Nrf2 Inhibits GAPDH/Siah1 Axis to Reduce Inflammatory Reactions and Proliferation of Microglia After Simulating Spinal Cord Injury.

OBJECTIVE: To explore the effect of nuclear factor erythroid 2-related factor 2 (Nrf 2) on microglial inflammatory response and proliferation after spinal cord injury (SCI) through the glyceraldehyde phosphate dehydrogenase (GAPDH) / Seven in absentia homolog 1 (Siah 1) signaling pathway. METHODS: Human microglia HMC3 was induced by lipopolysaccharide (LPS) to establish a SCI cell model. Microglia morphology after LPS stimulation was observed by transmission electron microscope (TEM), and cellular Nrf2, GAPDH/Siah1 pathway expression and cell viability were determined. Subsequently, the Nrf2 overexpression plasmid was transfected into microglia to observe changes in cell viability and GAPDH/Siah1 pathway expression. RESULTS: Microglia, mostly amoeba-like, were found to have enlarged cell bodies after LPS stimulation, with an increased number of cell branches, highly expressed Nrf2, GAPDH and Siah1, and decreased cell viability (P<0.05). Up-regulating Nrf2 inhibited the GAPDH/Siah1 axis, decreased inflammatory responses, and enhanced activity in post-SCI microglia (P<0.05). CONCLUSION: Up-regulating Nrf2 expression can reverse the inflammatory reaction of microglia after LPS stimulation and enhance their activity by inhibiting the GAPDH/Siah1 axis.

Value of contrast-enhanced magnetic resonance imaging-T2WI-based radiomic features in distinguishing lung adenocarcinoma from lung squamous cell carcinoma with solid components >8 mm.

Background: Radiomics is one of the research frontiers in the field of imaging and has excellent diagnostic performance. However, there is a lack of magnetic resonance imaging (MRI)-based omics studies on identifying pathological subtypes of lung cancer. Here we explored the value of the contrast-enhanced MRI-T2-weighted imaging (T2WI)-based radiomic analysis in distinguishing adenocarcinoma (Ade) from squamous cell carcinoma (Squ) with solid components >8 mm. Methods: A retrospective analysis was performed of a total of 71 lung cancer patients who undergoing contrast-enhanced MRI and computed tomography (CT) before treatment, and the nodules had solid components ≥8 mm in our center from January 2020 to September 2021. All enrolled patients were divided into Squ and Ade groups according to the pathological results. In addition, the two groups were randomly divided into training set and validation set in a ratio of about 7:3. Radiomics software was used to extract the relevant radiomic features. The least absolute shrinkage and selection operator (Lasso) was used to screen radiomic features that were most relevant to lung cancer subtypes, thus calculating the radiomic scores (Rad-score) and constructing the radiomic models. Multivariate logistic regression was used to combine relevant clinical features with Rad-score to form combined model nomograms. The receiver operating characteristic (ROC) curves. the area under the ROC curve (AUC), the decision curve analysis (DCA) and the DeLong's test were used to evaluate the clinical application potentials. Results: The sensitivity and specificity of the clinical model based on smoking was 75.0% and 93.8%. The AUC of the constructed magnetic resonance (MR)-Rad model for differentiating the pathological subtypes of lung cancer was 0.8651 in the validation sets. The AUC of the CT-Rad model in the validation set were 0.9286. The combined model constructed by combining clinical features and Rad-score had AUC of 0.8016, for identifying the 2 pathological subtypes of lung cancer in the validation set. There was no significant difference in diagnostic performance between MR-Rad model and CT-Rad model (P>0.05). Conclusions: The MR-Rad model has a diagnostic performance similar to that of CT-Rad model, while the diagnostic performance of the combined mode was better than the single MR model.

Surface modulation of extracellular vesicles with cell-penetrating peptide-conjugated lipids for improvement of intracellular delivery to endothelial cells.

