MHY1485 promotes adriamycin sensitivity in HepG2 cells by inhibiting autophagy.

MHY1485 is an mTOR activator that inhibits the autophagy process by inhibiting the fusion between autophagosomes and lysosomes. This study aimed to explore the role and mechanism of MHY1485 in hepatocellular carcinoma (HCC) and to provide an in-depth understanding of the mechanisms of autophagy regulation in relation to adriamycin (ADM) resistance, as well as the development of a molecularly targeted autophagy-modulating approach. Here, ADM was used to treat HepG2 cells and construct an ADM-resistant cell model. The HepG2/ADM cell line and HepG2 cells were treated with MHY1485 and ADM, respectively, and the proliferation and apoptosis of HCC cells were detected using CCK8, clone formation, flow cytometry, and 5-ethynyl-2'-deoxyuridine staining assays. Ki-67, mTOR phosphorylation, and LC3A expression were detected by IF staining; the expression or phosphorylation levels of autophagy-related proteins (i.e., GLUT1, PGI, PFK, END, and MTHFD2) and apoptosis-related proteins (caspase-3, caspase-8, and caspase-9) were detected by qPCR and western blotting. The number of autophagosomes was determined by monodansylcadaverine staining. Our results showed that MHY1485 can inhibit the proliferation and growth of liver cancer cells, and that MHY1485 combined with ADM can effectively inhibit the tolerance of HepG2/ADM cells to ADM and enhance the efficacy of ADM. The results of the detection of the autophagy-related protein LC3A also indicated that MHY1485 activates mTOR and can affect the phosphorylation level of ULK1, inhibit autophagy, and enhance the sensitivity of liver cancer cells to adriamycin. In summary, MHY1485 can enhance the sensitivity of adriamycin-resistant cells to adriamycin by activating mTOR and blocking the autophagy process in cells; therefore, mTOR may become a potential target for the treatment of liver cancer.

Nanoparticles Mediated circROBO1 Silencing to Inhibit Hepatocellular Carcinoma Progression by Modulating miR-130a-5p/CCNT2 Axis.

Background: Circular RNAs (circRNAs) are becoming vital biomarkers and therapeutic targets for malignant tumors due to their high stability and specificity in tissues. However, biological functions of circRNAs in hepatocellular carcinoma (HCC) are still not well studied. Methods: Gene Expression Omnibus (GEO) database and qRT-PCR were used to evaluate expression of circROBO1 (hsa_circ_0066568) in HCC tissues and cell lines. CCK-8, colony formation, EdU staining, flow cytometry for cell cycle analysis, and xenograft model assays were performed to detect the circROBO1 function in vitro and in vivo. RNA pull-down, RNA immunoprecipitation (RIP), and Luciferase reporter assays were used to investigate the relationship among circROBO1, miR-130a-5p, and CCNT2. More importantly, we developed nanoparticles made from poly lactic-co-glycolic acid (PLGA) and polyethylene glycol (PEG) chains as the delivery system of si-circROBO1 and then applied them to HCC in vitro and in mice. Results: circROBO1 was obviously upregulated in HCC tissues and cell lines, and elevated circROBO1 was closely correlated with worse prognosis for HCC patients. Functionally, knocking down circROBO1 significantly suppressed HCC cells growth in vitro and in mice. Mechanistically, circROBO1 acted as a competing endogenous RNA to downregulate miR-130a-5p, leading to CCNT2 expression upregulation. Furthermore, miR-130a-5p mimic or CCNT2 knockdown reversed the role of circROBO1 overexpression on HCC cells, which demonstrated that circROBO1 promoted HCC development via miR-130a-5p/CCNT2 axis. In addition, we developed nanoparticles loaded with si-circROBO1, named as PLGA-PEG (si-circROBO1) NPs, which significantly prevented the proliferation of HCC cells, and did not exhibit apparent toxicity to major organs in vivo. Conclusion: Our findings firstly demonstrate that circROBO1 overexpression promotes HCC progression by regulating miR-130a-5p/CCNT2 axis, which may serve as an effective nanotherapeutic target for HCC treatment.

[Effect of Flax lignans on apoptosis of growth plate chondrocytes in rats].

