Abstract Objectives: Lung cancer was one of the most common malignancies around the world. It has great significance in to search for the mechanism of occurrence and development of lung cancer. LIM Domain Binding protein 2 (LDB2) belongs to the LIM-domain binding family, it can be used as a binding protein that combined with other transcription factors to form the transcription complex for regulating the expression of target genes. The expression of microRNA-96-5p (miR-96-5p) has been investigated in various tumors. The aim of this study is to investigate the potential role of LDB2 and miR-96-5p in lung cancer. Methods: Real-time quantitative PCR was applied to detect the expression of LDB2 and miR-96-5p. The proliferation, invasion, and metastasis of H1299 cells were analyzed by CCK8, transwell, and wound healing assay after LDB2 or miR-96-5p transfection. Luciferase activities assay and western blot were used to reveal the targeted regulation between LDB2 and miR-96-5p. Results: Here the authors found LDB2 was down-regulated in lung cancer tissues and negatively correlated with miR-96-5p expression, it could promote or inhibit the proliferation, invasion and metastasis of H1299 cells after LDB2 knockdown or overexpression and regulate the expression of cyclinD1, MMP9, Bcl-2, and Bax via ERK1/2 signaling pathway. Furthermore, miR-96-5p exerted its function by directly binding to 3′-UTR of LDB2 and regulating expression of LDB2. miR-96-5p could promote the proliferation, invasion, and metastasis of H1299 cells. Conclusion: These findings demonstrate that LDB2 can act as a new regulator to inhibit cell proliferation, invasion, and metastasis via the ERK1/2 signaling pathway, and miR-96-5p may be a potential promising molecular by targeting LDB2 in lung cancer.

ABSTRACT Salt-tolerant ‘Chuanqiao No.1’ and salt-sensitive ‘Chuanqiao No.2’ Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) were used as materials. Effects of ethephon on physiological characteristics of Tartary buckwheat under salt stress were studied by different concentrations of ethephon (ETH) under 150 mM NaCl stress. Ethephon treatment could improve seed germination rate of both varieties under salt stress, and the effect of ethephon treatment on salt-sensitive variety was better. Ethephon treatment could improve physiological characteristics of Tartary buckwheat seedlings under salt stress, and appropriate concentrations of ETH root treatment in salt-tolerant and salt-sensitive variety were 1.0 and 1.5 mM, and for ETH spraying were 0.4 and 0.6 mM. The effect of ETH spraying on improving physiological characteristics of Tartary buckwheat under salt stress was better than that of ETH root treatment. Salt tolerance gene FtNHX1 reached its maximum expression level at 24 h, which was increased by 277.77% and 251.17% in salt-tolerant and salt-sensitive variety compared with salt stress under ETH root treatment, while under ETH spraying treatment, which was increased by 232.39% and 190.91% in salt-tolerant and salt-sensitive variety compared with salt stress. Salt tolerance gene FtSOS1 reached its maximum expression levels at 12 h, which was increased by 60.94% and 98.75% in salt-tolerant and salt- sensitive variety compared with salt stress under ETH root treatment, while under ETH spraying treatment, which was increased by 35.98% and 67.63% in salt-tolerant and salt-sensitive variety compared with salt stress.

Abstract To observe effect and safety of different doses of ketorolac tromethamine in patient controlled intravenous analgesia (PCIA) after hip arthroplasty. 60 hip arthroplasty patients injected with 0.5 mg/kg ketorolac tromethamine before anesthesia were randomly divided into group A, B and C in which the patients were injected 2, 3 and 4 mg/kg ketorolac tromethamine after operation respectively. Compared with group A, MAP and HR in group B and group C were lower at T1 (3 h after operation) and T2 (6 h after operation). In group A, MAP and HR at T3 (24 h after operation) and T4 (48 h after operation) were lower than at T1. Compared with group A, NRS scores of group B and C were significantly lower during the analgesia. Compared with group A, ramesay sedation scores in group B and group C were significantly higher at T1 and T2; Harris hip scores of group B and group C were significantly higher at T3 and T4; the total number of pushing PCA pump and dezocine dosage were significantly decreased in group B and group C. Preemptive analgesia combined with postoperative analgesia on patients after hip replacement was appropriate.

