摘要
ObjectiveTo investigate the induction of ferroptosis by polyphyllin Ⅱ (PPⅡ) in hepatocellular carcinoma HepG2 cells and its underlying mechanism. MethodThe effect of PPⅡ (0, 1.5, 3.0, 4.5, 6.0, 9.0, 18.0 mg·L-1) on the in vitro proliferation of HepG2 cells was assessed using the methyl thiazolyl tetrazolium (MTT) assay. Colony formation ability of HepG2 cells was evaluated through a colony formation assay. Cell migration ability was assessed via a scratch assay. Lactate dehydrogenase (LDH) content in HepG2 cells was measured using a kit. Reactive oxygen species (ROS) levels in HepG2 cells were observed using a fluorescence inverted microscope. Malondialdehyde (MDA), glutathione (GSH), and free Fe2+ content in HepG2 cells were detected using respective kits. The mitochondrial ultrastructure in HepG2 cells was observed by transmission electron microscopy. The expression of ferroptosis-related proteins p53, solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), long-chain acyl-CoA synthetase 4 (ACSL4), and transferrin receptor 1 (TFR1) in HepG2 cells was detected using Western blot. ResultCompared with the control group, the PPⅡ treatment groups showed significantly decreased survival rate of HepG2 cells in a dose-dependent manner (P<0.01), significantly reduced number of cell colonies (P<0.01), significantly shortened scratch healing distance, inverse correlation of the migration distance with drug concentration (P<0.01), significantly increased LDH leakage in cells (P<0.01), significantly enhanced relative fluorescence intensity of intracellular ROS, and significantly increased accumulation of lipid peroxide MDA (P<0.01), decreased intracellular GSH content with increasing drug concentration (P<0.01), and significantly enhanced fluorescence intensity of FeRhoNox-1 in cells (P<0.01). Moreover, cells exhibited vacuolation, and mitochondria showed significant shrinkage with reduced or even disappeared cristae. Compared with the results in the control group, the expression of p53, ACSL4, and TFR1 proteins significantly increased, while the expression of SLC7A11 and GPX4 proteins significantly decreased in the PPⅡ treatment groups (P<0.05). ConclusionIn summary, PPⅡ induces ferroptosis in HepG2 cells by regulating the p53/SLC7A11/GPX4 signaling axis, promoting ACSL4 expression and Fe3+ uptake, leading to an imbalance in the antioxidant system.
摘要
Ferroptosis, a new form of programmed cell death different from apoptosis, necrosis, and autophagy, is closely associated with a variety of physiological and pathological processes. Iron-mediated accumulation of reactive oxygen species is the main inducement of ferroptosis, the mechanism of which is related to intracellular lipid metabolism, iron metabolism, and antioxidant defense pathways. Multiple signaling axes and regulators jointly regulate the occurrence and disruption of ferroptosis. Studies have demonstrated that ferroptosis regulates the growth and proliferation of tumor cells. Inducing ferroptosis in tumor cells can control the growth, metastasis, and multi-drug resistance of tumors. Therefore, the effect and mechanism of ferroptosis on tumor cells have become a hot topic in anti-cancer research. As the research advances, a variety of ferroptosis inducers has been used in the clinical chemotherapy for cancers and demonstrate significant efficacy. Accordingly, the development of ferroptosis-inducing anticancer drugs has become a new research direction for tumor treatment. Some active ingredients such as lycorine, oleanolic acid, dihydroartemisinin, pseudolaric acid B, and ophiopogonin B of Chinese medicines can induce ferroptosis in tumor cells via lipid metabolism, iron metabolism, system Xc-, and GPX4/GSH to regulate the development of tumors, demonstrating a promising prospect in clinical treatment. Based on the theory of the mechanism of ferroptosis, this paper reviews the research progress in ferroptosis induced by active ingredients of Chinese medicines in tumor cells and describes the metabolic regulatory network of ferroptosis from signaling pathways and regulatory factors, providing new strategies for applying active ingredients of Chinese medicines in the treatment of tumors.
