Identification and validation of UBE2B as a prognostic biomarker promoting the development of esophageal carcinomas.

Introduction: Ubiquitination is a crucial biological mechanism in humans, essential for regulating vital biological processes, and has been recognized as a promising focus for cancer therapy. Our objective in this research was to discover potential enzymes associated with ubiquitination that may serve as therapeutic targets for individuals with esophageal carcinoma (ESCA). Methods: To identify genes linked to the prognosis of ESCA, we examined mRNA sequencing data from patients with ESCA in the TCGA database. Further investigation into the role of the candidate gene in ESCA was conducted through bioinformatic analyses. Subsequently, we carried out biological assays to assess its impact on ESCA development. Results: Through univariate Cox regression analysis, we identified Ubiquitin Conjugating Enzyme E2 B (UBE2B) as a potential gene associated with the prognosis of ESCA. UBE2B exhibited significant upregulation and was found to be correlated with survival outcomes in ESCA as well as other cancer types. Additionally, UBE2B was observed to be involved in various biological pathways linked to the development of ESCA, including TNF-a signaling via NF-κB, epithelial-mesenchymal transition, inflammatory response, and hypoxia. Moreover, immune-related pathways like B cell activation (GO: 0042113), B cell receptor signaling pathway (GO: 0050853) and B cell mediated immunity (GO:0019724) were also involved. It was found that high expression of UBE2B was correlated with the increase of several kinds of T cells (CD8 T cells, Th1 cells) and macrophages, while effector memory T cell (Tem) and Th17 cells decreased. Furthermore, UBE2B showed potential as a prognostic biomarker for ESCA, displaying high sensitivity and specificity. Notably, proliferation and migration in ESCA cells were effectively suppressed when the expression of UBE2B was knocked down. Conclusions: To summarize, this study has made a discovery regarding the importance of gaining new insights into the role of UBE2B in ESCA. UBE2B might be an oncogene with good ability in predicting and diagnosing ESCA. Consequently, this discovery highlights the feasibility of targeting UBE2B as a viable approach for treating patients with ESCA.

The Application of Deep Brain Stimulation for Parkinson's Disease on the Motor Pathway: A Bibliometric Analysis across 10 Years.

BACKGROUND AND OBJECTIVE: Since its initial report by James Parkinson in 1817, Parkinson's disease (PD) has remained a central subject of research and clinical advancement. The disease is estimated to affect approximately 1% of adults aged 60 and above. Deep brain stimulation, emerging as an alternative therapy for end-stage cases, has offered a lifeline to numerous patients. This review aimed to analyze publications pertaining to the impact of deep brain stimulation on the motor pathway in patients with PD over the last decade. METHODS: Data were obtained from the Web of Science Core Collection through the library of Huazhong University of Science and Technology (China). The search strategy encompassed the following keywords: "deep brain stimulation", "Parkinson's disease", "motor pathway", and "human", from January 1, 2012, to December 1, 2022. Additionally, this review visualized the findings using the Citespace software. RESULTS: The results indicated that the United States, the United Kingdom, Germany, and China were the primary contributors to this research field. University College London, Capital Medical University, and Maastricht University were the top 3 research institutions in the research area. Tom Foltynie ranked first with 6 publications, and the journals of Brain and Brain Stimulation published the greatest number of relevant articles. The prevailing research focal points in this domain, as determined by keywords "burst analysis", "encompassed neuronal activity", "nucleus", "hyper direct pathway", etc. CONCLUSION: This study has provided a new perspective through bibliometric analysis of the deep brain stimulation therapy for treating patients with PD, which can shed light on future research to advance our comprehension of this particular field of study.

Clinical outcomes of endoscopic resection for the treatment of esophageal gastrointestinal stromal tumors: a ten-year experience from a large tertiary center in China.

