ACADL suppresses PD-L1 expression to prevent cancer immune evasion by targeting Hippo/YAP signaling in lung adenocarcinoma.

Lung cancer is the leading cause of cancer-related death. Cancer immune evasion is a key barrier in the treatment of lung cancer and the development of effective anticancer therapeutics. Long-chain Acyl-CoA dehydrogenase (ACADL), a key enzyme that regulates ß-oxidation of long-chain fatty acyl-CoAs, has been found to act as a tumor suppressor in cancers. However, the role of ACADL in lung adenocarcinoma (LUAD) has not been explored. In the current study, we find that ACADL functions as a tumor suppressor in LUAD to inhibit proliferation and enhanced chemotherapeutic drug-induced apoptosis. Interestingly, ACADL prevents tumor immune evasion by suppressing PD-L1 expression in LUAD. ACADL is critical for Hippo/YAP pathway-mediated PD-L1 regulation. Moreover, YAP activation is essential for ACADL suppression of PD-L1 transcription. In addition, ACADL increases the protein stability and kinase activity of LATS kinase to inhibit YAP activation and PD-L1 transcription. Furthermore, we show that ACADL expression is positively correlated with a better OS and FP in LUAD. Our data reveals that ACADL could be a promising target for regulating Hippo/YAP pathway to prevent tumor immune evasion in LUAD.

Percutaneous vertebroplasty by percutaneous bi-level bilateral puncture for treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fractures with dense bone bands / 中华创伤骨科杂志

Objective:To evaluate the efficacy of percutaneous vertebroplasty (PVP) by percutaneous bi-level bilateral puncture in the treatment of type ⅡA acute symptomatic osteoporotic thoracolumbar fracture (ASOTLF) with dense bone bands.Methods:From March 2017 to March 2018, 65 patients (65 vertebrae) with type ⅡA ASOTLF with dense bone bands were treated at Department of Orthopaedic Trauma, Qingdao Central Hospital. They were 25 males and 40 females, with an age of (71.6±8.4) years. The time from injury to operation was (3.5±0.7) d. They were divided into 2 groups according to different treatments. In the observation group of 31 cases (31 vertebrae), PVP was conducted by percutaneous bi-level bilateral puncture; in the control group of 34 cases (34 vertebrae), PVP was conducted by percutaneous single-level bilateral puncture. The 2 groups were compared in terms of operation time, volume of polymethyl methacrylate injected, incidence of bone cement leakage, incidence of vertebral refracture, diffusive distribution of bone cement in the vertebral body, and visual analogue scale (VAS) and Oswestry dysfunction index (ODI) at postoperative 1 day and 6 months.Results:There was no significant difference in the preoperative general data between the 2 groups, showing comparability ( P>0.05). Operations were completed uneventfully in all the 65 patients. All the 65 patients were followed up for (8.2±1.2) months. There was no significant difference between the 2 groups in operation time, incidence of bone cement leakage or incidence of vertebral refracture for each vertebral body injured ( P>0.05). The volume of polymethyl methacrylate injected [(6.64±0.93) mL] and the excellent and good rate of diffusive distribution of bone cement in the vertebral body [87.1% (27/31)] in the observation group were significantly larger than those in the control group [(4.36±0.79) mL and 64.7% (22/34)] ( P<0.05). The VAS scores at postoperative 1 day and 6 months [2 (1, 2) and 1 (1, 2)] and ODIs at postoperative 1 day and 6 months (23.7%±1.6% and 18.8%±1.4%) in the observation group were significantly lower than those in the control group [2 (2, 3) and 2 (2, 2); 26.9%±4.2% and 22.1%±3.3%] ( P<0.05). The VAS scores and ODIs at postoperative 1 day and 6 months in all patients were significantly lower than those before operation ( P<0.05). Compression symptoms of the spinal cord and nerve root were observed in none of the patients. Conclusion:In the treatment of type ⅡA ASOTLF with dense bone bands, PVP by percutaneous bi-level bilateral puncture is more effective than the traditional PVP by percutaneous single-level bilateral puncture, and will not increase bone cement leakage or vertebral refracture.

