RNF43 mutation as a predictor of immunotherapeutic efficacy in colorectal cancer.

RNF43 is a tumor suppressor for various cancers and is considered to drive carcinogenesis when mutated. However, the correlation between RNF43 mutation and colorectal cancer (CRC) immunotherapy remains unreported. We evaluated the role of RNF43 using publicly available data from the Cancer Genome Atlas (TCGA) and Memorial Sloan Kettering Cancer Center (MSKCC). In addition, further analysis was performed on an internal validation cohort (hcohort). The mutant profiles of RNF43 were analyzed in 873 Chinese CRC patients. The relationship between clinical pathologic features and RNF43 were analyzed using the two-sided chi-squared test or the Fisher exact test. Clinicopathologic characteristics were associated with overall survival using Cox regression and the Kaplan-Meier method. We found that RNF43 mutation was significantly associated with high TMB and high MSI score (all p-values < 0.05) in the MSKCC cohort. Additionally, RNF43 mutation was found to be enriched in MSI instability. Kaplan-Meier survival analysis revealed that patients with RNF43 mutation had better OS compared to RNF43 wild-type (not reached vs. 13 months, HR, 0.12; 95% CI 0.03 to 0.49; P = 0.0034). However, no association was observed between RNF43 and OS in the TCGA cohort (HR, 1.83; 95% CI 0.66 to 5.07; P = 0.2479). Our CRC hcohort confirmed the significance of RNF43 mutation in predicting better clinical outcomes, including ORR (45% vs. 21%, P = 0.0468). RNF43 mutation correlated with a high tumor mutation burden (P < 0.001). The mutation frequency of RNF43 in CRC patients was 8.4% (73/873); RNF43 G659Vfs*41 was found to be the most frequent mutation site. In patients with RNF43 mutations, TP53, KRAS, and TGFBR2 were genes with a high frequency of mutations. Compared with RNF43 wild-type patients, those with RNF43 mutations had a higher TMB score and a greater proportion of MSI-H, but no difference in PD-L1 expression. Moreover, the content of immune-related B cells, CD8+ T cells, neutrophils, and dendritic cells was higher in the RNF43 mutant group than in the wild-type group. Our results suggest that RNF43 mutation may correlate with better OS in CRC patients receiving PD-1/PD-L1 inhibitors. The exact mechanisms underlying RNF43 require further investigation.

Immune regulation in gastric adenocarcinoma is linked with therapeutic efficacy and improved recovery.

Adenocarcinomas are one of the most common histological types of gastric cancer. It has been ranked fifth among common cancers and is the third among death causing cancers worldwide. The high mortality rate among patients with gastric cancer is because of its silent evolution, genetic heterogeneity, high resistance to chemotherapy as well as unavailability of highly effective therapeutic strategy. Until now a number of several treatment strategies have been developed and are being practiced such as surgery, chemotherapy, radio therapy, and immunotherapy, however, further developments are required to improve the treatment responses and reduce the side effects. Therefore, novel personal therapeutic strategies based on immunological responses should be developed by targeting different check points and key immune players. Targeting macrophages and related molecular elements can be useful to achieve these goals. In this minireview, we discuss the available treatment options, molecular underpinnings and immunological regulations associated with gastric adenocarcinoma. We further describe the possible check points and immunological targets that can be used to develop novel therapeutic options.

Identification of natural compound garcinone E as a novel autophagic flux inhibitor with anticancer effect in nasopharyngeal carcinoma cells.

CONTEXT: Current chemotherapeutic drugs cannot meet the treatment needs of patients with nasopharyngeal carcinoma (NPC), so urgent action is needed to discover novel chemotherapeutic agents. Our previous study revealed that garcinone E (GE) inhibited the proliferation and metastasis of NPC, suggesting that the compound might display promising anticancer activity. OBJECTIVE: To examine the mechanism underlying the anti-NPC activity of GE for the first time. MATERIALS AND METHODS: For MTS assay, NPC cells were treated with 2.5-20 µmol/L GE or dimethyl sulfoxide for 24, 48, and 72 h. Colony formation capacity, cell cycle distribution, and in vivo xenograft experiment of GE were assessed. MDC staining, StubRFP-sensGFP-LC3 observation, LysoBrite Blue staining, and immunofluorescence examined the autophagy of NPC cells after GE exposure. Western blotting, RNA-sequencing, and RT-qPCR measured protein and mRNA levels. RESULTS: GE suppressed cell viability with an IC50 of 7.64, 8.83 and 4.65 µmol/L for HK1, HONE1 and S18 cells. GE inhibited colony formation and cell cycle, increased autophagosome number, and inhibited the autophagic flux partially by blocking lysosome-autophagosome fusion, and repressed S18 xenograft growth. GE dysregulated the expression of autophagy- and cell cycle-related proteins such as Beclin-1, SQSTM1/p62, LC3, CDKs, and Cyclins. Bioinformatics GO and KEGG pathway enrichment analysis of RNA-seq showed that autophagy was enriched in differentially expressed genes upon GE treatment. DISCUSSION AND CONCLUSION: GE acts as an autophagic flux inhibitor, which may have potential chemotherapeutic use for NPC treatment and may have an application in basic research to explore the mechanisms of autophagy.

