Gossypol acetic acid regulates leukemia stem cells by degrading LRPPRC via inhibiting IL-6/JAK1/STAT3 signaling or resulting mitochondrial dysfunction.

BACKGROUND: Leukemia stem cells (LSCs) are found to be one of the main factors contributing to poor therapeutic effects in acute myeloid leukemia (AML), as they are protected by the bone marrow microenvironment (BMM) against conventional therapies. Gossypol acetic acid (GAA), which is extracted from the seeds of cotton plants, exerts anti-tumor roles in several types of cancer and has been reported to induce apoptosis of LSCs by inhibiting Bcl2. AIM: To investigate the exact roles of GAA in regulating LSCs under different microenvironments and the exact mechanism. METHODS: In this study, LSCs were magnetically sorted from AML cell lines and the CD34+CD38- population was obtained. The expression of leucine-rich pentatricopeptide repeat-containing protein (LRPPRC) and forkhead box M1 (FOXM1) was evaluated in LSCs, and the effects of GAA on malignancies and mitochondrial function were measured. RESULTS: LRPPRC was found to be upregulated, and GAA inhibited cell proliferation by degrading LRPPRC. GAA induced LRPPRC degradation and inhibited the activation of interleukin 6 (IL-6)/janus kinase (JAK) 1/signal transducer and activator of transcription (STAT) 3 signaling, enhancing chemosensitivity in LSCs against conventional chemotherapies, including L-Asparaginase, Dexamethasone, and cytarabine. GAA was also found to downregulate FOXM1 indirectly by regulating LRPPRC. Furthermore, GAA induced reactive oxygen species accumulation, disturbed mitochondrial homeostasis, and caused mitochondrial dysfunction. By inhibiting IL-6/JAK1/STAT3 signaling via degrading LRPPRC, GAA resulted in the elimination of LSCs. Meanwhile, GAA induced oxidative stress and subsequent cell damage by causing mitochondrial damage. CONCLUSION: Taken together, the results indicate that GAA might overcome the BMM protective effect and be considered as a novel and effective combination therapy for AML.

Anti-PD-1/PD-L1 therapy for colorectal cancer: Clinical implications and future considerations.

Colorectal cancer (CRC) is the third most prevalent cancer in the world. The PD-1/PD-L1 pathway plays a crucial role in modulating immune response to cancer, and PD-L1 expression has been observed in tumor and immune cells within the tumor microenvironment of CRC. Thus, immunotherapy drugs, specifically checkpoint inhibitors, have been developed to target the PD-1/PD-L1 signaling pathway, thereby inhibiting the interaction between PD-1 and PD-L1 and restoring T-cell function in cancer cells. However, the emergence of resistance mechanisms can reduce the efficacy of these treatments. To counter this, monoclonal antibodies (mAbs) have been used to improve the efficacy of CRC treatments. mAbs such as nivolumab and pembrolizumab are currently approved for CRC treatment. These antibodies impede immune checkpoint receptors, including PD-1/PD-L1, and their combination therapy shows promise in the treatment of advanced CRC. This review presents a concise overview of the use of the PD-1/PD-L1 blockade as a therapeutic strategy for CRC using monoclonal antibodies and combination therapies. Additionally, this article outlines the function of PD-1/PD-L1 as an immune response suppressor in the CRC microenvironment as well as the potential advantages of administering inflammatory agents for CRC treatment. Finally, this review analyzes the outcomes of clinical trials to examine the challenges of anti-PD-1/PD-L1 therapeutic resistance.

LncRNAs in colorectal cancer: Biomarkers to therapeutic targets.

Colorectal cancer (CRC) is the third leading cause of cancer-related death in men and women worldwide. As early detection is associated with lower mortality, novel biomarkers are urgently needed for timely diagnosis and appropriate management of patients to achieve the best therapeutic response. Long noncoding RNAs (lncRNAs) have been reported to play essential roles in CRC progression. Accordingly, the regulatory roles of lncRNAs should be better understood in general and for identifying diagnostic, prognostic and predictive biomarkers in CRC specifically. In this review, the latest advances on the potential diagnostic and prognostic lncRNAs as biomarkers in CRC samples were highlighted, Current knowledge on dysregulated lncRNAs and their potential molecular mechanisms were summarized. The potential therapeutic implications and challenges for future and ongoing research in the field were also discussed. Finally, novel insights on the underlying mechanisms of lncRNAs were examined as to their potential role as biomarkers and therapeutic targets in CRC. This review may be used to design future studies and advanced investigations on lncRNAs as biomarkers for the diagnosis, prognosis and therapy in CRC.