摘要
Paridis Rhizoma possesses the functions of clearing heat and detoxifying, alleviating swelling and relieving pain, cooling the liver and calming the convulsion. Saponins are the main active components of Paridis Rhizoma. Studies have shown that total saponins in Paridis Rhizoma have obvious inhibitory effect on solid tumors such as breast cancer, lung cancer, gastric cancer, and liver cancer and non-solid tumors such as leukemia. The saponins may exert the anti-tumor effects by inhibiting the proliferation, migration, and invasion of tumor cells, regulating cell cycle, inducing apoptotic and non-apoptotic death pathways, and regulating metabolism and tumor microenvironment. Furthermore, total saponins in Paridis Rhizoma showed anti-inflammatory, antioxidant, antimicrobial, hemostatic, and uterus-contracting activities. At the same time, they may induce apoptosis of normal cells, inflammation and oxidative stress, and metabolic disorders. In recent years, the reports of liver injury, reproductive injury, gastrointestinal injury, hemolysis, and other adverse reactions caused by total saponins in Paridis Rhizoma have been increasing. Pharmacokinetic studies have shown that there are significant differences in the metabolism of total saponins in Paridis Rhizoma administrated in different ways. Injection has a fast clearance rate, while oral administration may have hepatoenteric circulation. Meanwhile, due to the low solubility and activation of P-glycoprotein (P-gp) molecular pump, the prototype absorption, intestinal permeability, and recovery rate of total saponins in Paridis Rhizoma are poor, which affects the bioavailability. The bioavailability can be improved to some extent by preparing new dosage forms or new drug delivery systems with advanced technology. This paper reviews the pharmacological effect, pharmacokinetics, and adverse reactions of Rhizoma Paridis total saponins by searching the China National Knowledge Infrastructure (CNKI), VIP, and Web of Science with ''Rhizoma Paridis total saponins'' as the keywords, hoping to provide references for the research, development, and clinical application of such components.
摘要
Paridis Rhizoma, a traditional valuable Chinese herbal medicine, has the functions of clearing heat and removing toxin, relieving edema and pain, cooling liver and calming convulsion, which can be used to treat various diseases such as mumps, abscess, burn, bleeding, and tumor. It has been used in folk medicine for a long time and is the main raw material of various Chinese patent medicines such as Gongxuening Capsules and Yunnan Baiyao. Polyphyllin Ⅰ, an isospirostanol saponin and one of the main active components in Paridis Rhizoma, is distributed in the rhizome, pericarp, and leaves of Paris polyphylla. With high polarity, polyphyllin Ⅰ is mainly extracted by n-butanol extraction and macroporous adsorption resin chromatography, separated by silica gel column chromatography and preparative high performance liquid chromatography, and purified with the combination of methods. With anti-tumor, anti-inflammatory, antibacterial, and anti-virus effects, it is generally employed to treat liver cancer, lung cancer, gastric cancer and other cancers as well as arthritis, influenza, sore toxin, and bacterial infection. However, polyphyllin Ⅰ may cause stomach irritation, hemolysis, liver damage, kidney damage, heart damage, and other adverse reactions. Pharmacokinetic studies show that it has problems such as low bioavailability and poor intestinal absorption and permeability, which affect the clinical application of polyphyllin Ⅰ. This paper summarizes the research on the plant sources, extraction and separation methods, pharmacological effects, adverse reactions, and pharmacokinetics of polyphyllin Ⅰ in recent years, which is expected to provide a reference for the rational clinical application and other in-depth research work of polyphyllin Ⅰ.
