Effect of dura mater on enhancement of cranial osteogenesis in rats / 中国组织工程研究

BACKGROUND:The dura mater and skull are physically and functionally related,although there have been few investigations on primary extraction of dura mater and cranial cells,as well as co-culture of the two.The use of primary cells to investigate the influence of the dura mater on the skull is novel,and it is hoped that it may give a theoretical foundation for therapeutic therapy. OBJECTIVE:Rat dura mater and cranial bone cells were retrieved in situ to observe the influence of dura mater on cranial bone proliferation and differentiation,as well as to get a basic knowledge of the involvement of Twist1 in this process. METHODS:The enzyme digestion method was used in conjunction with the tissue block method to extract dural cells and cranial osteoblasts from rats within three days of birth.Immunofluorescence staining was used to identify the extracted cells,and alizarin red staining was used to identify and evaluate cranial osteoblasts and their mineralization ability.After co-culturing dural cells and cranial osteoblasts,real-time PCR was utilized to identify the expression of genes associated to cranial osteoblast proliferation and osteogenesis,as well as Twist1. RESULTS AND CONCLUSION:(1)Morphology:The retrieved dural cells had morphological traits similar to fibroblasts,while the osteoblasts were spindle-shaped.(2)Cell identification:immunofluorescence staining revealed that extracted dural cells expressed high levels of vimentin and cranial osteoblasts expressed high levels of alkaline phosphatase;cranial osteoblasts were stained with alizarin red 28 days after osteogenic induction,and obvious mineralized nodules were observed.(3)Real time PCR detection showed that the co-culture group had higher levels of PCNA,alkaline phosphatase,and RUNX2 mRNA expression than the control group(P＜0.01);however,Twist1 mRNA expression was lower(P＜0.01).(4)The findings showed that the primary extracted cranial osteoblasts had a high mineralization capacity,and that the dura mater was a key factor in promoting cranial growth and development and osteogenic differentiation,with Twist1 playing a key role in this process.

Mechanism of TFF3-dependent TWIST1 Upregulating TRIB3 to Promote Colorectal Cancer Metastasis / 肿瘤防治研究

Objective To investigate the effects of TFF3 overexpression on the proliferation, migration, and invasion ability of colorectal cancer HT29 cells and the mechanisms involved in cancer metastasis. Methods HT29 cells were transfected with pIRES2-TFF3, and the expression levels of mRNAs and proteins related to TFF3 gene, TWIST1/TRIB3 signaling pathway, and epithelial-mesenchymal transition (EMT) were detected by qRT-PCR and Western blot. The proliferation, migration, and invasion ability of HT29 cells were detected by the CCK-8, cell scratch, and Transwell assays. Changes in cell morphology after TFF3 overexpression were observed through transmission electron microscopy. Results After the HT29 cells were transfected with pIRES2-TFF3, the expression levels of TFF3 mRNA and protein significantly increased (P<0.01); cell proliferation, migration, and invasion were significantly enhanced (P<0.01); and the expression of related genes, such as TWIST1, TRIB3, Vimentin, and Snail were significantly upregulated. By contrast, the expression of E-cadherin significantly decreased (P<0.05). Various changes in cell morphology were observed after TFF3 overexpression, such as decrease in cell junctions, loss of cilia, formation of pseudopodia, and increase in fusiform cells. Conclusion TFF3 overexpression may promote EMT in colorectal cancer cells through the Twist1/TRIB3 signaling pathway, increase their metastatic potential, and accelerate the malignant progression of colorectal cancer.

Influence of compound Huangbai liquid on wound healing in rats with skin ulcer by activating Notch1/Twist1 axis / 中国免疫学杂志

Objective:To explore the influence and possible mechanism of compound Huangbai liquid on wound healing in rats with skin ulcer.Methods:A rat model of skin ulcer was constructed and randomly separated into model group,compound Huang-bai liquid low-dose,medium-dose and high-dose groups(compound Huangbai liquid L,M,H groups)and erythromycin ointment group,10 rats/group;another 10 rats were randomly collected as the normal group.Compound Huangbai liquid L,M,H groups were given 2.5 ml,5 ml and 10 ml of compound Huangbai liquid,respectively;the erythromycin ointment group was given erythromycin ointment to spread evenly on the wound surface using cotton swabs;the model group and normal group were not given drug treatment.At 0 d,3 d,7 d and 14 d of administration,the wound healing of the rats were recorded by taking pictures,and the wound healing rate were calculated.On the 14th day of administration,the pathological changes and collagen deposition in the wounds of rats were determined by HE and Masson staining methods;the levels of IL-1β,TNF-α,nitric oxide(NO),transforming growth factor-β(TGF-β)and hydroxyproline in wound tissue were determined by kits;and the expressions of Notch1 and Twist1 proteins in wound tissue were determined by Western blot.Results:On the 7th and 14th day of administration,compared with model group,the wound healing rate of the rats in the compound Huangbai liquid L,M,H groups and the erythromycin ointment group were significantly improved(P<0.05),and the inflammatory cell infiltration and other phenomena in rats were significantly improved on 14 days of administration,and the deposition of collagen in the wound surface of rats was also significantly increased.On the 14th day of administration,com-pared with the normal group,the levels of IL-1β,TNF-α,NO,hydroxyproline and TGF-β in the wound tissues of the model group were significantly increased(P<0.05),the protein expressions of Notch1 and Twist1 were significantly decreased(P<0.05);com-pared with model group,the levels of IL-1β,TNF-α and NO in the compound Huangbai liquid L,M,H groups and erythromycin oint-ment group were significantly decreased(P<0.05),the levels of hydroxyproline and TGF-β and the protein expressions of Notch1 and Twist1 were significantly increased(P<0.05).Conclusion:Compound Huangbai liquid can promote wound healing in rats with skin ulcers by activating the Notch1/Twist1 axis.

