protein 1 (Egr-1),NGFI-A binding protein 2 (Nab2) and caveolin 1 (Cav-1) in normal skin,flat-cicatrix and hypertrophic scar,and explore its role in the formation of hypertrophic scar.Methods The expression of Egr-1,Nab2 and Cav-1 protein in 9 normal skin tissues,8 flat-cicatrix tissues and 9 hypertrophic scar tissues were examined with immunohistochemistry SP method and were analyzed statistically.Results The expression of Egr-1 in epidermal cells of hypertrophic scar was significantly higher than that in normal skin and flat scar tissue.The expression of Egr-1 increased in the course of scar proliferation.The distribution patterns of Nab2 were different from Egr-1.The expression of Egr-1 was increased,while expression of Nab2 was decreased.The expression of Cav-1 in normal skin and flat-cicatrix was significantly higher than that in hypertrophic scar.Conclusions The expression of Egr-1,Nab2 and Cav-1 is closely related to the formation of hypertrophic scar,and the up-regulated expression of Egr-1 and the deficient expression of Nab2 and Cav-1 may be the indicators of the progress of formation of hypertrophic scar.

Objective To investigate the expression of early growth response protein 1 (Egr-1),NGFI-A binding protein 2 (Nab2) and caveolin 1 (Cav-1) in normal skin,flat-cicatrix and hypertrophic scar,and explore its role in the formation of hypertrophic scar.Methods The expression of Egr-1,Nab2 and Cav-1 protein in 9 normal skin tissues,8 flat-cicatrix tissues and 9 hypertrophic scar tissues were examined with immunohistochemistry SP method and were analyzed statistically.Results The expression of Egr-1 in epidermal cells of hypertrophic scar was significantly higher than that in normal skin and flat scar tissue.The expression of Egr-1 increased in the course of scar proliferation.The distribution patterns of Nab2 were different from Egr-1.The expression of Egr-1 was increased,while expression of Nab2 was decreased.The expression of Cav-1 in normal skin and flat-cicatrix was significantly higher than that in hypertrophic scar.Conclusions The expression of Egr-1,Nab2 and Cav-1 is closely related to the formation of hypertrophic scar,and the up-regulated expression of Egr-1 and the deficient expression of Nab2 and Cav-1 may be the indicators of the progress of formation of hypertrophic scar.

OBJECTIVETo understand the epidemiological profiles of hand, foot and mouth disease (HFMD) and the major enteroviruses causing the epidemics of HFMD in Shanghai from 2010 to 2014.

METHODThe city-wide surveillance data between 2010 and 2014 were used to analyze the epidemiologic characteristics of the HFMD outbreaks in Shanghai. The annual incidence of HFMD was estimated based on the 2010 Shanghai Census data.

RESULTFrom 2010 to 2014, the reported HFMD cases were 41 080, 37 323, 51 172, 42 198, and 65 018, respectively; the severe cases (case-severity ratio) were 469 (1.14%), 456 (1.22%), 318 (0.62%), 104 (0.25%) and 248 (0.38%), respectively. Based on Shanghai census data by the end of 2010, the attack rates of HFMD in Shanghai were 0.16%-0.28% in the entire population. In terms of the proportion of HFMD cases and severe cases in the specific population, male accounted for 59.62%-61.48% and 62.26%-73.08%, migrant population accounted for 51.86%-62.40% and 72.01%-80.38%; children aged 1.0-1.9 years comprised the highest proportion, up to 22.70%-27.00% and 32.08%-36.40%. HFMD peaked from April to July, in parallel with the peak circulation of enterovirus (EV) 71, and a small peak usually occurred in autumn and winter. All the critically severe and fatal cases were caused by EV71. The detection rates of EV71 and Coxsackievirus A (CA) 16 were 73.08%-88.09% and 1.12%-2.90% in severe HFMD cases, 19.75%-48.74% and 2.02%-23.69% in uncomplicated inpatients, and 16.78%-40.08% and 8.36%-33.39% in mild community cases, respectively. The detection rates of CA6 and CA10 in the mild community cases in 2014 were 18.38% and 1.43%, respectively. In 2013 non-EV71 and non-CA16 enteroviruses comprised 74.86% in the community cases.

CONCLUSIONThe annual HFMD outbreaks occurred in Shanghai during 2010-2014. Children under 5 years of age, migrant population and male were the major susceptible population. EV71 and CA16 were the predominant pathogens causing the epidemics of HFMD except in 2013, and CA6 was prevalent in the community cases in 2014. The major peak season of HFMD usually overlapped with the peak of EV71 circulation and the majority of severe HFMD cases were associated with EV71 infection.

Child ; China ; epidemiology ; Disease Outbreaks ; Enterovirus A, Human ; Female ; Hand, Foot and Mouth Disease ; epidemiology ; Humans ; Incidence ; Male ; Prevalence ; Seasons

Objective To choose one protocol that can quickly ,safely and effectively provide amount enough of bone marrow derived mesenchymal stem cells(MSCs) for use of clinical or experimental test through comparison of their growth characteristics and growth factors levels in culture solution .Methods Cells extracted from bone marrow of C57BL/C mice respectively underwent two different isolation protocols :whole bone marrow adherent culture(WBMAC) or gradient density separation(GDS);characteris‐tic surface antigens of MSCs were identified by flow cytometry on cells isolated in different ways ;the distinct growth curve of pri‐mary stem cells cultured in vitro described their different proliferation rate;levels of vascular endothelial growth factor(VEGF) and stromal cell‐derived factor‐1α(SDF‐1α) in culture medium were detected by ELISA .Results Primary MSCs obtained by WBMAC proliferated at higher speed and exhibited shorter growth cycle than those separated by GDS ;on MSCs from both groups ,surface antigens CD29 were detected positively ,and antigens including CD31 ,CD34 and CD45 were assayed negatively ;concentration of VEGF and SDF‐1αin both two nutrient solution primarily keep at low levels ,comparatively ,level of VEGF and SDF‐1αin the di‐shes which contain MSCs by WBMAC was higher than the one in the dishes which contain MSCs by GDS .Conclusion MSCs ex‐tracted by WBMAC shows unimpaired cell function ,can build automatically more suitable microenviroment for their growth;this classic method was qualified for clinical and experimental use in a safe ,rapid ,effective way .