Abstract Background: Pulmonary hypertension (PH) is a frequent complication of systemic sclerosis (SSc) and is currently one of the primary causes of death in patients with this disease. We conducted a systematic review and meta-analysis to assess the association between PH and mortality in patients with SSc to verify trends in mortality in patients with SSc-associated PH. Methods: We searched the PubMed and Embase databases for published studies on SSc-associated PH from inception to May 2021. All cohort studies in which mortality and/or survival for SSc-associated PH were reported were included in the analysis. The outcome parameters were pooled and analyzed using a random-effects model via generic inverse-variance weighting in conventional and cumulative meta-analysis. Results: The literature search identified 1161 citations, and the full texts of 54 studies were examined. Sixteen articles, with a total of 7857 patients with SSc and 1140 patients with SSc-associated PH, were included in the metaanalysis. Patients with SSc-associated PH had a higher pooled risk of mortality than patients with SSc without PH (risk ratio = 3.12; 95% confidence interval: [2.44, 3.98]). Conclusions: This meta-analysis revealed a higher mortality in patients with SSc-associated PH. PH was a significant predictor of death in patients with SSc. Thus, early diagnosis and treatment of PH are important in patients with SSc.

Abstract Genetically modified soybean strains are trade monitoring object by China. Currently, the genetically modified soybean MON89788 has been approved in China to be imported as processing raw material. Therefore, there is an urgent need to establish a method to quantitatively assess MON89788. This study used droplet digital PCR technology to quantitatively detect MON89788. The results showed that the genomic DNA concentration and its copy number of genetically modified soybeans showed a certain linear relationship. The formula was y = 2.5967x + 3.1437, where R2 = 0.999. When the same 4.5% genetically modified sample was contained, the results of the droplet digital PCR ratio and linear methods were 5.88% and 3.599%, respectively. Comparing the two results showed that the droplet digital PCR ratio method has a larger error than the linear relationship method. The novel droplet digital PCR linear relationship method established in this study has high sensitivity and specificity, and thus is a good prospect for quantitative research.

RESUMO Objetivo: Este estudo teve como objetivo avaliar o efeito da cobertura contínua do olho doente de pacientes com coriorretinopatia serosa central por 48 horas. Métodos: Este estudo retrospectivo, caso-controle, incluiu 32 pacientes com coriorretinopatia serosa central, dos quais 17 receberam tratamento de cobertura contínua por 48 horas no olho doente com gaze médica como grupo de tratamento e 15 foram acompanhados como grupo de observação. Todos os pacientes não receberam nenhum outro tratamento ou medicamento. O logaritmo do ângulo mínimo de resolução da acuidade visual melhor corrigida (LogMar), a espessura macular da retina e o valor médio da raiz quadrada da densidade da amplitude no primeiro anel do eletroretinograma multifocal foram examinados antes e após o tratamento por 48 horas, respectivamente. Resultados: Após o tratamento contínuo, a acuidade visual melhor corrigida pela escala logMar foi de 0, 31 ± 0, 18 no grupo de tratamento e 0, 56 ± 0, 37 no grupo de observação (p=0, 019). A espessura macular da retina foi de 461 ± 43 µm no grupo de tratamento e 498 ± 50 µm no grupo de observação (p=0,032). O valor médio da raiz quadrada da densidade de amplitude no primeiro anel do eletroretinograma multifocal foi de 32,5 ± 5,3 nV/deg2 no grupo com cobertura e foi de 26,6 ± 4,3 NV/deg2 no grupo de observação (p=0,002). Conclusões: O tratamento de cobertura contínua no olho doente, durante 48 horas, apresentou efeitos positivos na acuidade visual melhor corrigida, na espessura e na função macular da retina no tratamento da coriorretinopatia serosa central.

