In this study, a novel polyurethane membrane, modified by superfine silk-fibroin powder, was prepared for small-diameter vascular grafting. Scanning electron microscopy, transmission electron microscopy, and histological examination were applied to evaluate histocompatibility of this polyurethane membrane. The polyurethane membrane was compared with polytetrafluoroethylene material. A pseudomembrane and gap formed between polytetrafluoroethylene and the surrounding tissues, and no cells infiltrated or grew into the polytetrafluoroethylene material. On the contrary, superfine silk-fibroin powder/polyurethane blend membrane merged tightly with the surrounding tissues without gaps, and cells infiltrated and grew into the material. Moreover, the negative effects of superfine silk-fibroin powder/polyurethane blend membrane on cells were less than those of its polytetrafluoroethylene counterpart. Our findings indicated that the superfine silk-fibroin powder/polyurethane blend membrane has better histocompatibility than polytetrafluoroethylene membrane. It is concluded that the superfine silk-fibroin powder/polyurethane blend membrane is a promising biomaterial for small-diameter prosthesis.

ibility than polytetrafluoroethylene mem-brane. It is concluded that the superfine silk-fibroin powder/polyurethane blend membrane is a prom-ising biomaterial for small-diameter prosthesis.

OBJECTIVETo evaluate the biomechanical effects of intracellular changes on the voltage-gated sodium channels (VGSCs) on trigeminal ganglion neuron (TRGN).

METHODSTRGN cells were acutely isolated from the neonatal SD rats. The voltage-dependent currents of the VGSCs on these neurons were elicited and analyzed by whole-cell patch-clamp recordings and the intracellular anisotonicity stimuli was established by adjusting the content of pipette solution. The effects of hypo-(260 mOsm) and hypertonic (350 mOsm) osmolarity on the activation and inactivation kinetics of VGSCs on TRGN were evaluated, compared with the normal intracellular environment.

RESULTSThe results demonstrated that intracellular hypotonic stimuli could influence both the activation and inactivation characteristics of VGSCs currents, including the membrane potential at half inactivation (V0.5) of the G-V and inactivation curves had obvious statistics significance (P<0.05) between hypotonicity (260mOsm) and isotonicity (306mOsm). However, only inactivation properties changed under intracellular hypertonic effects, including inactivation rate and k value.

CONCLUSIONIt suggests that the kinetics of VGSCs on TRGN can be modulated both by intracellular hypo- and hypertonic with different characteristics.

Animals ; Cells, Cultured ; Membrane Potentials ; Neurons ; Osmolar Concentration ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Sodium Channels ; Trigeminal Ganglion