Betulinic Acid Improves Cardiac Function in Septic Rats Through AKT / mTOR and AKT / AMPK -modulated Autophagy / 中国生物化学与分子生物学报

Betulinic acid (BA) exerts protective effects on organs in septic animals. However, whether BA can improve cardiac function in sepsis and the underlying mechanism remain unclear. Here, male Sprague-Dawley rats were pretreated with BA (25 mg/ kg/ d, i. g.) for 5 days and then intraperitoneally injected with lipopolysaccharide (LPS, 10 mg/ kg). The rats were anesthetized to determine transthoracic echocardiography using a high-resolution imaging system for small animals after they were treated with LPS for 6 h. Histopathologic alterations were examined by HE staining. Myocardial injury markers (cTnI and CK-MB) and inflammatory factors (TNF-α, IL-1β and IL-6) in the serum were measured by the enzyme-linked immunosorbent assay. Autophagy-related proteins (p62 and LC3 Ⅱ) and AKT-modulated autophagy pathways in the myocardium were determined by Western blotting. Pretreatment with BA markedly improved left ventricular ejection fraction (EF) and fraction shortening (FS) (P<0. 05), improved myocardial histomorphology, and significantly inhibited cTnI, CK-MB, TNF-α, IL-1β and IL-6 (P<0. 05) in the septic rat serum. BA markedly decreased p62 (P<0. 01), increased LC3 Ⅱ (P< 0. 001), and significantly down-regulated p-AKT (Thr308), p-AMPKα (Ser485/ 491), p-mTOR (Ser2448) and p-S6K (Thr389) (P<0. 05), while markedly up-regulated p-AMPKα (Thr172) and pULK1 (Ser317) (P<0. 01) in septic rat hearts. The findings indicate that BA can attenuate sepsis-induced myocardial dysfunctions associated with down-regulating autophagy inhibiting pathways mediated by AKT/ mTOR and AKT/ AMPK pathways.

Prognosis of traumatic spinal cord injury in children: Follow-up of 86 patients / 中华创伤杂志(英文版)

PURPOSE@#The long-term situation of children with spinal cord injury (SCI) was investigated, and suggestions for helping them better return to the society were provided.@*METHODS@#SCI patients less than 18 years old hospitalized in Beijing Boai Hospital from January 2011 to December 2020 were retrospectively analyzed. Information including motor function, complications, characteristic changes, self-care abilities, school attendance and social participation were collected by telephone interview and electronic questionnaire. All the answers were statistically analyzed.@*RESULTS@#A total of 86 cases were enrolled, 77 girls and 9 boys, with a median injury age of 6 years and 2 months. The follow-up time was 3-130 months. The main cause of trauma in these children was sport injury (66.3%), the thoracic spinal cord was involved the most (91.9%), and complete SCIs accounted for the majority (76.7%). In terms of complications, children with complete SCIs were more likely to have urinary incontinence, constipation and characteristic changes (p < 0.05); whereas the incomplete SCIs often have spasticity (p < 0.05). As to the daily living abilities, children with incomplete lumbar SCIs were more capable to accomplish personal hygiene, transfer, and bathing independently than those with complete injuries, or cervical/thoracic SCIs, respectively (p < 0.05). Moreover, children older than 9 years care more able to dress and transfer independently than the youngers (p < 0.05). Wheelchair users accounted for 84.9% and more than half of them were able to propel wheelchair independently, and those who move passively in wheelchairs were mostly introverted kids (p < 0.05). Almost all (93.8%) children with incomplete injuries were able to walk independently. Most (79.1%) children continued to attending school, and 41.9% participated in interest classes. Unfortunately, 67.4% of the children spent less time playing with their peers than before the injury.@*CONCLUSION@#SCIs impair physical structures and function of children, affect their independence in daily living, and restrict school attendance and social interaction. Comprehensive rehabilitation after injury is a systematic work. Medical staff and caregivers should not only pay attention to neurological function, but also help them improve self-care abilities. It is also important to balance rehabilitation training and school work and social participation.

NDR1 activates CD47 transcription by increasing protein stability and nuclear location of ASCL1 to enhance cancer stem cell properties and evasion of phagocytosis in small cell lung cancer.

