INHBA regulates Hippo signaling to confer 5-FU chemoresistance mediated by cellular senescence in colon cancer cells.

Colon cancer has become a global public health challenge, and 5-Fluorouracil (5-FU) chemoresistance is a major obstacle in its treatment. Chemoresistance can be mediated by therapy-induced cellular senescence. This study intended to investigate mechanisms of INHBA (inhibin A) in 5-FU resistance mediated by cellular senescence in colon cancer. Bioinformatics analysis of INHBA expression in colon cancer tissues, survival analysis, and correlation analysis of cellular senescence markers were performed. The effects of INHBA on the biological characteristics and 5-FU resistance of colon cancer cells were examined through loss/gain-of-function and molecular assays. Finally, a xenograft mouse model was built to validate the mechanism of INHBA in vivo. INHBA was upregulated in colon cancer and was significantly positively correlated with cellular senescence markers uncoupling protein 2 (UCP-2), matrix metalloproteinase-1 (MMP-1), dense and erect panicle 1 (DEP1), and p21. Cellular senescence in colon cancer mediated 5-FU resistance. Downregulation of INHBA expression enhanced 5-FU sensitivity in colon cancer cells, inhibited cell proliferation, promoted apoptosis, increased the proportion of cells in G0/G1 phase, and it resulted in a lower proportion of senescent cells and lower levels of the cellular senescence markers interleukin 6 (IL-6) and interleukin 8 (IL-8). Analysis of whether to use the pathway inhibitor Verteporfin proved that INHBA facilitated colon cancer cell senescence and enhanced 5-FU chemoresistance via inactivation of Hippo signaling pathway, and consistent results were obtained in vivo. Collectively, INHBA conferred 5-FU chemoresistance mediated by cellular senescence in colon cancer cells through negative regulation of Hippo signaling.

Correlation of immune inflammatory indices and nutritional risk index with prognosis in patients with non-small cell lung cancer.

OBJECTIVE: To investigate the relationship of lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR) and nutritional risk index (NRI) with the prognosis of non-small cell lung cancer (NSCLC). METHODS: The clinical data of 400 NSCLC patients undergoing surgery at Shaoxing Shangyu Hospital of Traditional Chinese Medicine from January 2019 to June 2022 were collected for this retrospective analysis. The optimal cutoff values for NLR, PLR, LMR and NRI were determined using receiver operating characteristic (ROC) curves. The patients were grouped according to the optimal cutoff values, and the clinicopathological characteristics were compared between groups. The Kaplan-Meier survival curve and Cox risk model were used to identify the independent risk factors affecting the prognosis of patients with NSCLC. A nomogram risk prediction model was constructed and its effectiveness was verified. RESULTS: ROC curve analysis revealed that the area under the curve (AUC) values for NLR, PLR, LMR and NRI in predicting overall survival of NSCLC patients were 0.827, 0.753, 0.719 and 0.770, respectively. The optimal cutoff values for NLR, PLR, LMR and NRI were 2.49, 126.32, 3.02 and 89, respectively. Survival analysis found that the survival time was shorter in patients with NLR>2.49, PLR>126.32, LMR>3.02 and NRI≤89. Results from Cox model indicated that TNM staging, NLR>2.49, LMR>3.02, NRI≤89, surgical method, intraoperative blood loss, postoperative complication, and adjuvant chemotherapy were risk factors affecting the prognosis of NSCLC patients. A nomogram was constructed based on the results of multivariate analysis. The AUC of the nomogram was 0.967 (95% CI: 0.943-0.992) and 0.948 (95% CI: 0.874-1) in the training set and the test set, respectively. The C-index was 0.90 and 0.89, respectively. The calibration curve demonstrated good agreement between the predicted values of the nomogram and the actual observed values. CONCLUSION: NLR, LMR and NRI are significant predictors of the prognosis of patients with NSCLC. NLR>2.49, LMR>3.02, and NRI≤89 are risk factors for the prognosis of NSCLC patients.

Eukaryotic Elongation Factor 1Alpha-2 (EEF1A2) Participates in the Progression of Gastric Cancer via Interaction with Heat Shock Protein B8 (HSPB8).

