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Extended circulation of anticancer nanodrugs in blood stream is essential for their clinical applications. However, administered nanoparticles are rapidly sequestered and cleared by cells of the mononuclear phagocyte system (MPS). In this study, we developed a biomimetic nanosystem that is able to efficiently escape MPS and target tumor tissues. The fabricated nanoparticles (TM-CQ/NPs) were coated with fibroblast cell membrane expressing tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL). Coating with this functionalized membrane reduced the endocytosis of nanoparticles by macrophages, but increased the nanoparticle uptake in tumor cells. Importantly, this membrane coating specifically induced tumor cell apoptosis via the interaction of TRAIL and its cognate death receptors. Meanwhile, the encapsulated chloroquine (CQ) further suppressed the uptake of nanoparticles by macrophages, and synergized with TRAIL to induce tumor cell apoptosis. The vigorous antitumor efficacy in two mice tumor models confirmed our nanosystem was an effective approach to address the MPS challenge for cancer therapy. Together, our TM-CQ/NPs nanosystem provides a feasible approach to precisely target tumor tissues and improve anticancer efficacy.
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RNA therapeutics inhibit the expression of specific proteins/RNAs by targeting complementary sequences of corresponding genes, or synthesize proteins encoded by the desired genes to treat genetic diseases. RNA-based therapeutics are categorized as oligonucleotide drugs (antisense oligonucleotides, small interfering RNA, RNA aptamers), and mRNA drugs. The antisense oligonucleotides and small interfering RNA for treatment of genetic diseases have been approved by the FDA in the United State, while RNA aptamers and mRNA drugs are still in clinical trials. Chemical modifications are applied to RNA drugs, such as pseudouridine modification of mRNA, to reduce immunogenicity and improve the efficacy. The secure and effective delivery systems like lipid-based nanoparticles, extracellular vesicles, and virus-like particles are under development to address stability, specificity, and safety issues of RNA drugs. This article provides an overview of the specific molecular mechanisms of 11 RNA drugs currently used for treating genetic diseases, and discusses the research progress of chemical modifications and delivery systems of RNA drugs.
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Objective:To investigate the clinical efficacy of induced membrane technique in the staged treatment of adult chronic hematogenous osteomyelitis (CHOM) of long bone.Methods:The clinical data were retrospectively analyzed of the 22 adult patients with CHOM of long bone who had been admitted to the 920th Hospital, Joint Logistics Support Force of PLA from January 2016 to December 2019. There were 18 males and 4 females, aged from 16 to 56 years (average, 31.81 years). Their disease duration ranged from 0.6 to 42.0 years, averaging 18.4 years. By the Cierny-Mader anatomical classification, 4 cases were type Ⅰ, 6 cases Type Ⅲ, and 12 cases type Ⅳ. In the first stage, the bone defects were filled with antibiotic bone cement after thorough debridement. In the second stage when the infection had been controlled, the bone defects were repaired with bone grafts after removal of the bone cement. Bone healing time and complications were followed up. The treatment effects were evaluated by comparisons of the infection control indexes [including clinical manifestations like local redness, swelling, pus, and pain, and blood white blood cell count, C-Reactive protein (CRP), and erythrocyte sedimentation rate (ESR) as well] before the primary surgery, before the secondary surgery and at the last follow-up.Results:The volumes of the bone defects after stage-one debridement ranged from 54 cm 3 to 176 cm 3 (mean, 90.9 cm 3). All patients were followed up for 20 to 51 months (mean, 30.1 months) after surgery. All bone defects healed after 4 to 11 months (mean, 6.6 months). Postoperatively, infection developed at the bone extraction site of the posterior superior iliac spine in 3 cases and pain was observed at the donor site in one case, but the conditions were relieved after symptomatic treatment. Fracture and plate breakage occurred at the bone defect site in one case who had fallen down 7 months after operation, but responded to reoperation. The last follow-up revealed such symptoms as redness, swelling and pus discharge in none of the patients. The white blood cell count [(5.70 ± 1.57) × 10 9/L and (5.65 ± 1.58) × 10 9/L], CRP [(7.56 ± 2.57) mg/L and (7.25 ± 3.83) mg/L] and ESR [(9.64 ± 2.90) mm/h and (10.55 ± 5.23) mm/h] before the secondary surgery and at the last follow-up were significantly lower than those before the primary surgery [(8.24 ± 2.18) × 10 9/L, (49.54 ± 19.56) mg/L, and (42.68 ± 13.77) mm/h] (all P < 0.05). However, there were no significant differences between the indexes before the secondary surgery and at the last follow-up ( P > 0.05). Conclusion:In the staged treatment of adult CHOM of long bone, the induced membrane technique can effectively control infection, achieve repair of bone defects, and reduce complications.
