Abstract The AMC-HN-8 cell line and the primary human laryngeal epithelial cell lines were utilized in this work to explore the molecular mechanism of miR-548-3p regulating the gene DAG1 to induce the occurrence and malignant transformation of laryngeal carcinoma. Non-coding RNA miR-548- 3p overexpression plasmid, interference plasmid and blank plasmid were constructed, and the plasmids were transfected into AMC-HN-8 cells, respectively. Meanwhile, a non-transfected plasmid group and a human laryngeal epithelial primary cell group were set up. Five groups of cells were named as NC (Normal control), Model, Ov-miR-548-3p, Sh-miR-548-3p and Blank-plasmid group. The luciferase reporter experiment was used to analyze the regulation characteristics of hsa-miR-548-3p on dystrophin-associated glycoprotein 1 (DAG1). Immunofluorescence was used to analyze the relative expression characteristics of the protein DAG1. The cell cloning experiment was used to analyze the proliferation characteristics of AMC-HN-8. The scratch healing test was used to analyze the migration ability of AMC-HN-8. The transwell test was used to analyze the invasion ability of AMC-HN-8. The RT-PCR was used to analyze the expression level of miR-548-3p. Western blot experiments were used to analyze the expression of protein DAG1, laminin α2 (LAMA2) and utrophin (UTRN). The luciferase report experiment and immunofluorescence test found that the expression of DAG1 and miR-548-3p are positively correlated. Cell cloning, scratching and migration experiments identified that the activity of laryngeal cancer cells was positively correlated with the expression of DAG1. The results of Western blot analysis further strengthened the above conclusions. Through carrying out research on the cellular levels, our work has demonstrated that miR-548-3p regulated the content of protein DAG1, and then further induced malignant transformation of laryngeal carcinoma.

Resumen En este trabajo se utilizaron la línea celular AMC-HN-8 y la línea celular epitelial laríngea humana primaria, para explorar el mecanismo molecular regulador del miR-548-3p sobre el gen DAG1 para inducir la aparición y la transformación maligna del carcinoma laríngeo. Se construyeron el plásmido de sobreexpresión de miR-548-3p de ARN no codificante, el plásmido de interferencia y el plásmido en blanco, y los plásmidos se transfectaron en células AMCHN-8 respectivamente. Mientras tanto, se establecieron un grupo de plásmidos no transfectados y un grupo de células primarias epiteliales laríngeas humanas. Se nombraron cinco grupos de células como NC (control normal), modelo, OvmiR-548-3p, Sh-miR-548-3p y grupo de plásmido en blanco. El experimento indicador de luciferasa se utilizó para analizar las características de regulación de hsa-miR-548-3p en la glicoproteína 1 asociada a distrofina (DAG1). Se utilizó inmunofluorescencia para analizar las características de expresión relativa de la proteína DAG1. El experimento de clonación celular se utilizó para analizar las características de proliferación de AMC-HN-8. Se utilizó la prueba de cicatrización por rascado para analizar la capacidad de migración de AMC-HN-8. La prueba de transwell se utilizó para analizar la capacidad de invasión de AMCHN-8. Se utilizó RT-PCR para analizar el nivel de expresión de miR-548-3p. Se usó un experimento de transferencia Western (Western blot) para analizar las expresiones de la proteína DAG1, laminina α2 (LAMA2) y utrofina (UTRN). El experimento de reporte de luciferasa y la prueba de inmunofluorescencia encontraron que la expresión de DAG1 y miR-548-3p están positivamente correlacionadas. Los experimentos de clonación celular, rascado y migración, identificaron que la actividad de las células cancerosas de laringe se correlacionó positivamente con la expresión de DAG1. Los resultados del análisis de transferencia Western fortalecieron aún más las conclusiones anteriores. A través de la investigación a nivel celular, nuestro proyecto ha demostrado que miR-548-3p regula el contenido de la proteína DAG1 y luego induce la transformación maligna del carcinoma de laringe.