ABSTRACT Purpose: To assess the effect of continuously covering the sick eye affected with central serous chorioretinopathy for 48 h. Methods: This retrospective, case-control study involved 32 central serous chorioretinopathy patients categorized in the treatment group composed of 17 sick eye that received continuous covering treatment for 48 h with a medical gauze and the observation group composed of 15 of these patients who were followed up. None of the patients received any other treatments or medications. The logarithm of the minimal angle of resolution (logMAR) best-corrected visual acuity, macular retinal thickness, and the root mean square value of the amplitude density in the first ring of multifocal electroretinogram were examined before and after the 48-h treatment. Results: After the continuous treatment, the logMAR best-corrected visual acuities were 0.31 ± 0.18 and 0.56 ± 0.37 in the treatment and observation groups, respectively (p=0.019). The macular retinal thicknesses were 461 ± 43 µm and 498 ± 50 µm in the treatment and observation groups, respectively (p=0.032). The root mean square values of the amplitude density in the first ring of multifocal electroretinogram were 32.5 ± 5.3 nV/deg2 and 26.6 ± 4.3 nV/deg2 in the treatment and observation groups, respectively (p=0.002). Conclusions: The continuous application of the covering treatment for 48 h on the sick eye showed positive outcomes with respect to the best-corrected visual acuity, macular retinal thickness, and macular retina functions in the treatment of central serous chorioretinopathy.

Resumo Fundamento: A doença cardiovascular é a principal causa de morte em todo o mundo. A apoptose mediada por hipóxia em cardiomiócitos é uma das principais causas de distúrbios cardiovasculares. O tratamento com a proteína do fator de crescimento endotelial vascular (VEGF, do inglês vascular endothelial growth factor) foi testado, mas as dificuldades operacionais limitaram seu uso. Entretanto, com os avanços da terapia gênica, aumentou o interesse na terapia gênica baseada no VEGF em doenças cardiovasculares. No entanto, o mecanismo preciso pelo qual a reposição de VEGF resgata os danos pós-hipóxia em cardiomiócitos não é conhecido. Objetivos: Investigar o efeito da expressão de VEGF121 pós-hipóxia utilizando cardiomiócitos de ratos neonatos. Métodos: Cardiomiócitos isolados de ratos neonatos foram utilizados para estabelecer um modelo in vitro de lesão cardíaca induzida por hipóxia. O efeito da superexpressão de VEGF, isolado ou em conjunto com inibidores de moléculas pequenas que têm como alvo os canais de cálcio, receptores sensíveis ao cálcio (CaSR, do inglês calcium-sensitive receptors) e calpaína, no crescimento e proliferação celular em lesão de cardiomiócitos induzidos por hipóxia, foram determinados com ensaio de MTT, coloração TUNEL, coloração com Anexina V/PI, lactato desidrogenase e atividade da caspase. Para análise estatística, um valor de p<0,05 foi considerado significativo. Resultados: Verificou-se que o efeito do VEGF121 foi mediado por CaSR e calpaína, mas não foi dependente dos canais de cálcio. Conclusões: Nossos resultados, mesmo em um ambiente in vitro, estabelecem as bases para uma validação futura e testes pré-clínicos da terapia gênica baseada em VEGF em doenças cardiovasculares.

Abstract Background: Cardiovascular disease is the major cause of death worldwide. Hypoxia-mediated apoptosis in cardiomyocytes is a major cause of cardiovascular disorders. Treatment with vascular endothelial growth factor (VEGF) protein has been tested but operational difficulties have limited its use. However, with the advancements of gene therapy, interest has risen in VEGF-based gene therapy in cardiovascular disorders. However, the precise mechanism by which VEGF replenishment rescues post-hypoxia damage in cardiomyocytes is not known. Objectives: To investigate the effect of post-hypoxia VEGF121 expression using neonatal rat cardiomyocytes. Methods: Cardiomyocytes isolated from neonatal rats were used to establish an in vitro model of hypoxia-induced cardiac injury. The effect of VEGF overexpression, alone or in combination with small-molecule inhibitors targeting calcium channel, calcium sensitive receptors (CaSR), and calpain on cell growth and proliferation on hypoxia-induced cardiomyocyte injury were determined using an MTT assay, TUNEL staining, Annexin V/PI staining, lactate dehydrogenase and caspase activity. For statistical analysis, a value of P<0.05 was considered to be significant. Results: The effect of VEGF121 was found to be mediated by CaSR and calpain but was not dependent on calcium channels. Conclusions: Our findings, even though using an in vitro setting, lay the foundation for future validation and pre-clinical testing of VEGF-based gene therapy in cardiovascular diseases.