OBJECTIVE: The critical roles of eukaryotic elongation factor 1alpha-2 (EEF1A2) and heat shock protein B8 (HSPB8) in the carcinogenesis and progression of cancers have been well documented. However, the regulatory role of EEF1A2/HSPB8 in the development of gastric cancer (GC) have not been fully understood. This study was aimed at clarifying the biological effects of EEF1A2/HSPB8 on the malignant behaviors of GC cells and to investigate the molecular mechanism underlying the involvement of EEF1A2/HSPB8 in GC. METHODS: In the present work, expression differences of EEF1A2 and HSPB8 in GC cells were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blot assay. Cell counting kit -8 (CCK-8), 5-ethynyl-2'-deoxyuridine (EdU) staining, wound healing and transwell assays were employed to detect the proliferation, migration and invasion of GC cells. In addition, tube formation assay was adopted to assess in vitro angiogenesis of HUVECs incubated with the conditioned media (CM) of GC cells. Moreover, the interaction between EEF1A2 and HSPB8 was predicted from BioGrid database and analyzed through co-immunoprecipitation (Co-IP). RESULTS: The present research revealed that EEF1A2 and HSPB8 were highly expressed in GC cell lines. EEF1A2 knockdown markedly suppressed the proliferation, migration and invasion of GC cells as well as in vitro angiogenesis. Furthermore, it was verified that EEF1A2 interacted with HSPB8 and positively regulated HSPB8 expression. Overexpression of HSPB8 reversed the suppressive effects of EEF1A2 knockdown on GC cell proliferation, migration, invasion and in vitro angiogenesis. CONCLUSION: In conclusion, EEF1A2 could act as an oncogene in the development of GC via promoting HSPB8 expression.

Management guideline for the off-label use of medicine in China (2021).

BACKGROUND: Off-label drug use embodies a thorough clinical diagnosis and evaluation of treatment needs and should not be confused with unreasonable drug use, but it also faces potential risks with drug safety and legal issues. RESEARCH DESIGN AND METHODS: We first established a guideline working group. Following the guideline development process recommended by the World Health Organization Handbook and the Chinese Medical Association, the key questions were determined through literature searches of PubMed, CNKI (Chinese National Knowledge Infrastructure) and other databases. Both the evidence and the clinicians' diagnosis and treatment workload were considered to formulate the initial recommendations. Finally, two rounds of Delphi surveys and one expert seminar were organized to determine the final recommendations of this guideline. Meanwhile, we graded the recommendations based on the body of evidence. RESULTS: We determined nine questions and proposed a total of 23 recommendations regarding the definition of off-label use of drugs, applicable circumstances, classification of evidence, informed consent, legal basis, adverse drug reaction monitoring and evaluation, management procedure, responsibilities and obligations of different stakeholders, medical insurance reimbursement, and the national approval system. CONCLUSIONS: This guideline standardized clinical off-label drug use and provided suggestions and references for the management of off-label drug use.

Associations of Clinical and Molecular Characteristics with the Response to Immune Checkpoint Blockade in Advanced Gastric Cancers.

Purpose: Immunotherapy provides a new treatment option for advanced gastric cancer (AGC). This study aims to explore the response markers of immunotherapy in AGCs. Methods: Next-generation sequencing was performed on 44 AGC patients who received immune checkpoint inhibitors and the associations between their outcomes after combination immunotherapy, and the clinicopathological/molecular characteristics were analyzed. Results: The current study cohort had a median progression-free survival (PFS) of 5.9 months, an overall survival (OS) of 12.1 months, and an objective response rate (ORR) of 36.4%. Through multivariable analysis of the clinical characteristics, primary tumor resection (HR = 2.66, 95% CI: 1.06-6.70, p=0.037) and increased proportion of lymphocytes after combination immunotherapy (HR = 0.40, 95% CI: 0.16-0.99, p=0.048) were revealed as independent predictors for patient outcomes. All the 18 patients who underwent genetic profiling were microsatellite-stable with a median TMB of four mutations per Mb. ATM alterations, PI3K pathway mutations, increased TMB, and positive PD-L1 expression were associated with the increased trend of PFS and ORR. According to the combination of baseline lymphocyte count, ATM mutation, TMB status, and PD-L1 expression, patients were stratified into higher- and lower-risk groups, with the lower-risk group showing improved PFS (HR = 4.7e-10, 95% CI: 0-inf, p=0.02) and ORR (75% vs. 0%, p=0.007). Conclusion: Several highly relevant potential biomarkers predictive of immunotherapy response in AGC patients have been identified in this research.

