Tanshinone IIA induces ER stress and JNK activation to inhibit tumor growth and enhance anti-PD-1 immunotherapy in non-small cell lung cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) remains at the forefront of new cancer cases, and there is an urgent need to find new treatments or improve the efficacy of existing therapies. In addition to the application in the field of cerebrovascular diseases, recent studies have revealed that tanshinone IIA (Tan IIA) has anticancer activity in a variety of cancers. PURPOSE: To investigate the potential anticancer mechanism of Tan IIA and its impact on immunotherapy in NSCLC. METHODS: Cytotoxicity and colony formation assays were used to detect the Tan IIA inhibitory effect on NSCLC cells. This research clarified the mechanisms of Tan IIA in anti-tumor and programmed death-ligand 1 (PD-L1) regulation by using flow cytometry, transient transfection, western blotting and immunohistochemistry (IHC) methods. Besides, IHC was also used to analyze the nuclear factor of activated T cells 1 (NFAT2) expression in NSCLC clinical samples. Two animal models including xenograft mouse model and Lewis lung cancer model were used for evaluating tumor suppressive efficacy of Tan IIA. We also tested the efficacy of Tan IIA combined with programmed cell death protein 1 (PD-1) inhibitors in Lewis lung cancer model. RESULTS: Tan IIA exhibited good NSCLC inhibitory effect which was accompanied by endoplasmic reticulum (ER) stress response and increasing Ca2+ levels. Moreover, Tan IIA could suppress the NFAT2/ Myc proto oncogene protein (c-Myc) signaling, and it also was able to control the Jun Proto-Oncogene(c-Jun)/PD-L1 axis in NSCLC cells through the c-Jun N-terminal kinase (JNK) pathway. High NFAT2 levels were potential factors for poor prognosis in NSCLC patients. Finally, animal experiments data showed a stronger immune activation phenotype, when we performed treatment of Tan IIA combined with PD-1 monoclonal antibody. CONCLUSION: The findings of our research suggested a novel mechanism for Tan IIA to inhibit NSCLC, which could exert anti-cancer effects through the JNK/NFAT2/c-Myc pathway. Furthermore, Tan IIA could regulate tumor PD-L1 levels and has the potential to improve the efficacy of PD-1 inhibitors.

Inhibition of Ca2+ induced Macrophage Oxidative Stress Cascade in Mice With Ulcerative Colitis.

Context: In the context of ulcerative coloproctitis (UC), a chronic and non-specific inflammatory condition of the colon and rectum with an elusive etiology, the therapeutic potential of G protein-coupled receptor 109a (GPR109a) has gained prominence. GPR109a is expressed in various cell types including colonic epithelium, adipocytes, neutrophils, and macrophages, positioning it as a promising candidate for pharmacological intervention in colitis. Objective: the mechanistic role of GPR109a in a mouse model of UC and to assess its capacity to mitigate the disease's progression. Design: The research team performed an animal study. Setting: Conducted by a research team in the biomedical setting of the People's Hospital of Dongxihu District in Wuhan, Hubei province of China. Animals: This animal study engaged 16 specific pathogen-free (SPF) male BALB/c mice, aged between 4 to 6 weeks and weighing 20-24 grams each. Outcome Measures: (1) analysis of rectal lesions via hematoxylin-eosin (HE) staining, (2) the use of transmission electron microscopy to examine suborganelle spatial relationships, (3) Western blot analysis to evaluate a spectrum of protein expressions such as phosphorylated IP3R, p-PERK, p-IRE-1α, mitofusins, NADPH oxidases, TNF-α, and IL-1ß, and (4) determination of intracellular calcium concentrations to gauge intestinal barrier function impairment involving specific calcium pathway signals. Results: Results from the study painted a picture of significant infiltration by inflammatory cells within the mucosal and submucosal layers, compounded by pronounced endoplasmic reticulum expansion. Abundant interfaces between the ER and the mitochondria formed multiple structures known as mitochondria-associated membranes or MAMs. When evaluated under calcium-rich conditions, the protein levels mentioned earlier were notably elevated in the GPR109a knockdown (KD) group versus the infection control (NC) counterparts. However, this differential expression disappeared under calcium-depleted conditions or when treated with 2-aminoethyl diphenyl borinate (2-APB). Additionally, fluorescence intensity assays showed marked suppression in the GPR109a-KD plus lipopolysaccharide (LPS) group compared to the control. Conclusions: Drawing these observations to a close, the study concluded that GPR109a has an inhibitory effect on the advancement of ulcerative coloproctitis in mice. This is mediated through the activation of a calcium-induced oxidative stress cascade within macrophages, delineating a promising therapeutic pathway for the management of this inflammatory disease.

