Ocean is a unique and excellent resource that provides a diverse array of intriguing natural products. Marine natural products have demonstrated significant and extremely potent biological activities and have captured the attention of natural products chemists in the past few decades. It is increasingly recognized that a wealth of fascinating natural products and novel chemical entities will play a dominant role in the discovery of useful leads for the development of pharmaceutical agents and provide useful probes to lead to breakthroughs in a variety of life-science fields. This article focused on the research progress of chemistry of marine natural products in recent five years.

Objective To investigate the expressions of TESTIN gene and Caspase-3 protein and thier relations with the clinicopathological features and prognosis of esophageal squamous cell carcinomas(ESCC). Methods The expressions of TESTIN and Caspase-3 in 50 ESCC tissues and paracancerous tissues (＞5 cm apart form the ESCC tissue) were detected by immunohistochemistry.The expression of TESTIN in 65 matched-pairs of ESCC tissues was detected by Western blotting and RT-PCR. The association of TESTIN and Caspase-3 with clinicopathological features and prognosis of ESCC were analyzed. Results The immunohistochemistry examination showed that the positive rates of TESTIN protein [30. 0% (15/50)] and Caspase-3 protein [24.0% ( 12/50)] were significantly lower in ESCC tissues than those in paracancerous tissues [(84. 0% (42/50) and 94. 0% (47/50),respectively, P＜0.01)]. The mRNA level of TESTIN was down-regulated in ESCC tissues (P＜0.05). Whereas the protein level of TESTIN was down-regulated in 45 (69.2%) of 65 ESCC tissues in comparison with paracancerous tissues. TESTIN expression was positively correlated with the differentiation of ESCC, but not associated with gender, age, tumor size, TNM stage and lymph node metastasis. There was a positive correlation between TESTIN and Caspase-3 in protein expressions (P＜0. 05). Patients with negative expression of TESTIN had lower survival rate compared to those with positive expression (P＜0. 05). Conclusions The positive relation between low-expression of TESTIN and Caspase-3 implicates that both are involved in the development of esophageal squamous cell carcinogenesis, and TESTIN might be a novel ESCC marker with prognostic significance.

Aim To detect the antitumor activities of five sesquiterpene lactones from the roots of Inula helenium.Method Using MTT assay to evaluate the antitumor activities of five sesquiterpene lactones to the cell lines of U251SP,T-98,HLE,MM1-CB,HMV-1 and KT,and to explore the relationship between structure and activities.Results Isoalantolactone exhibited significant anti-growth activities to U251SP,HLE and MM1-CB,while other compounds demonstrated weak or moderate anti-growth activities to cell lines even in the 50% inhibition concentration≤100 μmol·L~(-1).Conclusion Isoalantolactone exhibits significant anti-growth activities to three cell lines and its anti-tumor activitiesve close relationship with the structures,α,β-unsaturated five-membered lactone ring is necessary for the activities,and saturated lactone or 5,10-seco and A,B-ring become a ten-membered macro-ring leading to the loss or decrease of anti-growth activity.

Objective To observe the relationship between plasma level of homocysteine(HCY) and ischemic stroke in young and middle-aged adults,explore the clinical signficance concerning the occurrence and development of ischemic stroke in young and middle-aged adults.Methods The plasma homocysteine level of 132 young and middle-aged adults patients with ischemic and 86 control peoples were measured by means of enzymatic cycling assay.Results The plasma homocysteine level in ischemic stroke was higher than that in control group( P ＜ 0.01 ).The higher the plasma homocysteine level,the larger infarcted focus in acute ischemic stroke group of young and middleaged adults.The plasma homocysteine level and the infarcted focus was positively linearly correlated.Conclusion Hyperhomocysteinemia is a risk factor of ischemic stroke among young and middle-aged adults.The higher the plasma homocysteine level,the larger infarcted focus in acute ischemic stroke group.The plasma homocysteine level can reflect the size of the infarcted focus and the degree of disease,and the plasma homocysteine level should serve as a kind of regular examination or as an index intervention can be taken,all of which are great importance to prevent ischemic stroke and reduce its morbilily.

