AIM: To investigate the effects of antitumor drug paclitaxel(PTX)on the proliferation, apoptosis, cell cycle, cell morphology, and related protein expression of Müller cells, and to evaluate its potential toxicity to the retina.METHODS:Müller cells were cultured in vitro and divided into two groups: control group(normal medium)and PTX group. Retinal Müller cells were treated with different concentrations of PTX(0.005, 0.05, 0.5 and 5mg/L)for varying durations(12, 24, 36, 48 and 72h). The CCK8 method was used to assess the effects of different concentrations of PTX and treatment duration on the proliferation Müller cells. Flow cytometry was employed to investigate the impact of different concentrations of PTX on Müller cells apoptosis and cell cycle arrest. Immunofluorescence was used to observe morphological changes in Müller cells. The effects of PTX on the expression of apoptosis-related proteins and aquaporins were analyzed by Western blot and qRT-PCR.RESULTS: PTX exhibits the ability to inhibit the proliferation of Müller cells when cultured in vitro. The efficacy of this inhibition was found to be dependent on both the concentration of the drug and the duration of the stimulation. Higher concentrations of the drug and longer stimulation times resulted in a weaker ability of the cells to proliferate. Additionally, PTX also induces apoptosis in Müller cells, with increased drug concentrations and longer stimulation times leading to higher apoptosis rates. Flow cytometry analysis demonstrates that PTX arrests Müller cells in the G2-M phase of the cell cycle. Moreover, there is a distinct change in cell morphology, with a shift from the typical appearance characterized by clear and slender fibrous structures to a rounder morphology, accompanied by a significant decrease in cell numbers. Further, our findings reveal that there is a transient increase in the expression of cytoinflammatory factors following drug treatment compared to the control group. However, discontinuation of drug stimulation can alleviate this heightened expression. In treated cells, the expression of the CA XIV protein is upregulated compared to the control group, while the expression of vascular endothelial growth factor(VEGF)is downregulated(P<0.05). Additionally, the levels of inflammatory factors in the PTX group are significantly higher than those in the control group(P<0.05), suggesting that PTX has the potential to disrupt the retinal barrier function.CONCLUSION: PTX affects the proliferation and apoptosis of Müller cells, with the effects dependent on stimulation duration and drug concentration. In addition, PTX blocks the Müller cell cycle at the G2-M phase and alters cell morphology, leading to a transient upregulation of inflammatory factors and affecting the integrity of the retinal barrier. These findings indicate the potential toxicity of the antitumor drug PTX to the retina.

To investigate the crucial role of particle size in the biological effects of nanoparticles, a series of mesoporous silica nanoparticles (MSNs) were prepared with particle size gradients (50, 100, 150, 200 nm) with the traditional Stober method and adjusting the type and ratio of the silica source. The correlation between toxicity and size-caused biological effects were then further examined both in vitro and in vivo. The results indicated that the prepared MSNs had a uniform size, good dispersal, and ordered mesoporous structure. Hemolytic toxicity was found to be independent of particle size. At the cellular level, MSNs with smaller particle sizes were more readily internalized by cells, which initiated to more intense oxidative stress, therefor inducing higher cytotoxicity, and apoptosis rate. In vivo studies demonstrated that MSNs primarily accumulated in the liver and kidneys of mice. Pharmacokinetic analysis revealed that larger MSNs were eliminated more efficiently by the urinary system than smaller MSNs. The mice's body weight monitoring, blood tests, and pathological sections of major organs indicated good biocompatibility for MSNs of different sizes. Animal welfare and the animal experimental protocols were strictly consistent with related ethics regulations of Zhejiang Chinese Medical University. Overall, this study prepared MSNs with a particle size gradient to investigate the correlation between toxicity and particle size using macrophages and endothelial cells. The study also examined the biosafety of MSNs with different particle sizes in vivo and in vitro, which could help to improve the safety design strategy of MSNs for drug delivery systems.