BACKGROUNDS: Esophageal gastrointestinal stromal tumors (E-GISTs) are extremely rare and surgical resection is the recommended approach. However, surgical resection usually causes severe trauma that may result in significant postoperative morbidity. Endoscopic resection (ER) has developed rapidly in recent years and has been widely used in gastrointestinal lesions. Nevertheless, the feasibility and efficacy of ER in the management of E-GISTs are unknown. METHODS: Retrospective data were collected from January 2011 to December 2020 in a large tertiary center of China. Twenty-eight patients with E-GISTs treated by ER were included in the study. RESULTS: Of the 28 patients, there were 21 males and 7 females, with a median age of 55 years (40-70 years). The median tumor size was 15 mm (5-80 mm). The technical success rate was 100% (28/28), while the en bloc resection rate was 96.4% (27/28). The median operation time was 35 min (10-410 min). Sixteen (57.2%) tumors were categorized into very low risk group, six (21.4%) into low risk group, and six (21.4%) into high risk group. Pathologists carefully examined margins of each lesion. There were 11 lesions (39.3%) determined as R0 resection and 17 lesions (60.7%) as R1 resection with positive margins. The median hospital stay was 2 days (range, 1-8 days). One patient suffered from hydrothorax and required drainage, leading to a major adverse event rate of 3.6% (1/28). There was no conversion to surgery, and no death occurred within 30 days after the procedure. Imatinib was given to two patients after ER under multidisciplinary team surveillance. During follow-up (median of 54 months, 9-122 months), no recurrences or metastasis were observed. CONCLUSION: ER is safe and effective for E-GISTs and might become an optional choice in the future. Multicenter, prospective, large samples with long-term follow-up studies are still needed.

Single-cell transcriptomic analysis deciphers key transitional signatures associated with oncogenic evolution in human intramucosal oesophageal squamous cell carcinoma.

BACKGROUND AND AIMS: The early diagnosis and intervention of oesophageal squamous cell carcinoma (ESCC) are particularly important because of the lack of effective therapies and poor prognosis. Comprehensive research on early ESCC at the single-cell level is rare due to the need for fresh and high-quality specimens obtained from ESD. This study aims to systematically describe the cellular atlas of human intramucosal ESCC. METHODS: Five paired samples of intramucosal ESCC, para-ESCC oesophageal tissues from endoscopically resected specimens and peripheral blood mononuclear cells were adopted for scRNA-seq analysis. Computational pipeline scMetabolism was applied to quantify the metabolic diversity of single cells. RESULTS: A total of 164 715 cells were profiled. Epithelial cells exhibited high intra-tumoural heterogeneity and two evolutionary trajectories during ESCC tumorigenesis initiated from proliferative cells, and then through an intermediate state, to two different terminal states of normally differentiated epithelial cells or malignant cells, respectively. The abundance of CD8+ TEX s, Tregs and PD1+ CD4+ T cells suggested an exhausted and suppressive immune microenvironment. Several genes in immune cells, such as CXCL13, CXCR5 and PADI4, were identified as new biomarkers for poor prognosis. A new subcluster of malignant cells associated with metastasis and angiogenesis that appeared at an early stage compared with progressive ESCC was also identified in this study. Intercellular interaction analysis based on ligand-receptor pairs revealed the subcluster of malignant cells interacting with CAFs via the MDK-NCL pathway, which was verified by cell proliferation assay and IHC. This indicates that the interaction may be an important hallmark in the early change of tumour microenvironment and serves as a sign of CAF activation to stimulate downstream pathways for facilitating tumour invasion. CONCLUSION: This study demonstrates the changes of cell subsets and transcriptional levels in human intramucosal ESCC, which may provide unique insights into the development of novel biomarkers and potential intervention strategies.

Comparison of safety and short-term outcomes between endoscopic and laparoscopic resections of gastric gastrointestinal stromal tumors with a diameter of 2-5 cm.

BACKGROUND AND AIM: Developments of endoscopic techniques brought the possibility of endoscopic resection for gastrointestinal stromal tumors (GISTs) of larger sizes. We aim to compare safety and short-term outcomes between endoscopic and laparoscopic resections of gastric GISTs with a diameter of 2-5 cm. METHODS: This is a single-center, retrospective cohort study. The clinical data, perioperative conditions, and the adverse events of patients who underwent endoscopic or laparoscopic resection for gastric GIST of 2-5 cm in Zhongshan Hospital, Fudan University, from January 2016 to December 2020 were retrospectively reviewed. RESULTS: A total of 346 patients were reviewed; 12 patients who failed to accomplish the planned procedure were excluded; 182 underwent laparoscopic resection; and 152 underwent endoscopic resection. Significant differences exist in the tumor size between the laparoscopic group (3.43 ± 0.86 cm) and the endoscopic group (2.78 ± 0.73 cm) (P < 0.01). Compared with laparoscopic resection, endoscopic resection was associated with faster recovery (P < 0.01), shorter hospital stays (P < 0.01), and lower cost (P < 0.01). The incidence of Clavien-Dindo grade II-V adverse events in the endoscopic group (3/152) was significantly lower than that in the laparoscopic group (12/182) (P = 0.04). After a propensity score matching analysis, the endoscopic group showed similar incidences of complications with the laparoscopic group, while the advantages over laparoscopic resection in postoperative hospital stay, time to first oral intake, and hospitalization expenses remained significant (P < 0.01). CONCLUSIONS: Endoscopic resection is a safe and cost-effective method for 2-5 cm of gastric GISTs compared with laparoscopic resection.

