PARP inhibition leads to synthetic lethality with key splicing-factor mutations in myelodysplastic syndromes.

BACKGROUND: Splicing factors are frequently mutated in patients with myelodysplastic syndromes and acute myeloid leukaemia. Recent studies have revealed convergent molecular defects caused by splicing factor mutations, among which R-loop dysregulation and resultant genome instability are suggested as contributing factors to disease progression. On the other hand, understanding how mutant cells survive upon aberrant R-loop formation and genome instability is essential for developing novel therapeutics. METHODS: The immunoprecipitation was performed to identify R-loops in association with PARP1/poly-ADP-ribosylation. The western blot, immunofluorescence, and flow cytometry assays were used to test the cell viability, cell cycle arrest, apoptosis, and ATM activation in mutant cells following the treatment of the PARP inhibitor. The Srsf2(P95H) knock-in murine hematopoietic cells and MLL-AF9 transformed leukaemia model were generated to investigate the potential of the PARP inhibitor as a therapy for haematological malignancies. RESULTS: The disease-causing mutations in SRSF2 activate PARP and elevate the overall poly-ADP-ribosylation levels of proteins in response to R-loop dysregulation. In accordance, mutant cells are more vulnerable to the PARP inhibitors in comparison to the wild-type counterpart. Notably, the synthetic lethality was further validated in the Srsf2(P95H) knock-in murine hematopoietic cell and MLL-AF9 leukaemia model. CONCLUSIONS: Our findings suggest that mutant cells antagonise the genome threat caused by R-loop disruption by PARP activation, thus making PARP targeting a promising therapeutic strategy for myeloid cancers with mutations in SRSF2.

Anesthetic managements, morbidities and mortalities in retroperitoneal sarcoma patients experiencing perioperative massive blood transfusion.

Objective: Given high risks of major bleeding during retroperitoneal sarcoma(RPS) surgeries, severe complications and deaths are common to see perioperatively. Thus, effective anesthetic management is the key point to ensuring the safety of patients. This study aimed to introduce anesthesia management and mortalities in RPS patients receiving massive blood transfusions during surgeries. Methods: Records of RPS surgeries under general anesthesia from January 2016 through December 2021 were retrospectively retrieved from our database. Patients who received massive blood transfusions (MBT) exceeding 20 units in 24h duration of operations were finally included in this study. Demographics, modalities of anesthesia management, blood loss, transfusion, peri-anesthesia biochemical tests as well as morbidities and mortalities were collected. Risk factors of postoperative 60d mortality were determined through logistic regression in uni-and multi-variety analysis using the statistics software STATA 17.0. Results: A total of 70 patients (male 31) were included. The mean age was 50.1 ± 15.8 years. All patients received combined resections of sarcoma with involved organs under general anesthesia. Mean operation time and anesthesia time were 491.7 ± 131.1mins and 553.9 ± 132.6mins, respectively. The median intraoperative blood loss was 7000ml (IQR 5500,10000ml). Median red blood cells (RBC) and fresh frozen plasma (FFP) transfusion were 25.3u (IQR 20,28u), and 2400ml (IQR 2000,3000ml), respectively. Other blood products infusions included prothrombin complex concentrate (PCCs), fibrinogen concentrate (FC), platelet(plt) and albumin(alb) in 82.9% (58/70), 88.6% (62/70), 81.4% (57/70) and 12.9% (9/70) of patients. The postoperative severe complication rate(Clavien-Dindo grade≥3a) was 35.7%(25/70). A total of 7 patients (10%) died during the postoperative 60-day period. BMI, volumes of crystalloid infusion in anesthesia, and hemoglobin and lactate levels at the termination of operation were found significantly associated with postoperative occurrence of death in univariate analysis. In logistic multivariate analysis, extended anesthesia duration was found associated with postoperative venous thrombosis embolism (VTE) and severe complication. The lactate level at the immediate termination of the operation was the only risk factor related to perioperative death (p<0.05). Conclusion: RPS patients who endure MBT in surgeries face higher risks of death postoperatively, which needs precise and effective anesthesia management in high-volume RPS centers. Increased blood lactate levels might be predictors of postoperative deaths which should be noted.

