Molecular mechanism of miR-1914-3p targeting ARL4C to inhibit the invasion and proliferation of renal cancer cells / 中国医师杂志

Objective:To explore the molecular mechanism of microRNA (miRNA, miR)-1914-3p regulating the expression of ARL4C and affecting the invasion and proliferation of renal cancer cells.Methods:Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression level of miR-1914-3p in tumor tissues and adjacent tissues of 53 renal cancer patients, 4 types of renal cancer cell lines (ACHN, OS-RC-2, 786-O, A498) and normal proximal renal tubular epithelial cell line (HK-2). The nonsense sequence (NC) and miR-1914-3p mimic were transiently transfected into renal cancer cells with the lowest miR-1914-3p expression by liposome method, namely the NC group and miR-1914-3p group. qRT-PCR was used to detect the expression level of miR-1914-3p in transfected cells. Transwell invasion test and cell counting kit-8 (CCK-8) were used to detect the invasion and proliferation ability of each group of cells. Bioinformatics software and dual luciferase gene report experiment were used to predict and test the targeted regulation mechanism of miR-1914-3p on target genes. qRT-PCR and Western blot was conducted to analyze the target gene expression level in cells of each group.Results:The expression level of miR-1914-3p in renal cancer tissue was significantly lower than that in adjacent tissues ( P<0.01). The expression level of miR-1914-3p in renal cancer cell lines was significantly lower than that in HK-2 cell lines ( P<0.01), and the expression of miR-1914-3p in OS-RC-2 cells was the lowest ( P<0.01). The expression of miR-1914-3p in the NC group and the miR-1914-3p group were (1.04±0.17) and (11.40±0.91), respectively. The expression level of miR-1914-3p in the miR-1914-3p group was significantly increased ( P<0.01), indicating that the transfection was successful. Overexpression of miR-1914-3p can significantly inhibit the invasion ( P<0.01) and proliferation ( P<0.05) of renal cancer OS-RC-2 cells. Dual luciferase gene report experiment indicated that the target gene of miR-1914-3p may be ADP-ribosylation factor-like 4C (ARL4C); miR-1914-3p can significantly inhibit the luciferase activity of wild-type ARL4C-3′UTR ( P<0.01). Overexpression of miR-1914-3p decreased the expression of ARL4C mRNA and protein in OS-RC-2 cells ( P<0.01), and decreased the expression of cell invasion phenotype proteins (Snail, Slug) and cell proliferation phenotype proteins (Mcm2, Mcm7) ( P<0.01). Conclusions:miR-1914-3p is low-expressed in renal cell carcinoma. It inhibits the invasion and proliferation of renal cell carcinoma OS-RC-2 cells through targeted interference with the expression of the oncogene ARL4C, and participates in the occurrence and development of renal cell carcinoma.

The Joubert Syndrome Gene arl13b is Critical for Early Cerebellar Development in Zebrafish / 神经科学通报·英文版

Joubert syndrome is characterized by unique malformation of the cerebellar vermis. More than thirty Joubert syndrome genes have been identified, including ARL13B. However, its role in cerebellar development remains unexplored. We found that knockdown or knockout of arl13b impaired balance and locomotion in zebrafish larvae. Granule cells were selectively reduced in the corpus cerebelli, a structure homologous to the mammalian vermis. Purkinje cell progenitors were also selectively disturbed dorsomedially. The expression of atoh1 and ptf1, proneural genes of granule and Purkinje cells, respectively, were selectively down-regulated along the dorsal midline of the cerebellum. Moreover, wnt1, which is transiently expressed early in cerebellar development, was selectively reduced. Intriguingly, activating Wnt signaling partially rescued the granule cell defects in arl13b mutants. These findings suggested that Arl13b is necessary for the early development of cerebellar granule and Purkinje cells. The arl13b-deficient zebrafish can serve as a model organism for studying Joubert syndrome.

The Joubert Syndrome Gene arl13b is Critical for Early Cerebellar Development in Zebrafish / 神经科学通报·英文版

Joubert syndrome is characterized by unique malformation of the cerebellar vermis. More than thirty Joubert syndrome genes have been identified, including ARL13B. However, its role in cerebellar development remains unexplored. We found that knockdown or knockout of arl13b impaired balance and locomotion in zebrafish larvae. Granule cells were selectively reduced in the corpus cerebelli, a structure homologous to the mammalian vermis. Purkinje cell progenitors were also selectively disturbed dorsomedially. The expression of atoh1 and ptf1, proneural genes of granule and Purkinje cells, respectively, were selectively down-regulated along the dorsal midline of the cerebellum. Moreover, wnt1, which is transiently expressed early in cerebellar development, was selectively reduced. Intriguingly, activating Wnt signaling partially rescued the granule cell defects in arl13b mutants. These findings suggested that Arl13b is necessary for the early development of cerebellar granule and Purkinje cells. The arl13b-deficient zebrafish can serve as a model organism for studying Joubert syndrome.