Small cell lung cancer (SCLC) is one of the most malignant types of lung cancer. Cancer stem cell (CSC) and tumor immune evasion are critical for the development of SCLC. We previously reported that NDR1 enhances breast CSC properties. NDR1 might also have a role in the regulation of immune responses. In the current study, we explore the function of NDR1 in the control of CSC properties and evasion of phagocytosis in SCLC. We find that NDR1 enhances the enrichment of the ALDEFLUORhigh and CD133high population, and promotes sphere formation in SCLC cells. Additionally, NDR1 upregulates CD47 expression to enhance evasion of phagocytosis in SCLC. Furthermore, the effects of NDR1 enhanced CD47 expression and evasion of phagocytosis are more prominent in CSC than in non-CSC. Importantly, NDR1 promotes ASCL1 expression to enhance NDR1-promoted CSC properties and evasion of phagocytosis in SCLC cells. Mechanically, NDR1 enhances protein stability and the nuclear location of ASCL1 to activate the transcription of CD47 in SCLC. Finally, CD47-blocking antibody can be used to target NDR1 enhanced CSC properties and evasion of phagocytosis by suppressing EGFR activation in SCLC. In summary, our data indicate that NDR1 could be a critical factor for modulating CSC properties and phagocytosis in SCLC.

NDR1 increases NOTCH1 signaling activity by impairing Fbw7 mediated NICD degradation to enhance breast cancer stem cell properties.

BACKGROUND: The existence of breast cancer stem cells (BCSCs) causes tumor relapses, metastasis and resistance to conventional therapy in breast cancer. NDR1 kinase, a component of the Hippo pathway, plays important roles in multiple biological processes. However, its role in cancer stem cells has not been explored. The purpose of this study was to investigate the roles of NDR1 in modulating BCSCs. METHODS: The apoptosis was detected by Annexin V/Propidium Iodide staining and analyzed by flow cytometry. BCSCs were detected by CD24/44 or ALDEFLUOR staining and analyzed by flow cytometry. The proliferation ability of BCSCs was evaluated by sphere formation assay. The expression of interested proteins was detected by western blot analysis. The expression of HES-1 and c-MYC was detected by real-time PCR. Notch1 signaling activation was detected by luciferase reporter assay. Protein interaction was evaluated by immunoprecipitation. Protein degradation was evaluated by ubiquitination analysis. The clinical relevance of NDR1 was analyzed by Kaplan-Meier Plotter. RESULTS: NDR1 regulates apoptosis and drug resistance in breast cancer cells. The upregulation of NDR1 increases CD24low/CD44high or ALDEFLUORhigh population and sphere-forming ability in SUM149 and MCF-7 cells, while downregulation of NDR1 induces opposite effects. NDR1 increased the expression of the Notch1 intracellular domain (NICD) and activated the transcription of its downstream target (HES-1 and c-MYC). Critically, both suppression of Notch pathway activation by DAPT treatment or downregulation of Notch1 expression by shRNA reverses NDR1 enhanced BCSC properties. Mechanically, NDR1 interactes with both NICD or Fbw7 in a kinase activity-independent manner. NDR1 reduces the proteolytic turnover of NICD by competing with Fbw7 for NICD binding, thereby leading to Notch pathway activation. Furthermore, NDR1 might function as a hub to modulate IL-6, TNF-α or Wnt3a induced activation of Notch1 signaling pathway and enrichment of breast cancer stem cells. Moreover, we find that the elevation of NDR1 expression predictes poor survival (OS, RFS, DMFS and PPS) in breast cancer. CONCLUSION: Our study revealed a novel function of NDR1 in regulating BCSC properties by activating the Notch pathway. These data might provide a potential strategy for eradicating BCSC to overcome tumor relapses, metastasis and drug resistance.

Long-term follow-up ultrasonography surveillance in a large cohort of patients with papillary thyroid carcinoma.