Four case reports of left anterior descending restenosis treated via the internal mammary artery: A literature review.

Objective: To analyse four cases of intervention via the internal mammary artery-anterior descending branch and provide and summarise the clinical treatment experience. Methods: The clinical data of four patients with distal restenosis of a left anterior descending artery (LAD) anastomosis after left internal mammary artery (LIMA)-LAD bypass surgery, who were admitted to the Gansu Institute of Cardiovascular Diseases between March 2013 and April 2022, were retrospectively analysed and reviewed together with the relevant literature. Results: Among the four patients, one was treated with intracoronary stenting via the internal mammary artery route, two were treated with intracoronary drug-coated balloon dilation (one of whom underwent fractional flow reserve [FFR] testing), and two underwent FFR testing (one of whom had a negative test result until the end of the procedure and continued to take medication during follow-up; the other patient had a positive result and further interventions). There were no deaths or postoperative complications in the group, and the patients were followed up for 4 months to 9 years, with good long-term outcomes. Conclusion: Percutaneous coronary intervention (PCI) via the internal mammary artery route is safe and effective, and patients with anastomotic distal stenosis or anastomotic stenosis of LAD bypass anastomosis may be considered for PCI via the internal mammary artery route.

Fibrolipomatous Hamartoma of the Median Nerve: A Clinical Report of One Case and Review of the Literature.

BACKGROUND: The ultrasound imaging findings and clinical treatment procedure of a case of fibrolipomatous hamartoma of the median nerve are reported in this paper. The disease diagnosis was mainly dependent on magnetic resonance, and rarely described by ultrasound. This paper introduces the imaging manifestations of median neurofibromatoid hamartoma under ultrasound. CASE PRESENTATION: This case is a middle-aged female with pituitary adenoma. Fibrolipomatous hamartoma, as a rare benign fibrofatty tumor, is characterized by the slow proliferation of mature adipocytes and fibrous tissue around and inside the peripheral nerves, eventually leading to the fusiform enlargement of nerve fiber bundles. DISSCUSSION: This paper discusses the value and application of color doppler ultrasound in the diagnosis of FLH to provide a reference for the clinical diagnosis and treatment of this disease in the future and reviews the literature on this disease. CONCLUSION: Because fibrolipomatous hamartoma is very rare, it is very important to improve the understanding and diagnosis of this disease.

Acupuncture Enhances Gastrointestinal Motility and Improves Autonomic Nervous Function in Patients with Septic Gastrointestinal Dysfunction.

Gastrointestinal dysfunction (GD) is a common complication after endotoxemia, which can further aggravate the progress of infection. Acupuncture uses metal needles of different shapes and techniques to stimulate specific points on the human body, which are effective in treating various diseases, including gastrointestinal diseases. We aimed at exploring the clinical effect of acupuncture on the recovery of visceral sensation, proximal gastric compliance, and autonomic nervous function in patients with septic GD. A total of 73 sepsis with GD patients were selected using modified single section ultrasonography combined with clinical symptoms in the First People's Hospital of Lanzhou City during 2019. The participants were randomly allocated to routine-treatment (control group) and study group receiving acupuncture. The indexes before and after treatment included gastric residue, gastric dilatation, pressure and volume, gastric compliance, autonomic nerve function, APACHE II score, and infection index were measured and compared. Before treatment, there was no significant difference in the basic information of the two groups, including gastric volume and pressure, gastric residue, gastric compliance, autonomic nerve function, and APACHE II score. After treatment, the maximum gastric volume and pressure, gastric residue, and APACHE II score of the two groups were significantly improved (P < 0.05). In addition, the maximum gastric volume and pressure of the study group were significantly higher, while gastric residual, autonomic nerve function, and APACHE II were significantly lower than those of the control group (P < 0.05). However, our results showed that acupuncture did not further reduce inflammatory markers, including white blood cells, C-reactive protein, and procalcitonin. To sum up, on the basis of basic treatment, the application of acupuncture can further improve the clinical symptoms of GD in patients with sepsis, enhance gastrointestinal motility, and improve autonomic nervous function, which is worthy of clinical application and promotion.