Three new indole alkaloid derivatives from Fissistigma oldhamii Levl. and their anti-inflammatory effects.

Three new indole alkaloid derivatives, fissindoalkas A-C (1-3) together with one known biogenetically related polysubstituted indole alkaloid (4) were isolated from the roots of Fissistigma oldhamii (Hemsl.) Merr. The structures of compounds 1-4 were elucidated using comprehensive spectroscopic methods. The inhibitory activities of compounds 1-4 against nitric oxide (NO) production induced by lipopolysaccharide (LPS) were evaluated in vitro using mouse macrophage RAW264.7 cells. Compounds 2 and 3 showed potent inhibitory activities on NO production with IC50 values of 2.52 ± 0.18 and 2.33 ± 0.16 µM. These results indicate that the discovery of indole alkaloid derivatives, from the roots of F. oldhamii, which show significant anti-inflammatory properties, could be of great importance to the research and for the development of novel natural anti-inflammatory agents.

Hyperbaric oxygen treatment on keloid tumor immune gene expression / 中华医学杂志(英文版)

BACKGROUND@#Hyperbaric oxygen treatment (HBOT) has been demonstrated to influence the keloid recurrence rate after surgery and to relieve keloid symptoms and other pathological processes in keloids. To explore the mechanism of the effect of HBOT on keloids, tumor immune gene expression and immune cell infiltration were studied in this work.@*METHODS@#From February 2021 to April 2021, HBOT was carried out on keloid patients four times before surgery. Keloid tissue samples were collected and divided into an HBOT group (keloid with HBOT before surgery [HK] group, n = 6) and a non-HBOT group (K group, n = 6). Tumor gene expression was analyzed with an Oncomine Immune Response Research Assay kit. Data were mined with R package. The differentially expressed genes between the groups were compared. Hub genes between the groups were determined and verified with Quantitative Real-time PCR. Immune cell infiltration was analyzed based on CIBERSORT deconvolution algorithm analysis of gene expression and verified with immunohistochemistry (IHC).@*RESULTS@#Inflammatory cell infiltration was reduced in the HK group. There were 178 upregulated genes and 217 downregulated genes. Ten hub genes were identified, including Integrin Subunit Alpha M (ITGAM), interleukin (IL)-4, IL-6, IL-2, Protein Tyrosine Phosphatase Receptor Type C (PTPRC), CD86, transforming growth factor (TGF), CD80, CTLA4, and IL-10. CD80, ITGAM, IL-4, and PTPRC with significantly downregulated expression were identified. IL-10 and IL-2 were upregulated in the HK group but without a significant difference. Infiltration differences of CD8 lymphocyte T cells, CD4 lymphocyte T-activated memory cells, and dendritic resting cells were identified with gene CIBERSORT deconvolution algorithm analysis. Infiltration levels of CD4 lymphocyte T cell in the HK group were significantly higher than those of the K group in IHC verification.@*CONCLUSION@#HBOT affected tumor gene expression and immune cell infiltration in keloids. CD4 lymphocyte T cell, especially activated memory CD4+T, might be the key regulatory immune cell, and its related gene expression needs further study.