The mechanism of FAK/Twist1 signal pathway in the closure of cranial suture / 安徽医科大学学报

Objective@#To investigate the function of FAK / Twist1 signaling pathway during craniosynostosis closure.@*Methods@#Ten days old rats were divided into a control group (n = 50) and a rotation group (n = 50) . Both of them were made a approximately 0. 5 cm circle bone window at the midpoint of the lambdoid suture of the rat.The bone flaps were left free without damaging the dura mater. The bone flaps in the control group were repositioned in situ , and the bone flaps in the rotation group were rotated 180 ° and repositioned 3 weeks later. Then the experiments were performance as followed :open field test , measurement of body weight , head circumference , bone flap area , and thickness of bone flap in the two groups , observation of cranial suture closure by microscopy and HE staining , FAK / Twist1 expression determined by Western blot , real⁃time PCR , and immunohistochemical staining in the bone flap and dure , respectively. @*Results@#The cranial sutures was completely closed in the rotation groupand that was open in the control group through detecting by microscopic examination and HE staining. The thickness of the bone flap in the derotation group was greater than that in the control group , with statistical significance (P < 0. 01) . There were no significant differences between two groups in head circumference , weight , bone flap area , and operative area. The results of behavioral test showed that after the closure of cranial suture , the acsion of FAK was significantly increased in the calvaria and dura as well as Twsit1 was significantly decreased in the dura in rotating group measuring by Western blot , real⁃time PCR , and immunohistochemical staining (P < 0. 05) .@*Conclusion@#FAK/Twist1 may play an important role in craniosynostosis after rotation.

Overexpression of lncRNA-MEG3 inhibits endometrial cell proliferation and invasion via miR-21-5p/DNMT3B/Twist

Abstract Recent studies have found that lncRNA-MEG3(MEG3) plays an important role in the development of EMs (Endometriosis), but the specific mechanism needs to be further explored. This study aimed to investigate the effect of MEG3 on the proliferation, invasion of EMs cells. The authors used RT-qPCR to detect the expression of MEG3 and miR-21-5p in EMs tissues and hESCs cells, MTT and Transwell to detect cell proliferation and invasion, western blotting assay to detect the expression of DNMT3B and Twist, MSP to detect the methylation of Twist. The present study's detection results showed that MEG3 was lowly expressed in EMs tissues and hESCs cells, and overexpression of MEG3 could down-regulate miR-21-5p and inhibit endometrial cell proliferation and invasion. In addition, overexpression of MEG3 upregulated the expression of DNMT3B and promoted the methylation of TWIST. In conclusion, the present findings suggest that MEG3 is downregulated in EMs tissues, and overexpression of MEG3 can promote the activity of DNA methyltransferase DNMT3B by downregulating miR-21-5p, thereby promoting the methylation of Twist, downregulating Twist level to inhibits hESCs cells proliferation and invasion.

Estudo da imunoexpressão de proteínas envolvidas na transição epitélio-mesênquima em tumores de glândula salivar / Study of the immunoexpression of proteins involved in the epithelial-mesenchymal transition in salivary gland tumors

Os tumores de glândula salivar (TGS) apresentam notável complexidade clínica e biológica, razão para a qual muitos estudos investigam os eventos envolvidos na sua progressão. Uma das dinâmicas envolvidas na invasão tumoral de diversos tipos de carcinomas é a transição epitélio-mesênquima (TEM). Neste processo, as células epiteliais sofrem transição para um estado mesenquimal móvel, favorecendo a invasão e metástase. Sendo assim, esta pesquisa analisou a expressão imuno-histoquímica de E-caderina, Twist1, Snail1, α-SMA, metaloproteinases de matriz 9 (MMP-9) e Vimentina (VM) em 90 casos de TGS, correlacionando-os entre si e com parâmetros clinicopatológicos. Foram selecionados 20 casos de Adenoma pleomórfico (AP), 20 casos de Carcinoma mucoepidermoide (CME), 20 casos de Carcinoma adenoide cístico (CAC), 10 casos de Adenocarcinoma polimorfo (ACP), 10 casos de Carcinoma epitelial-mioepitelial (CEME) e 10 casos de Carcinoma ex-adenoma pleomórfico (CexAP). A análise de E-caderina, Twist1, Snail1 foi realizada em parênquima tumoral sendo observado o percentual de células positivas (PP), com escores variando de 0 a 4, e a intensidade de expressão (IE), cujos escores variaram de 0 a 3. A avaliação de MMP-9 foi realizada em parênquima e estroma tumoral, também avaliando-se a PP e a IE, ambos baseados em escores que variaram de 0 a 3. A marcação para α-SMA e VM foi analisada em região de estroma tumoral. Células positivas para α-SMA foram contabilizadas em 10 campos, obtendo-se, então a média. A VM foi avaliada de forma qualitativa, utilizando-se 4 escores de acordo com a IE e se a marcação é difusa ou focal. Os dados obtidos foram analisados no software Statistical Package for Social Science, GraphPad Prism e STATA. O nível de significância de 5% foi adotado para os testes estatísticos. Foi verificada menor imunomarcação de E-caderina nos APs em relação às neoplasias malignas de glândula salivar (NMGS). Observou-se baixa imunoexpressão de Twist1 e Snail1 em APs. Em relação a expressão nuclear do Twist1, constatou-se maior expressão nas neoplasias malignas quando comparadas aos APs. Ainda, Twist1 em núcleo foi correlacionado à expressão citoplasmática de E-caderina nas NMGS. No que concerne aos parâmetros clinicopatológicos, esta proteína se relacionou estatisticamente com maiores chances de óbito. Foi evidenciada baixa imunoexpressão de Snail1 entre as NMGS. No entanto, na análise dos CACs, foi verificada maior expressão nuclear na variante sólida em relação às demais. A expressão de MMP-9 em parênquima demonstrou correlação positiva com Twist1 citoplasmático e Snail1nuclear nas NMGS. A MMP-9 também apresentou correlação positiva na comparação da sua imunoexpressão em região de parênquima e de estroma. A VM se apresentou como um biomarcador a ser considerado na avaliação clínica dos pacientes, já que esta apresentou relação significativa com tamanho do tumor (T3-T4) e maior frequência de óbito. Ademais, a alta expressão desta proteína se apresentou como um fator preditivo independente para piores taxas de sobrevida global (SG). A avaliação dos demais fatores clinicopatológicos apresentou estágios clínicos avançados como indicador de valor prognóstico independente para menores taxas de SG, enquanto que para a sobrevida livre da doença, estes foram a localização em glândula salivar menor e presença de metástase à distância. Os resultados deste estudo sugerem que o processo de TEM pode estar relacionado ao estágio de diferenciação celular em APs e à progressão tumoral nas NMGS. Ressalta-se, também, maior participação de Twist1 e MMP-9 no cenário da TEM em tumores malignos de glândula salivar, além da possibilidade de utilização da VM como indicador de valor prognóstico (AU).