摘要
As a rare Chinese medicinal material, Paridis Rhizoma is mainly distributed in Yunnan, Guangxi, and Guizhou in southwestern China, with the effect of clearing heat and detoxifying, alleviating edema and relieving pain, cooling liver and tranquilizing mind. It is particularly effective for injuries from falls, fractures, contusions and strains, snake bites, cold wind-induced convulsion, and other diseases, which has been used for more than 2 000 years. According to modern research, polyphyllin Ⅱ, one of the main active components of Paridis Rhizoma, belongs to diosgenin in structure. It has the anti-tumor, anti-inflammatory, antiviral, antibacterial, immune-regulating, antioxidant, and multidrug resistance-reversing activities, showing good application prospect. Especially, the anti-tumor effect of polyphyllin Ⅱ has attracted wide attention, and the mechanism is inhibiting proliferation, migration, and invasion of tumor cells, inducing cell cycle arrest, apoptosis, and autophagy, suppressing angiogenesis, and modulating tumor microenvironment. However, the pharmacokinetic results show that polyphyllin Ⅱ has low bioavailability in vivo due to the low solubility, poor absorption, unsatisfactory distribution, and slow metabolism, which limit the clinical application. In recent years, there has been an explosion of research on the adverse reactions of polyphyllin Ⅱ, such as the strong hemolytic activity and obvious cytotoxicity to liver, kidney, myocardium and cardiovascular cells. Thus, papers were retrieved from "CNKI", "VIP", "Wanfang Data", "PubMed", "Web of Science", and "Elsevier SD" with "Paris saponin Ⅱ", "Polyphyllin Ⅱ" as the main keywords, and the pharmacological activities and mechanisms, pharmacokinetics, and adverse reactions were summarized. The findings are expected to serve as a reference for the in-depth research, development, and utilization of polyphyllin Ⅱ.
摘要
As arsenic widely exists in nature and has been used in the pharmaceutical preparations, the traditional Chinese medicine(TCM) with arsenic include realgar(As_2S_2 or As_4S_4), orpiment(As_2S_3), and white arsenic(As_2O_3). Among the above representative medicine, the TCM compound formulas with realgar are utilized extensively. Just in Chinese Pharmacopoeia(2020 edition), there are 37 Chinese patent medicines including realgar. The traditional element analysis focuses on the detection of the total amount of elements, which neglects the study on the speciation and valence of elements. The activity, toxicity, bioavailability, and metabolic pathways of arsenic in vivo are closely related to the existence of its form, and different forms of arsenic have different effects on organisms. Therefore, the study on the speciation and valence of arsenic is of great importance for arsenic-containing TCMs and their compound formulas. This paper reviewed four aspects of the speciation and valence of arsenic, including property, absorption and metabolism, toxicity, and analytical assay.
Subject(s)
Arsenic/analysis , Arsenicals/analysis , Sulfides , Arsenic Trioxide , Medicine, Chinese Traditional , Drugs, Chinese Herbal/analysis , Biological Products摘要
Fermented Chinese medicine has long been used. Amid the advance for preservation of experience, the connotation of fermented Chinese medicine has been enriched and improved. However, fermented Chinese medicine prescriptions generally contain a lot of medicinals. The fermentation process is complicated and the conventional fermentation conditions fail to be strictly controlled. In addition, the judgment of the fermentation end point is highly subjective. As a result, quality of fermented Chinese medicine is of great difference among regions and unstable. At the moment, the quality standards of fermented Chinese medicine are generally outdated and different among regions, with simple quality control methods and lacking objective safe fermentation-specific evaluation indictors. It is difficult to comprehensively evaluate and control the quality of fermented medicine. These problems have aroused concern in the industry and also affected the clinical application of fermented Chinese medicine. This article summarized and analyzed the application, quality standards, and the modernization of fermentation technology and quality control methods of fermented Chinese medicine and proposed suggestions for improving the quality standards of the medicine, with a view to improving the overall quality of it.
Subject(s)
Medicine, Chinese Traditional , Reference Standards , Quality Control , Fermentation摘要
Objective:By retrieving published patents on novel coronavirus related technologies, we analyzed the differences in patent applicants, target areas, patent quality, etc. in different countries and regions, and learned from the patent layout of relevant key innovation entities in countries and regions with relatively mature patent systems, such as Europe and the United States, to provide a reference for the subsequent writing and layout of related Chinese patents.Methods:" Novel coronavirus pneumonia, nCov pneumonia, novel coronavirus, nCov, COVID-19, 2019-nCoV, SARS-CoV-2" were searched as key words in the PatSnap Patent Database, with a limitation of public date before September 30, 2020. Based on the bibliometric method, this study analyzed applicants, regional distribution, patent quality, patent subject, etc.Results:4 541 patents on novel coronavirus related technology were obtained through search and filtering, whose applicants are mainly from China, which is also the major, region of applications. However, in an analysis of patent quality regarding novel coronavirus related technologies, the number of claims of US patent applications reached 44.48, and the number of patents in the same family reached 5.21, while in China, the numbers were just 9.27 (claims) and 1.45 (patents of the same family).Conclusions:China outperformed other countries in terms of the number of patent applications for novel coronavirus related technologies, but fell rather far behind the US in terms of patent quality and layout. Therefore, it is necessary to introduce and train professional patent management talents, improve the patent management system, raise awareness of patent legal protection, write high-quality legal documents, attach importance to the global market of patent technology, and strengthen the internationalization of patent layout, further promote industry-university-research cooperation, strengthen exchanges and cooperation among research institutions, pharmaceutical enterprises, hospitals and other institutions, pay attention to the combination of scientific research and clinical, prevention and control practices, and promote the healthy development of coronavirus research.