China ; Dysferlin/genetics* ; Humans ; Muscular Dystrophies, Limb-Girdle/genetics* ; Mutation

To investigate the potential molecular mechanism of the combination of Platycodonis Radix and Lilii Bulbus with the homology of medicine and food in the treatment of pneumonia by means of network pharmacology and in vitro verification experiment. Under the condition of bioavailability(OB)≥30% and drug-like(DL)≥0.18, the active components of Platycodonis Radix and Lilii Bulbus were screened in TCMSP database; the prediction targets of active components were searched from TCMSP, DrugBank and other databases, and the potential targets of pneumonia were obtained through GeneCards and OMIM database. The common targets were obtained by the intersection of drug and disease targets. The PPI network of common targets was constructed by STRING 11.0, and the core targets were obtained by topological analysis. Then the core targets received GO and KEGG analysis with use of WebGestalt and Metascape. The "component-target-pathway" network was constructed with the help of Cytoscape 3.7.1 software, and the component-target molecular docking verification was carried out with Discovery Studio 2016 software. Finally, the core targets and pathways were preliminarily verified in vitro. In this study, 12 active components were screened, 225 drug prediction targets and 420 potential diseases targets were obtained based on data mining method, and 14 core targets were obtained by topological analysis, including TNF, MMP9, AKT1, IL4 and IL2. The enrichment results of GO and KEGG showed that "Platycodonis Radix and Lilii Bulbus" drug pair may regulate inflammation, cell growth and metabolism by acting on 20 key signaling pathways such as TNF and IL-17, thereby exerting anti-pneumonia effects. The results of molecular docking showed that 12 active components had good binding ability with 14 core targets. In vitro experiment results showed that the core components of "Platycodonis Radix and Lilii Bulbus" drug pair could inhibit the expression of MMP9 and TNF-α by regulating TNF signal pathway. This study confirmed the scientificity and reliability of the prediction results of network pharmacology, and preliminarily revealed the potential molecular mechanism of the compatibility of Platycodonis Radix and Lilii Bulbus in the treatment of pneumonia. It provides a novel insight on systematically exploring the mechanism of the compatible use of Platycodonis Radix and Lilii Bulbus, and has a certain reference value for the research, development and application of new drugs.
Drugs, Chinese Herbal ; Humans ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Pneumonia/drug therapy* ; Reproducibility of Results

Objective:To quickly analyze and identify the components in raw and wine-processed products of Polygonatum cyrtonema （PC） dried rhizomes by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry （UPLC-Q-TOF-MS）， and then find out the differential components before and after processing. Method:The ACQUITY UPLC BEH C₁₈ column （2.1 mm×100 mm， 1.8 μm） was used with 0.1% formic acid aqueous solution-acetonitrile as the mobile phase for gradient elution at a flow rate of 0.25 mL·min^-1. Electrospray ionization was selected for collection and detection in positive and negative ion modes， and the data were analyzed by PeakView 1.2.0.3. According to the retention time， accurate relative molecular weight and fragmentation ion information provided by MS， and combined with the reference substance and literature， the components were identified. After normalized treatment， the MS data of each sample were analyzed with principal component analysis （PCA） and orthogonal partial least squares-discriminant analysis （OPLS-DA）， and then the differential components before and after processing were screened according to the principle that variable importance in the projection （VIP） value was >1. Result:A total of 38 components were identified from raw and wine-processed products of PC dried rhizomes， including 15 steroidal saponins， 6 alkaloids， 3 flavonoids， 2 amino acids， 2 organic acids and 10 others. The results of PCA and OPLS-DA showed that there were significant differences in the contents of components in PC dried rhizomes before and after processing， and 16 differential components such as kingianoside Z， disporopsin and linoleic acid were screened. Conclusion:UPLC-Q-TOF-MS technique can accurately and comprehensively identify the components in PC dried rhizomes， these components are mainly steroidal saponins， flavonoids and alkaloids. It takes a great difference in the contents of components before and after processing， and transformation of the same category components is the main reason for the differences of raw and wine-processed products， which will provide reference for the researches on material basis and processing chemistry of PC dried rhizomes.