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Hippo signaling pathway is highly conservative in evolution. MST1/2, LATS1/2, and the effector protein YAP/TAZ are the core members of this signaling pathway in mammalian cells. There have been many studies on YAP/TAZ and its downstream, however, the upstream regulatory factors of the Hippo signaling pathway remain unclear, and become one of the hot research directions of this pathway at present. In addition, Hippo signaling pathway can cross-talk with other signaling pathways such as Wnt and Notch signaling pathways, and plays an important role in controlling organ size, maintaining tissue homeostasis, and promoting tissue repair and regeneration. Abnormal Hippo signaling pathway may lead to the occurrence of a variety of tumors, especially gastrointestinal cancers such as liver cancer, colorectal cancer and gastric cancer. The abnormal expression of its members in gastrointestinal cancers is related to cancer cell proliferation, apoptosis, invasion and migration. Hippo signaling pathway is vital for liver repair and regeneration. Its inactivation will lead to the occurrence of primary liver cancer. The mechanism of YAP in liver cancer mainly depends on TEAD-mediated gene transcription. Hippo signaling pathway is also important for maintaining intestinal homeostasis, and its imbalance can lead to the occurrence and recurrence of colorectal cancer. In primary and metastatic gastric cancer, the expression of YAP/TAZ is significantly up-regulated, but the specific molecular mechanism is unclear. This article summarizes the recent progress on Hippo signaling pathway and its upstream regulatory factors, its roles in the development of gastrointestinal cancers and related molecular mechanisms; and also discusses the future research directions of Hippo signaling pathway.
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Humanos , Apoptosis , Fisiología , Proliferación Celular , Fisiología , Neoplasias Gastrointestinales , Transducción de SeñalRESUMEN
OBJECTIVE： To establish a method for content determination of total polyphenols from Gastrodia elata， and to optimize the purification technology of macroporous resin. METHODS： The content of total polyphenols from G. elata was determined by Folin-ciocaileu colorimetry. Using the absorption and desorption performance as index， 4 kinds of macroporous adsorption resins were selected by static adsorption and desorption tests. The flow rate and mass concentration of the sampling solution， volume fraction of eluent， eluent flow rate and eluent volume were investigated by dynamic adsorption and desorption tests. The purification technology of macroporous resin was optimized. RESULTS： The linear range of gallic acid was 4-32 μg/mL （r＝0.999 9）. RSDs of precision， stability and repeatability tests were all less than 2％. The recovery rate of the sample was 95.51％-102.94％（RSD＝2.54％，n＝6）. D301 macroporous resin had strong static adsorption and desorption ability from G. elata polyphenols. The optimal purification technology included that the sample solution flow rate 2 BV/h； the sample solution mass concentration 4 mg/mL； the elution solvent 70％ ethanol； the elution flow rate was 3 BV/h， and the eluent volume 5 BV. The content of total polyphenols from G. elata optimized by the optimal purification technology was 0.381 mg/g. CONCLUSIONS： Established method is sensitive and stable. The optimized purification technology is stable and feasible.
