Abstract Human periodontal ligament stem cells (hPDLSCs) are promising cells for dental and periodontal regeneration. Objective This study aimed to develop novel alginate-fibrin fibers that encapsulates hPDLSCs and metformin, to investigate the effect of metformin on the osteogenic differentiation of hPDLSCs, and to determine the regulatory role of the Shh/Gli1 signaling pathway in the metformin-induced osteogenic differentiation of hPDLSCs for the first time. Methodology CCK8 assay was used to evaluate hPDLSCs. Alkaline phosphatase (ALP) staining, alizarin red S staining, and the expression of osteogenic genes were evaluated. Metformin and hPDLSCs were encapsulated in alginate-fibrinogen solutions, which were injected to form alginate-fibrin fibers. The activation of Shh/Gli1 signaling pathway was examined using qRT-PCR and western blot. A mechanistic study was conducted by inhibiting the Shh/Gli1 pathway using GANT61. Results The administration of 50 μM metformin resulted in a significant upregulation of osteogenic gene expression in hPDLSCs by 1.4-fold compared to the osteogenic induction group (P < 0.01), including ALP and runt-related transcription factor-2 (RUNX2). Furthermore, metformin increased ALP activity by 1.7-fold and bone mineral nodule formation by 2.6-fold (P<0.001). We observed that hPDLSCs proliferated with the degradation of alginate-fibrin fibers, and metformin induced their differentiation into the osteogenic lineage. Metformin also promoted the osteogenic differentiation of hPDLSCs by upregulating the Shh/Gli1 signaling pathway by 3- to 6- fold compared to the osteogenic induction group (P<0.001). The osteogenic differentiation ability of hPDLSCs were decreased 1.3- to 1.6-fold when the Shh/Gli1 pathway was inhibited, according to ALP staining and alizarin red S staining (P<0.01). Conclusions Metformin enhanced the osteogenic differentiation of hPDLSCs via the Shh/Gli1 signaling pathway. Degradable alginate-fibrin hydrogel fibers encapsulating hPDLSCs and metformin have significant potential for use in dental and periodontal tissue engineering applications. Clinical Significance Alginate-fibrin fibers encapsulating hPDLSCs and metformin have a great potential for use in the treatment of maxillofacial bone defects caused by trauma, tumors, and tooth extraction. Additionally, they may facilitate the regeneration of periodontal tissue in patients with periodontitis.

Resumo O presente artigo analisa a jurisprudência do Supremo Tribunal Federal (STF) sobre políticas para prevenção e tratamento da covid-19. Com o auxílio da ciência de dados e da inteligência artificial, foi realizada a coleta sistemática e exaustiva da jurisprudência da Corte sobre esse assunto. As decisões coletadas foram organizadas e classificadas nos seguintes temas: distanciamento social; medidas diversas de combate à pandemia; uso de máscara; requisição administrativa; leitos de Unidade de Terapia Intensiva (UTI); vacinação; e acesso a dados. As decisões dentro de cada um desses temas foram analisadas para entender quais questões foram decididas pelo STF e como foram julgadas. A organização e a análise de 253 decisões nos permitem chegar, com nuances e detalhes, a algumas conclusões gerais sobre o papel da Corte em matéria de medidas para prevenção e tratamento da covid-19: parte significativa das decisões foi monocrática e em sede de liminar, o STF buscou proteger a capacidade de entes estaduais responderem à covid-19, foram manifestadas opiniões críticas à atuação do governo federal, em diversos casos houve explícita sobreposição entre questões formais de competência federativa e avaliações substantivas sobre a melhor política, e houve hesitação do STF em determinar a realização de medidas muito complexas para o combate à pandemia de covid-19. (STF covid19. covid19 covid 19. 19 covid-19 artificial assunto social máscara administrativa UTI (UTI) vacinação julgadas 25 chegar detalhes liminar covid19, 19, federal política covid1 1 covid-1 (UTI 2 covid-

Abstract This article analyses the case-law of the Brazilian Supreme Federal Court (STF) in relation to the policies for the prevention and treatment of Covid-19. With the support of data science and artificial intelligence, the STF´s decisions were systematically and exhaustively collected. These decisions were organized and classified into the following topics: social distancing, miscellaneous responses to Covid-19, face covering, requisition of private property, ICU beds, vaccination, and access to information. The organization and analysis of 253 judgments allow us to draw, with nuance and in detail, the following conclusions: an important share of the STF´s decisions was preliminary injunctions and made by a single Justice, the Court sought to protect the capacity of local governments to respond to the pandemic, some decisions show a critical attitude of the Court in relation to the federal government, in many cases there is an explicit overlap between the formal assessment of which federal entity (the federal government, state governments or municipal governments) has the authority to decide an issue and the substantive analysis of which policy should be implemented, and the STF has been hesitant to order complex measures for responding to the Covid-19 pandemic. caselaw case law (STF Covid19. Covid19 Covid 19. 19 intelligence STFs s collected topics distancing Covid19, 19, covering property beds vaccination information 25 draw detail conclusions Justice pandemic government implemented Covid-1 Covid1 1 2 Covid-