Platycodonis Radix, which was first recorded in the Agriculture God's Canon of Materia Medica. It is a multi-functional drug with a wide range of applications. The processing of Platycodonis Radix has been recorded as early as in the Jin dynasty, and has a long history of processing. Today, in addition to the washing, cutting and stir-frying with honey, there have also been more than 20 kinds of processing methods, such as stir-frying with wine, stir-frying with bran, stir-frying with Lilii Bulbus juice and so on. The ancients believed that Platycodonis Radix could enhance the effect of diffusing the lung, promoting pharynx and relieving cough by processing. In terms of the chemical compositions in Platycodonis Radix, more than 100 compositions, like triterpenoid saponins, flavonoids, phenols, sterols, polysaccharides and polyacetylenes, have been isolated and identified from it. Among them, triterpenoid saponins are the essential compositions. In addition, Platycodonis Radix has the pharmacological effects of expectorant, antitussive, anti-inflammatory, anti-tumor, etc. The medicinal ingredients of Platycodonis Radix are mainly triterpenoid saponins and polysaccharides. Among them, triterpenoid saponins have diverse biological activities, which lead it to be one of the hotspots of current researches. Platycodonis Radix has a good role in promoting lung and removing phlegm. After being processed, its medicinal effects are enhanced. It is complex and diverse in compositions of Platycodonis Radix so that has rich pharmacological activities. On the basis of sorting out the literature, this paper discusses the processing history, chemical composition and pharmacological effect of Platycodonis Radix, in order to provide reference for the special processing and modern research of Platycodonis Radix. Furtherly, it provides a theoretical basis for the research of its processing mechanism and quality control.

Objective: Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear. Methods: A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( Results: Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks. Conclusion Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.
Adult ; Aged ; COVID-19/virology* ; China/epidemiology* ; Comorbidity ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Severity of Illness Index ; Treatment Outcome

OBJECTIVE: To study the pharmacokinetic characteristics, clinical effect, and safety of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in children with acute lymphoblastic leukemia (ALL). METHODS: A prospective study was performed on children with ALL who cyclophosphamide, cytarabine, and 6-mercaptopurine were used for consolidation therapy. PEG-rhG-CSF (PEG-rhG-CSF group) or rhG-CSF (rhG-CSF group) was injected after chemotherapy. The plasma concentration of PEG-rhG-CSF was measured, and clinical outcome and safety were observed for both groups. RESULTS: A total of 17 children with ALL were enrolled, with 9 children in the PEG-rhG-CSF group and 8 children in the rhG-CSF group. In the PEG-rhG-CSF group, the peak concentration of PEG-rhG-CSF was 348.2 ng/mL (range 114.7-552.0 ng/mL), the time to peak was 48 hours (range 12-72 hours), and the half life was 14.1 hours (range 11.1-18.1 hours). The plasma concentration curve of PEG-rhG-CSF was consistent with the mechanism of neutrophil-mediated clearance. Compared with the rhG-CSF group, the PEG-rhG-CSF group had a significantly shorter median time to absolute neutrophil count (ANC) recovery (P<0.05). There were no significant differences between the two groups in ANC nadir, incidence rate of febrile neutropenia, duration of grade IV neutropenia, incidence rate of infection, and length of hospital stay. No bone pain or muscle soreness was observed in either group (P>0.05). CONCLUSIONS The pharmacokinetic characteristics of PEG-rhG-CSF in children with ALL receiving consolidation chemotherapy are consistent with the mechanism of neutrophil-mediated clearance, with a short half life and fast recovery of ANC, and there are no significant differences in safety between PEG-rhG-CSF and rhG-CSF.
Child ; Granulocyte Colony-Stimulating Factor/therapeutic use* ; Humans ; Neutropenia ; Polyethylene Glycols ; Precursor Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Prospective Studies ; Recombinant Proteins