摘要
Thalidomide and its derivatives have been used in the treatment of myelodysplastic syndrome (MDS) because of their anti-angiogenic and immunomodulatory effects. In recent years, some studies have found that thalidomide and its derivatives not only showed significant efficacy in lower-risk MDS patients with del (5q), but also showed advantages in non-del (5q) MDS patients. In addition, the discovery of its molecular targets and new substrates makes it possible to develop a new generation of immunomodulatory drugs (IMiDs) and to design IMiDs-based proteolysis-targeting chimeras. In this review, the new progress in mechanism and clinical application of thalidomide and its derivatives were summarized briefly, so as to provide a more scientific, reasonable and effective scheme to the treatment of MDS.
Subject(s)
Humans , Immunomodulating Agents , Myelodysplastic Syndromes/drug therapy , Thalidomide/therapeutic use摘要
This review made a systematic textual research on the historical evolution and changes of the origin, producing areas and quality, harvest time and processing methods, and cultivation technique of Poria in famous classical formulas from the Catalogue of Ancient Famous Classical Formulas (the First Batch) by referring to the literature of ancient materia medica and medical books combining with the modern literature in CNKI, Wanfang Data and other databases, which could provide reference for the development and research of the famous classical formulas containing Poria. According to the research, the origin of Poria in ancient times was the dried sclerotia of Poria cocos. The producing area of Poria is widely distributed in China, mainly produced in Anhui, Hubei, Henan and Yunnan provinces. The quality of Poria from Yunnan is better, and the yield is larger in Anhui and Hubei provinces. The quality evaluation of Poria is basically the same from ancient to modern, which has the characteristics of large block, heavy weight, thin and wrinkled skin, and white and delicate cross section. The harvesting period of Poria is usually in August of the lunar calendar. The initial processing has undergone the evolution from drying in shade in the ancient time to drying in shade after sweating in the modern time. From ancient times to the present, Poria has been processed by cutting to prepare the Fuling lumps and Fuling slices. The cultivation technique has experienced the evolution process of "wild Poria-inoculation of live pine root-basswood cultured". At present, Poria is mainly cultivated by artificial basswood with mature technical methods and abundant yields, which can meet the research and development needs of the famous classical formulas.
摘要
Objective: To investigate the variation regularity and influencing factors of cortical auditory evoked potential (CAEP) evoked by pure tone, syllable and tone stimuli in cochlear implant (CI) children. Methods: Cortical auditory evoked potential (CAEP) responses were collected from 46 CI children in the sound field. Pure tones with frequencies of 1 kHz and 2 kHz were used as the standard and the deviant respectively in the pure tone stimulation condition. The Chinese Mandarin tokens/ba/-/pa/and/ba1/-/ba4/pairs were used as the stimuli respectively in the syllable and tone stimulation condition. The latency, amplitude and presence rate of P1 and mismatch negative(MMN) were obtained and the correlation between the difficulty of auditory task, the age of hearing month, the duration of severe-profound hearing loss, the wearing history of hearing aid before CI, the hearing threshold of the better ear before CI and the latency and amplitude of P1 and MMN were analyzed. All statistical analyses and figures were conducted using SPSS 25.0. Results: The P1 presence rate of pure tone, syllable and tone group was 100% (17/17), 100% (13/13) and 75.0% (12/16), respectively, with significant difference (χ²=8.214, P=0.016). There was significant difference between pure tone group and tone group (χ²=4.836, P=0.028), but no significant difference between pure tone group and syllable group, syllable group and tone group. The MMN presence rate of pure tone, syllable and tone group was 94.1% (16/17), 84.6% (11/13) and 62.5% (10/16), respectively, but no significant difference among the three groups with different auditory tasks(χ²=0.066, P=0.066). No significant difference was observed among the three groups of different auditory tasks in the latency and amplitude of P1 and MMN. Multiple linear regression analysis showed that the latency of P1 was positively correlated with the difficulty of auditory task and the hearing threshold of the better ear before CI, and negatively correlated with hearing age and the history of hearing aid before CI. The latency of MMN was positively correlated with the difficulty of auditory task, and negatively correlated with hearing age. Conclusions: The P1 presence rate of pure tone auditory task is significantly higher than that of tone auditory task. The difficulty of auditory task, hearing age, the history of hearing aid before CI, and the hearing threshold of the better ear before CI has significant effects on the P1 latency. The difficulty of auditory task and hearing age has significant effects on the MMN latency.
