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Background: Multi-organ metastasis has been the main cause of death in patients with Gastric cancer (GC). The prognosis for patients with metastasized GC is still very poor. Long noncoding RNAs (lncRNAs) always been reported to be closely related to cancer metastasis., Methods: In this paper, the aberrantly expressed lncRNA CADM2-AS1 was identified by lncRNA-sequencing in clinical lymph node metastatic GC tissues. Besides, the role of lncRNA CADM2-AS1 in cancer metastasis was detected by Transwell, Wound healing, Western Blot or other assays in vitro and in vivo . Further mechanism study was performed by RNA FISH, Dual-luciferase reporter assay and RT-qPCR. Finally, the relationship among lncRNA CADM2-AS1, miR-5047 and NOTCH4 in patient tissues was detected by RT-qPCR., Results: In this paper, the aberrantly expressed lncRNA CADM2-AS1 was identified by lncRNA-sequencing in clinical lymph node metastatic GC tissues. Besides, the role of lncRNA CADM2-AS1 in cancer metastasis was detected in vitro and in vivo . The results shown that overexpression of the lncRNA CADM2-AS1 promoted GC metastasis, while knockdown inhibited it. Further mechanism study proved that lncRNA CADM2-AS1 could sponge and silence miR-5047, which targeting mRNA was NOTCH4. Elevated expression of lncRNA CADM2-AS1 facilitate GC metastasis by up-regulating NOTCH4 mRNA level consequently. What's more, the relationship among lncRNA CADM2-AS1, miR-5047 and NOTCH4 was further detected and verified in metastatic GC patient tissues., Conclusions: LncRNA CADM2-AS1 promoted metastasis in GC by targeting the miR-5047/NOTCH4 signaling axis, which may be a potential target for GC metastasis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Zhang, Zhao, Zhou, Zhao, Geng, Zhang, Sun and Chen.)

Neddylation is the writing of monomers or polymers of neural precursor cells expressed developmentally down-regulated 8 (NEDD8) to substrate. For neddylation to occur, three enzymes are required: activators (E1), conjugators (E2), and ligators (E3). However, the central role is played by the ubiquitin-conjugating enzymes E2M (UBE2M) and E2F (UBE2F), which are part of the E2 enzyme family. Recent understanding of the structure and mechanism of these two proteins provides insight into their physiological effects on apoptosis, cell cycle arrest and genome stability. To treat cancer, it is therefore appealing to develop novel inhibitors against UBE2M or UBE2F interactions with either E1 or E3. In this evaluation, we summarized the existing understanding of E2 interaction with E1 and E3 and reviewed the prospective of using neddylation E2 as a pharmacological target for evolving new anti-cancer remedies., (© 2023. Springer Nature America, Inc.)

In October-November 2020, the phytoplankton and the aquatic environment from 62 sites in the mainstream of the Qinhe River and the largest tributary of the Qinhe River (Danhe River) in the Jincheng region were investigated to clarify the spatial pattern of phytoplankton communities and their driving factors. A total of 7 phyla and 47 species of phytoplankton were identified in the Qinhe River basin and were composed of Cryptophyta, Chlorophyta, Pyrrophyta, Chrysophyta, Bacillariophyta, Cyanophyta, and Crytophyta. Six dominant species in the Qinhe River included: Chlorella vulgaris, Cryptomonas erosa, Chroomonas acuta, Cyclotella stelligera, Chlorococcum , and Euglena viridis . Six dominant species in the Danhe River included: C. erosa, Frustulia vulgaris, E. viridis, C. vulgaris, Trachelomonas oblonga Lemm, and C. stelligera . The Shannon-Wiener diversity index ( H' ) varied from 0.35 to 3.15, with a mean value 1.40. The Pielou evenness index ( J ) varied from 0.24 to 1.00, with a mean value of 0.68. H' values in the Qinhe River were higher than those in the Danhe River. J values were relatively low in the middle reaches of the Qinhe River and middle-low reaches of the Danhe River. The results in the Qinhe River through a canonical correspondence analysis (CCA) showed that the percent of forest land at a 300 m buffer was the driving factor of Chlorococcum in Chlorophyta, and nitrate, total phosphorus, and the percent of forest land at the 300 m buffer were the driving factors of E. viridis . Cyclotella stelligera was mainly influenced by the percent of urban land and water temperature, whereas C. vulgaris, C. erosa , and C. acuta were mainly influenced by the percent of farmland and residential land at the 300 m buffer. The results in the Danhe River via CCA showed that C. erosa and C. stelligera were mainly influenced by pH and sulfate, E. viridis was mainly influenced by the percent of urban land and grass land, T. oblonga Lemm was mainly influenced by chloride and the percent of forest land, F. vulgaris was mainly influenced by water temperature and the percent of farmland, and C. vulgaris was mainly influenced by ammonia and the percent of farmland.