Preparation of Biochar with Developed Mesoporous Structure from Poplar Leaf Activated by KHCO3 and Its Efficient Adsorption of Oxytetracycline Hydrochloride.

The effective removal of oxytetracycline hydrochloride (OTC) from the water environment is of great importance. Adsorption as a simple, stable, and cost-effective technology is regarded as an important method for removing OTC. Herein, a low-cost biochar with a developed mesoporous structure was synthesized via pyrolysis of poplar leaf with potassium bicarbonate (KHCO3) as the activator. KHCO3 can endow biochar with abundant mesopores, but excessive KHCO3 cannot continuously promote the formation of mesoporous structures. In comparison with all of the prepared biochars, PKC-4 (biochar with a poplar leaf to KHCO3 mass ratio of 5:4) shows the highest adsorption performance for OTC as it has the largest surface area and richest mesoporous structure. The pseudo-second-order kinetic model and the Freundlich equilibrium model are more consistent with the experimental data, which implies that the adsorption process is multi-mechanism and multi-layered. In addition, the maximum adsorption capacities of biochar are slightly affected by pH changes, different metal ions, and different water matrices. Moreover, the biochar can be regenerated by pyrolysis, and its adsorption capacity only decreases by approximately 6% after four cycles. The adsorption of biochar for OTC is mainly controlled by pore filling, though electrostatic interactions, hydrogen bonding, and π-π interaction are also involved. This study realizes biomass waste recycling and highlights the potential of poplar leaf-based biochar for the adsorption of antibiotics.

Endoscopic treatment and management of rectal neuroendocrine tumors less than 10 mm in diameter.

Rectal neuroendocrine tumors (rNETs) measuring less than 10 mm in diameter are defined as small rNETs. Due to the low risk of distant invasion and metastasis, endoscopic treatments, including modified endoscopic mucosal resection, endoscopic submucosal dissection, and other transanal surgical procedures, are effective. This review article proposes a follow-up plan according to the size and histopathology of the tumor after operation.

Systematic characterisation of the effective components of five Callicarpa species with UPLC-Q-TOF-MS and evaluation of their anti-hyperuricaemic activity.

Callicarpa kwangtungensis (C. Kw), C. macrophylla (C. Ma), C. nudiflora (C. Nu), C. formosana (C. Fo), and C. kochiana (C. Ko) were medicinal plant resource in China. In this study, the UPLC/Q-TOF-MS analysis was performed and 151 compounds were identified. PCA analysis metabolic profiles of C. Nu, C. Ko and C. Kw leaves differ significantly from the other two Callicarpa species, while C. Fo and C. Ma share similar chemical constituents. OPLS-DA highlight with an S-plot indicated that there are 14 robust known chemical markers enabling the differentiation between these five Callicarpa plants. C. Ma, C. Nu, and C. Fo leaves extracts treatment effectively reversed the body weight loss, uric acid and creatinine content, hepatic XOD activity, kidney, liver, and ankle tissues injury and inflammation induced by potassium oxonate in hyperuricemia mice. While Ko and C. Kw leaves extracts treatment showed less improvement in hyperuricemia mice.

Small molecules as potential biomarkers of early gastric cancer: A mass spectrometry imaging approach.

BACKGROUND AND AIMS: Early detection of gastric cancer is an effective way to decrease the associated mortality. Efficient methods are needed to provide more sensitive biomarkers for detection of early gastric cancer (EGC). MATERIALS AND METHODS: We performed liquid extraction-electrosonic spray ionization mass spectrometry imaging and immunofluorescent staining of enzymes related to lipid synthesis on cancerous and paracancerous tissues obtained from six EGC patients and investigated the serum lipid profiles. Areas under the receiver operating characteristic curves were used to determine the diagnostic accuracy of putative biomarkers. RESULTS: Five lipids detected in the positive ion mode and seven in negative ion mode displayed higher relative intensities in the cancerous than the paracancerous tissues. Seven lipids detected in the positive ion mode and seven in the negative ion mode displayed lower relative intensities in the cancerous than the paracancerous regions. Phosphatidylcholine (34:3) and phosphatidylcholine (36:1) showed higher relative intensities in the cancerous than in the paracancerous tissues and showed higher relative intensities in the serum of EGC patients than healthy controls. Phosphatidylcholine (32:0) showed lower relative intensities in the cancerous than in the paracancerous tissues and lower relative intensities in the serum of EGC patients than healthy controls. The areas under the receiver operating characteristic curves of serum phosphatidylcholine (32:0) and phosphatidylcholine (34:3) for identifying EGC patients were 1.000 and 0.978, respectively. CONCLUSIONS: Regions associated with EGC showed different lipid distributions and enzyme expression from the paracancerous regions. Serum Phosphatidylcholine (32:0) and phosphatidylcholine (34:3) are potential biomarkers for discriminating between EGC patients and healthy controls.