OBJECTIVE: To compare the effects of different concentrations of linolenin on inhibiting apoptosis of chondrocytes in the growth plate, and to screen the optimal concentration of linolenin, so as to provide theoretical support for delaying epiphyseal closure and promoting long bone growth in rats. METHODS: Two 4-week-old male SD rats (SPF grade) with a body mass of 80 g were selected. The growth plate cartilage of rat tibia and femur was dissected and isolated in vitro to obtain growth plate chondrocytes for culture. The chondrocytes were observed and identified by inverted phase contrast microscope and typeⅡ collagen immunofluorescence test, and then 20 ng/ml IL-1ß was used to induce apoptosis of growth plate chondrocytes as model group, and added with 1, 10, 20, 40 µM linolenin as the experimental group, and 5 µM letrozole as the positive control group. The cells were cultured for 24 and 48 hours respectively. The drug promoted cell proliferation was observed by MTT method, and the drug inhibited cell apoptosis was detected by flow cytometry. RESULTS: Contents 1, 10, 20, 40 µM could promote cell proliferation in varying degrees, and the principle was that the drug inhibits IL-1ß induced chondrocyte apoptosis in the growth plate, and the optimal concentration of drugs to inhibit apoptosis was 20 µM. CONCLUSION: The appropriate concentration of linseed lignans can significantly inhibit the apoptosis of chondrocytes in the growth plate of rats, and the optimal drug concentration is 20 µM. It provides possibility for delayed bone closure and longer growth time to promote bone growth during development.

The therapeutic prospects of N-acetylgalactosamine-siRNA conjugates.

RNA interference has become increasingly used for genetic therapy following the rapid development of oligonucleotide drugs. Significant progress has been made in its delivery system and implementation in the treatment of target organs. After a brief introduction of RNA interference technology and siRNA, the efficiency and stability of GalNAc-siRNA conjugates are highlighted since several oligonucleotide drugs of GalNAc have been approved for clinical use in recent years. The structure and features of GalNAc-siRNA conjugates are studied and the clinical efficiency and limitations of oligonucleotide-based drugs are summarized and investigated. Furthermore, another delivery system, lipid nanoparticles, that confer many advantages, is concluded, includ-ing stability and mass production, compared with GalNAc-siRNA conjugates. Importantly, developing new approaches for the use of oligonucleotide drugs brings hope to genetic therapy.

Insight Into the Prospects for RNAi Therapy of Cancer.

RNA interference (RNAi), also known as gene silencing, is a biological process that prevents gene expression in certain diseases such as cancer. It can be used to improve the accuracy, efficiency, and stability of treatments, particularly genetic therapies. However, challenges such as delivery of oligonucleotide drug to less accessible parts of the body and the high incidence of toxic side effects are encountered. It is therefore imperative to improve their delivery to target sites and reduce their harmful effects on noncancerous cells to harness their full potential. In this study, the role of RNAi in the treatment of COVID-19, the novel coronavirus disease plaguing many countries, has been discussed. This review aims to ascertain the mechanism and application of RNAi and explore the current challenges of RNAi therapy by identifying some of the cancer delivery systems and providing drug information for their improvement. It is worth mentioning that delivery systems such as lipid-based delivery systems and exosomes have revolutionized RNAi therapy by reducing their immunogenicity and improving their cellular affinity. A deeper understanding of the mechanism and challenges associated with RNAi in cancer therapy can provide new insights into RNAi drug development.

Enhancement of oral bioavailability of magnolol by encapsulation in mixed micelles containing pluronic F127 and L61.

OBJECTIVES: We aimed to prepare novel magnolol-loaded mixed micelles (MAG-M) by pluronic F127 and L61 to overcome the challenges of magnolol's poor solubility and then further improve its oral bioavailability. METHODS: Magnolol-loaded mixed micelles containing pluronic F127 and L61 were prepared by an organic solvent evaporation method. Physicochemical, transport experiment across Caco-2 cell monolayers and pharmacokinetic studies were performed to characterize MAG-M and to determine the final improvement of the oral bioavailability. KEY FINDINGS: The MAG-M solution was transparent and colourless with average size, polydispersity index and zeta potential of 228.0 ± 2.1 nm, 0.298 ± 0.012 and -0.89 ± 0.02 mV. The micelle solution has a higher EE% and DL% of 81.57 ± 1.49% and 27.58 ± 0.53%, respectively. TEM result showed that the morphology of MAG-M was homogeneous and spherical shape. The dilution stability of MAG-M was no significant change in particle size and entrapment efficiency. MAG was demonstrated a sustained-release behaviour after encapsulated in micelles. MAG permeability across a Caco-2 cell monolayer was enhanced, and the pharmacokinetics study of MAG-M showed a 2.83-fold increase in relative oral bioavailability compared with raw MAG. CONCLUSIONS: The mixed micelles containing pluronic F127 and L61 as drug delivery system provided a well strategy for resolving the poor solubility and bioavailability problems of MAG.

Therapeutic effect of minimally invasive decompression combined with impaction bone grafting on osteonecrosis of the femoral head.