[Effects of Clostridium butyricum on serum uric acid and inflammatory mediators in rats with hyperuricemia].

OBJECTIVE: To investigate the effects of intragastric administration of Clostridium butyricum in regulating serum uric acid, lipopolysaccharides (LPS), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in rats with hyperuricemia rats. METHODS: Forty SD rats were randomized into 4 equal groups, namely the normal control group, hyperuricemia model group, benzbromarone (3 mg/kg daily) intervention group and live Clostridium butyricum group (1.5×107 CFU/day). Except for those in the control group, the rats were subjected to intragastric administration of yeast extract and oteracil potassium once daily for 12 weeks to induce hyperuricemia with corresponding treatments. The changes in serum uric acid, lipopolysaccharides , IL-6 and TNF-α in each group were detected. RESULTS: The serum level of uric acid was significantly higher in rats fed with high-purine diet than in the control rats (P<0.01), demonstrating the successful establishment of hyperuricemia models. In rats with hyperuricemia, serum uric acid level was positively correlated with the levels of LPS, IL-6 and TNF-α, and their serum levels decreased significantly and progressively with time in Benzbromarone group and Clostridium butyricum group. Benzbromarone was more effective in decreasing serum uric acid in the rats, while Clostridium butyricum produced a stronger effect in down-regulating the inflammatory mediators. CONCLUSION: Chronic inflammatory reaction exists in rats with hyperuricemia. Intragastric administration of Clostridium butyricum can effectively decrease serum uric acid level and inhibit the inflammatory cytokines, and thus contributes to immune homeostasis in the intestines.

Plasmid-based Survivin shRNA and GRIM-19 carried by attenuated Salmonella suppresses tumor cell growth.

Persistent activation of Survivin and its overexpression contribute to the formation, progression and metastasis of several different tumor types. Therefore, Survivin is an ideal target for RNA interference mediated-growth inhibition. Blockade of Survivin using specific short hairpin RNAs (shRNA) can significantly reduce prostate tumor growth. RNA interference does not fully ablate target gene expression, owing to the idiosyncrasies associated with shRNAs and their targets. To enhance the therapeutic efficacy of Survivin-specific shRNA, we employed a combinatorial expression of Survivin-specific shRNA and gene associated with retinoid-interferon-induced mortality-19 (GRIM-19). Then, the GRIM-19 coding sequences and Survivin-specific shRNAs were used to create a dual expression plasmid vector and were carried by an attenuated strain of Salmonella enteric serovar typhimurium (S. typhimurium) to treat prostate cancer in vitro and in vivo. We found that the co-expressed Survivin-specific shRNA and GRIM-19 synergistically and more effectively inhibited prostate tumor proliferation and survival, when compared with treatment with either single agent alone in vitro and in vivo. This study has provided a novel cancer gene therapeutic approach for prostate cancer.

Functional expression of spider neurotoxic peptide huwentoxin-I in E. coli.

The coding sequence of huwentoxin-I, a neurotoxic peptide isolated from the venom of the Chinese spider Ornithoctonus huwena, was amplified by PCR using the cDNA library constructed from the spider venom glands. The cloned fragment was inserted into the expression vector pET-40b and transformed into the E. coli strain BL21 (DE3). The expression of a soluble fusion protein, disulfide interchange protein (DsbC)-huwentoxin-I, was auto-induced in the periplasm of E. coli in the absence of IPTG. After partial purification using a Ni-NTA column, the expressed fusion protein was digested using enterokinase to release heteroexpressed huwentoxin-I and was further purified using RP-HPLC. The resulting peptide was subjected to gel electrophoresis and mass spectrometry analysis. The molecular weight of the heteroexpressed huwentoxin-I was 3750.69, which is identical to that of the natural form of the peptide isolated from spider venom. The physiological properties of the heteroexpressed huwentoxin-I were further analyzed using a whole-cell patch clamp assay. The heteroexpressed huwentoxin-I was able to block currents generated by human Na(v1.7) at an IC50 of 640 nmole/L, similar to that of the natural huwentoxin-I, which is 630 nmole/L.

[Expressions of survivin and GRIM-19 in prostate cancer].