Abstract Purpose: To investigate the impact of Ramipril (RAM) on the expressions of insulin-like growth factor-1 (IGF-1) and renal mesangial matrix (RMM) in rats with diabetic nephropathy (DN). Methods: The Sprague Dawley rats were divided into normal control (NC) group (n = 12), DN group (n = 11), and DN+RAM group (n = 12). The ratio of renal weight to body weight (RBT), fasting blood glucose (FBG), HbA1c, 24-h urine protein (TPU), blood urea nitrogen (BUN), creatinine (Cr), renal pathological changes, the levels of IGF-1, fibronectin (FN), type IV collagen (Col-IV), and matrix metalloproteinases (MMP)-2 were compared among the groups. Results: Compared with NC group, the RBT, FBG, HbA1c, TPU, BUN, Cr, and RMM in DN group were significantly increased (P < 0.05), the IGF-1, FN, and Col-IV were significantly upregulated (P < 0.05), while MMP was significantly downregulated (P < 0.05). Compared with DN group, the indexes except for the FBG and HbA1c in DN+RAM group were significantly improved (P < 0.05), among which IGF-1 exhibited significant positive correlation with TPU(r=0.937), FN(r=0.896) and Col-IV(r=0.871), while significant negative correlation with MMP-2 (r=-0.826) (P<0.05). Conclusion: RAM may protect the kidneys by suppressing IGF-1 and mitigating the accumulation of RMM.

RESUMO Racional: É importante, mas difícil de se tratar fístula anal complexa devido à alta taxa de recorrência e de incontinência pós-operatória. A ligadura do trajeto da fístula interesfincteriana (LIFT) - um novo procedimento cirúrgico com a vantagem de evitar a incontinência anal - tem taxa de sucesso variável entre 57-94,4%. Objetivo: Avaliar os resultados em longo prazo do procedimento cirúrgico LIFT modificado - ligadura do trato interesfincteriano com fístula - para tratar fístula complexa anal. Métodos: Análise retrospectiva de 62 casos de fístula complexa no ânus tratados com abordagem modificada de LIFT (incisão curva na pele do canal anal; sutura em bolsa realizada em torno da fístula; as fístulas residuais removidas em um túnel) e teve tempo de acompanhamento de mais de um ano. A condição geral pré-operatória dos pacientes, a eficácia pós-operatória e a função anal foram comparadas. Resultados: A mediana de idade dos participantes foi de 34 anos, e 43 (69,4%) dos casos eram de homens. Quarenta e um (66,1%) casos eram de fístula transesfincteriana alta, quatro (6,5%) de fístula intra-esfincteriana alta e 17 (27,4%) de fístula anal anterior em mulheres. A mediana da duração do acompanhamento foi de 24,5 meses (12-51). A taxa de sucesso no final do acompanhamento foi de 83,9% (52/62). A pressão anorretal e a Incontinência Fecal da Cleveland Clinic Florida (CCF-FI) avaliadas três meses antes e após a operação não encontraram alterações aparentes. Conclusões: Comparado com o LIFT, o LIFT modificado reduz notavelmente a falha pós-operatória e a taxa de recorrência de fístula complexa com resultados aceitáveis em longo prazo.

ABSTRACT Background: It is important but difficult to treat complex fistula-in-ano due to the high recurrent rate and following incontinence. Ligation of the intersphincteric fistula tract (LIFT), a novel surgical procedure with the advantage of avoiding anal incontinence, has a variable success rate of 57-94.4 %. Aim: To evaluate the long-term outcomes of modified LIFT operative procedure - ligation of intersphincteric fistula tract - to treat complex fistula-in-ano. Methods: Retrospective analysis of 62 cases of complex fistula-in-ano. The group was treated with the modified approach of LIFT (curved incision was made in the anal canal skin; purse-string suture was performed around the fistula; the residual fistulas were removed in a tunnel-based way) and had a follow-up time of more than one year. Patient´s preoperative general condition, postoperative efficacy and their anal function were compared. Results: The median age of the participants was 34, and 43 (69.4%) cases were male. Forty-one (66.1%) cases were of high transsphincteric fistula, four (6.5%) cases of high intrasphincter fistula, and 17 (27.4%) cases of anterior anal fistula in female. The median follow-up duration was 24.5 (range, 12-51) months. The success rate in the end of follow-up was 83.9% (52/62). The anorectal pressure and Cleveland Clinic Florida Fecal Incontinence (CCF-FI) evaluated three months before and after the operation did not find apparent changes. Conclusions: Compared with LIFT, the modified LIFT remarkably reduces postoperative failure and the recurrence rate of complex fistula with acceptable long-term outcomes.