The aim of this study was to observe the therapeutic effect of minimally invasive decompression combined with impaction bone grafting on osteonecrosis of the femoral head. A total of 67 patients underwent minimally invasive lightbulb decompression combined with impaction bone grafting. The therapeutic effect was evaluated according to Harris scores, and fluoroscopic and magnetic resolution imaging results at different time points. The Harris score was significantly increased after operation. The fineness rate was 85.4%. Postoperative disease progression was found in nine patients with the progression rate of 14.63%. The average necrotic area percentage was noticeably reduced 6 months, 1 year, and 2 years after operation. The one-year postoperative percentage showed a significant difference compared with the preoperative one. Minimally invasive lightbulb decompression combined with impaction bone grafting can achieve a satisfactory curative effect on ONFH. This method has the advantages of small trauma, thorough decompression, and good bone implantation.

[Internal fixation of spinal column by posterior approach and extraperitoneal bilateral focal debridement for treatment of tuberculosis of lumbosacral vertebral body].

OBJECTIVE: To evaluate the application value of internal fixation of spinal column by posterior approach and extraperitoneal bilateral focal debridement for the treatment of tuberculosis of lumbosacral vertebral body. METHODS: From March 2000 to February 2005, 16 cases of spinal tuberculosis in L3-S1 were treated with internal fixation of spinal column by posterior approach and extraperitoneal bilateral focal debridement. Sixteen cases included 11 males and 5 females, and the age was 21-56 years. The locations of spinal tuberculosis were L3 in 4 cases, L4 in 6, Ls in 4, and S1 in 2. The course of disease averaged 13 months (range 6 months to 6 years). The ESR of all cases was greater than 20 mm/h (average 40 mm/h); WBC was normal in 14 cases, and a little high in 2 cases. The X-ray picture showed narrow intervertebral space in 5 cases, compressed body of vertebra in 7 cases, and destroyed 2 consecutive vertebra and its sclerotin was conduplicate in 1 case. All cases were abscess in major psoas muscle. The CT showed destruction of bone, abscess-formation and dead bone in body of vertebra of 16 cases. The abscess were found in vertebral canal in 5 cases and dura mater of spinal cord and spinal nerve root crushed in 2 cases. The MRI showed destruction of bone, abscess-formation and hibateral abscess in major psoas muscle in 11 cases. The abscess were found in vertebral canal and dura mater of spinal cord and spinal nerve root crushed in 6 cases. The therapy of paid was treated after operation in all the cases. RESULTS: Incision healed by first intention in 16 cases; and disruption of wound occurred and healing was achieved after symptomatic treatment in 1 case. Sixteen cases were followed up 2-5 years (29 months on average). Among all the cases, anaesthesia in double thigh was found in 3 cases, adynamia of dorsiflexion in foot in 1 case, gatism in 1 case; after 2 weeks the symptoms were vanished. Indirect hernia of fold inguen were found in 1 case after 2 years, and the patient refused operation for age and was no longer followed up. Fuzzy, ensic and acerb in eyes were found in 6 cases, hearing disturbance were found in 2 cases, and symptoms were vanished after medication adjustment. Low-grade fever and lumbar myalgia were found in 1 case and cured after staying in bed and medication adjustment. All ESR were normal, synostosis of lumbosacral vertebral body was found in 13 cases, kyphosis in 1 case. CONCLUSION: Internal fixation of spinal column by posterior approach and extraperitoneal bilateral focal debridement is safe and available way for the treatment of tuberculosis of lumbosacral vertebral body, it can save the times of operation, shorten period and enhance effect of treatment.

[Repair of defects in chest wall with thoracico-abdominal skin flap and muscle flap of musculus rectus abdominis].

OBJECTIVE: To evaluate the application value of repairing the defects of the chest wall with the thoracico-abdominal skin flap and the muscle flap of the musculus rectus abdominis. METHODS: From January 2002 to June 2005, five patients with defects in the chest wall underwent the prosthesis with the thoracico-abdominal skin flap and the muscle flap of the musculus rectus abdominis under general anesthesia. Focal cleaning was performed first; then, the skin flap was designed and taken (15 cm x 10 cm); and finally, the defects of the chest wall were repaired with the muscle flap of the musculus rectus abdominis. RESULTS: Of the 5 patients, 4 had the flap healing by the first intention, and 1 had the delayed healing, with no complication. The skin flap had a good appearance, without edema or pigmentation. The X-ray examination showed that the shadow of the sternal sequestrum disappeared. There was no recurrence or complication during the follow-up for 1-3 years (average, 18 mon). CONCLUSION: The repairing of the defects in the chest wall with the thoracico-abdominal skin flap and the muscle flap of the musculus rectus abdominis is a simple and effective surgical treatment for defects of the chest wall around the sternum, and this kind of treatment is worth applying extensively in clinical practice.