ZhenQi FuZheng formula inhibits the growth of colorectal tumors by modulating intestinal microflora-mediated immune function.

Zhenqi Fuzheng formula (ZQFZ), of which the main ingredients are Astragalus membranaceus and Ligustrum lucidum, has immune system regulatory functions and potential anti-tumor bioactivity. The inhibition of colorectal tumor growth by ZQFZ was analyzed in inflammatory cells and B6/JGpt-Apcem1Cin(MinC)/Gpt (ApcMin/+) mice. ZQFZ exhibited anti-inflammatory activity by decreasing the phosphorylation of nuclear factor-kappa B (NF-κB) pathway-related proteins in lipopolysaccharide-induced RAW264.7 cells. After 56 days of treatment, ZQFZ alleviated the progression of colorectal cancer (CRC) and increased the body weight and thymic index values of the ApcMin/+ mice. An analysis of the intestinal microflora showed that ZQFZ affected the abundance of certain immune-related bacteria, which may explain its immunomodulatory effects. Moreover, the percentages of T cells and NK cells in peripheral blood were significantly increased and 15 immune-related cytokines were regulated in serum or the colon or both. ZQFZ upregulated the levels of CD4 and CD8 in the spleen and colorectal tumors and decreased the expression levels of cytotoxic T-lymphocyte-associated protein 4 and programmed death-ligand 1 in colorectal tumors. ZQFZ promoted an anti-tumor immune response and inhibited the occurrence and development of CRC by regulating the immune system. This study provides the experimental basis for the application of ZQFZ as a therapeutic agent for CRC.

An alternative splicing isoform of wheat TaYRG1 resistance protein activates immunity by interacting with dynamin-related proteins.

Wheat (Triticum aestivum) is a commercially important crop and its production is seriously threatened by the fungal pathogen Puccinia striiformis f. sp. tritici West (Pst). Resistance (R) genes are critical factors that facilitate plant immune responses. Here, we report a wheat R gene NB-ARC-LRR ortholog, TaYRG1, that is associated with distinct alternative splicing events in wheat infected by Pst. The native splice variant, TaYRG1.6, encodes internal-motif-deleted polypeptides with the same N- and C-termini as TaYRG1.1, resulting in gain of function. Transient expression of protein variants in Nicotiana benthamiana showed that the NB and ARC domains, and TaYRG1.6 (half LRR domain), stimulate robust elicitor-independent cell death based on a signal peptide, although the activity was negatively modulated by the CC and complete LRR domains. Furthermore, molecular genetic analyses indicated that TaYRG1.6 enhanced resistance to Pst in wheat. Moreover, we provide multiple lines of evidence that TaYRG1.6 interacts with a dynamin-related protein, TaDrp1. Proteome profiling suggested that the TaYRG1.6-TaDrp1-DNM complex in the membrane trafficking systems may trigger cell death by mobilizing lipid and kinase signaling in the endocytosis pathway. Our findings reveal a unique mechanism by which TaYRG1 activates cell death and enhances disease resistance by reconfiguring protein structure through alternative splicing.

Induction of T helper 17 cell response by interleukin-7 in patients with primary cutaneous melanoma.