OBJECTIVE: This study aimed to investigate the predictive factors as well as the time and age course of recurrence/persistence in a large cohort of postoperative patients with papillary thyroid carcinoma (PTC) based on the long-term ultrasonography (US) follow-up data. METHODS: Between January 2007 and December 2016, 3106 patients underwent surgery for PTC and at least two postoperative US follow-up examination over more than three years. Tumor recurrence/persistence was confirmed based on the follow-up US data and histopathological results. Univariate and multivariate analyses were performed to evaluate the predictive factors of tumor recurrence/persistence. Kaplan-Meier survival analysis was used to evaluate the recurrence-/persistence-free survival curve based on the US results. RESULTS: A total of 321(10.3%) patients developed tumor recurrence/persistence during 54.3 months of mean follow-up (range 36-135 months), including 268(83.5%) cases of lymph node recurrence/persistence, 37 (11.5%) cases of non-lymph node recurrence/persistence, and 16(5%) cases of both types. Recurrence/persistence was observed using US examination at a mean interval of 23.6 ± 21.6 months (range 1-135 months) after surgery and peak incidence was observed 1-2 years after initial treatment. Younger (20-30 years old) and older (70-80 years old) patients had a higher proportion of tumor recurrence/persistence. Multifocality, advanced T and advanced N stages were independent risk factors of tumor recurrence/persistence. CONCLUSION: Tumor recurrence/persistence of PTC usually occurs during the early postoperative period. For patients with multifocal cancer, advanced T and N stage, the US surveillance examination should be cautiously performed, especially in younger and older patients.

Core prescriptions in treatment of edema by traditional Chinese medicine masters and mechanism prediction / 中国中药杂志

The core prescriptions and formulation characteristics in the treatment of edema by traditional Chinese medicine(TCM) masters were analyzed through data mining and their mechanisms were explored by network pharmacology. We collected journal reports on the treatment of edema by TCM masters in three sessions from China National Knowledge Infrastructure(CNKI) and constructed a database by Traditional Chinese Medicine Inheritance Support System 3.0. The prescriptions in the case studies were analyzed by association rules and k-means clustering. The chemical components and targets of Chinese medicines in core prescriptions were collected through TCMSP and TCMID. Edema-related targets were collected from DrugBank and GeneCards. The protein-protein interaction(PPI) network was constructed by STRING and the core targets were screened out. FunRich 3.1.3 was used to enrich the expression sites of core prescriptions. Metascape was used to perform Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment analysis of intersection targets. Cytoscape 3.6.0 was used to visualize the "Chinese medicine-active ingredient-core target-pathway" network. The results showed that 315 pieces of medical records in the treatment of edema by TCM masters were obtained and five core prescriptions were analyzed by association rules and k-means clustering. Core prescription 1 contained Poria, Atractylodis Macrocephalae Rhizoma, Astragali Radix, Alismatis Rhizoma, Glycyrrhizae Radix et Rhizoma, and Codonopsis Radix, involving 166 chemical components and 1 125 targets. Core prescription 2 contained Astragali Radix, Salviae Miltiorrhizae Radix et Rhizoma, Poria, Chuanxiong Rhizoma, Paeoniae Radix Rubra, and Angelicae Sinensis Radix, involving 138 chemical components and 1 112 targets. Core prescription 3 contained Poria, Salviae Miltiorrhizae Radix et Rhizoma, Astragali Radix, Atractylodis Macrocephalae Rhizoma, Alismatis Rhizoma, and Coicis Semen, involving 126 chemical components and 1 121 targets. Core prescription 4 contained Poria, Forsythiae Fructus, Atractylodis Macrocephalae Rhizoma, Imperatae Rhizoma, Cicadae Periostracum, and Coicis Semen, involving 58 chemical components and 820 targets. Core prescription 5 contained Poria, Atractylodis Macrocephalae Rhizoma, Astragali Radix, Alismatis Rhizoma, Trionycis Carapax, and Dioscoreae Rhizoma, involving 68 chemical components and 919 targets. The core targets of core prescriptions included AKT1, ALB, CASP3, MAPK3, EGFR, SRC, MAPK1, and TNF. The potential targets of core prescriptions in the treatment were highly expressed in the stomach, bladder, lung, and kidney. KEGG pathways were enriched in inflammation and cell cycle pathways, especially the inflammation-relation pathways. The therapeutic effect of core prescriptions on edema is presumedly achieved by tonifying the spleen, draining water, activating blood, and benefiting Qi to resist inflammation and regulate the immune system. This study is expected to provide references for the summary of TCM masters' experience and new drug development.