Targeting epigenetics and lncRNAs in liver disease: From mechanisms to therapeutics.

Early onset and progression of liver diseases can be driven by aberrant transcriptional regulation. Different transcriptional regulation processes, such as RNA/DNA methylation, histone modification, and ncRNA-mediated targeting, can regulate biological processes in healthy cells, as well also under various pathological conditions, especially liver disease. Numerous studies over the past decades have demonstrated that liver disease has a strong epigenetic component. Therefore, the epigenetic basis of liver disease has challenged our knowledge of epigenetics, and epigenetics field has undergone an important transformation: from a biological phenomenon to an emerging focus of disease research. Furthermore, inhibitors of different epigenetic regulators, such as m6A-related factors, are being explored as potential candidates for preventing and treating liver diseases. In the present review, we summarize and discuss the current knowledge of five distinct but interconnected and interdependent epigenetic processes in the context of hepatic diseases: RNA methylation, DNA methylation, histone methylation, miRNAs, and lncRNAs. Finally, we discuss the potential therapeutic implications and future challenges and ongoing research in the field. Our review also provides a perspective for identifying therapeutic targets and new hepatic biomarkers of liver disease, bringing precision research and disease therapy to the modern era of epigenetics.

Risk Prediction for Locoregional Recurrence in Epidermal Growth Factor Receptor-Mutant Stage III-pN2 Lung Adenocarcinoma after Complete Resection: A Multi-center Retrospective Study.

Background: This study aimed to develop a predictive model based on the risk of locoregional recurrence (LRR) in epidermal growth factor receptor (EGFR)-mutant stage III-pN2 lung adenocarcinoma after complete resection. Methods: A total of 11,020 patients with lung surgery were screened to determine completely resected EGFR-mutant stage III-pN2 lung adenocarcinoma. Patients were excluded if they received preoperative therapy or postoperative radiation therapy (PORT). The time from surgery to LRR was recorded. Clinicopathological variables with statistical significance predicting LRR in the multivariate Cox regression were incorporated into the competing risk nomogram. Patients were then sub-grouped based on different recurrence risk as a result of the nomogram. Results: Two hundred and eighty-eight patients were enrolled, including 191 (66.3%) with unforeseen N2 (IIIA1-2), 75 (26.0%) with minimal/single station N2 (IIIA3), and 22 (7.6%) with bulky and/or multilevel N2 (IIIA4). The 2-year overall cumulative incidence of LRR was 27.2% (confidence interval [CI], 16.3%-38.0%). IIIA4 disease (hazard ratio, 2.65; CI, 1.15-6.07; P=0.022) and extranodal extension (hazard ratio, 3.33; CI, 1.76-6.30; P<0.001) were independent risk factors for LRR and were incorporated into the nomogram. Based on the nomogram, patients who did not have any risk factor (low-risk) had a significantly lower predicted 2-year incidence of LRR than those with any of the risk factors (high-risk; 4.6% vs 21.9%, P<0.001). Conclusions: Pre-treatment bulky/multilevel N2 and pathological extranodal extension are risk factors for locoregional recurrence in EGFR-mutant stage III-pN2 lung adenocarcinoma. Intensive adjuvant therapies and active follow-up should be considered in patients with any of the risk factors.

Shenqi Fuzheng Injection Ameliorates Radiation-induced Brain Injury.

Shenqi Fuzheng injection (SFI) has been confirmed to be able to alleviate brain injury in mice. This study examined the brain-protective effect of SFI on patients after cranial radiation. Lung cancer patients with brain metastasis were randomly assigned to two groups. The SFI group received cranial radiation in combination with SFI. The control group received cranial radiation alone. The changes in cognitive function were evaluated pre- and post-radiation against the Mini-Mental State Exam (MMSE), Montreal Cognitive Assessement (MoCA), Zung Self-Rating Depression Scale (SDS) and Zung Self-Rating Anxiety Scale (SAS). The changes in inflammatory factors, such as TGF-ß1, TNF-α and IL-10, were also detected before, during and after radiation (15Gy/5F). The results showed that 6 months after cranial radiation, the total scores on the MMSE and MoCA scales of the patients decreased, especially memory ability. The control group experienced a more evident decline, the memory ability being the greatest. TGF-ß1 and TNF-a increased shortly after radiation and decreased one month later, and the change was more conspicuous in SFI group than in control group. IL-10 increased after radiation and stayed at a high level one month later in both groups, the level being higher in the SFI group than in the control group. Our study indicated that cognitive functions, especially memory ability, were impaired after cranial radiation. SFI could alleviate radiation-induced brain injury by regulating inflammatory factors.