Interleukin (IL)-7 plays a vital role in proliferation and activation of T cells, however, its signaling through CD127 is impaired in T cells in cancers and chronic infections. The mechanisms underlying T helper 17 (Th17) cell responses by IL-7 in melanoma remain not fully understood. The aim of this study was to assess the effect of IL-7 signaling on Th17 responses in patients with primary cutaneous melanoma. Healthy and primary cutaneous melanoma donors were selected for this study of Th17 cell function. IL-17+CD4+ Th17 cells and CD127 expression on Th17 cells were determined by flow cytometry. Cytokine level was measured by ELISA. Peripheral and tissue-infiltrating CD4+ T cells were isolated using magnetic beads, and then stimulated with IL-7 and/or signal transducer and activator of transcription 5 inhibitor. Activated signaling molecules were analyzed by flow cytometry. Peripheral and tumor-infiltrating Th17 cells percentage was decreased, while peripheral IL-7 level was also reduced in melanoma patients. There was no significant difference of CD127 expression on Th17 cells between melanoma patients and controls. Antiapoptotic protein Bcl-2 was downregulated, whereas proapoptotic protein-activated caspase-3 was upregulated in peripheral and tissue-infiltrating Th17 cells in melanoma patients. Higher concentration of IL-7 (10 ng/mL), but not lower IL-7 concentration (1 ng/mL), promoted Bcl-2 expression and decreased caspase-3 expression in Th17 cells in melanoma patients. Inhibition of signal transducer and activator of transcription 5 resulted in the downregulation of Bcl-2 while upregulation of caspase-3 in Th17 cells. The present data suggested that reduced IL-7 responsiveness might be insufficient for Th17 activation in patients with primary cutaneous melanoma.

Ultrasound-targeted microbubble destruction enhances the anti-tumor action of miR-4284 inhibitor in non-small cell lung cancer cells.

MicroRNAs (miRNAs/miRs) are known to be involved in various human cancer types. Ultrasound-targeted microbubble destruction (UTMD) may improve the transfection efficiency of exogenous genes into target tissues and organs, thereby improving cancer treatment. In the present study, the role of miR-4284 in non-small cell lung cancer (NSCLC) was investigated and the effect of UTMD-mediated inhibition of miR-4284 on tumor progression was further analyzed. The expression of miR-4284 in NSCLC cells and tissues was detected by reverse transcription-quantitative PCR. UTMD-mediated inhibition of miR-4284 was achieved by co-transfection of microvesicles and miR-4284 inhibitors into NSCLC cells. A Cell Counting Kit-8 assay was used to determine NSCLC cell proliferation, and the migration and invasion of NSCLC cells were examined by Transwell assays. Compared with that in the control group, the expression of miR-4284 was increased in NSCLC tissues and cells. Knockdown of miR-4284 in NSCLC cells inhibited cell proliferation, migration and invasion. UTMD improved the transfection efficiency of miR-4284 inhibitors in NSCLC cells, resulting in more significant inhibition of tumor cell proliferation, migration and invasion. In conclusion, the results indicated that the expression of miR-4284 was increased in clinical samples and cell lines of NSCLC and that knockdown of miR-4284 inhibited the proliferation, migration and invasion of tumor cells. UTMD-mediated miR-4284 inhibition further promoted this effect, indicating that this technique may represent a novel strategy for the treatment of NSCLC.

Ultrasonographic characteristics of thyroid nodule rupture after microwave ablation: Three case reports.

RATIONALE: Thyroid nodule rupture is a rare complication after microwave ablation (MWA). The ultrasonographic characteristics, clinical course, treatment, or prognosis of thyroid nodule rupture after ablation have not been systematically summarized. Three cases with thyroid nodule rupture after MWA were reported in this study, including the characteristic ultrasound images before ablation and after rupture. Meanwhile, we investigated the etiology, diagnosis, treatment and prevention of the rupture. These findings can provide references for the future clinical practice. PATIENTS CONCERNS: All 3 patients were pathologically diagnosed as benign thyroid nodules by core needle biopsy and then received 1 session of MWA. DIAGNOSES: Fourteen days to 1 month after MWA later, all 3 patients presented with abrupt neck pain and swelling, and 1 of them had a fever. Ultrasound examinations shared common features that the rupture of thyroid capsule and a soft-tissue mass with unclear margin in front of the thyroid gland, which connected with the post-ablation nodule. Three patients were diagnosed as thyroid nodule ruptures. INTERVENTIONS: All 3 patients received conservative management after the ruptures. With the treatment of intravenous antibiotics for 1 week, the neck swelling of patients 1 and 2 both disappeared. The aggravation of neck swelling was found in patient 3. Ultrasonography of the neck revealed irregular fluid echo in the soft-tissue mass, suggesting abscess formation. Aspiration and irrigation were performed. The neck swelling regressed gradually over another 2 weeks with the treatment of antibiotics. Two months after ablation, ultrasound examination showed that the mass had completely disappeared. OUTCOMES: None of the 3 patients underwent open surgery due to thyroid nodule rupture. At 1-year follow-up, the volume reduction rate of thyroid nodules in 3 patients were as follows: 100%, 98.1% and 90.7%. LESSONS: Nodule rupture is a rare but severe complication after MWA of the thyroid nodules. The diagnosis can be confirmed by clinical symptoms and ultrasound examination, and most nodule ruptures could be cured with conservative treatment. Grasping the characteristics of ultrasound imaging during the course of disease, and dynamically assessing course of disease progression by ultrasonography could avoid unnecessary imaging examinations or invasive procedures.