Exosomes (diameter 30-200 nm) are a subtype of extracellular vesicles secreted by cells containing DNA, microRNA (miRNA), and proteins. Exosomes are expected to be valuable as a means of delivering drugs or functional miRNAs in treatment of diseases. However, the delivery of exosomes is not sufficiently effective, even though exosomes have intrinsic delivery functions. Cell-penetrating peptides (CPPs) are short peptide families that facilitate cellular intake of molecules and vesicles. We previously reported that the modification of cells, and liposomes with CPP-conjugated-lipids, CPPs conjugated with poly (ethylene glycol)-conjugated phospholipids (PEG-lipid), that induce adhesion by CPPs, can be useful for cell-based assays and harvesting liposomes. In this study, we aimed to modulate the exosome surface using Tat peptide (YGRKKRRQRRR)-PEG-lipids to improve intracellular delivery to endothelial cells. We isolated and characterized exosomes from the medium of HEK 293 T cell cultures. Tat conjugated PEG-lipids with different spacer molecular weights and lipid types were incorporated into exosomes using fluorescein isothiocyanate labeling to optimize the number of Tat-PEG-lipids immobilized on the exosome surface. The exosomes modified with Tat-PEG-lipids were incubated with human umbilical vein endothelial cells (HUVECs) to study the interaction. Tat conjugated with 5 kDa PEG and C16 lipids incorporated on the exosome surface were highly detected inside HUVECs by flow cytometry. Fluorescence was negligible in HUVECs for control groups. Thus, Tat-PEG-lipids can be modified on the exosome surface, improving the intracellular delivery of exosomes.

Impact of spontaneous liposome modification with phospholipid polymer-lipid conjugates on protein interactions.

Liposome surface coating has been studied to avoid the immunological responses caused by the complement system, and alternative materials to poly(ethylene glycol) (PEG) have been explored recently since the production of anti-PEG IgM antibodies has been found in humans. We previously reported a liposome coating with poly(2-methacryloyloxyethyl phosphorylcholine) (poly(MPC))-conjugated lipids (PMPC-lipids) and demonstrated its protective effect on blood protein interactions. Here, we attempted to modify the liposome surface by exogenously adding PMPC-lipids, which were spontaneously incorporated into the outer membrane via hydrophobic interactions. The polymerization degree of the PMPC segment was regulated from 10 to 100. The incorporated ratio of PMPC-lipid increased with a decrease in the degree of PMPC polymerization. Due to surface modification with PMPC-lipids, increase in the length of the PMPC-chain increased the size of the liposomes. The modified liposomes were kept stable for 14 d in terms of their size, polydispersity, and surface properties, where approximately 70% of PMPC-lipids were incorporated into the liposome surface. We demonstrated that liposome surface modification with PMPC-lipids can inhibit protein adsorption when exposed to serum, regardless of the degree of polymerization of PMPC. In addition, the PMPC-lipid modified surface was not recognized by the anti-PEG IgM antibody, whereas PEG-lipid was recognized by the antibody. Thus, we successfully fabricated an inert liposome surface via spontaneous modification with PMPC-lipids, where only the outer bilayer surface was modified. This technique can be available for full loading of water-soluble active pharmaceutical ingredient inside the modified liposome.

Microwave resonant absorption of SARS-CoV-2 viruses.

Low power microwave can effectively deactivate influenza type A virus through the nonthermal structure-resonant energy transfer effect, at a frequency matching the confined-acoustic dipolar mode frequency of the virus. Currently, aerosol is considered the major route for SARS-CoV-2 transmission. For the potential microwave-based sterilization, the microwave-resonant frequency of SARS-CoV-2 must be unraveled. Here we report a microwave absorption spectroscopy study of the SARS-CoV-2 and HCoV-229E viruses through devising a coplanar-waveguide-based sensor. Noticeable microwave absorption can be observed, while we identified the resonant frequencies of the 1st and 2nd dipolar modes of SARS-CoV-2 virus as 4 and 7.5 GHz respectively. We further found that the resonant frequencies are invariant to the virus titer, and we also studied the microwave absorption of HCoV-229E in weak acidity medium to simulate the common pH value in fluid secretion. Our results suggest the possible radiation frequency for the recently proposed microwave sterilization devices to inactivate SARS-CoV-2 virus through a nonthermal mechanism so as to control the disease transmission in the post-pandemic era.

Genomic Alteration Spectrum of Non-Small Cell Lung Cancer Patients in East-China Characterized by Tumor Tissue DNA and Cell-Free DNA.