Salivary gland tumors (SGTs) present remarkable clinical and biological complexity; therefore, many studies investigate the events involved in their progression. One of the dynamics involved in the tumor invasion of different types of carcinomas is the epithelial-mesenchymal transition (EMT). In this process, epithelial cells undergo a transition to a mobile mesenchymal state, favoring invasion and metastasis. Therefore, this research analyzed the immunohistochemical expression of E-cadherin, Twist1, Snail1, α-SMA, vimentin (VM) and matrix metalloproteinase 9 (MMP-9) in 90 SGTs cases; correlations among the biomarkers, as well as between the biomarkers and clinicopathological parameters were made. We selected 20 cases of pleomorphic adenoma (PA), 20 cases of mucoepidermoid carcinoma (MEC), 20 cases of adenoid cystic carcinoma (ACC), 10 cases of polymorphous adenocarcinoma (PAC), 10 cases of epithelial-myoepithelial carcinoma (EMC) and 10 cases of carcinoma ex-pleomorphic adenoma (CXPA). E-cadherin, Twist1, and Snail1 were analyzed in tumor parenchyma, observing the percentage of positive cells (PP) using scores ranging from 0 to 4, and the expression intensity (EI), whose scores were ranged from 0 to 3. The evaluation of MMP-9 was performed in tumor parenchyma and stroma, also evaluating PP and IE, both based on scores that ranged from 0 to 3. The labeling for α-SMA and VM was analyzed in stromal cells. Positive cells for α-SMA were counted in 10 fields and the mean was calculated. VM was evaluated qualitatively, using 4 scores according to EI and whether the labeling was diffuse or focal. Obtained data were analyzed using Statistical Package for Social Science, GraphPad Prism, and STATA software. The significance level of 5% was adopted for the statistical tests. Patients were mostly female, with a mean age of 49.8 years; the major salivary glands were the most affected anatomical site, mainly the parotid gland. A lower E-cadherin immunostaining was verified in PAs in comparison to malignant neoplasms of salivary glands (MNSGs). Low immunoexpression of Twist1 and Snail1 was observed in PAs. Regarding the nuclear expression of Twist1, it was found greater expression in malignant neoplasms than in PAs. Furthermore, Twist1 in the nucleus was correlated with cytoplasmic expression of E-cadherin in MNSGs. Regarding clinicopathological parameters, this protein was statistically related to higher chances of death. Low immunoexpression of Snail1 was evidenced among the MNSGs. However, in the analysis of CACs, greater nuclear expression was observed in the solid variant compared to the others. Expression of MMP-9 in parenchyma showed a positive correlation with cytoplasmic Twist1 and Snail1nuclear in MNSGs. MMP-9 also showed a positive correlation when comparing its immunoexpression in the parenchyma and the stroma. VM was presented as a biomarker to be considered in the clinical evaluation of patients since it showed a significant correlation between greater tumor size and a higher frequency of death. Furthermore, the high expression of this protein appeared as an independent predictive factor for worse overall survival (OS) rates. The evaluation of the rest of the clinicopathological factors showed advanced clinical stages as an indicator of independent prognostic value for lower rates of OS. For disease-free survival, these indicators were the location in the minor salivary gland and the presence of distant metastasis. Our results suggest that the EMT may be related to myoepithelial differentiation in PAs and tumor progression in MNSGs. Also, Twist1 and MMP-9 appear to play a greater role in the scenario of EMT in MNSGs; finally, VM might be used as a prognostic value indicator (AU).

Surface EMG Analysis on Pre-Activation and Co-Contraction of Knee and Ankle Joint in Brush-Knee Twist-Step / 医用生物力学

Objective To analyze muscle pre-activation and surface electromyography (sEMG) characteristics of knee and ankle joints of long-term Tai Chi practitioners during brush-knee twist-step and normal walking, and explore the neuromuscular control strategies of Tai Chi to prevent falls. Methods Vicon motion capture system, Kistler force plate, and Noraxon sEMG system were synchronously used to collect the EMG signals of the rectus femoris, biceps femoris, tibialis anterior muscle, lateral head of gastrocnemius and body posture information during brush-knee twist-step and normal walking. The pre-activation and co-contraction of knee and ankle joints were calculated by integrated EMG of the rectus femoris/biceps femoris, tibial anterior/lateral gastrocnemius muscles. Results Compared with normal walking, the average time of brush-knee twist-step in four phases was significantly increased. There was a significant difference in the percentage of time in four phases. The knee joint co-contraction level and pre-activation level decreased, and the ankle joint co-contraction level and pre-activation level increased. Conclusions Long-term Tai Chi exercises may increase the activation level of the muscles around knee joints and enhance the synergy in muscle groups to help stabilize the joint. The results provide references for rehabilitation assessment and training of neuromuscular control disorders.