The traditional medicinal plant, and endangered species Aristolochia delavayi (Aristolochiaceae) is an endemic species in China and occurs in the warm and dry areas along the Jinsha river. It is also a specific host of the larvae of Byasa daemonius , a vulnerable butterfly. In this study, 15 pairs of polymorphic microsatellite primers of A. delavayi were designed and screened based on the Simple Sequence Repeats (SSR) loci found by using the results of genome skimming. Based on these 15 SSR markers, the genetic diversity and structure of 193 individuals from ten natural populations were analyzed in detail. In comparison to other endemic and endangered plants in the region, the population of A. delavayi possess a relatively high genetic diversity ( H e = 0.550 , I  = 1.112). AMOVA analysis showed that 68.4% of the total genetic diversity was within populations and 31.6% of the variation occurred among populations. There was a significant genetic differentiation among natural populations of A. delavayi detectable, with low gene flow ( N m = 0.591). This might be attributed to geographical barriers and limited seed dispersal . To test the isolation by distance (IBD), we performed Mantel test, which showed a significant correlation between the geographic and genetic distances. In order to cope with the possible biases caused by IBD, we additionally performed Bayesian genetic cluster analyses and principal coordinate analysis (PCoA). The final cluster analysis revealed three groups with distinct geographical distribution. Habitat fragmentation and limited gene flow between these populations may be the main reasons for the current genetic structure. For conservation of this species, we suggest to divide its populations into three protection management units, with subsequent focus on the Yongsheng and Luquan populations which experienced a genetic bottleneck event in the past., Competing Interests: The authors declare that there are no conflicts of interest., (© 2020 Kunming Institute of Botany, Chinese Academy of Sciences. Publishing services by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.)

Background: Lifestyle and risk factor management may improve outcomes in patients with atrial fibrillation (AF). We aim to evaluate the prevalence of modifiable risk factors and how these factors impact clinical outcomes in patients with AF., Methods and Results: Data on 17 898 AF cohort patients with AF enrolled between 2011 and 2016 was analyzed. A healthy lifestyle was defined as not smoking, not drinking, a healthy body mass index (BMI), untreated total cholesterol less than 200 mg/dL, untreated blood pressure (BP) less than 120/80 mm Hg, and untreated fasting plasma glucose (FPG) less than 100 mg/dL. The association between risk factors and risk of the composite endpoint of all-cause mortality and nonfatal ischemic stroke were assessed using Cox proportional hazards regression model. Only 4.0% of patients achieved a healthy lifestyle. In multivariate analysis, current smoking, a low BMI, not well-controlled FPG were independently and significantly associated with higher risk of all-cause mortality and nonfatal ischemic stroke, with corresponding hazard ratio (HR) estimates 1.22 (95% confidence interval [CI], 1.00-1.47), HR = 1.72 (95% CI, 1.34-2.20), and HR = 1.25 (95% CI, 1.06-1.46), respectively. High BP was also associated with higher risk with the outcomes (HR = 1.15, 95% CI, 1.00-1.34). Compared with patients with no risk factor, those who failed to maintained or achieved optimal risk factor control had a progressively higher risk of death and nonfatal ischemic stroke (HR for 1 risk factor = 1.44; 95% CI, 1.07-1.92; and more than 2 risk factors =  1.75; 95% CI, 0.99-3.09)., Conclusions: Maintenance of well-controlled risk factors may substantially lower the risk of death and ischemic stroke in patients with AF., (© 2019 Wiley Periodicals, Inc.)