Background: The impact of enteral nutrition (EN) on surgical risk in Crohn's disease (CD) patients suffering from spontaneous intra-abdominal abscess (IAA) was evaluated. Methods: CD patients diagnosed with spontaneous IAA from 2008 to 2015 were included in the study. The impact of EN on surgical risk was evaluated using both univariate and multivariate analyses. Results: A total of 87 patients were enrolled, 66 (75.9%) were male. The mean age at the development of an abscess was 30.2 ± 10.1 years and the median duration of illness from CD diagnosis until the development of an abscess was three (2-6) years. After a median follow-up of 1.9 (1.1-2.9) years, surgical intervention was performed in 42 patients (48.3%). Patients treated with EN were less likely to require surgical intervention (26.1% vs 56.3%, p = 0.01). Multivariate analysis showed that EN was an independent protective factor for the risk of surgery with a hazard ratio of 0.27 (95% confidence interval: 0.11-0.65, p = 0.004) after adjusting for abdominal pain, history of abdominal surgery, concomitant intestinal stenosis and prior use of antibiotics within three months. Conclusions: Surgical intervention is common for CD patients with IAA. Appropriate application of EN may help obviate the need for surgical treatment.

Abstract Background Cimicifuga racemosa is one of the herbs used for the treatment of climacteric syndrome, and it has been cited as an alternative therapy to estrogen. Apart from hectic fevers, dyspareunia and so on, dry mouth also increase significantly after menopause. It has not yet been reported whether C. racemosa has any impact on the sublingual gland, which may relate to dry mouth. In an attempt to determine this, we have compared the effects of estrogen and C. racemosa on the sublingual gland of ovariectomized rats. Results HE staining showed that the acinar cell area had contracted and that the intercellular spaces were broadened in the OVX (ovariectomized rats) group, while treatment with estradiol (E2) and iCR (isopropanolic extract of C. racemosa) improved these lesions. Transmission electron microscopy showed that rough endoplasmic reticulum expansion in mucous and serous acinar epithelial cells and apoptotic cells was more commonly seen in the OVX group than in the SHAM (sham-operated rats) group. Mitochondria and plasma membrane infolding lesions in the striated ducts were also observed. These lesions were alleviated by both treatments. It is of note that, in the OVX + iCR group, the volume of mitochondria in the striated duct was larger than in other groups. Immunohistochemical staining showed that the ratio of caspase-3 positive cells was significantly increased in the acinar cells of the OVX group compared with the SHAM group (p < 0.05); and the MA (mean absorbance) of caspase-3 in the striated ducts also increased (p < 0.05). Estradiol decreased the ratio of caspase-3 positive cells and the MA of caspase-3 in striated ducts significantly (p < 0.05). ICR also reduced the ratio of caspase-3 positive cells and the MA in the striated ducts (p < 0.05), but the reduction of the MA in striated ducts was inferior to that of the OVX + E2 group (p < 0.05). Conclusion Both estradiol and iCR can inhibit subcellular structural damage, and down-regulate the expression of caspase-3 caused by ovariectomy, but their effects were not identical, suggesting that both drugs confer a protective effect on the sublingual gland of ovariectomized rats, but that the specific location and mechanism of action producing these effects were different.

Sugarcane white leaf (SCWL) is an important disease caused by a phytoplasma. In Baoshan, Yunnan, China, SCWL was firstly observed in 2012, and has extended its area of occurrence to 600 hm² . Up to 52% of the plants may become diseased in a field and even complete loss of cane yield may result in the heavily infected fields, posing a serious threat to Yunnan sugar industry. To ascertain the causal agent of suspected SCWL disease in Yunnan, nested PCR using two sets of phytoplasma primer pairs (MOLX/MLOY and P1/ P2) was used to successfully amplify a genomic region of the 16S ribosomal DNA (16S rDNA) from 36 suspected SCWL samples. On the basis of sequencing, phylogenetic analysis and nucleotide alignments of 17 nested PCR products randomly selected from positive samples, identical fragments of 210bp in length were obtained that could be clustered into the 6Sr group XI (Rice Yellow Dwarf group) and shared 100% identity with the 16S-23S intergenic spacer region (ISR) of a member of this group, the SCWL phytoplasma (GenBank: HQ917068), and 99.52% with Sugarcane grassy shoot phytoplasma, of the same group. These results indicate that the SCWL disease in Baoshan is caused by a phytoplasma of the 6Sr group XI.