Síndrome de Saethre-Chotzen: a propósito de un caso / Saethre-Chotzen syndrome: a case report

El síndrome de Saethre-Chotzen es un síndrome malformativo craneofacial caracterizado por una sinostosis de las suturas coronales y alteraciones de extremidades. Tiene una prevalencia de 1 de cada 25 000-50 000 recién nacidos vivos. Se presenta el caso de un neonato sin antecedentes de interés con alteraciones craneofaciales al nacer. Ante los rasgos fenotípicos del paciente, se realizó una tomografía axial computada craneal, que mostró la fusión parcial de la sutura coronal y evidenció la presencia de huesos wormianos en localización metópica y lambdoidea derecha. Con la sospecha clínica de síndrome malformativo craneofacial, se solicitó análisis del exoma dirigido, que confirmó que el paciente era portador heterocigoto de la variante patogénica c.415C>A, que inducía un cambio de prolina a treonina en la posición 139 del gen TWIST1, responsable del síndrome. La presencia de huesos wormianos, hallazgo no descrito hasta ahora en la literatura, amplía la variabilidad fenotípica conocida de este síndrome.

The Saethre-Chotzen syndrome is a craniofacial malformation syndrome characterized by synostosis of coronal sutures and limb anomalies. The estimated prevalence of this syndrome is 1 in 25 000-50 000 live births. We present a case report of a neonate, without relevant family history, who presented craniofacial alterations at birth. Given the phenotypic features, a cranial computed tomography scan was performed, showing partial fusion of the coronal suture, evidencing the presence of wormian bones in the metopic and right lambdoid location. With the clinical suspicion of craniofacial malformation syndrome, an analysis of the directed exome was requested confirming that the patient is a heterozygous carrier of the pathogenic variant c.415C>A, which induces a change of proline to threonine at position 139 of the TWIST1 gene, responsible for Saethre-Chotzen syndrome.The presence of wormian bones, a finding not described so far in the literature, extends the well-known phenotypic variability of this syndrome.

Analysis and Diagnosis on Backhand Twist Technique of Leading Men’s Player of National Table Tennis Team / 医用生物力学

Objective To make analysis and diagnosis on backhand twist technique used by player A, who is the leading men’s player of national table tennis team, so as to provide references for improving his backhand twist technique. Methods The three-dimensional kinematics test and analysis were used. The backhand twist techniques of player A and those of player B who has good backhand twist techniques were compared by quantitative data and picture analysis. Results At the stage of swinging racket backward, the racket swing amplitude, shoulder angle and wrist angle of player A were significantly smaller than those of player B. The roll angle of trunk of player A was significantly larger than that of player B. At the stage of swinging and hitting the ball, the shoulder angle, elbow angle of player A were significantly bigger than those of player B, while the increasing amplitude of shoulder angle and elbow angle, as well as the changing amplitude in roll angle of trunk of Lin Guoyuan were significantly smaller than those of player B. At the stage of swinging racket forward, the shoulder angle and elbow angle of player A were significantly bigger than those of player B. The increasing amplitude of shoulder angle and elbow angle, as well as the roll angle of trunk and its changing amplitude of player A were significantly smaller than those of player B. Conclusions The racket swing amplitude of player A was smaller, and the distance between the racket and the ball of player A was close at the end of swinging racket backward stage. During swinging and hitting the ball stage, the hitting point was far from the body, the shoulder joint was not stable enough to support, so that the wrist was used more. The center of gravity was not enough to force forward, and the outburst power was not concentrated. At swinging racket forward stage, player A’s braking was not active enough, which affected the stability of hitting the ball. At hitting the ball stage, the torsion of the trunk was smaller, and the waist power was not concentrated. On the basis of unaffecting the forehand outburst power, player A should slightly adjust his backhand twisting technique, or appropriately increase the the racket swing amplitude and torsion of the body. In the process of hitting the ball, the sequence of outburst power was the waist, the forearm and the wrist.

Expression of Twist1 and Twist2 in hypopharyngeal carcinoma and its relationship with clinicopathological features / 中国医师杂志

Objective:To investigate the relationship between the expression of Twist1 and Twist2 protein and with clinicopathological features in hypopharyngeal carcinoma.Methods:The clinical and pathological data of 58 patients with hypopharyngeal carcinoma who underwent surgery in Liaoyang Central Hospital from May 2014 to September 2019 were retrospectively analyzed. The general demographic and pathological data of the patients were collected, and the expression of Twist1 and Twist2 protein in the hypopharyngeal cancer tissues and adjacent tissues were detected by immunohistochemistry. The patients were followed up to record their survival. The relationship between protein expression of Twist1 and Twist2 and clinical pathological stages and prognosis was analyzed.Results:Immunohistochemistry showed that the positive expression rates of Twist1 and Twist2 protein in 58 patients with hypopharyngeal cancer were significantly higher than those in adjacent tissues ( P<0.05). The positive expression rates of Twist1 and Twist2 in patients with T stage T 3+ T 4 and clinical stage Ⅲ+ Ⅳ were significantly higher than those in patients with T 1+ T 2 and Ⅰ+ Ⅱ ( P<0.05). The positive expression rates of Twist1 and Twist2 in patients with lymph node metastasis were significantly higher than those without lymph node metastasis, and the positive expression rates of Twist1 and Twist2 in low differentiation were significantly higher than those in high differentiation ( P<0.05). 52 patients were followed up postoperatively, the follow-up rate was 89.67%, among which 28 patients recurred (53.85%), and the 1-, 3-, and 5-year survival rates were 80.77%(42/52), 53.85%(28/52), and 21.15%(11/52), respectively. The survival rate of patients with positive expression of Twist1 and Twist2 protein was significantly lower than that of patients with negative expression ( P<0.05). Multiple analysis showed that recurrence, tumor stage, positive expression of Twist1 and Twist2 protein were independent risk factors for survival ( P<0.05). Conclusions:The expression of Twist1 and Twist2 protein in hypopharyngeal carcinoma is stronger than that in normal tissues. The expression of Twist1 and Twist2 protein is related to T stage, clinical stage, pathological tissue type and lymph node metastasis. Meanwhile, the high expression of Twist1 and Twist2 proteins is an independent factor affecting the prognosis of patients. They may be potential indicators to judge the prognosis of patients with hypopharyngeal cancer.