OBJECTIVE To investigate the neuroprotective effect and possible mechanisms of lute-olin-7-O-β-D-glucuronide (LGU) against focalcerebral ischemic injury. METHODS The focal cerebral ischemic injury model was established by middle cerebral artery occlusion (MCAO). Male Sprague Dawley rats were randomly divided into sham group,model group(MCAO),LGU group(0.24,0.72 and 2.16 mg·kg-1)and positive control group(Edaravone at 5 mg·kg-1).LGU was injected intravenously 30 min after MCAO.Neurological severity score,infarct volume and brain water content were detected 24 h after MCAO and the levels of Na+-K+ATPase,Ca2+ATPase,TNF-α and IL-1β were detected to explore the possible mechanisms.For the therapeutic time window test,LGU(0.72 mg·kg-1)was injected intrave-nously 0.5, 2, 4, 6, 8, 10 and 12 h respectively after MCAO. To evaluate motion behavior, LGU were injected intravenously 30 min after MCAO and once per day during detection period. The changes of motor coordination were detected by rotating rod method and grip strength analysis, and the changes of gaits were detected using DigiGait Imaging System. RESULTS LGU improved the neurological severity score, infarct volume ratio and brain water content. The therapeutic time window of LGU for cerebral infarction and brain edema was at least 6 h and for neurological dysfunction was 12 h.LGU also prolonged the latency on rotarod, increased the forelimb tension and improved 8 gait parameters, including stance duration,stride length,stance width,paw area,paw area variability,gait symmetry,ataxia coefficient and tau propulsion.Furthermore,LGU increased Na+-K+-ATPase and Ca2+-ATPase levels in the cortex and hippocampus in the ischemic side,reduced the levels of TNF-α and IL-1β in the serum. CONCLUSION LGU has a significant neuroprotective effect against cerebral ischemic injury via improving energy metabolism and reducing inflammation.

BACKGROUND: Poly(3-hydroxybutyrate-4-hydroxybutyrate) (P3HB4HB) is a kind of polymer material that can be completely degraded, has good film-forming property and physical properties, but has poor hydrophilicity. OBJECTIVE: To prepare P3HB4HB/polyvinyl alcohol (PVA) coaxial electrospun scaffolds, and to investigate the physical properties and biocompatibility of scaffolds in vitro. METHODS: We prepared P3HB4HB electrospun scaffold, PVA electrospun scaffold and P3HB4HB/PVA coaxial electrospun composite scaffold, and then detected the morphology and characterization, contact angle, and tensile mechanical properties of the scaffolds. Passage 4 bone marrow mesenchymal stem cells (BMSCs) from Sprague-Dawley rats were seeded on the three kinds of scaffolds. Cell adhesion rate was detected at 1, 3, 6 hours after seeding; cell proliferation was detect at 1, 3, 5, 7 days after seeding; and cell viability was observed fluorescence staining at 7 days after seeding. Passage 4 BMSCs were seeded onto the three kinds of scaffolds followed by 14 days of osteogenic and chondrogenic induction. Then, alizarin red staining and toluidine blue staining were used to verify BMSCs differentiation potentials. RESULTS AND CONCLUSION: (1) Scaffold morphology: Under the scanning electron microscope, the structure of the scaffold in each group was a three-dimensional interconnected network. The fiber diameters of P3HB4HB electrospun scaffold and P3HB4HB/PVA electrospun scaffold were homogeneous and ordered. The P3HB4HB/PVA scaffold showed an obvious core-shell structure under the transmission electron microscope. (2) Scaffold characterization: The tensile strength, tensile modulus and maximum stress of the P3HB4HB and P3HB4HB/PVA scaffolds were significantly higher than those of the PVA electrospun scaffold (P < 0.05). The contact angle of the P3HB4HB/PVA composite scaffold was less than 90°. (3) Cell adhesion rate was ranked as follows: PVA electrospun scaffold group >P3HB4HB/PVA composite scaffold group > P3HB4HB electrospun scaffold group (P < 0.05). (4) Proliferation and activity of cells: The cell proliferation of the P3HB4HB/PVA composite scaffold group was faster than that of the other two groups at 5 and 7 days (P < 0.05). There were more viable cells on the PVA electrospun scaffold and composite scaffold than on the P3HB4HB electrospun scaffold. (5) Cell differentiation: Osteogenesis and cartilage specific staining of the composite scaffold were stronger than those in the other two groups. Overall, the P3HB4HB/PVA coaxial electrospun scaffold has good biocompatibility and a certain mechanical strength.