Naringenin (NAR) is a major flavanone in citrus fruits that has multiple pharmacological attributes such as anticancer and antiatherogenic. This study aims to investigate the mechanism of NAR in high-fat-diet (HFD)-induced atherosclerosis (AS) in apolipoprotein E-knockout (ApoE-/-) mice. A HFD-induced AS ApoE-/- mouse model was established. The mice were treated with HFD, different doses of NAR and simvastatin (Simv). After drug treatment, the levels of total cholesterol (TC), triglyceride (TG), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), superoxide dismutase (SOD), and alanine aminotransferase (ALT) were determined. The expression of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) was detected using qRT-PCR and enzyme-linked immunosorbent assay. The plaque area of the aorta of AS mice was determined using oil red O staining. Western blot analysis was applied to measure the levels of autophagy-related proteins [protein 1 light chain 3B (LC3B), beclin 1, and p62]. The TC, TG, LDL-C, TNF-α, ALT, and MDA levels were significantly increased while the HDL-C, SOD, and GSH-Px levels were decreased in the HFD-induced AS ApoE-/- mice. NAR treatment reversed the expression of the above indicators in mice. After they were treated with different doses of NAR, the LC3B and beclin 1 levels were improved while the p62 protein level was decreased. This study suggested that NAR could promote cell autophagy to improve HFD-induced AS in ApoE-/- mice.

Fibrosis caused by the increase in extracellular matrix in cardiac fibroblasts plays an important role in the occurrence and development of atrial fibrillation (AF). The aim of this study was to investigate the role of hsa-miR-4443 in AF, human cardiac fibroblast (HCFB) proliferation, and extracellular matrix remodeling. TaqMan Stem-loop miRNA assay was used to measure hsa-miR-4443 expression in patients with persistent AF (n=123) and healthy controls (n=100). Patients with AF were confirmed to have atrial fibrosis by late gadolinium enhancement. At the cellular level, after hsa-miR-4443 mimic and inhibitor were transfected with HCFBs, proliferation, apoptosis, migration, and invasion were analyzed. Lastly, hsa-miR-4443-targeted gene and transforming growth factor (TGF)-β1/α-SMA/collagen pathway were evaluated by dual-luciferase reporter assay and western blot, respectively. In patients with AF, hsa-miR-4443 decreased significantly and collagen metabolism level increased significantly. Logistic regression analysis showed that low hsa-miR-4443 level was a risk factor of AF (P<0.001). The receiver operating characteristic curve revealed that hsa-miR-4443 was useful for predicting AF (area under the curve: 0.828, sensitivity: 0.71, specificity: 0.78, P<0.001). In HCFBs, hsa-miR-4443 targeted thrombospondin-1 (THBS1) and downregulated TGF-β1/α-SMA/collagen pathway. The inhibition of hsa-miR-4443 expression promoted HCFB proliferation, migration, invasion, myofibroblast differentiation, and collagen production. The significant reduction of hsa-miR-4443 can be used as a biomarker for AF. hsa-miR-4443 protected AF by targeting THBS1 and regulated TGF-β1/α-SMA/collagen pathway to inhibit HCFB proliferation and collagen synthesis.