Development and validation of a prediction model for treatment failure in peritoneal dialysis-associated peritonitis patients: a multicenter study / 南方医科大学学报

OBJECTIVE@#To develop and validate a risk prediction model of treatment failure in patients with peritoneal dialysis-associated peritonitis (PDAP).@*METHODS@#We retrospectively analyzed the data of patients undergoing peritoneal dialysis (PD) in 3 dialysis centers in Jilin Province who developed PDAP between January 1, 2013 and December 31, 2019. The data collected from the Second Hospital of Jilin University and Second Division of First Hospital of Jilin University) were used as the training dataset and those from Jilin Central Hospital as the validation dataset. We developed a nomogram for predicting treatment failure using a logistic regression model with backward elimination. The performance of the nomogram was assessed by analyzing the C-statistic and the calibration plots. We also plotted decision curves to evaluate the clinical efficacy of the nomogram.@*RESULTS@#A total of 977 episodes of PDAP were included in the analysis (625 episodes in the training dataset and 352 episodes in the validation dataset). During follow-up, 78 treatment failures occurred in the training dataset and 35 in the validation dataset. A multivariable logistic regression prediction model was established, and the predictors in the final nomogram model included serum albumin, peritoneal dialysate white cell count on day 5, PD duration, and type of causative organisms. The nomogram showed a good performance in predicting treatment failure, with a C-statistic of 0.827 (95% CI: 0.784-0.871) in the training dataset and of 0.825 (95% CI: 0.743-0.908) in the validation dataset. The nomogram also performed well in calibration in both the training and validation datasets.@*CONCLUSION@#The established nomogram has a good accuracy in estimating the risk of treatment failure in PDAP patients.

Folic acid modified Fe3O4 nanoclusters by a one-step ultrasonic technique for drug delivery and MR imaging.

Multifunctional nanoclusters based on Fe3O4 nanoparticles for magnetic resonance imaging (MRI) and drug delivery are reported here. At first, oleic acid (OA)-coated Fe3O4 nanoparticles were prepared. Then block copolymer Pluronic F127 or folic acid (FA) conjugated-Pluronic F127 was used to modify the hydrophobic nanoparticles to become hydrophilic Fe3O4@F127 nanoclusters via facile ultrasonic treatment. During this process, drug molecules can also be introduced into the nanoclusters and therefore the targeted drug delivery system was formed. Next, we verified the feasibility of the nanoclusters as drug delivery vehicles and magnetic contrast agents. The nanoclusters have an average size of 200 nm and remained stable in water for long periods. Folic acid-modified nanoclusters showed an enhanced intracellular uptake into HepG2 cells by using both cellular iron amount analysis and flow cytometry analysis. Besides, Fe3O4@F127@FA nanoclusters showed good compatibility in the tested concentration range and good sensitivity in T 2-weighted MRI. The magnetic nanoclusters combined with drug delivery properties have greatly increased the significance in the diagnosis and therapy of diseases, which are suitable for systematical administration of hydrophobic drugs and simultaneously MRI diagnosis.

An ultra-sensitive T 2-weighted MR contrast agent based on Gd3+ ion chelated Fe3O4 nanoparticles.

An ultra-sensitive T 2-weighted MR imaging contrast agent was prepared based on Fe3O4 nanoparticles and Gd3+ ions (Fe3O4@Gd). Amino modified Fe3O4 nanoparticles were conjugated to diethylenetriamine pentaacetic acid, and finally coordinated with Gd3+ ions. The nanoparticles had a uniform morphology with a size of 100 nm and a Gd/Fe mass ratio of 1/110. The r 2 (transverse relaxivity) of the Fe3O4 nanoparticles increased from 131.89 mM-1 s-1 to 202.06 mM-1 s-1 after coordination with Gd3+ ions. MR measurements showed that the aqueous dispersion of Fe3O4@Gd nanoparticles had an obvious concentration-dependent negative contrast enhancement. Hepatoma cells were selected to test the cytotoxicity and MR imaging effect. The application of Fe3O4@Gd nanoparticles as contrast agents was also exploited in vivo for T 2-weighted MR imaging of rat livers. All the results showed the effectiveness of the nanoparticles in MR diagnosis.