LINC01232 Sponges Multiple miRNAs and Its Clinical Significance in Pancreatic Adenocarcinoma Diagnosis and Prognosis.

BACKGROUND: Long noncoding RNAs have been demonstrated to play important roles in different kinds of human malignancy. The purpose of this study was to evaluate the diagnostic and prognostic value of long intergenic non-protein coding RNA 1232 (LINC01232) in patients with pancreatic adenocarcinoma (PAAD) and further explore the clinical significance of the potential miRNAs that might be sponged by LINC01232. METHODS: The potential target miRNAs that might be sponged by LINC01232 were analyzed using bioinformatics analysis. The Real-Time quantitative PCR was adopted to measure the relative expression of LINC01232 and target miRNAs in PAAD serum and tissue samples. The diagnostic and prognostic value of LINC01232 was evaluated using the receiver operating characteristic analysis and Kaplan-Meier survival analysis, respectively. RESULTS: LINC01232 expression was upregulated in PAAD serum and tissues and associated with patients' TNM stage. Serum LINC01232 expression had diagnostic value, and the high levels of LINC01232 could predict unfavorable prognosis in PAAD patients. miR-204-5p, miR-370-5p and miR-654-3p were proposed as 3 targets of LINC01232 in PAAD, and their decreased expression levels in PAAD patients showed certain clinical significance in diagnosis and prognosis. CONCLUSION: The data of this study revealed that LINC01232 expression is upregulated in PAAD serum and tissue samples with considerable diagnostic and prognostic significance. In addition, miR-204-5p, miR-370-5p and miR-654-3p may be sponged by LINC01232 in PAAD, which also show potencies in PAAD diagnosis and prognosis.

Dickkopf­1/cysteine­rich angiogenic inducer 61 axis mediates palmitic acid­induced inflammation and apoptosis of vascular endothelial cells.

Cardiovascular diseases (CVDs) are a major cause of mortality around the world, and the presence of atherosclerosis is the most common characteristic in patients with CVDs. Cysteine­rich angiogenic inducer 61 (CCN1) has been reported to serve an important role in the pathogenesis of atherosclerotic lesions. The aim of the present study was to investigate whether CCN1 could regulate the inflammation and apoptosis of endothelial cells induced by palmitic acid (PA). Dickkopf­1 (DKK1) is an important antagonist of the Wnt signaling pathway, which can specifically inhibit the classic Wnt signaling pathway. Firstly, the mRNA and protein expression levels of CCN1 were detected. Additionally, endothelial nitric oxide (NO) synthase (eNOS), DKK1, ß­catenin, and inflammation­ and apoptosis­associated proteins were measured. Detection of NO was performed using a commercial kit. The expression levels of inflammatory cytokines were assessed to explore the effect of CCN1 on PA­induced inflammation. TUNEL assay was used to detect the apoptosis of endothelial cells. The results revealed that PA upregulated the expression levels of CCN1, inflammatory cytokines and pro­apoptotic proteins in endothelial cells. PA decreased the production of NO, and the levels of phosphorylated­eNOS, whereas knockdown of CCN1 partially abrogated these effects triggered by PA. Furthermore, the Wnt/ß­catenin signaling pathway was activated in PA­induced endothelial cells; however, the levels of DKK1 were downregulated. Overexpression of DKK1 could reduce CCN1 expression via inactivation of the Wnt/ß­catenin signaling pathway. In conclusion, knockdown of CCN1 attenuated PA­induced inflammation and apoptosis of endothelial cells via inactivating the Wnt/ß­catenin signaling pathway.