Chrysin inhibited epithelial-mesenchymal transition of type Ⅱ alveolar epithelial cell by regulating NF-κB/Twist 1 signaling pathway / 中国中药杂志

This study aimed to assess whether chrysin(ChR) can inhibit epithelial-mesenchymal transition(EMT) of type Ⅱ alveolar epithelial cell and produce anti-pulmonary fibrosis effect by regulating the NF-κB/Twist 1 signaling pathway. Sixty rats were randomly divided into the control group, the bleomycin(BLC) group, BLC+ChR(50 mg·kg~(-1)) group and BLC+ChR(100 mg·kg~(-1)) group, with 15 rats in each group. The pulmonary fibrosis model was induced by intratracheal injection of BLC(7 500 U·kg~(-1)). Rats were orally administered with different doses of ChR after BLC injection for 28 days. The cells were divided into control group, TGF-β1 group(5 ng·mL~(-1)), and TGF-β1+ChR(1, 10, 100 μmol·L~(-1)) groups. The type Ⅱ alveolar epithelial cells were treated with TGF-β1 for 24 h, and then treated with TGF-β1 for 48 h in the presence or absence of different doses of ChR(1, 10 and 100 μmol·L~(-1)). The morphological changes and collagen deposition in lung tissues were analyzed by HE staining, Masson staining and immunohistochemistry. The mRNA and protein expression levels of collagen Ⅰ, E-cadherin, zonula occludens-1(ZO-1), vimentin, alpha smooth muscle actin(α-SMA), inhibitor of nuclear factor kappa B alpha(IκBα), nuclear factor-kappa B p65(NF-κB p65), phospho-NF-κB p65(p-p65) and Twist 1 in lung tissues and cells were detected by qPCR and Western blot, respectively. The animal experiment results showed that as compared with the BLC group, after administration of ChR for 28 days, bleomycin-induced pulmonary fibrosis in rats was significantly relieved, collagen Ⅰ expression in lung tissues was significantly reduced(P<0.05 or P<0.01), and EMT of alveolar epithelial cells was obviously inhibited [the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], concomitantly with significantly reduced IκBα and p65 phosphorylation level in cytoplasm and decreased NF-κB p65 and Twist 1 expression in nucleus(P<0.05 or P<0.01). The cell experiment results showed that different doses of ChR(1, 10 and 100 μmol·L~(-1)) significantly reduced TGF-β1-induced collagen Ⅰ expression(P<0.05 or P<0.01), significantly inhibited EMT of type Ⅱ alveolar epithelial cells[the expression levels of E-cadherin and ZO-1 were increased and the expression levels of vimentin and α-SMA were decreased(P<0.05 or P<0.01)], and inhibited IκBα and p65 phosphorylation in cytoplasm and down-regulated NF-κB p65 and Twist 1 expression in nucleus induced by TGF-β1(P<0.05 or P<0.01). The results suggest that ChR can reverse EMT of type Ⅱ alveolar epithelial cell and alleviate pulmonary fibrosis in rats, and its mechanism may be associated with reducing IκBα phosphorylation and inhibiting NF-κB p65 phosphorylation and nuclear transfer, thus down-regulating Twist 1 expression.

Biomechanical analysis of ankle-foot complex during a typical Tai Chi movement-Brush Knee and Twist Step / 生物医学工程学杂志

The purpose of this study is to analyze the biomechanics of ankle cartilage and ligaments during a typical Tai Chi movement-Brush Knee and Twist Step (BKTS). The kinematic and kinetic data were acquired in one experienced male Tai Chi practitioner while performing BKTS and in normal walking. The measured parameters were used as loading and boundary conditions for further finite element analysis. This study showed that the contact stress of the ankle joint during BKTS was generally less than that during walking. However, the maximum tensile force of the anterior talofibular ligament, the calcaneofibular ligament and the posterior talofibular ligament during BKTS was 130 N, 169 N and 89 N, respectively, while it was only 57 N, 119 N and 48 N during walking. Therefore, patients with arthritis of the ankle can properly practice Tai Chi. Practitioners with sprained lateral ligaments of the ankle joint were suggested to properly reduce the ankle movement range during BKTS.