Objective:To explore the neural mechanism of language dysfunction in patients with subacute stroke using functional near-infrared spectroscopy (fNIRS).Methods:Sixteen patients with non-fluent aphasia after subacute stroke (aphasia group), 16 patients with non-aphasia after stroke (non-aphasia group), and 16 healthy middle-aged and elderly subjects (control group) were enrolled into our study. The 6-min resting-state data of fNIRS were collected. Four language-related regions, Broca area, Wernicke area, dorso lateral prefrontal cortex (DLPFC), and supplementary motor area (SMA), were selected as regions of interest (ROIs), and the whole brain functional connection strength and functional connection strength in ROIs and between each two ROIs were analyzed by NirSpark software.Results:Compared with the control group (0.53±0.15) and non-aphasia group (0.47±0.12), the aphasia group had significantly decreased whole brain functional connection strength (0.29±0.14, P<0.05). Compared with the control group and non-aphasia group, the aphasia group had significantly decreased functional connection strength in the left Wernicke area, right Wernicke area, left Broca area, left SMA area, right SMA area and left DLPFC area ( P<0.05, FDR). Compared with the control group and non-aphasia group, the aphasia group had significantly decreased functional connection strength in the right Wernicke-left Wernicke area, right Wernicke-right Broca area, right Wernicke-left Broca area, right Wernicke-right DLPFC area, right Wernicke-left DLPFC area, right Wernicke-right SMA area, right Wernicke-left SMA area, left Wernicke-right Broca area, left Wernicke-left Broca area, left Wernicke-right DLPFC area, left Wernicke-left DLPFC, left Wernicke-right SMA area, left Wernicke-left SMA area, right Broca-left Broca area, right Broca-left DLPFC area, right Broca-right SMA area, right Broca-left SMA area, left Broca-right DLPFC area, left Broca-left DLPFC area, left Broca-right SMA area, left Broca-left SMA area, right DLPFC-left DLPFC area, right DLPFC-right SMA area, right DLPFC-left SMA area, left DLPFC-right SMA area, left DLPFC-left SMA area, and right SMA-left SMA area ( P<0.05, FDR). Conclusion:Abnormal functional connectivity strength of the whole brain and language-related key brain areas might be the neural mechanism of language dysfunction in patients with non-fluent aphasia after subacute stroke.

OBJECTIVETo evaluate the expression of FBXW7 in esophageal squamous cell carcinoma (ESCC) and to explore the correlation of FBXW7 expression with clinicopathologic features and prognosis of ESCC.

METHODSNinety cases of ESCC and twenty cases of tumor-adjacent normal tissues and forty intraepithelial neoplasia tissues were detected by immunohistochemistry methods. The expression of FBXW7 in 40 surgical ESCC tissues and tumor-adjacent normal tissues was detected by Western blotting and RT-PCR. The relationship between the expression of FBXW7, clinicopathological characteristics and prognosis was analyzed.

RESULTSThe positive rate of FBXW7 protein was significantly lower in the ESCC and high-grade intraepithelial neoplasia tissues than in the tumor-adjacent normal tissues and low grade intraepithelial neoplasia tissues (40.0% and 33.3% vs. 85.0% and 77.3%, χ² = 21.923, P < 0.001). The FBXW7 protein was down-regulated in ESCC tissues (P < 0.05), whereas the FBXW7 mRNA was down-regulated in ESCC tissues compared with that in the paracancerous tissues. The expression of FBXW7 was significantly correlated with TNM stage, degree of differentiation, invasion depth and lymph node metastasis (χ² =9.643, 14.908, 16.294, 10.222, respectively, P = 0.002, 0.001, 0.000, 0.001). In the ninety patients, the 5-year survival rates of cases with positive and negative expression of FBXW7 protein was 67.6% and 39.3%, respectively (χ² =6.699, P = 0.01).

CONCLUSIONSThe results of this study demonstrate that FBXW7 expression is significantly declined in ESCC and high grade intraepithelial neoplasia tissues, and is closely correlated with poor prognosis of this disease. FBXW7 as a tumor suppressor gene may play an important role in the carcinogenesis, development and metastasis of ESCC. These results suggest that FBXW7 may become a valuable marker for the severity and prognosis in ESCC.

Biomarkers, Tumor ; metabolism ; Blotting, Western ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Cell Cycle Proteins ; genetics ; Down-Regulation ; Esophageal Neoplasms ; diagnosis ; metabolism ; F-Box Proteins ; genetics ; F-Box-WD Repeat-Containing Protein 7 ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Staging ; Prognosis ; RNA, Messenger ; Survival Rate ; Ubiquitin-Protein Ligases ; genetics