Ultrasonic-microwave assisted extraction of total triterpenoid acids from Corni Fructus and hypoglycemic and hypolipidemic activities of the extract in mice.

Triterpene acids, the main component of Corni Fructus, could improve diabetes mellitus, for which the underlying hypoglycemic mechanism is still unclear, in patients. In this study, total triterpenoid acids were extracted by ultrasonic-microwave assisted extraction optimized by the response surface methodology. The extract was then purified with an X-5 macroporous resin, and the yield of total triterpenoid acids increased to 281.24 mg g-1 as compared with the 35.71 mg g-1 obtained by unassisted extraction. The contents of five components were determined by ultrafast performance liquid chromatography. In addition, the hypoglycemic and hypolipidemic activities of total triterpenoid acids in diabetic mice induced by streptozotocin and a high fat diet were studied. The results indicated that all parameters (oral glucose tolerance, insulin resistance and liver damage) related to diabetes were significantly improved by total triterpenoid acids. Furthermore, total triterpenoid acids significantly recovered the expression level of AMP-activated protein kinase and its downstream proteins, including acetyl-CoA carboxylase, carnitine palmityltransferase-1, peroxisome proliferator-activated receptor alpha, sterol regulatory element-binding protein 1c and fatty acid synthase. Altogether, total triterpenoid acids could ameliorate hyperlipidemia and hyperglycemia in diabetic mice, probably by activating the AMP-activated protein kinase-peroxisome proliferator-activated receptor signaling pathway and inhibiting the sterol regulatory element-binding protein 1c and fatty acid synthase signaling pathways. Therefore, total triterpene acids, isolated from Corni Fructus which is a prevailing health food, could be a functional food ingredient with therapeutic and commercial values.

Ultrasonographic characteristics of thyroid metastasis from clear cell renal cell carcinoma: A case report.

INTRODUCTION: Thyroid metastasis from clear cell renal cell carcinoma (ccRCC) is a very rare condition, and its ultrasonographic characteristics have not been summarized in the literature. We herein report a case of thyroid metastasis from ccRCC that occurred 11 years after the surgery and the ultrasonographic characteristics of it are described. PATIENT CONCERNS: A 57-year-old male patient was admitted to our hospital in September 2018 due to discomfort in the neck. No other abnormalities were found during laboratory examination of thyroid function. The previous medical history of the patient included a right nephrectomy for the treatment of ccRCC in June 2007. DIAGNOSIS: Ultrasound examinations revealed multiple thyroid nodules. After nephrectomy, there was no obvious abnormality in the right renal area. Computed tomography (CT) showed an oval lesion with slightly lower density in the right lobe of the thyroid, and the patient was initially diagnosed with nodular goiter. INTERVENTIONS: Bilateral partial thyroidectomy under general anesthesia was conducted. Intraoperative frozen pathological examination showed clear cell carcinoma in the right lobe of the thyroid gland. Therefore, total thyroidectomy and lymph node dissection in the central neck area were performed. OUTCOMES: The patient underwent surgical treatment and recovered successfully. The patient was followed up for 2 years with no further metastasis. CONCLUSION: Ultrasound examination is a safe and convenient screening method. For patients with a renal malignant tumor, if the ultrasound image of thyroid nodule is found to have the characteristics of malignant tumors, the occurrence of metastasis of renal cancer to the thyroid should be highly suspected. Core needle biopsy (CNB) and histopathological diagnosis should be conducted subsequently for early diagnosis.

The curative effect of synthetic treatment for refractory acute myocardial infarction.