[Preliminary Analysis of the Genetic Loci of SAG1 and SAG3, the Surface Antigens of Toxoplasma gondii, in HIV-positive People in Dali of Yunnan Province].

OBJECTIVE: To identify the genotypes of Toxoplasma gondii that infects HIV-positive people in Dali of Yunnan Province through analyzing the genetic loci of the surface antigens SAG1 and SAG3. METHODS: A total of 291 blood samples from HIV-positive cases were collected from the HIV/AIDS Prevention and Control Institution in Yunnan. Nested PCR was used to amplify SAGI and SAG3 genes in the blood samples. The products were digested with restriction enzymes Sau96 I, Hae II and Nci I, and sequenced. RESULTS: Of the 291 HIV-positive blood samples, 64 showed successful amplification of SAGI gene, and 42 of SAG3 gene, with product sizes of 390 bp and 225 bp, respectively. Enzymetic digestion of the PCR products resulted in fragments of 350 bp and 50 bp for SAGI, and -200 bp band for SAG3, consistent with RH, a particular type I strain of T. gondii. Sequencing of the SAG1 and SAG3 PCR products showed that their sequence identities with SAGI (Accession No. GQ253073) and SAG3 (Accession No. JX218225.1) of the type I strain of T. gondii were 99.98%-100% and 99.96% -99.98% respectively. CONCLUSION: The Toxoplasma gondii in HIV-positive cases in Dali of Yunnan Province is the type I strain of T. gondii.

Shenqi fuzheng injection attenuates irradiation-induced brain injury in mice via inhibition of the NF-κB signaling pathway and microglial activation.

AIM: Radiation-induced brain injury (RIBI) is the most common and severe adverse effect induced by cranial radiation therapy (CRT). In the present study, we examined the effects of the traditional Chinese medicine Shenqi Fuzheng Injection (SFI) on RIBI in mice, and explored the underlying mechanisms. METHODS: C57BL/6J mice were subjected to a single dose of 20-Gy CRT. The mice were treated with SFI (20 mL·kg(-1)·d(-1), ip) for 4 weeks. Morris water maze test was used to assess the cognitive changes. Evans blue leakage and a horseradish peroxidase (HRP) assay were used to evaluate the integrity of the blood-brain barrier (BBB). The expression of inflammatory factors and microglial activation in brain tissues were detected using RT-PCR, Western blotting and immunofluorescence staining. RESULTS: CRT caused marked reductions in the body weight and life span of the mice, and significantly impaired their spatial learning. Furthermore, CRT significantly increased the BBB permeability, number of activated microglia, expression levels of TNF-α and IL-1ß, and the levels of phosphorylated p65 and PIDD-CC (the twice-cleaved fragment of p53-induced protein with a death domain) in the brain tissues. Four-week SFI treatment (administered for 2 weeks before and 2 weeks after CRT) not only significantly improved the physical status, survival, and spatial learning in CRT-treated mice, but also attenuated all the CRT-induced changes in the brain tissues. Four-week SFI pretreatment (administered for 4 weeks before CRT) was less effective. CONCLUSION: Administration of SFI effectively attenuates irradiation-induced brain injury via inhibition of the NF-κB signaling pathway and microglial activation.

[Analysis of sites of Toxoplasma gondii SAG2 gene in blood of HIV-positive people in Lincang City, Yunnan Province].

OBJECTIVE: To preliminarily understand the genotype characteristics of Toxoplasma gondii in blood of HIV-positive persons in Lincang City, Yunnan Province. METHOD: Two segments of SAG2 gene of T. gondii from blood samples of HIV-positive persons in Lincang City were extracted and amplified by using the nested PCR method and the genotype was identified and compared with the standard strain (Type I) of Toxoplasma gondii. RESULTS: Thirty-five SAG2 genes (241 bp) and 35 SAG2 genes (221 bp) of T. gondii were amplified from 170 blood samples of the HIV-positive people, and 4 of each case were selected and digested with enzyme, then 2 aim gene fragments of each case were chosen and compared with the standard strain (Type I) of T. gondii. The digestion of SAG2 gene (241 bp) showed the genotype of the blood samples was Type I or Type II, and the digestion of SAG2 gene (221 bp) confirmed that the genotype was Type I. CONCLUSION: It is preliminarily confirmed that the genotype of T. gondii in blood of HIV-positive persons in Lincang City, Yunnan Province is Type I.