Objective:To investigate the effects of superoxide dismutase 1 (SOD1) mutations G41D and G41S on the cognitive behavior of mice.Methods:The recombinant adeno-associated virus (rAAV) which overexpressed human SOD1WT, SOD1G41S, SOD1G41D and the blank virus without the target gene were constructed, then they were stereotaxic injected into mice bilateral medial prefrontal cortex (mPFC) area respectively.According to the difference of injected virus, they were divided into CONTROL group, SOD1WT group, SOD1G41S group and SOD1G41D group ( n=16 in each group). One month later, open field test, Y-maze spontaneous alternation experiment, three box social interaction experiment and trace fear conditioning test were conducted to observe the effect of mutant gene on cognitive behavior of mice. Results:In the open field test, the movement distance of SOD1WT group((39.67±6.04)m)was significantly higher than that of SOD1G41D group((28.47±6.92)m, P=0.034). In the Y-maze spontaneous alternations experiment, the number of arm entries and actual alternations of arm entries of SOD1WT group((40.56±10.12)times, (32.63±8.19)times)and SOD1G41S group((36.75±9.43)times, (29.06±8.32)times)were significantly higher than those of SOD1G41D group((24.50±11.30)times, (18.38±9.09)times, P<0.05). In the three-compartment social experiment, there was no statistical difference between the residence time of SOD1G41D group in the area containing mouse ((279.08±134.94) s) and the empty metal cage area ((218.54±125.63) s) ( t=1.313, P=0.199). SOD1WT group and SOD1G41S group showed no statistical difference in the residence time in the regions of the unfamiliar mouse 1((253.07±55.60)s, (253.20±57.61)s) and the unfamiliar mouse 2 ((243.44±55.33) s, (239.76±67.49) s) ( P>0.05), and SOD1WT group and SOD1G41S group presented new social barrier.In the test stage of trace fear condition task, the percentage of freezing time of SOD1G41S group was significantly higher than that of other experimental groups and CONTROL group ( P<0.05). Conclusion:SOD1G41S and SOD1G41D have significantly changed the cognitive behavior of mice, and the two types of mutations at the same site have significant differences in the cognitive behavior changes.

Objective:To explore the value of intravascular optical coherence tomography (OCT) in evaluating carotid atherosclerotic stenosis, and compare the morphological characteristics of symptomatic and asymptomatic carotid atherosclerotic plaques.Methods:Patients diagnosed as carotid atherosclerotic stenosis and performed OCT in the Department of Neurology, Jinling Hospital, Medical School of Nanjing University from January 2017 to November 2019 were enrolled retrospectively. Digital subtraction angiography (DSA) and OCT were used to determine the degree of stenosis of the diseased vessels, and the plaque characteristics observed by OCT were recorded. Symptomatic carotid atherosclerotic stenosis is defined as a history of transient ischemic attack, amaurosis fugax, or stroke related to the blood supply area of the diseased vessel within 6 months before the carotid artery examination. The baseline clinical and imaging data of the symptomatic group and the asymptomatic group were compared. Multivariate logistic regression analysis was used to determine the independent risk factors for symptomatic carotid atherosclerotic stenosis. Results:A total of 56 patients were enrolled. DSA and OCT had good consistency in carotid artery diameter measurement ( r=0.93, P<0.001). When the OCT technique was used to evaluate the plaque properties, the consistency of inter-observer ( κ=0.96, P<0.001) and intra-observer ( κ=0.96, P<0.001) was higher. The proportions of patients with type Ⅵ plaque (66.7% vs. 34.5%; P=0.016) and macrophage infiltration (51.9% vs. 24.1%; P=0.032) in the symptomatic group were significantly higher than those in the asymptomatic group, while the proportion of patients with fibrotic plaque was significantly lower than that in the asymptomatic group (40.7% vs. 69.0%; P=0.034). Multivariate logistic regression analysis suggested that type Ⅵ plaques (odds ratio 13.798, 95% confidence interval 1.38-137.675; P=0.025) and macrophage infiltration (odds ratio 5.856, 95% confidence interval 1.405-24.406; P=0.015) were the independent risk factors for symptomatic carotid atherosclerotic stenosis. Conclusions:OCT can be used to evaluate the degree of vascular stenosis and plaque characteristics in patients with carotid atherosclerotic stenosis. The detection rate of complex plaques in patients with symptomatic carotid atherosclerotic stenosis is significantly higher. Type Ⅵ plaque and macrophage infiltration are the independent risk factors for symptomatic carotid atherosclerotic stenosis.