Objective:To investigate the effects of circular RNA (circRNA) BICD2 (circ-BICD2) on glutamine metabolism, cell proliferation, migration, invasion and apoptosis of oral squamous cell carcinoma (OSCC) cells and to further explore the possible mechanism.Methods:OSCC cells were purchased from Shanghai Institute of Cellular Cells, Chinese Academy of Sciences. Samples of OSCC tissues and corresponding adjacent tissues were collected from 35 OSCC patients who underwent surgical resection at the First Affiliated Hospital of Zhengzhou University from January 2016 to January 2018. Real-time quantitative PCR (RT-qPCR) and western blot were performed to detect the expression levels of circ-BICD2, miR-296-5p and transgelin2 (TAGLN2) in OSCC tissues and cells. Bioinformatics, dual luciferase report experiment and RNA immunoprecipitation (RIP) experiment were used to determine the targeting relationship between circ-BICD2 and miR-296-5p, miR-296-5p and TAGLN2. According to different transfection oligonucleotides or plasmids, CAL27 and SCC9 cells were divided into si-NC group (transfection si-NC), si-circ-BICD2 group (transfection si-circ-BICD2), si-circ-BICD2+anti-miR-NC group (transfected with si-circ-BICD2 and anti-miR-NC), si-circ-BICD2+anti-miR-296-5p group (transfected with si-circ-BICD2 and anti-miR-296-5p), miR-NC group (transfected with miR-NC), miR-296-5p group (transfected with miR-296-5p mimic), miR-296-5p+pcDNA (transfected with miR-296-5p mimic and pcDNA) and miR-296-5p+TAGLN2 group (transfected with miR-296-5p mimic and pcDNA-TAGLN2). Cell counting kit 8(CCK-8), colony formation experiment, flow cytometry, Scratch healing test and Transwell experiment were applied to detect OSCC cell viability, number of colonies, cycle distribution, apoptosis rate, migration rate and invasive cell numbers. Glutamine (gln) consumption, α-ketoglutaric acid (α-KG) production and adenosine triphosphate (ATP) concentration were also detected by the kit. The expression of cyclinD1 and Glutamine hydrolase (GLS1) proteins of CAL27 and SCC9 cells in each of the groups were detected by western blot. Twelve four-week-old clean BALB/c female nude mice were injected with a single-cell suspension of SCC9 cells into the axillary skin to establish a transplanted tumor model. Twelve transplanted tumor model mice were divided into sh-circ-BICD2 group and sh-NC group. Mice were sacrificed by cervical dislocation at 32 days after injection, the tumors were removed and the tumor weight was tested.Results:The expressions of circ-BICD2 (2.54±0.74) and TAGLN2 (1.86±0.15) were increased ( P<0.05), while the expression of MiR-296-5p was decreased in OSCC tissues ( P<0.05). Cell viability, clone formation numbers, migration rate, invasive cell numbers, S phase cell ratio, glu consumption, α-KG production, ATP concentration, expression of cyclinD1 and GLS1 proteins of OSCC cells were significantly reduced after interference with circ-BICD2 expression. The apoptosis rate, the proportion of cells in G0-G1 phase and the expression of miR-296-5p were significantly increased ( P<0.05). Inhibiting the expression of miR-296-5p coulld reverse the effect of interfering with circ-BICD2 on OSCC cell proliferation, migration, invasion, apoptosis and glutamine metabolism ( P<0.05). After overexpression of miR-296-5p, cell viability, clone formation red number, migration rate, number of invasive cells, S phase cell ratio, glu consumption, α-KG production, ATP concentration and expressions of cyclinD1, GLS1 and TAGLN2 proteins in OSCC cells were significant decrease. The rate of apoptosis and the proportion of cells in G0-G1 phase were significantly increased ( P<0.05). Compared with overexpression of miR-296-5p, cell viability, clone formation red number, migration rate, number of invasive cells, S phase cell ratio, glu consumption, α-KG production, ATP concentration, cyclinD1, GLS1 and TAGLN2 proteins in OSCC cells after overexpression of miR-296-5p and TAGLN2 were significantly increased, and the apoptosis rate and the proportion of cells in the G0-G1 phase were significantly decreased ( P<0.05). Compared with the sh-NC group, the tumor weights of mice in the sh-circ-BICD2 group were significantly reduced ( P<0.05). Conclusions:Circ-BICD2 was highly expressed in OSCC cells. Interfering with circ-BICD2 could inhibit the proliferation, migration and invasion of OSCC cells, glutamine metabolism and tumor growth and promote cell apoptosis, which might be relate to the regulation of miR-296-5p/TAGLN2 molecular axis.

Objective:To explore the mechanism of Yupingfeng Powder on patients with chronic obstructive pulmonary disease (COPD) through UHPLC-QE-MS combined with network pharmacology and molecular docking technology.Methods:UHPLC-QE-MS technology was used to detect the chemical composition of Yupingfeng Powder, and its targets were obtained using ChEMB and TCMIO databases; COPD treatment targets were obtained from TTD, GeneCards, SymMap, and DisGeNET databases, and the intersection of the two was taken. GO enrichment analysis and KEGG pathway analysis were performed on intersection targets. Top 20 metabolites with the most matching targets were selected, their corresponding targets with the top 20 KEGG pathway targets were merged, and potential targets for the treatment of COPD with Yupingfeng Powder were obtained. A target protein-target protein interaction network (PPI) was constructed, key targets of Yupingfeng Powder for the treatment of COPD were screened, and molecular docking was used for validation.Results:Under positive and negative ion modes, a total of 73 active components were identified in Yupingfeng Powder. The top three ranked in terms of degree were genistein, apigenin, and kaempferol. 449 targets of 73 active components and 3 455 disease targets were obtained from the database. The intersection of the two was obtained, resulting in 294 common targets. A total of 69 pathways were identified through KEGG pathway analysis, involved in the pathogenesis of cancer, non-small cell lung cancer signaling pathway, VEGF signaling pathway, T cell receptor (TCR) signaling pathway, arachidonic acid metabolism, Toll like receptor signaling pathway, etc. GO enrichment analysis obtained 2 647 biological processes, 126 cellular components, and 289 molecular functions, involving reactions to nutritional levels, extracellular stimuli, oxidative stress, etc. The top 20 KEGG pathways and the top 20 metabolites with the most matching targets correspond to a total of 281 targets. The key targets were TP53, STAT3, c-Jun, AKT1, and ESR1. The molecular docking results showed that genistein, apigenin, and kaempferol bound well with core targets TP53, STAT3, and c-Jun.Conclusion:Yupingfeng Powder may affect the levels of inflammation, immune regulation, and oxidative stress in COPD through multiple components, targets, and pathways, thereby affecting the progression of COPD.