[Comparation of Toxoplasma gondii separated from HIV-positive people and RH strain GRA6 gene].

OBJECTIVE: To comparatively analyze Toxoplasma gondii separated from HIV-positive people and RH strain GRA6 gene. METHODS: By using the nested PCR, the amplification of Dali HIV-positive blood samples and RH strains of Toxoplasma GRA6 genome was performed. The GRA6 gene amplification positive product was selected and the electrophoresis imaging was performed by being digested with the Mse I endonuclease, and the gene sequences were measured and analyzed. RESULTS: The GRA6 gene fragment (800 bp) was successfully amplified, and about 600 bp and 200 bp bands were got by Mse I. The sequencing results showed that T. gondii GRA6 gene positive samples had 2 nucleotide variation compared with T. gondii strain RH, namely 447 base pair at C becoming G, and 623 base pair at G becoming T. At 146 bp and 690 bp, the Mse I restriction sites (TTAA) were found. CONCLUSION: The preliminary judgment shows that the Dali HIV-positive T. gondii genotype is consistent with RH strain, belonging to genotype I.

[Advances in genetyping of Toxoplasma gondii].

Toxoplasma gondii is a nucleated cell obligate parasite, and can cause severe toxoplasmosis. The genetypes of Toxoplasma gondii isolates of different host infections have significant differences, and the pathogenicity and sensitivity to drugs of different genotypes of Toxoplasma gondii isolates are also significantly different. At present, the analysis of Toxoplasma gondii genotypes is performed by using PCR-RFLP, PCR of highly repetitive sequences, multiple PCR and nested PCR techniques, and the multiple loci of amplification are usually B1, SAG2, HSP70, GRA6 and other genetic markers. There are main 3 traditional genotypes of Toxoplasma gondii, and along with the deepening and extension of research, more and more genotypes are found. This paper reviews the advances in the research of Toxoplasma gondii genotypes.

[Genotypes of Toxoplasma gondii isolates from HIV positive patients in Yunnan Province].

One hundred and fifty serum samples of HIV positive patients were collected in western Yunnan Province from September 2011 to December 2012. Toxoplasma gondii B1 gene was amplified by nested PCR. Genotyping of T. gondii isolates were performed by restriction fragment length polymorphism (RFLP) with Pm1 I and Xho I. 13 samples were found positive with the B1 gene (530 bp) amplification and belonged to type I. The sequencing results showed that 4 T. gondii B1 gene positive samples were identical, with 3 nucleotide variation compared with T. gondii strain RH (type I) B1 gene (GenBank No. AF179871), and in the other sample a "G --> A" mutation at 230bp was detected. The results indicated that the genotype of Toxoplasma gondii in HIV positive patients in Yunnan Province is type I.

Sp1 inhibition-mediated upregulation of VEGF 165 b induced by rh-endostatin enhances antiangiogenic and anticancer effect of rh-endostatin in A549.

Recombinant human endostatin (rh-endostatin), a potential antiangiogenic agent, is used in non-small cell lung carcinoma treatment and represses vascular endothelial cell growth factor (VEGF) levels in tumor cell. However, precise affection of rh-endostatin on the proangiogenic VEGF isoforms (VEGF(165)) or antiangiogenic VEGF isoforms (VEGF(165)b) is not clear. We therefore tested the hypothesis that rh-endostatin could alter expression of these isoforms to regulate tumor growth. A549 cells were exposed to rh-endostatin, and the expression of VEGF(165) and VEGF(165)b was detected. The role of SP1 as a regulator of isoform expression was investigated. We then examined the anticancer and antiangiogenic efficacy of rh-endostatin in combination with exogenous VEGF(165)b against A549 cells, EA.HY 926 cells and xenograft model of human lung cancer. rh-Endostatin reduced VEGF(165) and induced VEGF(165)b as well as inhibited SP1 in A549 cells. SP1 inhibitor (betulinic acid) also developed those changes. VEGF(165)b-rh-endostatin combination was highly synergistic and inhibited growth, survival, and migration of A549 cells, VEGF-mediated VEGFR2 phosphorylation in EA.HY 926 cells, and tumor growth in xenograft model of human lung cancer. rh-Endostatin downregulates proangiogenic vascular endothelial growth factor A (VEGFA) isoform and upregulates antiangiogenic VEGFA isoform, possibly through inhibition of SP1. Furthermore, VEGF(165)b sensitizes A549 to rh-endostatin treatment and enhances the anticancer effect of rh-endostatin.