Introduction: From an oncologic perspective, genetic detection is becoming a frontline clinical test, used to identify actionable alterations for targeted therapy, monitor molecular clonal tumor evolution, indicate disease progression and prognosis, and predict medication efficacy and resistance. From analysis of both tumor tissue and cell-free DNA from a large cohort of non-small cell lung cancer patients in East-China, we characterized the full spectrum of genomic alterations. Methods: The study comprised 3000 unpaired samples including 1351 tumor tissue DNA (tDNA) and 1649 cell-free circulating tumor DNA (cfDNA) samples, from which 67 cancer-related genes were sequenced and the genetic alteration profiles were depicted. Integrative molecular analyses identified the frequently mutated genes, uncovered co-occurring somatic alterations, described the distribution of hotspot variants, analyzed the frequency of variant alleles, and notably distinguished actionable, novel variants. Results: The most commonly affected genes were EGFR, TP53, KRAS, CDKN2A, and PIK3CA in both tDNA and cfDNA samples. EGFR and CTNNB1, PIK3CA and PTEN, ERBB2 and SMO were found to have frequent co-occurring alterations in tDNA samples, while EGFR and SMO, KRAS and PDGFRA, PIK3CA and KDR were in cfDNA samples. A large number of primary druggable variants were identified in tDNA samples, while numerous drug-resistance variants, rare actionable variants, and non-EGFR actionable variants were detected in cfDNA samples. Novel variants were enriched in KDR, KIT, TP53, ABL1, FGFR1 in tDNA samples while the majority of novel variants were distributed in PDGFRA, EGFR, KIT, ROS1, BRCA2 in cfDNA samples. Variant allele frequency in tDNA samples was significantly (P < 0.001) higher than that in cfDNA samples. Conclusion: The results revealed considerable differences in the alteration characteristics between the two kinds of specimens. To date, this study represents the largest real-world investigation of its kind, derived from the largest number of patients in East-China. It reinforced and expanded the mechanism of molecular analysis of neoplastic genetic profiles.

Induction of Spontaneous Liposome Adsorption by Exogenous Surface Modification with Cell-Penetrating Peptide-Conjugated Lipids.

The use of amphiphilic molecules such as poly(ethylene glycol)-conjugated phospholipid (PEG-lipid) enables incorporation into liposome surfaces by exogenous addition as a result of the self-assembly with lipids. This technique can be applicable for manipulation of both liposomes and cells. In this study, we aimed to characterize Tat peptide (YGRKKRRQRRR)-conjugated PEG-lipids when used to exogenously surface modify liposomes (size: ca. 100 nm). We earlier reported that cells, which were surface modified with Tat peptides conjugated to PEG-lipids could attach spontaneously to material surfaces without any chemical modification. Here, we synthesized different types of Tat-PEG-lipids by combining PEG of different molecular weights (5 and 40 kDa) with different lipids with three acyl chains (myristoyl, palmitoyl, and stearoyl, respectively) and then studied the spontaneous adsorption of modified liposomes onto a substrate surface induced by the different Tat-PEG-lipids. The amount of adsorbed liposomes strongly depended on the number of incorporated Tat-PEG-lipid moieties: a decrease in both the PEG and the acyl chain lengths led to adsorption of higher amounts of liposomes. Furthermore, when a collagenase-cleavable amino acid sequence was inserted between the Tat sequence and the PEG segment, adsorbed liposomes could be harvested from the substrate by collagenase treatment with no difference in desorption efficiency between the different Tat-PEG-lipids. Thus, Tat-PEG-lipid can be a suitable tool for the manipulation of liposomes and cells.

Discussion of the experience and improvement of an enhanced recovery after surgery procedure for minimally invasive lobectomy: a cohort study.

BACKGROUND: It is of great significance to explore a path for expedited recovery from thoracic surgery for patients undergoing minimally invasive lobectomy to ensure their rapid and smooth recovery and to conserve medical resources. METHODS: We analyzed 629 cases from the Department of Thoracic Surgery, First Affiliated Hospital of Soochow University from January 2018 to January 2020. According to the length of postoperative stay (LOS) and perioperative management, the 629 patients were divided into group A [routine management group (RMG)], group B [rapidly recovery group (RRG), LOS >72 h], and group C (RRG, LOS ≤72 h). The t-test and chi-square test were used to compare the postoperative complications (PC), chest tube indwelling time (CTIT), LOS, postoperative opioid dosages (POD), and total costs (TC) of the 3 groups. RESULTS: Compared with the RMG, the LOS, PC, CTIT, POD, and TC of the RRG were statistically significantly ameliorated (P<0.05). When compared with group A, the PC (18.9% vs. 38.8%), LOS (2.74±0.80 vs. 5.70±1.10 d), CTIT (46.1±18.5 vs. 123.6±34.8 h), and TC (¥51,517±7,217 vs. ¥65,781±8,200) of group C were all decreased. Compared with group A, group B had more preoperative complications, less CTIT, but no significant difference in LOS, PC, and TC during hospital stay. Compared with group B, group C had less preoperative complications, and reduced LOS, CTIT, and TC. The POD in group A was significantly higher than that in groups B and C. CONCLUSIONS: Use of an enhanced recover after surgery (ERAS) protocol can reduce the postoperative complications and shorten the LOS. Some high-risk patients cannot be discharged within 72 h after surgery, but their incidence of grade II postoperative lung complications can be decreased and they can benefit from ERAS. Single intercostal nerve block, COX-2 selective inhibitor, and removal of the chest tube as early as possible can improve the ERAS. KEYWORDS: Enhanced recover after surgery (ERAS); video-assisted thoracic surgery (VATS); minimally invasive lobectomy.