The Role of REC8 in the Innate Immune Response to Viral Infection.

REC8 meiotic recombination protein (REC8) is a member of structural maintenance of chromosome (SMC) protein partners, which play an important role in meiosis, antitumor activity, and sperm formation. As the adaptor proteins of RIG-I-like receptor (RLR) signaling and cyclic GMP-AMP synthase (cGAS)-DNA signaling, the activity and stability of MAVS (mitochondrial antiviral signaling protein; also known as VISA, Cardif, and IPS-1) and STING (stimulator of interferon genes; also known as MITA) are critical for innate immunity. Here, we report that REC8 interacts with MAVS and STING and inhibits their ubiquitination and subsequent degradation, thereby promoting innate antiviral signaling. REC8 is upregulated through the JAK-STAT signaling pathway during viral infection. Knockdown of REC8 impairs the innate immune responses against vesicular stomatitis virus (VSV), Newcastle disease virus (NDV), and herpes simplex virus (HSV). Mechanistically, during infection with viruses, the SUMOylated REC8 is transferred from the nucleus to the cytoplasm and then interacts with MAVS and STING to inhibit their K48-linked ubiquitination triggered by RNF5. Moreover, REC8 promotes the recruitment of TBK1 to MAVS and STING. Thus, REC8 functions as a positive modulator of innate immunity. Our work highlights a previously undocumented role of meiosis-associated protein REC8 in regulating innate immunity. IMPORTANCE The innate immune response is crucial for the host to resist the invasion of viruses and other pathogens. STING and MAVS play a critical role in the innate immune response to DNA and RNA viral infection, respectively. In this study, REC8 promoted the innate immune response by targeting STING and MAVS. Notably, REC8 interacts with MAVS and STING in the cytoplasm and inhibits K48-linked ubiquitination of MAVS and STING triggered by RNF5, stabilizing MAVS and STING protein to promote innate immunity and gradually inhibiting viral infection. Our study provides a new insight for the study of antiviral innate immunity.

Oleanolic Acid Attenuates Morphine Withdrawal Symptoms in Rodents: Association with Regulation of Dopamine Function.

INTRODUCTION: Oleanolic acid (OA) has been shown to be useful for the treatment of mental disorders. METHODS: In this study, we investigated the effects of OA in animal models of spontaneous withdrawal and naloxone-precipitated withdrawal and evaluated the effects of OA on the acquisition, extinction, and reinstatement of morphine-induced conditioned place preference (CPP). RESULTS: OA significantly improved symptoms of withdrawal, and significantly reduced the acquisition and reinstatement of morphine-induced conditioned place preference. Moreover, OA significantly reduced the serum content of 5-hydroxy tryptamine (5-HT) and dopamine (DA) in a dose-dependent manner, and reduced norepinephrine (NE) and 5-HT content in the frontal cortex (PFC), while significantly increasing endorphin content in rats. OA also significantly reduced serum DA content in mice. CONCLUSION: These results indicate that OA can improve the withdrawal symptoms of rats and mice by regulating the DA system and suggest that OA may be useful in treatment of morphine addiction.

Topical gel based nanoparticles for the controlled release of oleanolic acid: design and in vivo characterization of a cubic liquid crystalline anti-inflammatory drug.

BACKGROUND: Oleanolic acid (OA) has multiple pharmaceutical applications including anti-inflammatory activity, but low permeability of the molecule limits its widespread use. METHODS: A cubic liquid crystalline nanoparticle (LCNP)-based gel was prepared as a potential topical delivery system for OA. The LCNP-based gel was optimized using rheological, drug release kinetic, and ex vivo permeation studies. RESULTS: The studies showed that the OA was trapped in the interior of the LCNP with a crystal form of Pn3m space. The optimized LCNP formulation performed well using in vitro release studies for up to 12 h (85.49 ± 0.21%). Ex vivo permeation studies showed that the LCNP-based gel formulation was superior to a standard gel formulation. The r2 value from the Peppas equation indicated good linearity, but showed irregular (non-Fickian) diffusion, suggesting that drug release was controlled by multiple processes. CONCLUSIONS: In this study, OA-loaded LCNPs were prepared by the precursor method, resulting in a well-characterized OA-LCNP gel preparation. The gel was shown to be effective in a rodent carrageenan-induced hind paw inflammation model with sustained efficacy after a single application.