An Overview of Methodology and Research Progress on Application of Serum Pharmacochemistry of Traditional Chinese Medicine / 中国实验方剂学杂志

Serum pharmacochemistry of traditional Chinese medicine(TCM) is a new subject based on the identification of migrating components in serum after oral administration of TCM.After more than 20 years of development,serum pharmacochemistry of TCM has been widely recognized and applied by researchers.Based on the references related with the serum pharmacochemistry from these years,the research summarized the methods of serum pharmacochemistry of TCM,for example,selection of experimental animals,preparation of gastric irrigation,design of drug administration scheme,method and time of adoption blood,pretreatment of serum containing drug,as well as analysis of serum sample.In addition,the paper will emphatically introduce the application of serum pharmacochemistryof TCM,which includes clarifying the substance basis of Chinese medicine,establishing the quality standard of TCM,and clarifying the compatibility of compound Chinese medicine.At the same time,we will through some existing problems to look forward to the future of serum pharmacochemistryof TCM,so that people can have a comprehensive understanding of serum pharmacochemistryof TCM,hoping to lay a foundation for the further application of this method in TCM research.

Feasibility study of QAMS for quantitative analysis of multiple structural types of ingredients in Atractylodis Rhizome by GC / 中国中药杂志

In this study， quantitative analysis of multi-components with single marker(QAMS) was established and validated to simultaneously determine four sesquiterpenoids(β-eudesmol， atractylon， atractylolideⅠ， atractylolide Ⅱ) in Atractylodis Rhizome based on the gas chromatographic method(GC). Using β-eudesmol as the contrast， the relative correctionfactors(RCF) of the other three sesquiterpenoids were determined by GC. Within the line arranges，the values of RCF of β-eudesmol to atractylon， atractylolideⅠand atractylolide Ⅱ were 0.823， 0.690 and 0.766， respectively. The RCF had a good reproducibility in various instruments， chromatographic columns. According to their RCF， we simultaneously determined four sesquiterpenoids in Atractylodis Rhizome only using one marker. The results of QAMS method were validated by comparing with that of internal standard method， and no obvious significant difference was found.

Curcumin temperature-sensitive in situ hydrogels for treatment of vaginal candidiasis / 国际药学研究杂志

Objective To investigate the therapeutic effect of curcumin in situ hydrogels for treatment of vaginal candidiasis. Methods In situ hydrogels were prepared using poloxamer 407 and 188 as the gel matrix.Gelation temperature,gelation time and rheological characteristics were determined.Mice models with vaginal candidiasis were established.The animals were divided into six groups:the healthy mice group,the model group,the positive control group(ketoconazole cream),curcumin-loaded in situ hydrogels of 0.025,0.10 and 0.20 g/ml.Before treatment and after the fourth and eighth treatments,the vaginal smears were collected and Gram-stained.The vagina washings fluid of mice were collected to determine DNA of Lactobacillus.On 8thday,the mice were sacrificed and the vaginal tissues were excised for pathological detection.Results The gelation temperature and time of in situ hydrogels were 29℃and 2′30″,respectively.Rheological profiles demonstrated that it showed good retention effect in vivo.Compared with the model group, curcumin hydrogels of 0.025,0.10 and 0.20 g/ml could inhibit the growth of Candida albicans,inhibit vaginal mucosal edema,inflam?matory hyperplasia,and decrease the expression of inflammatory factors including TNF-α(P<0.01)and IL-1β(P<0.01).Howev?er,curcumin hydrogels could not influence the growth of lactobacillus in the vagina.It indicated that curcumin hydrogels did not de?pend on lactobacillus to treat fungal vaginitis.Conclusion Curcumin in situ hydrogels could inhibit candidiasis growth,decrease ede?ma of vaginal mucosa and inflammatory hyperplasia,and reduce the expression of inflammatory factors.It provides a novel choice for the treatment of clinical vaginal candidiasis.

Effect of percutaneous intratumoral injection of lipiodol emulsion of chemotherapie agents on implanted VX2 tumor in rabbits / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the therapeutic effect of percutaneous intratumoral injection with lipiodol emulsion of chemotherapie agents (CALE) on implanted VX2 tumor in rabbits.</p><p><b>METHODS</b>Twelve New Zealand rabbits with implanted VX2 tumor (24 models) were divided into lipiodol group, chemotherapeutic agent group and CALE group with intratumoral injections of the corresponding agents. The pathological changes of all the lesions were observed and the expression of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were evaluated 7 days after the operation.</p><p><b>RESULTS</b>Compared with the lipiodol group and chemotherapie agent group, intratumoral injection of CALE resulted in the highest tumor necrosis rate and greatest tumor necrosis (P<0.01). The labeling indices of PCNA and VEGF expressions in CALE group were markedly lower than those in the other two groups (P<0.01).</p><p><b>CONCLUSION</b>Percutaneous intratumoral injection of CALE is an effective ablation approach for treatment of malignant solid tumors.</p>