Expert consensus on clinical application of Chinese herbal medicine decoction pieces (First Edition) / 中国中药杂志

Chinese herbal medicine decoction pieces(CHMDP), one of the main forms of traditional Chinese medicine(TCM) in clinic, have been widely used. However, the irrational use is increasingly serious due to the lack of the indicators for judging the rational use of CHMDP in medical institutions and the codes and standards for the clinical use of CHMDP. In order to regulate the rational clinical use of CHMDP, improve the clinical efficacy and ensure the drug safety for the patients, clinical pharmaceutical experts and clinical medical experts from 40 third-grade class-A hospitals nationwide were organized to give the "expert consensus on clinical application of CHMDP" in terms of prescription writing, combined use of drugs, use of special drugs, and drug use for special population. Detailed analysis and argumentation were conducted in accordance with the laws and regulations, Chinese Pharmacopoeia 2015 edition, Chinese Pharmacopoeia Code Notice for Clinical Use of Medicine, Administrative Regulations for Prescriptions, Administrative Specifications for Hospital Prescription Review(interim), and Chinese Traditional Medicine Prescription Format and Writing Specifications, as well as relevant project findings.

Change laws of water absorption in Chinese herbal pieces and prediction model of relative density for Chinese medicine decoction / 中国中药杂志

This study aims to explore the change laws of water absorption in Chinese herbal pieces and establish the prediction model of relative density for Chinese medicine compound decoction. Firstly, fitted equations of water absorption and decocting time was established by observing the change laws of water absorption in 36 kinds of Chinese herbal pieces in 12 groups(according to the drug-parts) with decocting time. The r value of the mineral group and other type group was 0.691 2 and 0.663 3, respectively. The r value of the remaining 10 groups was 0.802 2-0.925 4. All P values were less than 0.05(n=21). The formula of the amount of water added was optimized by combining the fitted equations with determined water absorption, and the liquid yield could be controlled in a range of 100%±10%. Secondly, it was determined that the liquid density tester could be used for the rapid determination of relative density of Chinese medicine decoction after methodological study and comparison with the pycnometer method. The linear regression equation between the corrected relative density(y) and extraction ratio(%, x) was built by measuring and analyzing the related parameters such as liquid yield, relative density and extraction ratio in 46 kinds of Chinese herbal pieces. The established equation was y=0.041 3x+1.003 7, r=0.930 9(P <0.01, n=46), with linear range of 1.94%-65.75%. Based on this, the prototype model for predicting relative density of Chinese medicine decoction was established, and the relative densities of 8 Chinese medicine decoctions were within the prediction interval of this model in verification. This study lays a foundation for database construction of Chinese medicine decoction, implementation of personalized decocting mode and rapid quality control of Chinese medicine decoction.

Emodin assay in Ampelopsis radix by RP-HPLC / 药学实践杂志

Objective To establish a method for Emodin assay in Ampelopsis radix, and to improve the quality control of Ampelopsis radix. Methods Reversed phase high performance liquid chromatography was used. The column: Apollo-C18 (4.6 mm×250 mm, 5 μm); mobile phase: methanol:0.2% phosphoric acid (85:15); flow rate: 1.0 ml/min; column temperature: room temperature; detection wavelength: 220 nm. Results With the specified chromatographic conditions, there was a good linear relationship for Emodin in Ampelopsis radix in the range of 0.124-3.968 μg/ml. The linear regression equation was Y= 53 962X–966.46, r=0.999 7. The average recovery was 99.7%. RSD=2.5% (n=9). Conclusion This method is accurate, sensitive and repeatable. It is suitable for the determination of emodin in Ampelopsis radix.

ACOX1 destabilizes p73 to suppress intrinsic apoptosis pathway and regulates sensitivity to doxorubicin in lymphoma cells.