The combination of mammaglobin A and TWIST-1 increases detection of circulating tumor cells in breast cancer / A combinação de mamaglobina A e TWIST-1 aumenta a detecção de células tumorais circulantes no câncer de mama

ABSTRACT Objectives: Breast cancer cells that are released into the bloodstream are called circulating tumor cells (CTCs). CTCs can express different genes, like TWIST-1 and mammaglobin A (MGA). The aims of this study were to analyze the expression of TWIST-1 and MGA in the blood of breast cancer patients to detect CTCs and to assess the association between the presence of CTCs and prognostic parameters of breast cancer. Methods: Prospective study. Total ribonucleic acid (RNA) from blood mononucleated cells was obtained from breast cancer patients (n = 36; age: 51.5 ± 12.5 years) and healthy donors (n = 14; age: 49.4 ± 9.4 years). Real-time polymerase chain reaction (RT-PCR) was performed to analyze the expression of TWIST-1 and MGA. Results: Patient carcinomas - ductal (86.7%), other types (13.3%). MGA gene expression was not detected in the donors' samples, while it was detected in 14% of the patient samples. Overexpression of TWIST-1 gene was observed in 17% of the patient samples. The combined analysis of both markers allowed the detection of CTCs in 27.8% of the samples, resulting in a significant (p < 0.05) sensitivity increase of detection. No significant associations (p > 0.05) were found between expression of the analyzed genes and the breast cancer prognostic factors. Conclusion: Combined analysis of TWIST-1 and MGA increased the sensitivity of CTCs detection compared to the single analysis of each gene. The detection of CTCs was not associated with known prognostic factors, suggesting that it is able to provide clinical information in addition to routine breast cancer clinicopathological parameters.

RESUMEN Objetivos: Las células de cáncer de mama liberadas al torrente sanguíneo se llaman células tumorales circulantes (CTCs). Las CTCs pueden expresar diferentes genes, como TWIST-1 y mamaglobina A (MGA). Los objetivos de este estudio fueron analizar la expresión de TWIST-1 y MGA en la sangre de pacientes con cáncer de mama (CM) para detectar CTCs y evaluar la asociación entre la presencia de CTCs y los parámetros pronósticos del CM. Métodos: Estudio prospectivo. Se obtuvo el ácido ribonucleico (ARN) de las células mononucleadas en la sangre de pacientes con CM (n = 36, edad: 51,5 ± 12,5 años) y donantes sanas (n = 14; edad: 49,4 ± 9,4 años). Se realizó reacción en cadena de la polimerasa en tiempo real (RT-PCR) para analizar la expresión de TWIST-1 y MGA. Resultados: Carcinoma ductal (86,7%), otros tipos (13,3%). No se detectó la expresión del gen MGA en las muestras de las donantes, pero en el 14% de las muestras de las pacientes. Se observó elevada expresión de TWIST-1 en el 17% de las muestras de pacientes con CM. El análisis combinado de ambos marcadores permitió detección de CTCs en el 27,8% de las muestras, resultando en un aumento significativo (p < 0,05) en la sensibilidad de detección. No se encontraron asociaciones significativas (p > 0,05) entre la expresión de los genes y los factores pronósticos. Conclusión: El análisis combinado de TWIST-1 y MGA aumentó la sensibilidad de detección de CTCs en comparación con el análisis de cada gen. La detección de CTCs no se asoció a factores pronósticos conocidos, sugiriendo que podría ofrecer informaciones clínicas adicionales a los parámetros clínico-patológicos de rutina del CM.

RESUMO Objetivos: As células cancerígenas da mama liberadas na corrente sanguínea são chamadas de células tumorais circulantes (CTCs). As CTCs podem expressar diferentes genes, como TWIST-1 e mamaglobina A (MGA). Os objetivos deste estudo foram analisar a expressão de TWIST-1 e MGA no sangue de pacientes com câncer da mama (CM) para detectar CTCs e avaliar a associação entre a presença de CTCs e os parâmetros prognósticos do CM. Métodos: Estudo prospectivo. O ácido ribonucleico (RNA) das células mononucleadas no sangue foi obtido de pacientes com CM (n = 36, idade: 51,5 ± 12,5 anos) e doadoras saudáveis (n = 14; idade: 49,4 ± 9,4 anos). Reação da cadeia da polimerase em tempo real (RT-PCR) foi realizada para analisar a expressão de TWIST-1 e MGA. Resultados: Carcinoma ductal (86,7%), outros tipos (13,3%). A expressão do gene MGA não foi detectada nas amostras das doadoras, mas foi observada em 14% das amostras das pacientes. Superexpressão de TWIST-1 foi observada em 17% das amostras dos indivíduos com CM. A análise combinada de ambos os marcadores permitiu a detecção de CTCs em 27,8% das amostras, resultando em um aumento significativo (p < 0,05) na sensibilidade da detecção. Associações significativas (p > 0,05) entre a expressão dos genes e os fatores prognósticos não foram encontradas. Conclusão: A análise combinada de TWIST-1 e MGA aumentou a sensibilidade da detecção de CTCs em comparação com a análise de cada gene. A detecção de CTCs não foi associada a fatores prognósticos conhecidos, sugerindo que ela pode fornecer informações clínicas adicionais aos parâmetros clinicopatológicos de rotina do CM.

Role of twist and podoplanin in partial epithelial-mesenchymal transition in oral squamous cell carcinoma

Abstract The aim of this study was to perform a comparative analysis of podoplanin (PDPN) and Twist immunoexpressions in lower lip and oral tongue squamous cell carcinomas (LLSCC and OTSCC, respectively). PDPN and Twist immunoexpressions were semi-quantitatively evaluated by analyzing the invasion front, the compressive areas, the large islands and nests and dissociated cells of the chosen carcinomas. Their statistical associations and correlations with clinical-pathological characteristics were verified by the Mann-Whitney and Spearman's test. Twist expression was low in both carcinomas, with <25% labeling on the invasive front. Significant differences were observed for LLSCC (p=0.032) and OTSCC (p=0.025) regarding PDPN immunoexpression in relation to the worst invasion patterns determined by a histological malignancy gradation system. Statistically significant negative correlations between PDPN membrane expression and general (r=-0.356, p=0.024) and cytoplasmic Twist expressions (r=-0.336; p=0.034) in LLSCC were also observed. Twist and PDPN are suggested to be associated to a more aggressive invasion pattern in both LLSCC and OTSCC cases but not related to the different biological behaviors on these anatomical sites. Also, it was seen that PDPN membrane expression is inversely related to general and cytoplasmic Twist expression in LLSCC cases.