Resumo Introdução e objetivos Os mecanismos de neurotoxicidade dos anestésicos locais são complexos. A apoptose e a autofagia são mecanismos altamente organizados que mantêm a homeostase celular durante o estresse. Estudos revelam que a ativação da autofagia atua como mecanismo de proteção in vitro. Não está claro se a autofagia também desempenha essa função in vivo. O objetivo deste estudo foi analisar o papel da autofagia na neurotoxicidade induzida por anestésico local e esclarecer o mecanismo dessa neurotoxicidade utilizando um modelo de injeção intratecal em ratos. Métodos Dezoito ratos Sprague‐Dawley machos adultos saudáveis foram divididos aleatoriamente em três grupos. Antes de receber a injeção intratecal de bupivacaína a 1%, cada rato recebeu injeção intraperitoneal de veículo ou rapamicina (1 mg.kg‐1) uma vez ao dia durante 3 dias. As alterações patológicas foram examinadas por coloração com Hematoxilina e Eosina (HE). A apoptose foi analisada por coloração com o método dUTP Nick‐End Labeling (TUNEL) mediado por TdT. A expressão de caspase‐3, Beclin1 e LC3 foram examinadas por coloração Imunohistoquímica (IHQ). A expressão de Beclin1 e LC3 e a razão LC3‐II/LC3‐I foram detectadas por análise de western blot. Resultados Após a injeção intratecal de bupivacaína, ocorreu lesão patológica nos neurônios da medula espinhal e os níveis de apoptose e caspase‐3 aumentaram. A ativação da autofagia causada pela rapamicina mitigou de forma expressiva as alterações patológicas e diminuiu os níveis de apoptose e caspase‐3, aumentando a expressão de LC3 e Beclin1 e a razão LC3‐II/LC3‐I. Conclusões O aumento da autofagia diminui a apoptose dependente da caspase‐3 e melhora a sobrevivência neuronal in vivo. A ativação da autofagia pode ser uma estratégia terapêutica potencial para a neurotoxicidade induzida por anestésicos locais.

Abstract Background and objectives The mechanisms by which local anesthetics cause neurotoxicity are very complicated. Apoptosis and autophagy are highly coordinated mechanisms that maintain cellular homeostasis against stress. Studies have shown that autophagy activation serves as a protective mechanism in vitro. However, whether it also plays the same role in vivo is unclear. The aim of this study was to explore the role of autophagy in local anesthetic-induced neurotoxicity and to elucidate the mechanism of neurotoxicity in an intrathecally injected rat model. Methods Eighteen healthy adult male Sprague-Dawley rats were randomly divided into three groups. Before receiving an intrathecal injection of 1% bupivacaine, each rat received an intraperitoneal injection of vehicle or rapamycin (1 mg.kg-1) once a day for 3 days. The pathological changes were examined by Haematoxylin and Eosin (HE) staining. Apoptosis was analysed by TdT-mediated dUTP Nick-End Labelling (TUNEL) staining. Caspase-3, Beclin1 and LC3 expression was examined by Immunohistochemical (IHC) staining. Beclin1 and LC3 expression and the LC3-II/LC3-I ratio were detected by western blot analysis. Results After bupivacaine was injected intrathecally, pathological damage occurred in spinal cord neurons, and the levels of apoptosis and caspase-3 increased. Enhancement of autophagy with rapamycin markedly alleviated the pathological changes and decreased the levels of apoptosis and caspase-3 while increasing the expression of LC3 and Beclin1 and the ratio of LC3-II to LC3-I. Conclusions Enhancement of autophagy decreases caspase-3-dependent apoptosis and improves neuronal survivalin vivo. Activation of autophagy may be a potential therapeutic strategy for local anaesthetic-induced neurotoxicity.

Abstract The effectiveness of magnesium (Mg) alloys to improve the capability of bone tissue generation may be severely diminished if the required mechanical properties are not provided. Here, the effort is directed to model the mechanical performance of severely plastically deformed biodegradable ZK60 Mg alloy in bone regeneration protocols. For this purpose, the effects of parallel tubular channel angular pressing (PTCAP) on yield strength (σ YS ), ultimate tensile strength (σ UTS ), and elongation to failure (δ) were addressed. Given the multifaceted variables of the PTCAP with nonlinear interactions, a precise determination of the mechanical properties requires a large number of experiments. Therefore, gene expression programming (GEP) and genetic programming (GP) models were proposed to achieve appropriate combinations of mechanical properties for bone implant purposes based on a rational hypothesis that for correlation coefficient (|R|) higher than 0.8, a strong correlation is established between the predicted and measured values. The results verified that the highest mechanical performance was achieved at the second pass of PTCAP, thus has a great potential to be the most promising candidate for biodegradable implant material. Besides, the proposed models were capable of precisely predicting the mechanical performance of the SPD-processed biodegradable ZK60 Mg.