Objective:To identify the morphological features and clinical significance of intra-vascular thrombus in carotid stenosis using optical coherence tomography (OCT).Methods:Twelve patients with carotid artery thrombosis detected by OCT assessment in Jinling Hospital between January 2017 and January 2020 were included. Serial area measurements within the athero-thrombotic target lesion were performed to evaluate the OCT-thrombus score, length, area and volume. The clinical data and plaque morphological features were also assessed.Results:Among the 12 patients demonstrating thrombus on OCT, eight patients presented with white thrombus, two patients presented with red thrombus, and another two patients displayed both white thrombus and red thrombus. OCT-thrombus scores were 1-32. The OCT-thrombus score was correlated to the OCT-thrombus volume ( ρ=0.739, P=0.006) and the thrombus length ( ρ=0.932, P<0.001). All lesions were presented with fibrous cap disruption, and 10 lesions were presented with thin-cap fibroatheroma. In view of the OCT findings, all patients received carotid balloon angioplasty and stent implantation. During an average follow-up of 14.2 months, none of the 12 patients had fatal stroke or recurrent ischemic stroke. Conclusions:OCT can be used to assess intra-carotid thrombus and its more detailed morphological characteristics, offering more possibilities in quantitative analysis of thrombus burden.

Objective To investigate the efficacy and safety of intravesical instillation of BCG vaccine in the prevention of early recurrence of middle and high risk non-muscle invasive bladder cancer.Methods From July 2015,patients with non-muscle invasive bladder cancer aged 18-75 years with informed consent were screened and underwent transurethral resection of bladder tumor (TURBT).Immediately intravesical instillation of epirubicin 50 mg was given postoperatively.After pathology was comfirmed,patients was enrolled in group 1 (BCG15) or group 2 (BCG 19) or the control group (epirubicin 18) randomly with SAS 9.3 software.Data of follow-up and Adverse event was collected and analyzed.Results By May 31,2019,531 patients were enrolled in the study.The drop-off rate was 20.1％.167 patients (143 males and 24 females)in group 1,172 patients (141 males and 31 females)in group2 and 84(75 males and 9 females) in the control group with follow-up data were analyzed.There were no significant differences in age,gender,BMI,ECOG score,risk stratification between the three groups (P =0.8641,P =0.2906,P =0.9384,P =0.6126).The median follow-up time makes no statistical difference between the groups (P =0.9251),12.0 (6.0,22.5) months,13.0 (6.0,22.3) months,and 13.0 (7.0,22.3) months.The median recurrence time of the three groups was 4.0 (3.0,6.0) months,4.5 (3.0,9.8) months,4.5 (3.0,8.8) months.There was no statistical difference between the three groups (P =0.2852).Risk stratification in the patients got no significant difference between the three groups (P ＞ 0.05).The 1-year recurrence-free survival rates were 80.0％ in the group 1 and 88.3％ in the group 2 and 73.7％ in the control group.The group 2 was superior to the group 1 and the control group (P =0.0281,P =0.0031).There was no significant difference between group 1 and control group (P =0.2951).There was no significant difference in the cumulative recurrence-free survival between the experimental group 1 and the experimental group 2,(95％ CI 0.80-2.43,P =0.2433).The cumulative recurrence-free survival in the group 1 and the group 2 was better than the control group (95 ％ CI 0.31-0.92,P =0.0266;95 ％ CI 0.20-0.65,P =0.0008).All the cases underwent instillation were analyzed for adverse events.The incidence of overall AE(adverse events) in group 1 was 68.5％ (152/222),the incidence of grade Ⅰ-Ⅱ AE was 53.2％ (118/222),the incidence of grade Ⅲ-Ⅳ AE was 15.3％ (32/222).The incidence of overall AE in the group 2 was 71.8％ (160/223),the incidence of grade Ⅰ-Ⅱ AE was 60.1％ (134/223),and the incidence of grade Ⅲ-Ⅳ AE was 11.7％ (26/223).The overall AE rate in the control group was 53.2％ (59/111),of which the incidence of grade Ⅰ-Ⅱ AE was 42.4％ (47/111),and the incidence of grade Ⅲ-Ⅳ AE was 10.8％ (12/111).There was no difference in the incidence of overall AE between the group 1 and the group 2 (P =0.4497).The incidence of AE in the two experimental groups was higher than that in the control group (P =0.0062,P =0.0008).There was no difference in the incidence of grade Ⅲ-Ⅳ AE between the three groups (P =0.3902).Conclusions BCG(19 instillation schedule) has a better effect on preventing recurrence after 1 year of bladder surgery,which is superior to epirubicin group.The long-term efficacy of BCG in preventing recurrence and the efficacy of different schedules need to be further followed up.The lower urinary tract symptoms,which are mainly urinary frequency,are one of the causes of case fallout and should be fouced in future.Compared with epirubicin,BCG perfusion does not increase the incidence of grade Ⅲ-Ⅳ adverse reactions,and is safe to use.