OBJECTIVE: To investigate the expressions of survivin and GRIM-19 in prostatic cancer tissue and their clinical implications. METHODS: We detected the expressions of survivin and GRIM-19 in the tissues of normal prostate (NP), benign prostate hyperplasia (BPH) and prostate cancer (PCa) using immunohistochemical staining, RT-PCR and Western blot, and processed the data by SPSS12. RESULTS: The positive rates of survivin expression were 6.25% , 18.18% and 90.62% in NP, BPH and PCa (P < 0.01), while those of GRIM-19 were 87.50%, 81.82% and 9.37% , respectively (P < 0.01). Semiquantitative RT-PCR and immunohistochemical staining showed that both survivin mRNA and survivin expressions were highly positive in PCa but negative in NP and BPH. Western blot exhibited that the survivin protein was expressed strongly in PCa but weakly in NP and BPH, while the GRIM-19 protein was expressed just contrariwise (P < 0.01). CONCLUSION: The expressions of survivin and GRIM-19 may be closely correlated with the pathogenesis of prostate cancer.

[Effects of a novel recombinant polypeptide from the sea anemone Anthopleura sp. on left ventricular function in dogs with acute cardiac insufficiency].

OBJECTIVE: To observe the effects of recombinant hk2a, a novel neurotoxin from the sea anemone Anthopleura sp., on left ventricular function of dogs with acute cardiac insufficiency. METHODS: Canine models of acute cardiac insufficiency were established by rapid ventricular pacing, in which the left ventricular ejection fraction (LVEF) was measured by Acuson ultrasound systems (Sequoia 512) at 0, 5, 15, 30 and 60 min, respectively, after intravenous injection of 30 microg/kg recombinant hk2a. The response of the canine models to hk2a treatments was observed in comparison with that of the dogs treated with Cedilanid (as positive control) and saline (as negative control). RESULTS: Intravenous injection of recombinant hk2a caused an immediate and significant increase in LVEF in the canine models of acute cardiac insufficiency (P<0.05), and the effect maintained for more than 30 min without significant effect on heart rate. Recombinant hk2a possessed such merits as quicker onset and greater potency in comparison with Cedilanid. CONCLUSION: Recombinant hk2a may significantly increase LVEF of the dogs with acute cardiac insufficiency.

[Effects of a novel recombinant polypeptide from the sea anemone Anthopleura sp. on left ventricular function in New Zealand rabbits with chronic congestive heart failure].

OBJECTIVE: To observe the effects of recombinant hk2a, a novel neurotoxin from the sea anemone Anthopleura sp. on the left ventricular function in New Zealand rabbits with chronic congestive heart failure(CCHF). METHODS: CHF model was established using New Zealand rabbits by abdominal aortic coarctation. The left ventricular ejection fraction (LVEF) was measured by Acuson ultrasound systems (Sequoia 512) at 0, 5, 15, 30, 45 and 60 min respectively after intravenous injection with a single dose of recombinant hk2a (20 microg/kg x b x w.). The response of the rabbits to hk2a treatment was observed in comparison with that of the control group. RESULTS: Recombinant hk2a immediately and significantly increased the left ventricular ejection fraction of the rabbit models of CCHF after intravenous injection (P<0.05), and its effects were maintained for more than 1 h without affecting the heart rate. CONCLUSION: recombinant hk2a significantly increases the left ventricular ejection fraction in New Zealand rabbits with CCHF.

[Effects of recombinant human interleukin-10 on tumor necorsis factor-alpha-induced adventitial fibroblast proliferation in vitro].

OBJECTIVE: To observe the effects of recombinant human interleukin-10 (rhIL-10) on adventitial fibroblast proliferation induced by tumor necrosis factor-alpha TNF-alpha in vitro. METHODS: In this controlled study, NIH/3T3 cells cultured in vitro were treated with TNF-alpha and recombinant human interleukin-10 (rhIL-10), respectively, and the cell proliferation was determined by non-radioactive MTS/PES assay. Cell cycle analysis was performed using flow cytometry. RESULTS: Compared with the control cells, TNF-alpha significantly stimulated NIH/3T3 cell proliferation, wherea. rhIL-10 had no such effect. With TNF-alpha stimulation, rhIL-10 at the dose as low as 1 ng/ml inhibited the proliferation of NIH/3T3 cells (P<0.05). The number of cells in G(0)/G(1) phase treated with both TNF-alpha and rhIL-10 was higher than those treated with TNF-alpha alone (P<<0.05), as shown by flow cytometry. CONCLUSION: rhIL-10 can significantly inhibit the proliferation of adventitial fibroblasts induced by TNF-alpha in vitro.