Analysis of the mechanism regulating the killing effect of EBNA1 on EBV-associated B-cell tumors.

In order to analysis of the mechanisms regulating EBNA1 killing of EBV associated B cell tumors, preparations were first made for EBV-associated B cells, and the cells were subsequently transformed. The killing effect of ebna1-28 T cells on EBV-positive B cell lymphoid tumor cells was detected using the FACS method. SF rats were also selected to analyze the inhibitory effect of ebna1-28t on transplanted tumors in nude mice with EBV-positive B cell lymphoma. Results showed that compared with the untransfected group, the expression of EBNA1 was higher in the empty plasmid SFG group, the recombinant plasmid rv-ebna1/car group compared with the empty plasmid SFG group, and the expression of EBNA1 was higher in the untransfected group compared with the empty plasmid SFG group, which was statistically significant (P < 0.05); As shown in Figure 1, in vitro studies found that, compared to the untransfected group, the empty plasmid SFG group, the recombinant plasmid rv-ebna1/car group showed better killing efficacy on Raji cells, and the recombinant plasmid rv-ebna1/car group showed better killing efficacy on Raji cells compared to the empty plasmid SFG group; The tumor volumes of the rats in group C were larger compared with those in groups A and B, and the tumor volumes of the rats in group A were smaller compared with those in group B. The tumor volumes of the rats in group C were larger compared with those of the rats in the three groups (P<0.05). In group C, the cells were more severely invaded, and the nuclei were damaged. In group B, cell invasion in tissues was mild in the nucleus. The infection of cells in the tissues of rats in group A was better compared to groups B and C. In vitro experiments found that inhibition of EBNA1 was able to kill EBV-positive B cell lymphoid tumor cells effectively. Animal experiments found that ebna1-28t was able to shrink the volume as well as tumor weight of transplanted tumors in nude mice with EBV-positive B cell lymphoma and played a better inhibitory role.

Anti-PD-1 Therapy-A Potential Treatment for Myocardial Metastasis From Nasopharyngeal Carcinoma: A Case Report.

Myocardial metastasis of nasopharyngeal carcinoma (NPC) is rarely reported in the literature. Some autopsy studies found metastases in more than 10% of cases with malignant neoplasm. However, patients are often diagnosed during the postmortem because myocardial metastasis is often asymptomatic, and its Cardiac complications tend to be severe and fatal. Patients with Cardiac metastases are often treated with chemotherapy or surgical intervention, although the prognosis is poor. Immunotherapy with anti-programmed cell death receptor-1 or ligand-1 (PD-1 or PD-L1) inhibitors has recently been reported to be therapeutically significant in multiple cancers, including melanoma, nonsmall cell lung cancer, and NPC, but the treatment of myocardial metastasis of NPC has not been reported. This study described the case of a 50-year-old male patient who presented initially with NPC and received radiotherapy as first-line therapy. For 20 years, he had recurrent Cardiac metastasis of NPC. The pathological examination suggested tPD-L1 expression. Therefore, off-label sintilimab (200 mg every 21 days) was administered. After 10 cycles of treatment, myocardial metastasis shrank and the enlarged mediastinal lymph nodes disappeared. This case report demonstrated that Cardiac metastasis of NPC expressing PD-L1 might have a sustained response to PD-L1 inhibitor-directed therapy.

Structure-Enhanced Mechanically Robust Graphite Foam with Ultrahigh MnO2 Loading for Supercapacitors.