The aim of this study was to group arteries of arterial arch at the superior margin of the abductor hallucis muscle according to their constitutes and provide anatomical basis for reverse bifolicated flap based on it. The constitute, track-way and distribution of the artery, which supply the medial pedis flap, the medial plantar and the medial tarsal flap, especial the arterial arch at the superior margin of the abductor hallucis muscle were observed and analyzed on 81 lower limbs cast specimens and 2 fresh feet specimens. According to their constitute, arterial arch at the superior margin of the abductor hallucis muscle can be classified into 3 types: i) type I, It was constituted mainly by the branch of anterior medial malleolus artery and (or) the medial tarsal artery. ii) type II, It was constituted mainly by the superficial branch of the medial plantar artery; iii) type II, It was constituted mainly by the branch of anterior medial malleolus artery and the branch of medial tarsal artery anastomose with the superficial branch of the medial plantar artery, which was divided into two subtypes according to the different anastomosis of the artery: Type III 1, the type of anastomosed directly was about 48.2%, type III 2, The type of anastomosed indirectly was about 24.1%. According to the constitute of arterial arch at the superior margin of the abductor hallucis muscle can be classified into three types: The type of anterior medial malleolus artery and medial tarsal artery, the type of superficial branch of the medial plantar artery and the type of mixed. For the type mixed, two subtypes can be classified according to the different anastomosis of the artery.

El objetivo fue clasificar las arterias de arco arterial del margen superior del músculo abductor del hálux según su constitución, y proporcionar una base anatómica para el colgajo bilobulado reverso basado en él. La constitución, recorrido y distribución de las arterias que suministran los colgajos pedicular medial, plantar medial y medial del tarso, en especial el arco arterial del margen superior del músculo abductor del hálux fueron observados y analizados en 81 modelos de miembros inferiores y 2 pies frescos. De acuerdo con su constitución, el arco arterial del margen superior del músculo abductor del hálux se pueden clasificar en 3 tipos: i) tipo I, constituido principalmentepor la rama de la arteria anterior del maléolo medial y/o la arteria medial del tarso. ii) tipo II, constituido principalmente por la rama superficial de la arteria plantar medial, iii) tipo III constituido principalmente por la rama de la arteria maleolar medial anterior y la rama de la anastomosis de la arteria tarsiana medial de la rama superficial de la arteria plantar medial, que se divide en 2 subtipos diferentes de acuerdo con el tipo de anastomosis: tipo III 1, una anastomosis directa que se observó en el 48,2%, y tipo III 2, una anastomosis indirecta observada en aproximadamente el 24,1% de los casos. Según la constitución, el arco arterial del margen superior del músculo abductor del hálux se puede clasificar en 3 tipos: el tipo de arteria anterior del maléolo medial y la arteria tarsiana medial, el tipo de rama superficial de la arteria plantar medial y el tipo mixto. En el tipo mixto, pueden ser clasificados 2 subtipos de acuerdo a la diferente anastomosis de la arteria.

Magnesium aluminate (MgAl2O4) spinel nanoparticles were synthesized by reverse microemulsion process in cyclohexane by using two kinds of surfactants, n-amyl alcohol as cosurfactant and mixture of aluminic/magnesic salt aqueous solution as basic reagents. The effects of surfactant types and titration methods on the morphologies and sizes of the MgAl2O4 nanoparticles were characterized by TEM, TGA-DTA, XRD, HR-TEM and FT-IR. TEM images show that the particles prepared by forward-titration method with SPAN-80/Triton X-100 compound emulsifier have uniform spherical shape and good monodispersity with an average size of 9.5 nm. However, the average size of the particles prepared by reverse-titration method was about 10 nm and some particles have irregular platelike appearance. The products prepared with NP-40 surfactant and forward-titration method were agglomerated with an average size of 13 nm. TGA and XRD results show that the reverse microemulsion method has dramatically lowered the calcination temperature of MgAl2O4 with a degree of 700 °C, and the precursor can transform to single spinel phase at 900 °C.