MicroRNAs (miRNAs), as post-transcriptional regulators, have been reported to be involved in the initiation and progression of various types of cancer, including gastric cancer (GC). The present study aimed to investigate the role of miR-383-5p in gastric carcinogenesis. Cell viability was analyzed using CCK-8 kit. Annexin V-fluorescein isothiocyanate/propidium iodide double staining was used to evaluate cell apoptosis. The expression levels of miR-383-5p and histone deacetylase 9 (HDAC9) mRNA in GC tissues and cell lines were analyzed using RT-qPCR. The protein expression of HDAC9 was detected by western blotting. We found that HDAC9 was up-regulated and miR-383-5p was down-regulated in GC tissues and cell lines. High HDAC9 expression or low miR-383-5p expression was closely related to poor prognosis and metastasis in GC patients. HDAC9 knockout or miR-383-5p mimics led to growth inhibition and increased apoptosis in AGS and SGC-7901 cells. More importantly, we validated that miR-383-5p as a post-transcriptional regulator inhibited HDAC9 expression and was inversely correlated with HDAC9 expression in GC tissues. miR-383-5p had the opposite effects to HDAC9 in gastric carcinogenesis. miR-383-5p played an important role in gastric carcinogenesis, and it is one of the important mechanisms to regulate oncogenic HDAC9 in GC, which might be helpful in the development of novel therapeutic strategies for the treatment of GC.

Abstract Purpose: To compare the safety, feasibility, and short-term clinical benefits of laparoscopic pylorus-preserving pancreaticoduodenectomy (L-PPPD) to open pylorus-preserving pancreaticoduodenectomy (O-PPPD) through retrospective matched cases. Methods: Web of Science, Cochrane, PubMed, CNKI were searched systematically identify studies published between January and December 2017 comparing L-PPPD to O-PPPD. The meta-analysis was performed by using Review Manager 5.3. Results: Two studies matched the selection criteria, including 108 (50%) cases of laparoscopic pylorus-preserving pancreaticoduodenectomy and 108(50%) cases of open pylorus-preserving pancreaticoduodenectomy. None of the included studies were randomized, which were both retrospective matched cases. There was no difference in the incidence of postoperative pancreatic fistula, blood loss, diet start and lymph nodes. However, L-PPPD has a shorter hospital stay (p=0.0003) and O-PPPD has a shorter operative time (p=0.02) and tend to decrease the delayed gastric emptying. Conclusions: The perioperative safety of laparoscopic surgery, which also has advantages of minimal invasion and shorter hospital stay, is comparable to that of open surgery. Laparoscopic surgery could be operated if the patients matched the indication and operation difficulty is not so great. However, blind pursuits of L-PPPD should be restrained because there is no essential difference between these two in terms of feasibility, safety and short-term complication.

An estradiol templated core-shell magnetic molecularly imprinted cryogel (MMIC) was synthesized to selectively extract and analyze estradiol in milk powder. The microsphere of Fe3O4@SiO2 was firstly prepared on Fe3O4 with ethyl silicate and 3-methyl propyl trimethoxy silane. Then the polymerization of α-methacrylic acid and ethylene glycol dimethyl acrylate was carried out on Fe3O4@SiO2 at −20 ºC for 24 h with estradiol as template. The morphology and microstructure of MMIC were investigated by transmission electron microscopy and infrared spectrogram. When used as chromatographic stationary phase, MMIC showed good selectivity and chromatographic performance for the template. Spiked milk powder samples were used to examine the adsorption capacity and selectivity of MMIC, giving recoveries of 92-101% and limit of detection 0.8-1.2 ng g−1. Experimental results indicate good reproducibility with relative standard deviation from 2.2 to 4.5% (n = 3) at spiked levels of estradiol 10, 50, and 100 ng g-1.