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Objective To evaluate surgical treatment of chronic tibial osteomyelitis of Cierny-Mader type Ⅳ with Ilizarov technique and lesion osteotomy. Methods From January 2010 to May 2016, 39 patients with chronic tibial osteomyelitis of Cierny-Mader type Ⅳ were treated at our center. They were 33 males and 6 females, 8 to 54 years of age (average, 33.8 years). After debridement and lesion osteotomy, the tibia was fixated with Ilizarov external fixator. Bone was transported to the bone defect after corticotomy was performed on the proximal and/or distal tibial metaphyses simultaneously. Bifocal corticotomy was per-formed in 11 cases, proximal corticotomy in 21 cases, and distal corticotomy in 7 cases. The transport began 3 to 5 days after operation at a speed of 0.5 to 1.0 mm/d initially. The speed was lowered according to the bone healing and pain. Radiographic examination was done every 2 weeks to observe transporting deviation and osteogenesis in the transporting area. The transporting was adjusted whenever any abnormality was observed. The bone transporting lasted for 50 to 130 days (average, 62.4 days). Results The patients were fol-lowed up for 11 to 49 months (average, 21 months). All the soft tissue wounds healed uneventfully and there was no relapse of osteomyelitis. The bone defects in the 32 cases were reconstructed primarily. Nonunion of fracture ends happened in 5 cases and nonunion of the bone lengthening zone in 2 cases. The 7 cases of nonunion were healed after secondary bone grafting. Malalignment happened in 5 cases, 4 of which responded to timely adjustment of the external fixation and one of which had to receive secondary bone grafting after failure in adjustment of the external fixation. Ankle joint dysfunction occurred in 7 cases, 5 of which re-sponded to functional exercise and 2 of which accepted joint dysfunction because they refused surgery after unsatisfactory functional exercise. Pin tract infection of different severities occurred in 9 cases, one of which was treated by replacement of the K-wires under local anesthesia and the other 8 of which responded to rein-forced dressing change. Conclusions Chronic tibial osteomyelitis of Cierny-Mader typeⅣcan be treated by Ilizarov technique and lesion osteotomy. However, the Ilizarov technique should be improved because of the risks of multiple complications which can be reduced significantly by strengthening postoperative instruction, nursing, and regular follow-up.
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Objective To investigate the chondrogenic feasibility of the human umbilical cord derived mesenchymal stem cells (hUCMSCs)as cartilage tissue engineering seed cells ,type Ⅱ collagen composite glycosaminoglycan scaffold as the cellular carrier and cell‐scaffold complex .Methods The type Ⅱ collagen composite glycosaminoglycan scaffolds was prepared .The pore diameter , porosity and hydrophilia of scaffold materials were observed and measured by electronic microscope .The corresponding histological analysis on the scaffold materials was performed .hUCMSCs of P3 generation were cultured and identified .The hUCMSCs suspen‐sion was inoculated in the type Ⅱ collagen composite glycosaminoglycan scaffold for conducting culture without adding inducer .The samples were taken out after 3 weeks and performed the toluidine blue and safranin O staining ,type Ⅱ collagen immunohistochemi‐cal staining and SEM scanning .Results hUCMSCs of P3 generation highly expressed the mesenchymal cell marker CD29 and CD105 ,while hardly expressed endothelial cells of CD34 and hematopoietic cell markers .The type Ⅱ collagen composite glycosami‐noglycan scaffold presented white porous foam like ,the porosity was (91 .8 ± 2 .17)% ,the average pore diameter was 110‐230 μm , which was homogeneously distributed and had interpenetration .The scaffold showed good hydrophilicity with the water absorption expansion rate of (213 .71 ± 1 .31)% .The scaffold staining of toluidine blue ,safranin O and type Ⅱ collagen was positive .The car‐tilage‐like tissues were observed ,and gradually increased in the surface of cell‐scaffold complex along with culture ,which were posi‐tive in Toluidine blue ,safranin O and type Ⅱ collagen staining ,the electronic microscopic observation displayed that the cells were actively proliferated in the scaffold ,closely adhered with the materials ,the cartilage‐like cells and a large number of peripheral colla‐gen fibers with zigzag connection could be seen .Conclusion Compositing hUCMSCs and type Ⅱ collagen composite glycosamin‐oglycan scaffold could construct tissue‐engineering cartilage in vitro without induction ,which lays a certain experimental foundation for the repair of cartilage damage .