With the fast bloom of flexible electronics and green vehicles, it is vitally important to rationally design and facilely construct customized functional materials with excellent mechanical properties as well as high electrochemical performance. Herein, by utilizing two modern industrial techniques, digital light processing (DLP) and chemical vapor deposition (CVD), a unique 3D hollow graphite foam (HGF) is demonstrated, which shows a periodic porous structure and robust mechanical properties. Finite element analysis (FEA) results confirm that the properly designed gyroidal porous structure provides a uniform stress area and mitigates potential structural failure caused by stress concentrations. A typical HGF can show a high Young's modulus of 3.18 MPa at a low density of 48.2 mg cm-3. The porous HGF is further covered by active MnO2 material with a high mass loading of 28.2 mg cm-2 (141 mg cm-3), and the MnO2/HGF electrode still achieves a satisfactory specific capacitance of 260 F g-1, corresponding to a high areal capacitance of 7.35 F cm-2 and a high volumetric capacitance of 36.75 F cm-3. Furthermore, the assembled quasi-solid-state asymmetric supercapacitor also shows remarkable mechanical properties as well as electrochemical performance.

Laboratory monitoring of rivaroxaban in Chinese patients with deep venous thrombosis: a preliminary study.

BACKGROUND: Rivaroxaban, a novel oral anticoagulant drug, is widely used in clinical practice. There is no standardized laboratory monitoring for rivaroxaban, and its plasma concentration in Chinese patients with deep vein thrombosis is unclear. The rivaroxaban concentrations in human plasma and determine the steady-state concentration of rivaroxaban in patients with deep vein thrombosis are needed. METHODS: An ultra-high-performance liquid chromatography with mass spectrometric detection method was developed. Chromatographic separation was performed on a Waters BEH C18 column with isocratic elution using a mobile phase composed of acetonitrile and water. Quantitation of the analytes was performed using positive ionization mode and mass transitions of m/z 437.3 → m/z 145.0 and m/z 440.1 → m/z 145.0 for rivaroxaban and the internal standard, respectively. Blood samples were collected at 0 h and 2 h after patients took rivaroxaban for 7 days or more. RESULTS: The method was validated over the concentration range of 0.5 ~ 400 ng•mL- 1 with a very low limit of quantification of 0.5 ng·mL- 1, and the intra- and inter-day precision (RSD%) were < 15%. The range of the steady state concentration in patients that took 15 mg rivaroxaban twice daily, 10 mg twice daily, 20 mg once daily, 15 mg once daily, and 10 mg once daily were 168.5 ~ 280.1 ng•mL- 1, 74.2 ~ 271.4 ng•mL- 1, 25.7 ~ 306.8 ng•mL- 1, 24.5 ~ 306.4 ng•mL- 1, and 15.4 ~ 229.2 ng•mL- 1, respectively. CONCLUSIONS: The plasma rivaroxaban concentration in patients who took 10 mg rivaroxaban twice daily fluctuated less than that in patients who took 20 mg rivaroxaban once daily. The plasma concentration can be used for therapeutic drug monitoring for rivaroxaban.

The interpretation of China national essential medicines list 2018.

Background: In 2018, China implemented the latest National Essential Medicines List (NEML) by enhancing the NEML 2012. The goal of our studies is to analyze the changes in the two lists and compared them with the 20th EML issued by WHO in 2017. And then provide suggestions for emerging problems.Method: The overall composition of the categories, specific drugs, characteristics, advantages and disadvantages of the lists were compared by descriptive analysis. The neuropsychiatric disorders system medicines and patented medicines were analyzed to illustrate the changes of NEML.Results: In 20th WHO-EML, the largest increase was the medicines used for children (13 to the core list and 12 to the complementary list). In 2018 NEML, rounding out the top were medicines used for cardiovascular system. Among the 120 new medicines, 30 new medicines were included in 2017 WHO-EML. Eleven patented medicines were new-added in NEML; however, 8 was not included in WHO-EML.Conclusion: China has a large population, and the territorial development is uneven. Although the essence of EMLs is a limited list, NEML should enlarge the choices properly. 2018 NEML provides a comprehensive coverage of diseases. Some of the medicines, including high-priced medicines that were not recommended by WHO.

Unique genomic profiles obtained from cerebrospinal fluid cell-free DNA of non-small cell lung cancer patients with leptomeningeal metastases.