Percutaneous intratumoral injection with pingyangmycin lipiodol emulsion for treatment of recurrent sacrococcygeal chordomas / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the effectiveness and safety of fluoroscopy-guided percutaneous intratumor injection of pingyangmycin lipiodol emulsion (PLE) in the management of recurrent sacrococcygeal chordomas.</p><p><b>METHODS</b>Seven patients with recurrent sacrococcygeal chordomas presenting with severe local pain with visual analogue score (VAS)≥8 received treatment sessions of fluoroscopy-guided percutaneous intratumor injection of PLE. The patients were followed up every 3 months after the last session to assess their clinical responses and observe the changes in the tumor size measured by computed tomography. The changes in the VAS, tumor necrosis and pain relief as well as the adverse events were recorded.</p><p><b>RESULTS</b>A total of 22 sessions of fluoroscopy-guided percutaneous intratumoral PLE injection was performed in these cases (3 or 4 sessions in each case). The total average pingyangmycin dose delivered was 48.0 mg and the average lipiodol dose was 40.0 ml in each case. Five patients showed low fever and vomiting 48 after the injection. During the follow-up (median time of 21.7 months, range 10-26 months), all the patients showed obviously reduced tumor size and VAS, and partial remission was achieved in 6 patients and stable disease (SD) in 1 patient. None of the patients had complications during the follow-up.</p><p><b>CONCLUSION</b>Fluoroscopy-guided percutaneous intratumoral injection of PLE can be effective and safe and may serve as a alternative for treatment of recurrent sacrococcygeal chordomas.</p>

Myocardial damage and change of mitochondrial Mn-superoxide dismutase activity in craniocerebral injured rats and the intervening effect of ginkgo biloba extract / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the myocardial damage and changes of myocardial mitochondrial Mn-superoxide dismutase (Mn-SOD) activity in craniocerebral injured rats and the effect of Ginkgo biloba extract (GBE) on them.</p><p><b>METHODS</b>Craniocerebral injured rats model was established by fluid-percussion and treated with GBE. The dynamical changes of electrocardiograph (ECG) in 24 h were monitored, the serum level of MB isoenzyme of creatine kinase (CK-MB) and the change of myocardial mitochondrial Mn-SOD activity as well as the pathologic changes of myocardium (HE staining) were observed.</p><p><b>RESULTS</b>The occurrence of ECG abnormality obviously increased in the injured rats, accompanied with increased serum CK-MB (P<0.05) and decreased myocardial Mn-SOD levels (P<0.05), and the Mn-SOD activity was negatively correlated with the level of CK-MB (r=-0.997, P<0.05). Pretreatment of GBE resulted in the decrease of ECG abnormality occurrence (P<0.01), serum CK-MB level (P<0.05), and degree of myocardial damage, as well as the increase of Mn-SOD activity in post-craniocerebral injured rats.</p><p><b>CONCLUSIONS</b>Craniocerebral injury can result in distinct myocardial damage, which is possibly correlated with the lowering of anti-oxidation stress level of myocardial cellular mitochondria. GBE possesses the protective effect on myocardial damage after craniocerebral injury.</p>

Quantification of isopimpinellin in root of Toddalia asiatica by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the content of isopimpinellin in root of Toddalia asiatica.</p><p><b>METHODS</b>A HPLC method was set up. Using Hypersil C18 column and methanol-water (70:30) as mobile phase, with the detection wavelength at 306 nm.</p><p><b>RESULT</b>The linear range of isopimpinellin was 0.004 20 approximately 0.420 microg. The average recovery was 99.7% and the RSD 2.8%.</p><p><b>CONCLUSION</b>The method is simple and accurate, with good reproducibility, and can be used as a quantitative analysis method for isopimpinellin.</p>