Subject(s)
Child , Humans , Acoustic Stimulation , Cochlear Implantation , Cochlear Implants , Evoked Potentials, Auditory , Hearing , Hearing Aids摘要
Aim To explore the regulatory mechanism of berberine in the high glucose (HG) -induced podocyte ferroptosis. Methods Western blot and R T - qPCR were used to detect the changes of G P X 4, PTGS2, ACSL4, podocin, and desmin at different time (0 h, 6 h, 12 h, 24 h, 36 h) of HG stimulation, and the relative expression of Nrf2, HO-1, GPX4 and podocin under HG after treated with berberine. The proliferation of podocytes stimulated by HG at different time points was observed by EdU staining. The therapeutic effect of berberine on podocyte damage was screened by CCK-8, and the effect of berberine on the level of oxidative stress in HG-induced podocytes were measured by fluorescence inverted microscope, GSH and GSSG kits. In addition, transmission electron microscopy was employed to observe the ultrastructural characteristics of podocytes in each group. Results The expression of podocin and GPX4 was significantly reduced at 24 h, and the mRNA levels of ACSL4 and PTGS2 were significantly up-regulated. Berberine could notably increase the expression of Nrf2, HO-1, GPX4 and podocin, and reduce the levels of PTGS2 and ACSL4. Moreover, berberine significantly improved the levels of ROS and GSH in podocytes under HG conditions, thereby alleviating membrane blistering, mitochondrial shrinkage and other morphological changes of HG-induced podocytes. Conclusions In this study, the number of podocytes decreases, which is a death mode different from autophagy and apoptosis, that is, ferroptosis. Berberine can alleviate the occurrence of this phenomenon by mediating the Nrf2/ H0-1/GPX4.
摘要
OBJECTIVE@#To establish a clinically applicable model of rapid identification of adverse drug reaction program (RiADP) for risk management and decision-making of clinical drug use.@*METHODS@#Based on the theory of disproportion analysis, frequency method and Bayes method, a clinically applicable RiADP model in R language background was established, and the parameters of the model were interpreted by MedDRA coding. Based on the actual monitoring data of FDA, the model was validated by the assessing hepatotoxicity of lopinavir/ritonavir (LPV/r).@*RESULTS@#The established RiADP model included four parameters: standard value of adverse drug reaction signal information, empirical Bayesian geometric mean value, ratio of reporting ratio and number of adverse drug reaction cases. Through the application of R language parameter package "phViD", the model parameters could be output quickly. After being encoded by MedDRA, it was converted into clinical terms to form a clinical interpretation report of adverse drug reactions. In addition, the evaluation results of LPV/r hepatotoxicity by the model were matched with the results reported in latest literature, which also proved the reliability of the model results.@*CONCLUSIONS@#In this study, a rapid identification method of adverse reactions based on post marketing drug monitoring data was established in R language environment, which is capable of sending rapid warning of adverse reactions of target drugs in public health emergencies, and providing intuitive evidence for risk management and decision-making of clinical drugs.