Lymphoma is one of the most curable types of cancer. However, drug resistance is the main challenge faced in lymphoma treatment. Peroxisomal acyl-CoA oxidase 1 (ACOX1) is the rate-limiting enzyme in fatty acid ß-oxidation. Deregulation of ACOX1 has been linked to peroxisomal disorders and carcinogenesis in the liver. Currently, there is no information about the function of ACOX1 in lymphoma. In this study, we found that upregulation of ACOX1 promoted proliferation in lymphoma cells, while downregulation of ACOX1 inhibited proliferation and induced apoptosis. Additionally, overexpression of ACOX1 increased resistance to doxorubicin, while suppression of ACOX1 expression markedly potentiated doxorubicin-induced apoptosis. Interestingly, downregulation of ACOX1 promoted mitochondrial location of Bad, reduced mitochondrial membrane potential and provoked apoptosis by activating caspase-9 and caspase-3 related apoptotic pathway. Overexpression of ACOX1 alleviated doxorubicin-induced activation of caspase-9 and caspase-3 and decrease of mitochondrial membrane potential. Importantly, downregulation of ACOX1 increased p73, but not p53, expression. p73 expression was critical for apoptosis induction induced by ACOX1 downregulation. Also, overexpression of ACOX1 significantly reduced stability of p73 protein thereby reducing p73 expression. Thus, our study indicated that suppression of ACOX1 could be a novel and effective approach for treatment of lymphoma. [BMB Reports 2019; 52(9): 566-571].

ß-Asarone increases doxorubicin sensitivity by suppressing NF-κB signaling and abolishes doxorubicin-induced enrichment of stem-like population by destabilizing Bmi1.

BACKGROUND: Lymphoma is one of the most common hematologic malignancy. Drug resistance is the main obstacle faced in lymphoma treatment. Cancer stem cells are considered as the source of tumor recurrence, metastasis and drug resistance. The ß-Asarone, a low-toxicity compound from the traditional medical herb Acorus calamus, has been shown to act as an anti-cancer reagent in various cancer types. However, the anti-cancer activities of ß-Asarone in lymphoma have not been shown. METHODS: Cell counting assay was used to evaluate Raji cell proliferation. CCK8 assay was used to evaluate the cell viability. Annexin-V/PI staining and flow cytometry analysis were used to evaluate apoptosis. ALDEFLUOR assay was used to evaluate the stem-like population. Luciferase reporter assay was used to examine the activation of NF-κB signaling. Western blot and polymerase chain reaction (PCR) were used to determine the expression of interested genes. RESULTS: We showed that ß-Asarone inhibited proliferation and induced apoptosis in Raji lymphoma cells in a dose-dependent manner. Additionally, ß-Asarone functioned as a sensitizer of doxorubicin and resulted in synergistic effects on inhibition of proliferation and induction of apoptosis when combined with doxorubicin treatment. Interestingly, we found that ß-Asarone also reduced the stem-like population of Raji lymphoma cells in a dose-dependent manner, and suppressed the expression of c-Myc and Bmi1. Importantly, ß-Asarone abolished doxorubicin-induced enrichment of the stem-like population. In the mechanism study, we revealed that ß-Asarone suppressed not only basal NF-κB activity but also Tumor necrosis factor α (TNF-α) induced NF-κB activity. Moreover, blocking NF-κB signaling inactivation was critical for ß-Asarone induced apoptosis and inhibition of proliferation, but not for the effect on ß-Asarone reduced stem-like population. In fact, ß-Asarone suppressed stem-like population by destabilizing Bmi1 via a proteasome-mediated mechanism. CONCLUSIONS: Our data suggested the application of ß-Asarone to lower the toxic effect of doxorubicin and increase the sensitivity of doxorubicin in clinical treatment. More importantly, our data revealed a novel role of ß-Asarone which could be used to eliminate stem-like population in lymphoma, implying that ß-Asarone might reduce relapse and drug resistance.

Facile fabrication of water-dispersible nanocomposites based on hexa-peri-hexabenzocoronene and Fe3O4 for dual mode imaging (fluorescent/MR) and drug delivery.