[Characteristics of fructification and seed ripening of Panax stipuleanatus].

Characteristics of fructification and seed ripening of Panax stipuleanatus were studied in this paper with a view to better understand the species' biological characteristics and promote cultivation. Dynamic of fructification and types of fruits were observed in a period of 80 days. The seeds' biological characteristics were evaluated by observing embryo's vitality and changes in form in a period of 225 d. The findings and conclusions were as follows: A plant produces an average of (38.7 +/- 6.5) seeds; Seeds become fully mature 80 days after setting fruit, and fruits grow more rapidly in 0-40 days with an increase of 1 500.83% in fresh weight. Three types of fruits were found, that is mono-seed type, bi-seeds type and triple-seeds type, and the proportion is 9.53%, 58.66%, 1.81%, respectively. The seeds are (5.16-6. 52) mm in length and (3.12-4.95) mm in width. The weight of 1000 seeds is (0.648 +/- 0.014) g with a moisture content of (55.23 +/- 0.49)%. The after-ripening of embryo in Panax stipuleanatus is morphological form, embryos develop 75 days after the seeds' detaching from mother plant and become mature in 165-180 days with the ending of the process of after-ripening. The vitality of embryo reaches 70% , and the length of embryo is (3.217 +/- 0.406) mm and the width (0.872 +/- 0.165) mm. The seeds will germinate in 195 d.

[Flowering biological characteristics of Panax stipuleanatus].

OBJECTIVE: To reveal the flowering characteristics of Panax stipuleanatus and provide theoretical basis for distant hybridization between P. stipuleanatus and P. notoginseng. METHOD: During the blossom of P. stipuleanatus, we observed and investigated blooming phenophase, growth dynamics of inflorescence, pollination and seed setting of its population and the flowering process and its period, the regularity of flowering and pollinating of the floret. Statistic analyses were carried out. RESULT AND CONCLUSION: The population florescence of P. stipuleanatus was about 60 d. The average florets quality of umbel was 44-47. The average natural pollination rates were 73.32%-95.39%. The average seed setting rates was 35.65%-51.76%. The highest growth periods of inflorescence are from March 25 to April 4. The diameter, the height and the length of its inflorescence in 10 d increased 44.65%, 42.19% and 106.25%, respectively. The whole stereotype period was the May beginning. The time that the floret from petal opening to withering it generally needed 36-48 h, 60 percent of the floret finished auther pollinating during the same day and 40 percent until the next day. The flowering and pollinating peak periods were 14:00-15:00, the flowering numbers was 28.48% of the total flowering amount and the pollinating numbers were 38.63% of the total pollinating amount. The high temperature (20-30 degrees C) and the low humidity (RH < 60%) were beneficial to flowering and pollinating.

[Primary studies on lack of nutritional elements of Sanqi].

The growth and development of Sanqi in the lack of nutritional elements had been done by the water culture. The results indicated that Sanqi plants had different symptoms in the lack of nutritional elements. According to the symptom the symptomatic list was made in this paper.

[Effects of fertilizing levels on output and quality of Pinellia ternate].

OBJECTIVE: To compare the effects of the different fertilizing levels on the output, the effective element, the content of heavy-metal, the grade and the ratio of wet to dry of Pinella ternate to obtain the best fertilize ratio on the planting. METHODS: The orthogonal desing method L9 (3(4)) was applied on the comparison between the different fertilization levels. The content of alkaloid was determined by UV spectrophotometer. The content of heavy-metal was determined by atom absorption. RESULTS: The factors that affected the output and the quality of Pinellia ternate were N > P > K. CONCLUSION: The best fertilizer combination is N 25 g/m2 ,P2O5 18 g/m2, K2O9 g/m2 and the application ratio is 2. 8: 2: 1 for them, in which the higher output and the content of alkaloid should be obtained.