Objective To determine the occurrence and clinical characteristics of ectopic pituitary adenoma (EPA) in China.Methods The study are done by searching systematically and comprehensively on major Chinese and English medical literature databases and conference papers before 2015,which are only limited to collected,summarized,sorted and analyzed EPA cases that reported by Chinese authors in English or Chinese occurred in China.Results ① Among the 86 Chinese articles and 27 English articles related to ectopic pituitary adenoma (EPA) gathered by the author,except for cases that have been confirmed as repeated reports,a total of 73 EPA cases were found.Of 70 cases with complete data,31 were male cases,accounting for 44.29％;39 were female cases,accounting for 55.71％;the ratio is 1:1.3.The frequency of EPA occurrence according to the location of the lesion,from high to low in turn,was sphenoid sinus (31 cases,42.47％),nasopharynx (7 cases,9.59％),suprasellar region (7 cases,9.59％),clivus (5 cases,6.85％),followed by the third ventricle,sphenoid sinus/clivus,nasal cavity,and the temporal lobe of the brain,with the same incidence of 4.11％.According to the functional properties of EPA,the frequency of different secreting hormones adenoma was PRL-ma(21 cases,28.77％),NF-ma (21 cases,28.77％),ACTH-ma (15 cases,20.55％),GH-ma (10 cases,13.70％),TSH-ma (2 cases,2.74％) and FSH-ma (1 cases.1.37％).Three cases of EPA were uncertain in their property due to lack of information.The incidence of PRL and nonfunctional tumors was the highest,which was different from what was reported in other countries.Among them,one case of EPA was in pineal region and one in parapharyngeal space,which was even more rare and were never reported abroad.(② Except for 3 cases with incomplete medical records,15 out of 70 cases of EPA were accompanied by empty sella,accounting for 21.43％,among which 11 (73.33％) cases involved the sphenoid sinus,and 3 (20％) cases involved clivus.The sphenoid sinus and clivus cases together accounted for 93.33％.(③ 29 out of the 69 cases of EPA with complete record were invasive pituitary adenomas,accounting for 42.03％ and including 1 case of pituitary adenocarcinoma,which accounted for 1.45％.(④ All cases were treated with surgery as the first choice,and some of them were treated with radiotherapy or drug therapy.Conc lusion Ectopic pituitary adenoma is extremely rare.By the end of 2015,the total number of cases reported in China is only 73,which are mostly located in the sphenoid sinus,suprasellar region and nasopharynx.In the functional classification,the most common types are still PRL adenoma and nonfunctional adenoma as in situ pituitary adenoma.42.03％ of EPA adenomas are invasive.Surgical resection of EPA is the first choice and some cases can be treated with radiotherapy and drug therapy.

To find out which biomaterial had the best biocompatibility, we compared the acute phase reaction of common biomaterials preparing for vascular grafts with the material of polyurethane modified by silk fibroin (SF-PU(1:1)). After transplanted the materials of dacron, polyterafluoroethylene (e-PTFE), polyurethane (PU), SF-PU(1:1) in rat muscle for one week, we studied the influence of different biomaterials on the histocompatibility by using rat acute toxicity test, test of local reaction in muscle, tissue section staining, WBC and PLT count. As a result, dacron had the worst histocompatibility. The other biomaterials had slight local inflammatory reaction. The WBC and PLT was nearly the same with the blank except dacron. e-PTFE, pure PU and SF-PU(1:1) had the better histocompatibility than traditional dacron. Especially SF-PU(1:1) had the best histocompatibility. Because of the better physical properties and histocompatibility of SF-PU( 1:1), the prospect of preparing small-diameter vascular grafts with SF-PU was cheerful.
Acute-Phase Reaction ; Animals ; Biocompatible Materials ; Blood Vessel Prosthesis ; Blood Vessel Prosthesis Implantation ; instrumentation ; Female ; Fibroins ; chemistry ; Implants, Experimental ; Macromolecular Substances ; chemistry ; Male ; Materials Testing ; methods ; Muscle, Skeletal ; physiology ; Polyurethanes ; chemistry ; Prosthesis Design ; Random Allocation ; Rats ; Rats, Sprague-Dawley