Background:  Anti-coagulant therapy satisfaction for patients with atrial fibrillation is a critical issue, which impacts on their treatment adherence and clinical outcomes. The disadvantages of long-term warfarin treatment are well-described, and novel oral anti-coagulants have become an alternative option., Materials and Methods:  We compared patient-reported treatment satisfaction with dabigatran versus warfarin in non-valvular atrial fibrillation (NVAF) patients in China. Treatment satisfaction was assessed using the Anti-Clot Treatment Scale (ACTS) questionnaire, which included a 12-item ACTS Burdens scale and a 3-item ACTS Benefits scale., Results:  Among 834 patients, 246 patients (29.5%) were taking dabigatran and the others were on warfarin. Propensity score matching was employed to identify 182 patient pairs with balanced baseline characteristics. The global ACTS Burdens score and the global ACTS Benefits score were comparable between the dabigatran and warfarin groups (44.86 ± 3.95 vs. 44.28 ± 3.51, p  = 0.423; 11.49 ± 2.92 vs. 11.42 ± 3.03, p  = 0.194, respectively). The monthly cost of dabigatran was significantly higher compared with that of warfarin due to a lack of insurance coverage (USD 176.78 ± 9.15 vs. USD 2.49 ± 0.76, p  = 0.000). The discontinuation rate of dabigatran was significantly higher than warfarin at the 6-month follow-up (33.5% vs. 19.2%, p  = 0.003). Adjusted logistic regression showed that dabigatran was associated with a significant greater odds of non-persistence (odds ratio: 2.13, 95% confidence interval: 1.27-3.59, p  = 0.004)., Conclusion:  Dabigatran therapy in patients with NVAF in China associated with no improvement in satisfaction and a higher discontinuation rate compared with warfarin therapy largely due to increased economic burden., Competing Interests: Dr. Chang-Sheng Ma received honoraria from Bristol-Myers Squibb (BMS), Pfizer, Johnson & Johnson, Boehringer-Ingelheim (BI) and Bayer for giving lectures. Dr. Jian-Zeng Dong also received honoraria from Johnson & Johnson for giving lectures. The other authors report no conflicts of interest., (Georg Thieme Verlag KG Stuttgart · New York.)

Mechoris ursulus (Roelofs) (Coleoptera: Attelabidae) is a pest weevil of Fagaceae oak trees in eastern Asia. The female has a distinct branch-cutting behavior in conjunction with oviposition in the acorns of its host plant. This study analyzed the factors influencing oviposition preference by carrying out continuous field surveys over the course of 2009-2010 and through laboratory rearing. The field survey showed that for both of the hosts, Cyclobalanopsis glaucoides Schottky (Fagales: Fagaceae) and Quercus franchetii Skan, M. ursulus preferred branches with fewer acorns and larger acorns on the same branch for oviposition. Laboratory rearing experiments showed offspring performance (i.e., survival rate and fresh weight of larvae) was significantly and positively correlated with acorn size. Preference for larger acorns could maximize the fitness of offspring by providing sufficient food source and space., (© The Author 2015. Published by Oxford University Press on behalf of the Entomological Society of America.)

Premise of the Study: Microsatellite primers were developed for Parthenium hysterophorus to investigate its genetic structure and genetic diversity., Methods and Results: Using the combined biotin capture method, 15 microsatellite primer sets were isolated and characterized. All markers showed polymorphism, and the number of alleles per locus ranged from two to nine across 60 individuals from two populations. The observed and expected heterozygosities ranged from 0.117 to 0.750 and from 0.182 to 0.835, respectively., Conclusions: These markers will be useful for investigating the invasion history of this weed globally and to help characterize its invasiveness.

Objective: To investigate the differenal protein expression profiles of ovarian tumor cell lines with distinct metastatic abilities., Methods: The ovarian cancer cell line HO8910 and HO8910pm, derived from same parental cells but exhibited different metastatic ability, were investigated by two-dimensional gel electrophoresis (2-DE)-MALDI-TOF-MS proteomic approach., Results: Thirty-nine proteins were detected by 2-DE to have expression disparity levels over 2 folds between two cell lines. Eighteen of them were identified by MALDI-TOF-MS. The proteins are involved in apoptosis, extra cellular matrix (ECM), cytoskeleton, growth factor, glycolysis, protein metabolism and immune system., Conclusion: The data are valuable for the identification of differentially expressed proteins involved in the biological behavior of human ovarian cancer.