Subject(s)
Humans , Databases, Pharmaceutical , Decision Making, Computer-Assisted , Drug Monitoring , Drug-Related Side Effects and Adverse Reactions , HIV Protease Inhibitors , Pharmacology , Liver , Lopinavir , Toxicity , Models, Statistical , Reproducibility of Results , Software , Reference Standards摘要
Objective To observe the hematological toxicity of 89SrCl2 in patients with multiple bone metastases of malignant tumors,and analyze the related-risk factors.Methods A total of 89 patients (63 males,26 females;age:(62.3±5.2) years) with multiple bone metastases and treated with 89SrCl2 were enrolled.Hematological data at 2 and 4 weeks after treatment with 89SrCl2 were analyzed.Common Terminology Criteria for Adverse Events (CTCAE) v4.03 was used to evaluate the hematological toxicity,and the influencing risk factors were analyzed.Logistic regression analysis was used to analyze the data.Results The incidences of grade Ⅰ-Ⅱ anemia,leukopenia and thrombocytopania at 2 and 4 weeks after treatment were 15.7%(14/89),18.0%(16/89),11.2%(10/89) and 18.0%(16/89),24.7%(22/89),18.0%(16/89),respectively.The incidences of grade Ⅲ-Ⅳ anemia,leukopenia and thrombocytopenia were 2.2%(2/89),0,0 and 2.2%(2/89),2.2%(2/89),3.4%%(3/89),respectively.Logistics multivariate analysis showed that the number of bone metastases and the Hb level before treatment were independent effect factors for hematological toxicity of 89SrCl2,with odds ratio (OR) values of 2.200(95% CI:1.269-3.841) and 0.961 (95% CI:0.932-0.991),respectively.Conclusions Serious hematological toxicity after 89SrCl2 treatment is rare.The number of bone metastases and the Hb level before treatment are independent effect factors for hematological toxicity.
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Objective: To investigate the effect of varying concentrations of polyethylene glycol(PEG)400 in receiving solution on in vitro transdermal test of drugs. Method: 5-Fluorouracil(5-FU) was selected as a model drug,by preparing different concentrations of PEG400-phosphate buffer solution(PBS) as the receiving solution,the receiving chamber did not add drug,the excised rat skins were treated with various additives for 12 h,then replaced by PBS and added the saturated model drug into the donor compartment to determine the transdermal parameters of the drug.Meanwhile,scanning electron microscopy(SEM) was employed to monitor the effect of PEG400 with different concentration on the stratum corneum of rat skin. Result: The 10%,15% and 40% PEG400-PBS groups had no significant effect on in vitro transdermal absorption parameters of the 5-FU.The steady transdermal rate and cumulative penetration rate of the drug in 20% and 30% PEG400-PBS groups were significantly higher than that in the PBS group(PPConclusion: In the rat skin transdermal test,the concentration of PEG400 in receiving solution should be controlled below 20%.
摘要
Background@#Collagen type IV (COL4)-related nephropathy includes a variety of kidney diseases that occur with or without extra-renal manifestations caused by COL4A3-5 mutations. Previous studies revealed several novel mutations, including three COL4A3 missense mutations (G619R, G801R, and C1616Y) and the COL4A3 chr:228172489delA c.4317delA p.Thr1440ProfsX87 frameshift mutation that resulted in a truncated NC1 domain (hereafter named COL4A3 c.4317delA); however, the mutation mechanisms that lead to podocyte injury remain unclear. This study aimed to further explore the mutation mechanisms that lead to podocyte injury.@*Methods@#Wild-type (WT) and four mutant COL4A3 segments were constructed into a lentiviral plasmid, then stably transfected into human podocytes. Real-time polymerase chain reaction and Western blotting were applied to detect endoplasmic reticulum stress (ERS)- and apoptosis-related mRNA and protein levels. Then, human podocytes were treated with MG132 (a proteasome inhibitor) and brefeldin A (a transport protein inhibitor). The human podocyte findings were verified by the establishment of a mus-Col4a3 knockout mouse monoclonal podocyte using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology.@*Results@#Our data showed that COL4A3 mRNA was significantly overexpressed in the lentivirus stably transfected podocytes. Moreover, the COL4A3 protein level was significantly increased in all groups except the COL4A3 c.4317delA group. Compared to the other test groups, the COL4A3 c.4317delA group showed excessive ERS and apoptosis. Podocytes treated with MG132 showed remarkably increased intra-cellular expression of the COL4A3 c.4317delA mutation. MG132 intervention improved higher ERS and apoptosis levels in the COL4A3 c.4317delA group. Mouse monoclonal podocytes with COL4A3 chr:82717932insA c.4852insA p.Arg1618ThrfsX4 were successfully acquired; this NC1-truncated mutation suggested a higher level of ERS and relatively remarkable level of apoptosis compared to that of the WT group.@*Conclusions@#We demonstrated that excessive ERS and ERS-induced apoptosis were involved in the podocyte injury caused by the NC1-truncated COL4A3 mutation. Furthermore, proteasome pathway intervention might become a potential treatment for collagen type IV-related nephropathy caused by a severely truncated COL4A3 mutation.