PURPOSE: To construct a new biomaterial-small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor, and to evaluate the new biomaterials for the reconstruction of abdominal wall defects.METHODS: Thirty six Sprague-Dawley rats were used in the animal experiments and randomly divided into three groups. The new biomaterial was constructed by combining small intestinal submucosa with gelatin hydrogel for basic fibroblast growth factor release. Abdominal wall defects were created in rats, and repaired using the new biomaterials (group B), compared with small intestinal submucosa (group S) and ULTRAPROTM mesh (group P). Six rats in each group were sacrificed at three and eight weeks postoperatively to examine the gross effects, inflammatory responses, collagen deposition and neovascularization.RESULTS: After implantation, mild adhesion was caused in groups B and S. Group B promoted more neovascularization than group S at three weeks after implantation, and induced significantly more amount of collagen deposition and better collagen organization than groups S and P at eight weeks after implantation.CONCLUSION: Small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor could promote better regeneration and remodeling of host tissues for the reconstruction of abdominal wall defects.

OBJECTIVES: The standard predictive factors of actuarial survival such as T and N stage become less important as patients live for more than 10 years after treatment of cancer. Reports of actual 10-year survivors of oesophageal squamous cell carcinoma (SCC) are rare, and demographic and clinicopathological factors associated with 10-year survival have not been well documented. In this research we evaluated factors predictive of actual, as opposed to actuarial, 10-year survival. METHODS: We retrospectively analysed 1 046 patients who had undergone oesophagectomy for oesophageal SCC. The demographic and clinicopathological characteristics of patients who were alive more than 10 years after oesophagectomy and those of patients who had died were compared. RESULTS: Univariate analysis showed that 18 factors differed significantly between the two groups. Based on logistic regression analysis, factors associated with 10-year survival were younger age, female gender, absence of dysphagia, a left transthoracic surgical approach, lower pathological T stage, and fewer metastatic lymph nodes. CONCLUSION: The independent positive predictors of actual as opposed to actuarial 10-year survival are younger age, female gender, absence of dysphagia, lower pathological T stage, and fewer metastatic lymph nodes.

The wolf spider Lycosa singoriensis is a large and venomous spider distributed throughout northwestern China. Like other spider venoms, the wolf spider venom is a chemical cocktail. Its protein content is 0.659 mg protein/mg crude venom as determined by the Lowry method. MALDI-TOF analysis revealed that the venom peptides are highly diverse and may be divided into three groups characterized by three independent molecular ranges: 2,000 to 2,500 Da, 4,800 to 5,500 Da and 7,000 to 8,000 Da, respectively. This molecular distribution differs substantially from those of most spider venoms studied so far. This wolf spider venom has low neurotoxic action on mice, but it can induce hemolysis of human erythrocytes. Furthermore, the venom shows antimicrobial activity against prokaryotic and eukaryotic cells.

Peptide toxins are usually highly bridged proteins with multipairs of intrachain disulfide bonds. Analysis of disulfide connectivity is an important facet of protein structure determination. In this paper, we successfully assigned the disulfide linkage of two novel peptide toxins, called HNTX-III and HNTX-IV, isolated from the venom of Ornithoctonus hainana spider. Both peptides are useful inhibitors of TTX-sensitive voltage-gated sodium channels and are composed of six cysteine residues that form three disulfide bonds, respectively. Firstly, the peptides were partially reduced by tris(2-carboxyethyl)-phosphine (TCEP) in 0.1 M citrate buffer containing 6 M guanidine-HCl at 40° C for ten minutes. Subsequently, the partially reduced intermediates containing free thiols were separated by reversed-phase high-performance liquid chromatography (RP-HPLC) and alkylated by rapid carboxamidomethylation. Then, the disulfide bonds of the intermediates were analyzed by Edman degradation. By using the strategy above, disulfide linkages of HNTX-III and HNTX-IV were determined as I-IV, II-V and III-VI pattern. In addition, this study also showed that this method may have a great potential for determining the disulfide bonds of spider peptide toxins.