The facile fabrication of multifunctional nanocomposites (Fe3O4/HBC@F127) consisting of superparamagnetic Fe3O4 nanoparticles and fluorescent organic hexa-peri-hexabenzocoronene (HBC) molecules incorporated in block copolymer diacylphospholipid-polyethyleneglycol F127 have been demonstrated for dual mode imaging (fluorescent/MR) and drug delivery. The obtained nanocomposites were water-dispersible, stable and biocompatible, as confirmed by dynamic light scattering (DLS) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Relativity measurements showed a T 2 relaxivity (r 2) of 214.61 mM-1 s-1, which may be used as T 2-weighted MR imaging agents. In vitro imaging studies indicated that the nanocomposites had good MR and fluorescence imaging effects with low cytotoxicity. Besides, the developed nanocomposites could also be applied as drug delivery vehicles. Doxorubicin (DOX) loaded Fe3O4/HBC@F127 nanocomposites significantly inhibited the growth of human hepatoma cells (HepG2). These findings suggested that the facile synthesized multifunctional nanocomposites may be used as a platform for dual mode imaging (both MR and fluorescence) and drug delivery.

Establishment and application of whole genome high-throughput sequencing of hepatitis B virus with different concentrations / 中华实验和临床病毒学杂志

Objective@#To establish a high-throughput sequencing method for a whole genome in different hepatitis B virus (HBV) concentrations.@*Methods@#Two method of amplicon-sequencing and direct sequencing without PCR amplification were used for library construction in the three plasmids, including the low HBV load sample, the moderate HBV load sample and the high HBV load sample. Whole genome sequencing was performed on Illumina MiSeq platform.@*Results@#There are significant differences in data yield between the two different library construction method. Only a few reads could be mapped to the HBV genome for direct sequencing. However, three samples were successfully amplified by the nested PCR amplification and amplicon-sequencing showed that all HBV samples had a good coverage and depth, which was not affected by HBV concentration. The alignment rate of HBV genome approached 80%. A total of 27 intra-host single nucleotide variations (iSNVs) were identified and 13 iSNVs were low-frequency mutation in three samples. Compared with the high HBV load sample, mutations in the reverse transcription (RT) region was more easily appeared in the low HBV sample and the moderate HBV load sample.@*Conclusions@#Integrating nested PCR with high-throughput sequencing to the HBV whole genome sequencing is not only a practical method to detect the infection of HBV with a high-sensitivity and accuracy, but also enables to detect the mutation in the infected patient with low HBV copy number.

Anesthesia management for orthotropic heart transplantation / 中华麻醉学杂志

Anesthesia was done for 36 patients undergoing orthotropic heart transplantation in Bei-jing Anzhen Hospital from April 2015 to November 2016. Anesthesia management for orthotropic heart transplantation and related problems were analyzed and investigated. Anesthesia management protocol for patients with end-stage heart disease was aimed at reducing fluctuation of hemodynamics and avoiding malig-nant arrhythmia. Anesthesia was induced by intravenously injecting diazepam 5-10 mg, etomidate 0. 2-0. 3 mg∕kg or ketamine 1 mg∕kg, sufentanil 1. 0-1. 5 μg∕kg or fentanyl 10-15 μg∕kg and rocuronium 0. 6 mg∕kg. Anesthesia was maintained by continuously infusing dexmedetomidine 0. 3-0. 5μg·kg-1 ·h-1 , ci-satracurium 10 mg∕h and sufentanil 0. 5-1. 0 μg·kg-1 ·h-1 . Pulmonary arterial pressure and donor heart function were monitored using the flow-directed pulmonary artery catheter. Dopamine, epinephrine and iso-prenaline were intravenously infused after cardiopulmonary bypass to maintain circulation stable. Nitroglyc-erin and prostacyclin were intravenously infused to decrease pulmonary arterial pressure. Immunosuppressive therapy was performed with methylprednisone, mycophenolate mofetil and cyclosporine∕FK506. Thirty-two patients were discharged from hospital, and 4 cases died. Among the 4 patients died, 1 patient died of pul-monary hypertension ( pulmonary arterial systolic pressure>67 mmHg) and right heart failure and, 1 patient showed difficulty in weaning from cardiopulmonary bypass and 2 patients died of refractory low cardiac outputand multi-organ failure. Anesthetic management for heart transplantation required an appreciation of the pathophysiological mechanism of heart failure. Invasive monitoring, steady anesthesia induction and mainte-nance, stable hemodynamics in the perioperative period and good donor heart protection were the keys to ensuring anesthesia management for orthotropic heart transplantation.