摘要
Objective@#To establish finite element models of skull, fronto-orbital advancement and fronto-orbital distraction osteogenesis of craniosynostosis, to analyze the mechanical characteristics of skull base and fronto-orbital operation area, so as to guide the later application of tractors.@*Methods@#One 6-year-old male patient with unilateral coronal synostosis was enrolled in October 2015. Three-dimensional (3D) computed tomography (CT) scan of skull was performed. DICOM data was imported into Mimics 17.0 for contour extraction and cranial 3D reconstruction. The skull model was processed by Mimics, Geomagic Studio 12.0, Hypermesh 12.0 and other software to establish a three-dimensional finite element model. The unilateral and bilateral fronto-orbital anterior osteotomy models were simulated respectively. The mechanical analysis was performed at point A in forehead area and point B in temporal area. Three different groups of traction forces were loaded: (1) 50 Newton for point A, 50 Newton for point B; (2) 80 Newton for point A and 50 Newton for point B; (3) 100 Newton for point A and 50 Newton for point B, to obtain the optimized traction force.@*Results@#Stress analysis was performed on established cranial finite element model, as well as unilateral and bilateral fronto-orbital advancement procedures. The stress distribution of the anterior and middle cranial fossae was found to be concentrated. After unilateral fronto-orbital advancement, the stress of anterior cranial fossa, especially the affected side, was decreased. The stress on both side in anterior cranial fossa was decreased after bilateral fronto-orbital advancement. After force was applied to point A and point B, the optimum deviation result at supraorbital notch point, midpoint of supraorbital margin, frontal temporal point and frontal zygomatic suture point in 3D (Deviation result of X value: -29.4%, -20.5%, -8.6%, -9.3%; Deviation result of Y value: 20.9%, 31.5%, 73.0%, 539.4%; Deviation result of Z value: 4.4%, 1.9%, 0.1%, 11.8%) demonstrated the application of traction force can inwardly, downwardly and forwardly move the bone flap. The optimized traction was 80 Newton at point A and 50 Newton at point B by preliminary assessment.@*Conclusions@#The finite element analysis of the fronto-orbital advancement can be used for more accurate preoperative simulation, to clarify the influence of fronto-orbital advancement on craniofacial morphology and development, as well as skull base. It also facilitates surgical decision and predicts the postoperative distraction vectors.
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To determine relative molecular weight of astragalus polysaccharides (APs), we used Shodex GS620 gel permeation chromatographic column and differential refraction detector (GPC-RI) with dextran as a reference. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and GPC combined with multi-angle laser light scattering detection (GPC-MALLS) were also used.GPC-RI measure showed four peaks of APs, with the Mw of 1 380 000, 231 000, 18 000, and 480, respectively. Three main peaks were found by GPC-RI-MALLS with the Mw as 1 148 000, 180 000, and 43 000, respectively. Strong signals in 155 000 and 18 000 were detected by MALDI-TOF-MS, which also indicated the sugar moieties of the APs as hexoses. From our study, we found that the GPC-RI method with universal correction is most suitable for Mw determination of the APs. Nevertheless, the three methods should be combined and contrasted with each other to obtain accurate information in research of polysaccharides from Chinese medicine.