[Formulation Optimization of Zuojin Floating-Bioadhesive Pellets by Central Composite Design-Response Surface Methodology].

OBJECTIVE: To optimize the formulation of Zuojin floating-bioadhesive pellets by the central composite design-response surface methodology (CCD-RSM). METHODS: In the formulation design using CCD-RSM, independent variables were the amounts of sodium bicarbonate (X1), HPMC (X2) and MCC (X3) as factors. Small pills roundness, 12 h floating rate and the percentages of in vitro cumulative releases at 2,6 and 12 h were dependent variables. Multilinear and quadratic model were used to estimate the relationship between the dependent and the independent variables. According to best model, the contour plots and RSM were drawn, and the optional formulation was selected. According to the optional formulation,the pellets were prepared and validated. RESULTS: The quadratic was the best fitting mode. Small pills roundness was 15.04 °. 12 h floating rate was 75.07%. Percentages of in vitro cumulation release at 2,6 and 12 h of the option formulation pellets was 27.01%, 70.00% and 84.61%, respectively. The actual value was close to the predicted value. Deviation was less than 5%. CONCLUSION: Quadratic model was preferred in the optimization of formulation due to the statistical confidence. The multi-objective simultaneous optimization of Zuojin floating pellet formulation can be achieved by the central composite design-response surface methodology.

[Investigation on Spray Drying Technology of Auricularia auricular Extract].

OBJECTIVE: To investigate the feasibility of spray drying technology of Auricularia auricular extract and its optimum process. METHODS: On the basis of single factor test, with the yield of dry extract and the content of polysaccharide as indexes, orthogonal test method was used to optimize the spray drying technology on the inlet air temperature, injection speed and crude drug content. Using ultraviolet spectrophotometry, thin layer chromatography(TLC) and pharmacodynamics as indicators, extracts prepared by traditional alcohol precipitation drying process and spray drying process were compared. RESULTS: Compared with the traditional preparation method, the extract prepared by spray drying had little differences from the polysaccharide content, TLC and the function of reducing TG and TC, and its optimum technology condition were as follows: The inlet air temperature was 180 °C, injection speed was 10 ml/min and crude drugs content was 0. 4 g/mL. CONCLUSION: Auricularia auricular extract by spray drying technology is stable and feasible with high economic benefit.

[Establishing quantitative model of naringin in citrus Grandis Exocarpium by near infrared spectroscopy].

OBJECTIVE: To establish the quantitative model of naringin in Citrus Grandis Exocarpium by near infrared reflectance spectroscopy(NIRS). METHODS: NIRSs of 54 Citrus Grandis Exocarpium samples were collected and pretreated by TQ Analyst 8.0 software with first derivative + Norris filter. The wavebands in 10,000-4000 cm(-1) were collected and 9 numbers of factors were used. The calibration model was built with partial least squares (PLS). RESULTS: The quantitative calibration model had good correlation coefficients (r = 0.9927) and low root mean square error of calibration (RMSEC = 0.0746). Root mean square error of prediction (RMSEP) was 0.282. The average rate of recovery of validation was 101.65% (n = 9). CONCLUSION: The quantitative calibration model is trustworthy and it can be used to predict naringin in Citrus Grandis Exocarpium accurately. This simple, fast and non-destructive advantages of NIRS can be used for quality control and exploitation of Citrus Grandis Exocarpium.

[Extraction technology optimization of huoxuehuayu granules].

OBJECTIVE: To optimize the extraction process of Huoxuehuayu granules. METHODS: To determine the activities of serum creatine kinase (CK) and lactate dehydrogenase (LDH) of experimental myocardial ischemia mice, in order to optimize the best extraction technology of Huoxuehuayu prescription. According to the pharmacological results,with composite score of extraction rate of tanshinone II(A) and dry extract rate as comprehensive evaluation indexes, orthogonal test was used to optimize the ethanol extraction technology by taking ethanol concentration, the amount of ethanol, extraction time and soaking time. With extraction rate of salvianolic acid B and dry extract yield as comprehensive evaluation indexes, effect of extraction times, the amount of water, and extraction time on water extraction technology were investigated by orthogonal test. RESULTS: Optimum ethanol extraction technology conditions were as follows: extracted once with ten times the amount of 80% ethanol, 1.5 h. Optimum water extraction technology conditions were as follows: extracted three times with eight times the amount of water, 1 h each time. The best ethanol precipitated technique was the relative density of the liquor to be condensed to 1.20 (60 degrees C), adding 95% ethanol to 80% of ethanol concentration and depositing for 24 h. CONCLUSIONS: Optimized extraction technology is simple, stable and feasible,which can settle foundation for molding technology of Huoxuehuayu granules.