BACKGROUND: This study aims to investigate the curative effect of synthetic treatment for refractory acute myocardial infarction (AMI). METHODS: A total of 76 patients with coronary AMI accompanied by shock, who were treated with combined therapy from August 1999 to April 2017, were included into this study. Sixty patients received emergency percutaneous coronary intervention (PCI). Among these patients, 39 patients received intra-aortic balloon counterpulsation (IABP), eight patients had failed PCI underwent emergency off-pump coronary artery bypass (E-OPCAB), and eight patients were treated by hybrid cardiac surgery. RESULTS: All patients were successfully rescued. However, two patients died afterward due to postoperative complications. CONCLUSIONS: For AMI patients complicated with shock, especially when emergency PCI fails or is difficult to perform, PCI + IABP, emergency E-OPCAB and hybrid cardiac surgery should be carried out, in order to achieve a good outcome and improve the success rate of rescue for this group of patients. KEYWORDS: Acute myocardial infarction (AMI); emergency percutaneous coronary intervention (PCI); intra-aortic balloon counterpulsation (IABP); emergency off-pump coronary artery bypass (E-OPCAB); hybrid cardiac surgery.

High expression of PRPS1 induces an anti-apoptotic effect in B-ALL cell lines and predicts an adverse prognosis in Chinese children with B-ALL.

Phosphoribosyl pyrophosphate synthetase 1 (PRPS1) is closely associated with a number of diseases; however, its influence in B-cell acute lymphoblastic leukemia (B-ALL) and the potential molecular mechanisms involved remain unclear. The present study aimed to evaluate the expression of PRPS1 in Chinese children with B-ALL and to investigate the mechanism of action of PRPS1 in this disease. A Cell Counting Kit-8 (CCK-8) assay was performed to examine the proliferation of B-ALL Sup-B15 and Raji cells, and flow cytometric analysis was conducted to determine the cell cycle distribution and rate of apoptosis. The mRNA and protein expression levels of PRPS1, MYC proto-oncogene, bHLH transcription factor, cyclin E1, B-cell lymphoma-2 (Bcl-2), cyclin dependent kinase 2 and caspase-3 were detected by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Elevated PRPS1 expression was associated with a high-risk stratification and poor prognosis in patients with B-ALL. Furthermore, overexpression of PRPS1 accelerated the growth of and inhibited apoptosis in Sup-B15 and Raji cells as well as increasing the expression of Bcl-2 to induce an anti-apoptotic effect in B-ALL cell lines. The results of the present study indicate that PRPS1 regulates multiple processes in B-ALL and may be an attractive therapeutic target.

Investigation of the immunomodulatory activity of Tricholoma matsutake mycelium in cyclophosphamide-induced immunosuppressed mice.

Tricholoma matsutake, a popular food and biopharmaceutical resource in Asia, possesses various pharmacological activities. Although T. matsutake mycelium (TM) may enhance immunity, previous studies, to the best of our knowledge, have been performed on normal animals or cells alone. The present study aimed to evaluate the immunomodulatory activity of TM at doses of 0.3, 1.0 and 2.0 g/kg in cyclophosphamide (CTX)­induced immunosuppressed mouse models. TM treatment for 2 weeks markedly improved the gain in bodyweight, increased organ indices, reduced hind paw swelling and positively regulated the cytotoxicity of natural killer cells and the proliferation of lymphocytes. These effects are similar to that of thymosin α1 (0.16 mg/kg) which served as the positive control. In CTX­induced immunosuppressed mice, TM demonstrated marked effects on the modulation of the production of immunoglobulin (Ig)G and IgA, and the levels of interleukin­2, 6, 10 and 12, interferon­α and γ and tumor necrosis factor­α in serum. Compared with CTX mice, the reduced activity of nuclear factor (NF)­κB in serum and spleen, and phosphorylation of inhibitor of NF­κB kinase α/ß in spleen were observed in TM­treated mice. Taken together, TM effectively improved immune function in immunosuppressed mice via modulation of ILs and inflammatory factors associated with the NF­κB signaling pathway.