摘要
In this paper, a new type of preparation for treatment of initial dry eye disease, thermosensitive in situ gel, was prepared using levocarnitine as a model drug. Poloxamer 407 and poloxamer 188 were used as the gel matrix, and sodium hyaluronate and sodium carboxymethylcellulose were used as bioadhesive materials. Gelation temperature was determined by a rotor method and the prescription was optimized by central composite design-response surface methodology. The pH value, viscosity value and gelation temperature of the optimal prescription were measured. The release of the drug in vitro was examined by dialysis membrane permeation, and retention time of the thermosensitive in situ gel preparation on the rabbit's ocular surface was observed by a slit lamp microscope. The results showed that the dosage of the poloxamer 407 and poloxamer 188 were 20.81% and 3.46%, respectively, and sodium hyaluronate was 0.02%, sodium carboxymethyl cellulose was 0.10% of the optimal formulation of levocarnitine thermosensitive in situ gel. The pH value was 6.90 ± 0.06 at room temperature and the viscosity value started to rise sharply at 27 ℃ of the optimal formulation. The gelation temperature of the optimal preparation before and after dilution by simulated tear fluid were (26.37 ± 0.06) ℃ and (33.57 ± 0.21) ℃, respectively. In the first 240 min, in vitro release rate per unit area of levocarnitine thermosensitive in situ gel was lower than that of solution (P<0.05), and after 600 min, the cumulative release rate of levocarnitine thermosensitive in situ gel could reach more than 80%. The retention time of the thermosensitive in situ gel preparation on rabbit's ocular surface reached about 25 min, at least 5 times as much as that of the solution. The animal experiment was conducted following the National Institutes of Health Guidelines for the use of experimental animals, and approved by the Ethics Committee of the Experimental Animal Center of Beijing University of Chinese Medicine. The levocarnitine thermosensitive in situ gel showed good characteristics and sustained release property and significantly improved the retention time of the drug on the rabbit's ocular surface.
摘要
BACKGROUND@#Collagen type IV (COL4)-related nephropathy includes a variety of kidney diseases that occur with or without extra-renal manifestations caused by COL4A3-5 mutations. Previous studies revealed several novel mutations, including three COL4A3 missense mutations (G619R, G801R, and C1616Y) and the COL4A3 chr:228172489delA c.4317delA p.Thr1440ProfsX87 frameshift mutation that resulted in a truncated NC1 domain (hereafter named COL4A3 c.4317delA); however, the mutation mechanisms that lead to podocyte injury remain unclear. This study aimed to further explore the mutation mechanisms that lead to podocyte injury.@*METHODS@#Wild-type (WT) and four mutant COL4A3 segments were constructed into a lentiviral plasmid, then stably transfected into human podocytes. Real-time polymerase chain reaction and Western blotting were applied to detect endoplasmic reticulum stress (ERS)- and apoptosis-related mRNA and protein levels. Then, human podocytes were treated with MG132 (a proteasome inhibitor) and brefeldin A (a transport protein inhibitor). The human podocyte findings were verified by the establishment of a mus-Col4a3 knockout mouse monoclonal podocyte using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) technology.@*RESULTS@#Our data showed that COL4A3 mRNA was significantly overexpressed in the lentivirus stably transfected podocytes. Moreover, the COL4A3 protein level was significantly increased in all groups except the COL4A3 c.4317delA group. Compared to the other test groups, the COL4A3 c.4317delA group showed excessive ERS and apoptosis. Podocytes treated with MG132 showed remarkably increased intra-cellular expression of the COL4A3 c.4317delA mutation. MG132 intervention improved higher ERS and apoptosis levels in the COL4A3 c.4317delA group. Mouse monoclonal podocytes with COL4A3 chr:82717932insA c.4852insA p.Arg1618ThrfsX4 were successfully acquired; this NC1-truncated mutation suggested a higher level of ERS and relatively remarkable level of apoptosis compared to that of the WT group.@*CONCLUSIONS@#We demonstrated that excessive ERS and ERS-induced apoptosis were involved in the podocyte injury caused by the NC1-truncated COL4A3 mutation. Furthermore, proteasome pathway intervention might become a potential treatment for collagen type IV-related nephropathy caused by a severely truncated COL4A3 mutation.