Resumo O objetivo deste estudo foi realizar uma análise comparativa das imunoexpressões de podoplanina (PDPN) e Twist em carcinomas de células escamosas de lábio inferior e língua oral (CCELI e CCELO, respectivamente). As imunoexpressões de PDPN e Twist foram avaliadas semi-quantitativamente através da análise do front invasivo, das áreas compressivas, das grandes ilhas e ninhos e das células dissociadas dos carcinomas escolhidos. Suas associações estatísticas e correlações com características clínico-patológicas foram verificadas pelos testes de Mann-Whitney e Spearman. A expressão de Twist foi baixa nos dois carcinomas, com marcação <25% no front invasivo. Diferenças significativas foram observadas para CCELI (p=0,032) e CCELO (p=0,025) em relação à imunoexpressão de PDPN em relação aos piores padrões de invasão determinados por um sistema de gradação histológica de malignidade. Também foram observadas correlações negativas estatisticamente significantes entre a expressão membranar de PDPN e as expressões geral (r=-0,356, p=0,024) e citoplasmática do Twist (r=-0,336; p=0,034) no CCELI. Sugere-se que o Twist e o PDPN estejam associados a um padrão de invasão mais agressivo nos casos de CCELI e CCELO, mas não relacionados aos diferentes comportamentos biológicos nesses sítios anatômicos. Também foi observado que a expressão membranar de PDPN está inversamente relacionada à expressão geral e citoplasmática de Twist em casos de CCELI.

Organoaxial volvulus of descending colon: a case report

“Colonic volvulus” refers to the twisting of colon, which most commonly involves sigmoid colon causing obstruction, ischemia and gangrene. But very rarely segment of descending colon can be involved. This is a case of 42 year old male with vomiting, abdomen pain and distension since one day, showing organoaxial volvulus of descending colon loop with a twist of mesentery.

Experimental study on the effect of curcumin on endometriosis in rats / 中国临床药理学与治疗学

AIM: To analyze the effect of curcumin on the expression of transcription factors Twist1, Twist2, E-cadherin and Syndecan-1 in rats with endometriosis. METHODS: Thirty SD rats were randomly divided into five groups: model group, low dose curcumin group (25 mg•kg-1•d-1), medium dose curcumin group (50 mg•kg-1•d-1), high dose curcumin group (100 mg•kg-1•d-1) and gestrinone group (0.5 mg•kg-1•d-1). There were 6 rats in each group, and 5 rats in sham operation group were set up as blank control group. The expression of Twist1, Twist2, E-cadherin and Syndecan-1 was detected by immunohistochemistry. RESULTS: Compared with the model group, the ectopic tissue volume of curcumin group and gestrinone group was significantly lower, the difference was statistically significant (P0.05). CONCLUSION: Curcumin can reduce the expression of Twist1, Twist2 and Syndecan-1, increase the expression of E-cadherin, and inhibit the growth of endometrium in the model rats, so as to achieve the purpose of treatment.

Performance of the TWIST Score in Patients with Testicular Torsion that Present to the Emergency Department / Desempeño del puntaje TWIST en pacientes con torsión testicular que se presentan al Departamento de Emergencias

Objective Testicular torsion (TT) is an emergency with an incidence of 1:1,500 in patients < 18 years old. Irreversible changes in the testicular parenchyma may happen. The Testicular Workup for Ischemia and Suspected Torsion (TWIST) questionnaire evaluates signs and symptoms to determine the risk of TT and improve the time of management. The aim of the present study was to compare the intraoperative findings of patients with TT with the result of the preoperative TWIST questionnaire. Methods A cohort of 33 pediatric patients that consulted to the emergency room was evaluated. The TWIST questionnaire was applied in the first approach. Imaging studies, time to the operating room (OR) and intraoperative findings were evaluated. Theoretical and real findings were compared. Results The median age was 13 years old (interquartile range [IQR] 10­15 years old). Edema and scrotal inflammation was the most frequent finding (42.4%), followed by testicular induration (21.2%), nausea and vomiting (15.2%), and horizontal testicle and absence of cremasteric reflex (9.1%). The TWIST stratification was 3% high-risk, 18.2% intermediate-risk and 78.8% low-risk. Testicular Doppler ultrasound was performed in 93.9% of the patients: vascular congestion was found in 21.9%. A total of 30.3% of the kids were taken to surgery after 163 minutes (±116.5). Intraoperative diagnosis of TT was confirmed in the high-risk patient, in 33.3% of the intermediate-risk, and in 50% of the low-risk. The receiver operating characteristic (ROC) curve showed an accuracy of 60% (p = 0.602). Conclusions The TWIST questionnaire in the first approach allows to take the high-risk patients im