ObjectiveTo explore the possible mechanism of Yupingfeng Granules （玉屏风散） in preventing and treating chronic obstructive pulmonary disease （COPD） from the perspective of “lung-gut axis”. MethodsThirty-two male Wistar rats were randomly divided into normal group，model group， roxithromycin group and Yupingfeng Granules group， with 8 rats in each group. Except for the normal group， the rat model of COPD was prepared by intratracheal instillation of lipopolysaccharide （LPS） combined with smoking for 12 weeks. Since the fifth week of modeling，the roxithromycin group and the Yupingfeng Granules group were given 31.5 mg/（kg·d） and 1.575 g/（kg·d） of corresponding drugs respectively by gavage，and normal group and model group were given 10 ml/（kg·d） physiolo-gical saline. Sample was collected 24 hours after the last administration. The pathological changes of lung tissue were observed using HE staining； Ultrahigh performance liquid chromatography quadrupole time of flight mass spectrometry （UHPLC-QTOFMS） was used to detect the differential metabolites in alveolar lavage fluid （BALF） in all groups but roxithromycin group；16S rDNA sequencing technology was used to detect the changes of intestinal flora， and the association analysis was conducted between the differential metabolites and the differential flora. ResultsCompared with the normal group， the model group showed an increase in goblet cells in the small bronchial wall， disappearance of the smooth muscle layer of the bronchial wall， and infiltration of inflammatory cells； compared with the model group， roxithromycin group showed slight alveolar interstital edema， and obviously reduced inflammatory cell， while no obvious alveolar interstital edema was observed in the Yupingfeng Granules group， showing a small amout of inflammatory cell infiltration. The results of the BALF differential metabolite screening showed that compared with the normal group， 12 substances were upregulated and 19 substances were downregulated in the model group； compared with the model group， 37 substances in the Yupingfeng Granules group were upregulated and 43 substances were downregulated KEGG analysis yielded a total of 2 metabolic pathways， glycerophospholipid metabolism， and unsaturated fatty acid biosynthetic metabolism； compared with the model group， choline， acetylcholine， glycerol-3-phosphate， glycerophosphate choline， palmitic acid， and arachidonic acid showed an upward trend， while stearic acid and docosahexaenoic acid showed a downward trend in Yupingfeng Granules group （P＜0.05）. The results of the intestinal flora showed that， there are 80 different species between the normal group and the model group， and 65 different species between the model group and Yupingfeng Granules group. Among the top 5 species with relative abundance levels，compared with the model group， the level of Prevotella_9，Ruminococcaceae_UCG-005，Ruminiclostridium_6 increase，and Lactobacillus，Bacteroides decrease（P＜0.05）.The results of the correlation analysis showed that， in the normal and model groups， arachidonic acid was negatively correlated with Oribacterium（r=-0.753，P＜0.01）； in the Yupingfeng Granules group and model group， stearic acid and Bacteroides（r=0.788）， Mycobacterium（r=0.826），［Eubacterium］_Ruminantium_Group（r=0.770） was positively correlated（P＜0.01）， Arachidic acid was negatively correlated with Roseiarcus（r=-0.779）， glycerol-3-phosphate was negatively correlated with Desulfovibrio（r=-0.758）， Arachidonic acid was negatively correlated with Oribacterium（r=-0.753）， and Palmitic acid was negatively correlated with Pseudolabs（r=-0.750，P＜0.01）. ConclusionYupingfeng Granules can affect the metabolism of BALF and the flora structure of intestinal microorganisms， and regulating the balance of “lung-gut axis” may be one of the mechanisms of Yupingfeng Granules in treatment of COPD.