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BACKGROUND:In animal experiments, transplantation of autologous nucleus pulposus cellscan effectively repair the intervertebral disk degeneration. However, nucleus pulposus cells have a poor ability of proliferation in vitro, which limits its application as seed cells in treatment of intervertebral disk disease. OBJECTIVE:To construct recombinant adeno-associated virus type-2 vector carrying human telomerase reverse transcriptase and observe the human telomerase reverse transcriptase mRNA expression in human nucleus pulposus cells in vitro. METHODS:After the plasmid pSNAV2.0-pRSV-hTERT was constructed and identified, recombinant adeno-associated virus type-2 vector carrying human telomerase reverse transcriptase were constructed, amplified and purified by AAVMaxTM package and purification system. The optimal multiplicity of infection for human nucleus pulposus cells was detected by recombinant adeno-associated virus type-2 vector carrying enhanced green fluorescent protein. According the optimal multiplicity of infection (5 × 104 v·g/cell), three different multiplicity of infection (1×104, 5×104, 1×105 v·g/cell) of recombinant adeno-associated virus type-2 vector carrying human telomerase reverse transcriptase were determined to transfect the first passage human nucleus pulposus cells in vitro. In control group, the cells were transfected with adeno-associated virus type-2 vector without human telomerase reverse transcriptase. At 1, 2, 4 weeks after transfection, mRNA expression of human telomerase reverse transcriptase in human nucleus pulposus cells were semi-quantitatively detected by RT-PCR. RESULTS AND CONCLUSION:The recombinant adeno-associated virus type-2 vector carrying human telomerase reverse transcriptase was successful y constructed, and the titer of the obtained vector was more than 2×1011 v·g/mL. The optimal multiplicity of infection was 5×104 v·g/cell. The mRNA expression of human telomerase reverse transcriptase in human nucleus pulposus cells could be detected in different multiplicity of infection (1×104, 5×104, 1×105 v·g/cell). At 2 weeks post-transfection, mRNA expression of human nucleus pulposus cells was the highest (P<0.05), as detected by semi-quantitative RT-PCR. Moreover, the stable and high mRNA expression of human telomerase reverse transcriptase could be detected at 4 weeks post-transfection. In control group, no human telomerase reverse transcriptase mRNA expression was found. The recombinant adeno-associated virus type-2 vector carrying human telomerase reverse transcriptase can be successful y constructed, and can mediate a stable mRNA expression of human telomerase reverse transcriptase in human nucleus pulposus cells. Our findings provide a novel strategy of enhancing the properties of nucleus pulposus cells.
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[Objective]To investigate the effect of selectively segmental cervical decompression through anterior approach in the treatment of senile segmental cervical spondylotic myelopathy. [Methods]Twenty-seven cases of senile segmental cervical spondylotic myelopathy experienced selectively segmental cervical decompression through anterior approach from 2004 to 2008 were reviewed retrospectively. Pre-and post-operative JOA scores were compared to evaluate the improvement of neurological function.[Results]Patients were followed up for an average of 19.7 months (ranged,18~27 months). No loosening or displacement was found in all cases. Postoperatively,ratio of JOA improvement was 56.1%,including excellent result in 8 cases (29.5%),good result in 11 cases (40.7%),fair result in 6 cases (22.2%),poor result in 2 cases (7.4%).[Conclusion]Selectively segmental cervical decompression through anterior approach is effective in the treatment of senile segmental cervical spondylotic myelopathy,giving consideration to patient's general physical condition and recovery of neurological function. Correct segments selection of preoperative decompression is the key point of result of postoperative restoration.
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Objective To analyze the simple sequence repeat(SSR)information in expressed sequence tag(EST)resource of Saruma henryi and lay a solid foundation for the development of EST-SSR markers in this species.Methods ESTs of S.henryi were downloaded from GenBank and used to perform the contig assembly using Sequencher 4.8.Uni-ESTs were obtained and screened for SSR-containing unigenes using SciRoKo 3.4.The distributing frequency of the EST-SSRs and the basic characteristics of motifs were analyzed.Results A total of 10 274 ESTs of S.henryi were retrieved and were assembled into 6 643 non-redundant Uni-ESTs with a total length of 5.11?106 bp.In all,the data mining yielded 1 408 SSR loci,which corresponded to 1 232 Uni-ESTs(18.55%).On average,EST-SSRs spanned 22.30 bp,and occurred every 3.63 kb in length.In S.henryi,mononucleotide repeats predominated with an occurrence frequency of 12.24%.Dinucleotide repeats followed with a frequency of 5.01%.The most frequent one was A/T among all the repeat motifs,then followed by AG/CT.Conclusion SSRs in ESTs of S.henryi display a relatively high level of occurrence frequency and show abundance of types.