Objetivo La torsión testicular (TT) es una emergencia con incidencia de 1:1.500 en pacientes menores de 18 años. Pueden ocurrir cambios irreversibles en el parénquima testicular. El cuestionario TWIST evalúa signos y síntomas para determinar el riesgo de TT y mejorar los tiempos de atención. El objetivo de este estudio fue comparar los hallazgos intraoperatorios de los pacientes con el resultado del TWIST preoperatorio. Métodos Se evaluó una cohorte de 33 pacientes pediátricos que consultaron al Departamento de Emergencias. Se aplicó el cuestionario TWIST en la primera aproximación. Estudios imagenológicos, tiempo de entrada a salas de cirugía y hallazgos intraoperatorios también fueron evaluados. Se compararon los hallazgos teóricos y reales. Resultados La mediana de edad fue de 13 años (rango intercuartil [RIQ]: 10­15). Edema e inflamación escrotal fueron los hallazgos principales, (42,4%) seguidos de induración testicular (21,2%), náusea y vómito (15,2%), y testículo horizontal y ausencia del reflejo cremastérico (9,1%). Estratificación TWIST: 3% alto riesgo, 18,2% intermedio, y 78,8% bajo. Se realizó Doppler testicular en 93,9% de los pacientes: se encontró congestión vascular en 21,9%. Se operaron 30,3% de niños tras 163 minutos (± 116,5). En el intraoperatorio se confirmó TT en el paciente de alto riesgo, en 33,3% de intermedio, y en 50% de bajo riesgo. La curva de característica operativa del receptor (COR) evidencia una exactitud de 60% (p = 0.602). Conclusiones El cuestionario TWIST en la primera aproximación es útil para operar inmediatamente pacientes de alto riesgo. Sin embargo, no ofrece un alto nivel de confianza para el diagnóstico de TT en pacientes de intermedio y bajo riesgo.

Overexpression of histone deacetylase 11 suppresses basal-like breast cancer cell invasion and metastasis / 南方医科大学学报

OBJECTIVE@#Histone deacetylase 11 (HDAC11) is a class Ⅳ member of histone deacetylase family, and its role in regulating cancer cell invasion and metastasis remains unclear. We aimed to investigate the role of HDAC11 in regulating the biological behaviors of basal-like breast cancer (BLBC) cells.@*METHODS@#We analyzed the expression of HDAC11 based on Gene Expression Omnibus (GEO) and the Cancer Genome Atlas (TCGA). The effects of HDAC11 on the cell invasion and metastasis were examined using Transwell assay and in a mouse model. The interaction between HDAC11 and Twist was detected with immunoprecipitation. We identified 2 as a target gene of Twist using promoter luciferase assay and chromatin immunoprecipitation assay.@*RESULTS@#HDAC11 was lowly expressed in BLBC cells. HDAC11 overexpression suppressed BLBC cell invasion and their metastasis in nude mice. Mechanistically, HDAC11 directly interacted with Twist protein, antagonized its pro-invasive function and repressed Twist-induced 2 gene transcription.@*CONCLUSIONS@#Our data suggest that HDAC11 acts as a negative modulator of invasion and metastasis of BLBC cells.

Effect of Twist on angiogenesis of nasopharyngeal cancer cells by regulating ERK signaling pathway / 中国病理生理杂志

AIM:To investigate the effect of Twist transcription factor on angiogenesis of nasopharyngeal cancer cells by regulating ERK signaling pathway in vitro.METHODS:Twist siRNA was transfected into the CNE2 cells with Lipofectamine 2000, and the CNE2 cell-conditioned medium (CM) was prepared, which was then used to treat human umbilical vein endothelial cells (HUVECs).The CCK-8 assay, Boyden chamber and capillary tube formation assay were used to evaluate the angiogenic activity of HUVECs.The mRNA expression of Twist in nasopharyngeal cancer cells was detected by real-time PCR.The protein levels of Twist, ERK, p-ERK, p38, p-p38, JNK, p-JNK and VEGF were detected by Western blot.The ERK, p38 and JNK signaling pathway inhibitors were employed to treat HUVECs, and their effect on the capillary tube formation of HUVECs was analyzed.RESULTS:The expression of Twist at mRNA and protein levels in the CNE2 cells was significantly inhibited after transfection with Twist siRNA (P<0.01).The abilities of proliferation, invasion and capillary tube formation of HUVECs were markedly suppressed after knockdown of Twist expression (P<0.01).The protein level of p-ERK was significantly elevated in the CNE2-siTwist cells, while VEGF expression was decreased.Knockdown of Twist did no change the levels of p-p38 and p-JNK.The ERK signaling pathway inhibitor partly abrogated the anti-angiogenic effect of Twist knockdown.CONCLUSION:Knockdown of Twist inhibits the angiogenesis of nasopharyngeal cancer cells in vitro partly through activation of ERK signaling pathway.

MicroRNA-10b promotes the proliferation and invasion of osteosarcoma cell lines by targeting twist gene / 局解手术学杂志

Objective To investigate the role of microRNA-10b (miR-10b) in the proliferation and invasion potential of osteosarcoma cell lines MG-63 and the exact underlying mechanism.Methods The expression level of miR-10b in human osteosarcoma tissue samples and adjacent normal bone tissues were detected by relative quantitative real-time PCR (qRT-PCR).miR-10b mimic and siRNA against Twist (Twist siRNA) were transfected into human osteosarcoma cell lines MG-63 respectively using lipofactamine 2000, and RT-PCR was used to detect the mRNA expression levels of miR-10b and Twist, and Twist protein expression level was detected by Western blot.The effect of miR-10b mimic and Twist siRNA on proliferation of MG-63 were detected by MTT[3- (4, 5-dimethylthiazol-2-yl) -5- (3-carboxymethoxyphenyl) -2- (4-sulfophenyl) -2 H-tetrazolium].The in-vitro cell invasion ability was determined by Transwell invasion assays after up-regulating miR-10b or knocking down of Twist.Results The expression levels of miR-10b was higher in human osteosarcoma tissue samples compared with adjacent normal bone tissues, the differences were extremely statistical significance (P＜0.01).miR-10b directly up regulated the mRNA and protein expression levels of Twist, the differences were significant (P＜0.05).In addition, miR-10b had enhanced the cell invasion and the proliferation (P＜0.05), whereas the proliferation and invasion ability of MG-63 which transfected by both miR-10b mimic and Twist siRNA were significantly reduced than that transfected by miR-10b mimic (P＜0.05).Conclusion miR-10b in MG-63 promotes the proliferation and invasion potential of human osteosarcoma cell lines MG-63, at least partly through the upregulation of Twist gene.