Tricholoma matsutake Aqueous Extract Induces Hepatocellular Carcinoma Cell Apoptosis via Caspase-Dependent Mitochondrial Pathway.

Tricholoma matsutake, one of widely accepted functional mushrooms, possesses various pharmacological activities, and its antitumor effect has become an important research point. Our study aims to evaluate the cytotoxicity activities of T. matsutake aqueous extract (TM) in HepG2 and SMMC-7721 cells. In in vitro experiments, TM strikingly reduced cell viability, promoted cell apoptosis, inhibited cell migration ability, induced excessive generation of ROS, and caused caspases cascade and mitochondrial membrane potential dissipation in hepatocellular carcinoma cells. In in vivo experiments, 14-day TM treatment strongly suppressed tumor growth in HepG2 and SMMC-7721-xenografted nude mice without influence on their body weights and liver function. Furthermore, TM increased the levels of cleaved poly-ADP-ribose polymerase (PARP), Bad, and Bax and reduced the expressions of B-cell lymphoma 2 (Bcl-2) in treated cells and tumor tissues. All aforementioned results suggest that caspase-dependent mitochondrial apoptotic pathways are involved in TM-mediated antihepatocellular carcinoma.

The optimization of Marasmius androsaceus submerged fermentation conditions in five-liter fermentor.

Using desirability function, four indexes including mycelium dry weight, intracellular polysaccharide, adenosine and mannitol yield were uniformed into one expected value (Da) which further served as the assessment criteria. In our present study, Plackett-Burman design was applied to evaluate the effects of eight variables including initial pH, rotating speed, culture temperature, inoculum size, ventilation volume, culture time, inoculum age and loading volume on Da value during Marasmius androsaceus submerged fermentation via a five-liter fermentor. Culture time, initial pH and rotating speed were found to influence Da value significantly and were further optimized by Box-Behnken design. Results obtained from Box-Behnken design were analyzed by both response surface regression (Design-Expert.V8.0.6.1 software) and artificial neural network combining the genetic algorithm method (Matlab2012a software). After comparison, the optimum M. androsaceus submerged fermentation conditions via a five-liter fermentor were obtained as follows: initial pH of 6.14, rotating speed of 289.3 rpm, culture time of 6.285 days, culture temperature of 26 °C, inoculum size of 5%, ventilation volume of 200 L/h, inoculum age of 4 days, and loading volume of 3.5 L/5 L. The predicted Da value of the optimum model was 0.4884 and the average experimental Da value was 0.4760. The model possesses well fitness and predictive ability.

Antifatigue Activity of Liquid Cultured Tricholoma matsutake Mycelium Partially via Regulation of Antioxidant Pathway in Mouse.

Tricholoma matsutake has been popular as food and biopharmaceutical materials in Asian countries for its various pharmacological activities. The present study aims to analyze the antifatigue effects on enhancing exercise performance of Tricholoma matsutake fruit body (ABM) and liquid cultured mycelia (TM) in mouse model. Two-week Tricholoma matsutake treatment significantly enhances the exercise performance in weight-loaded swimming, rotating rod, and forced running test. In TM- and ABM-treated mice, some factors were observed at 60 min after swimming compared with nontreated mice, such as the increased levels of adenosine triphosphate (ATP), antioxidative enzymes, and glycogen and the reduced levels of malondialdehyde and reactive oxygen species in muscle, liver, and/or serum. Further data obtained from western blot show that CM and ABM have strongly enhanced the activation of 5'-AMP-activated protein kinase (AMPK), and the expressions of peroxisome proliferator have activated receptor γ coactivator-1α (PGC-1α) and phosphofructokinase-1 (PFK-1) in liver. Our data suggest that both Tricholoma matsutake fruit body and liquid cultured mycelia possess antifatigue effects related to AMPK-linked antioxidative pathway. The information uncovered in our study may serve as a valuable resource for further identification and provide experimental evidence for clinical trials of Tricholoma matsutake as an effective agent against fatigue related diseases.