BACKGROUND: Leptomeningeal metastases (LM), associated with poor prognosis, are frequent complications of advanced non-small cell lung cancer (NSCLC) patients, especially in patients with epidermal growth factor receptor (EGFR) mutations. Due to limited access to leptomeningeal lesions, the mutational landscape of LM has not been comprehensively investigated in large cohorts and the underlining biology of LM remains elusive. Some studies have explored the potential of cerebrospinal fluid (CSF) in reflecting the molecular profile of LM but with limited number of patients enrolled. METHODS: In this study, we performed capture-based targeted sequencing using a panel consisting of 168 lung cancer-related genes on matched CSF and plasma samples from 72 advanced NSCLC patients with confirmed LM to interrogate the potential of CSF as a source of liquid biopsy. RESULTS: We revealed a rate of detection of 81.5% and 62.5% for CSF and plasma, respectively (p = 0.008). The maximum allelic fraction (MaxAF) was also significantly higher in CSF (43.6% vs. 4.6%) (p < 0.001). CSF, harboring a unique genomic profile by having a significant number of CSF-specific mutations, primarily copy number variations, is superior to plasma in reflecting the mutational profile of LM. Further pathway enrichment analysis revealed that most of CSF-specific mutations participated in pathways relevant to the tumorigenesis and the development of metastases. Moreover, our data also revealed that TP53 loss of heterozygosity (LOH) predominantly existed in CSF (p < 0.001). CONCLUSIONS: Collectively, we demonstrated that CSF provides a more comprehensive profile of LM than plasma in a large cohort, thus can be used as an alternative source of liquid biopsy for LM patients.

A stereo configuration-activity study of 3-iodo-4-(2-methylcyclohexyloxy)-6-phenethylpyridin-2(2H)-ones as potency inhibitors of HIV-1 variants.

3-Iodo-4-(2'-methylcyclohexyloxy)-6-phenethylpyridin-2(1H)-ones, as effective non-nucleoside reverse transcriptase inhibitors, were synthesized and resolved with different configurations. Biological results revealed that the trans-racemate 2b exhibited more potent activity than the cis-isomers. Noticeably, the trans-(S,S)-enantiomer 2e turned out to be significantly more potent than its counterpart enantiomer 2d against wild-type and double-mutant strains with high selectivity indexes.

[Synthesis of 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl)pyrimidine-2,4(1H,3H)-dione].

OBJECTIVE: To find the best synthesis method of 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl) pyrimidine-2,4(1H,3H)-dione e for observing the change of its biological activity after N-3 hydroxylation. METHODS: After trying some N-hydroxylation methods, the target compound was successfully synthesized via one-pot oxidizing process by sodium hydride (NaH) and 3-chloroperbenzoic acid(m-CPBA); the anti-HIV reverse transcriptase (RT) activity and integrase (IN) activity of the target compound was assayed via enzyme-linked immunesorbent assay (ELISA) and phosphorylation of DNA package method. RESULTS: The target compound could be obtained through the improved m-CPBA oxidative method by only one step, and the yield of the reaction could reach 60%-70%. And the structure of this compound was identified by 1H NMR, 13C NMR and MS; The activity result showed it added the anti-HIV IN activity after N-3 hydroxylation as well as retained the anti-HIV RT activity. CONCLUSION: The improved m-CPBA oxidative method is a convenient and efficient way to prepare the compound 6-benzyl-1-[(benzyloxy)methyl]-3-hydroxy-5-(hydroxymethyl)pyrimidine-2,4(1H,3H)-dione e which has both anti-HIV RT and IN activity.

Synthesis and biological evaluation of novel 2-arylalkylthio-5-iodine-6-substituted-benzyl-pyrimidine-4(3H)-ones as potent HIV-1 non-nucleoside reverse transcriptase inhibitors.

A novel series of 2-arylalkylthio-5-iodine-6-substitutedbenzyl-pyrimidine-4(3H)-ones (S-DABOs) 8a-x had been synthesized via an efficient method. Their biological activity against HIV virus and RT assay were evaluated. Some compounds, especially 8h, 8l and 8n, displayed promising activity against HIV-1 RT with IC50 values in a range of 0.41 µM to 0.71 µM, which were much better than that of nevirapine. Molecular modeling studies revealed that the binding mode would be affected via forming an additional hydrogen bond by incorporating an oxygen atom on the C-2 side chain. The biological activity was in accordance with the docking results.