Glioma is the most common malignant disease in the brain, and recurrence is the main cause of death from this disease. Tumor recurrence involves multiple steps, and requires the accumulation of the altered expression of many different proteins. Identification of the recurrence associated protein profile in glioma cell lines will be helpful in clarifying the molecular mechanisms underlying glioma recurrence. In this report, two glioma cell lines with distinct tumor forming ability in vitro and in vivo were chosen and the different protein expression patterns were analyzed by proteomics method. To confirm the utility of this method, we validated the differential expression of one protein, cathepsin D, by immunohistochemistry analysis. Forty-six proteins appeared differently between two cell lines and 18 of them were identified. These 18 are involved in cell proliferation, DNA replication, protein synthesis, invasion, angiogenesis and neurotrophic factor. All of these molecules are important in tumor growth, and a subset of them may be related to glioma recurrence. These findings may contribute to the discovery of new diagnostic markers and therapeutic targets of glioma.

Objective: To conduct a proteomic analysis of human colorectal carcinoma cell lines LS174T and SW480 by two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS)., Methods: The total proteins of human colorectal carcinoma cell lines LS174T and SW480 were separated with 2-DE using immobilized pH gradient strips and analyzed by MALDI-TOF-MS to obtain peptide mass fingerprints (PMFs). Proteins were identified by using Mascot software to search protein databases., Results: Good resolution 2-DE maps were obtained. Some proteins including immunoglobulin heavy chain variable region (IgVH) and co-stimulatory molecule B7-1 were identified. IgVH and B7-1 were confirmed by electrospray ionization tandem spectrometry (ESI-MS/MS) and immunocytochemistry., Conclusion: There are IgVH and B7-1 expressions in human colorectal carcinoma cell lines LS174T and SW480. Results obtained will help to elucidate the mechanisms of tumor immune escape.

Objective: To investigate the distribution of regulatory T-lymphocytes in the splenocytes cocultured with syngeneic low-immunogenic tumor cells, as compared with that of highly-immunogenic tumor cells, to investigate the mechanism underlining tumor evasion., Methods: Three different immunogenic tumor cells were cocultured with syngeneic splenocytes individually to mimic cancer immunity in vitro. The proliferation response of splenocytes was measured by thymidine incorporation. The distribution of TR cells, CD4(+) IFN-gamma (+) T cells and CD4(+) IL-10(+) T cells were analyzed by flow cytometry. The secretion of IFN-gamma and IL-10 in supernatants was measured by ELISA assay., Results: The stimulation Index of splenocytes cocultured with syngeneic highly-immunogenic H22 or FBL3 was much higher than that of poorly immunogenic melanoma D5. In each group, stimulation Index of splenocytes cocultured with allogeneic tumor cells was higher than that of the corresponding tumor immunity model. In addition, compared with those of highly-immunogenic tumors, there were more TR, CD4(+)IL-10(+) and less CD4 (+)IFN-gamma(+) T cells in the splenocytes, and higher IL-10 and lower IFN-gamma levels in the supernatant of the splenocytes stimulated with low-immunogenic D5 cells., Conclusion: Poorly-immunogenic tumor cells can induce the proliferation of TR cells, which may play an important role in tumor evasion.

Aim: To conduct the proteomic analysis of human colorectal carcinoma cell line, SW480 by using two-dimensional electrophoresis (2-DE) and matrix-assisted laser desorption /ionization-time of flight mass spectrometry (MALDI-TOFMS)., Methods: The total proteins of human colorectal carcinoma cell line, SW480 were separated with 2-DE by using immobilized pH gradient strips and visualized by staining with silver nitrate. The gel images were acquired by scanner and 2-DE analysis software, Image Master 2D Elite. Nineteen distinct protein spots were excised from gel randomly and digested in gel by TPCK-trypsin. Mass analysis of the tryptic digest peptides mixture was performed by using MALDI-TOF MS. Peptide mass fingerprints (PMFs) obtained by the MALDI-TOF analysis were used to search NCBI, SWISS-PROT and MSDB databases by using Mascot software., Results: PMF maps of all spots were obtained by MALDI-TOF MS and thirteen proteins were preliminarily identified., Conclusion: The methods of analysis and identification of protein spots of tumor cells in 2-DE gel with silver staining by MALDI-TOF MS derived PMF have been established. Protein expression profile of SW480 has been obtained. It is demonstrated that a combination of proteomics and cell culture is a useful approach to comprehend the process of colon carcinogenesis.

Aim: To examine the humoral and cellular immunoresponses induced by HPV18 L1-E6 and L1-E7 chimeric gene DNA vaccines in mice., Methods: 54 BALB/c mice were divided into 9 groups randomly, and then vaccinated with various recombinant plasmids(pVAX1-L1-E6M3 or pVAX1-L1-E7M3) and immune adjuvants (pLXHDmB7-2 or LTB) through different administration routes (intramuscular or intranasal). After the third inoculation, blood samples were taken to measure specific antibody, and footpad swelling test was used to detect delayed-type hypersensitivity(DTH). Mice were killed and spleens were taken to prepare single spleen cell suspension for lymphocyte proliferation assay and CD4(+)/CD8(+), IFN-gamma(+) or IL-4(+) T cells double staining FACS assay. And then the pathological changes of the swollen footpad were observed., Results: Compared with control, significant immunoresponses were observed in different experimental groups. The level of specific serum IgG against HPV in experiment groups was much higher than that of control group, and intramuscular immunization group had the highest antibody level. The footpad from immunized mice injected with virus-like particles was swollen and harden, and a large amounts of mononuclear cells can be seen in the footpad tissues. Intramuscular immunization groups were superior to intranasal immunization groups in DTH response, splenocyte proliferation and CD8(+) IFN-gamma(+) cell number, while CD4(+) IL-4(+) cell number was higher in intranasal immunization groups. The immunization groups using pLXHDmB7-2 as adjuvant were superior to other groups in immune responses., Conclusion: The recombinant plasmids could induce strong humoral and cellular immunoresponses in mice. Costimulatory molecule B7-2 could enhance the immune responses against HPV18 induced by the L1-E6 and L1-E7 DNA vaccines when used as an adjuvant.

Aim: To purify preliminary recombinant human TNF-alpha mutein 471 and detect its bioactivity on the basis of the TNF-alpha mutein 471 expressed in prokaryotic express system., Methods: The expression of recombinant human TNF-alpha mutein 471 in engineering bacteria strains E.coil was induced under the condition of optimal fermentation and expression. After cultured E.coil cells were collected and broken by using an ultrasonic disintegrator, the TNF-alpha mutein 471 existed in the form of inclusion body was extracted and purified, and then the effects of denaturation and protein concentration on protein folding were examined. The bioactivities of wild type TNF-alpha and the TNF-alpha mutein 471 were detected by MTT colorimetry., Results: The TNF-alpha mutein 471 was folded and polymerized successfully to from a trimer with bioactivity under the condition of proper denaturation and renaturation. The cytotoxic activity of the TNF-alpha mutein 471 to the L929 cells was 15 times as much as wild type TNF-alpha., Conclusion: The TNF-alpha mutein 471 expressed in prokaryotic expression system possesses significantly bioactivity after renaturation, which lays the foundation for further animal experiment and clinical experimental researches.

Aim: To construct an eukaryotic expression plasmid pcDNA3.1/hIL-18 and express it in mammalian cells., Methods: cDNA encoding mature hIL-18 was cleavaged by enzyme digestion from mesomeric clone vector pGEM-T/hIL-18 and inserted into an eukaryotic expression plasmid pcDNA3.1 to construct a recombinant expression plasmid pcDNA3.1/hIL-18. Then the constructed plasmid was transfected into COS-7 and Rlc310 cells by liposome-mediated gene transfer method. hIL-18 expressed in transfected COS-7 and Rlc310 cells was detected by immunohistochemical staining and level of hIL-18 mRNA in transfected Rlc310 cells was assayed by RT-PCR., Results: A recombinant eukaryotic expression plasmid pcDNA3.1/hIL-18 was successfully constructed and expressed transiently in COS-7 cells and stably in Rlc310 cells., Conclusion: The construction and expression of pcDNA3.1/hIL-18 have been achieved successfully, which lays a foundation for further research on anti-tumor effect of IL-18.