Screening for mutation hotspots in Bardet–Biedl syndrome patients from India

Background & objectives: Bardet–Biedl syndrome (BBS) is a genetically heterogeneous autosomal recessive disorder characterized by multiple organ defects involving retina, kidney, liver and brain. Disease-causing mutations in BBS genes narrowed down by homozygosity mapping in small consanguineous and non-consanguineous pedigrees were reported in 80 per cent of the study population. This study was aimed to screen these genes (BBS3, BBS10) and specific exons of BBS genes (BBS1, BBS5, MKKS, BBS9, BBS11 and BBS12) for recurrent mutations in a selected sample of BBS patients. Methods: The recurrent mutations in BBS genes were screened in the BBS affected individuals by PCR based direct sequencing. The pathogenicity of the observed mutations were confirmed by co-segregation analysis, screening of healthy unrelated controls and in silico analysis. Results: In the 64 BBS patients (44 males, 20 females) were studied, mutations were predominant in BBS10 and ARL6 genes; the c.272T>C; p.(I91T) mutation in ARL6 gene was a recurrent mutation. One novel non-sense mutation c.425T>G; p(L142*) was obtained in BBS5 gene (family BSI-31). Interpretation & conclusions: BBS10 gene mutations clustered in exon 2 of the gene suggesting the exon as a probable hotspot for mutations in Indian population. A cost- and time-effective strategy for the molecular diagnosis of BBS was designed based on these results.

Research progress on the role ofARL2 in the development of tumo / 中国肿瘤生物治疗杂志

@#ADP-核糖基化样因子2（ADP ribosylation factor-like protein 2，ARL2）是一种小型GTP结合蛋白，隶属于RAS超家族 中的ARF家族，广泛存在于真核细胞中，在分子结构上高度保守。ARL2参与调节微管动力学，维持细胞形态和极性；调控线粒 体功能，包括线粒体形态、运动和线粒体融合等。多种肿瘤中存在ARL2表达异常，并且改变肿瘤细胞中ARL2表达会影响肿瘤 细胞的形态、增殖和侵袭能力，影响肿瘤细胞对化疗药物的敏感性、细胞周期分布，甚至诱导细胞凋亡。本文拟对ARL2的目前研 究现状及其在肿瘤中的研究进展作一综述。

Relationship of the Arl8a with TLR4-TRIF pathway in dendritic cells / 中华微生物学和免疫学杂志

Objective To explore the relationship of the ADP-ribosylation factor-like 8A (Arl8a)with TLR4-TRIF-GEFH1 -RhoB pathway in dendritic cells(DCs). Methods DCs were prepared from wildtype and TRIF-knockout (TRIFKO) mice. After LPS stimulation, the cells were collected for cDNA amplification. Real-time PCR method was used to detect Arl8a mRNA levels. DCs from wild-type mice were transfected with guanine nucleotide-exchange factors H1 ( GEFH1 ) small interference RNA ( siRNA ), Arl8a mRNA levels were examined with or without LPS stimulation. Then RhoB mRNA expression was analyzed in DCs transfected with the siRNA of GEFH1 and Arl8a gene, respectively. Results LPS induced the up-regulation of Arl8a mRNA in DCs from control mice but not in DCs from TRIFKO, indicating that LPS-mediated up-regulation of Arl8a was suppressed in TRIFKO DCs. In addition, siRNA of GEFH1 significantly suppressed the LPS-mediated up-regulation of Arl8a mRNA, RNAi of Arl8a and GEFH1 significantly decreased RhoB mRNA level in DCs after LPS stimulation ( P ＜ 0. 01 ). Conclusion The expression of Arl8a is involved in the TLR4-TRIF pathway in DCs, and Arl8a is closely associated with GEFH1 and RhoB at transcriptional level.

The study by using the computerized tomography imaging in order to access to mandibular foramen while inferior alveolar nerve anesthesia

PURPOSE: This study was performed to provide an anatomical information of the mandibular ramus for the successful inferior alveolar nerve block. Three dimensional images were reconstructed from the computerized tomography (CT) and the anatomical evaluation of the mandibular ramus was done. MATERIALS AND METHODS: Sixty-four patients who had been taken the facial CT scans from 2000, Jan to 2003, June was selected. The patients who had the anterior or posterior teeth misssing, edentulous ridge, and jaw fracture were excepted. In the occulusal plane, the lingual surface angle (LSA) between the mid-sagittal plane and the mandibular molar lingual surface from the 2nd premolar to the 2nd molar, the inner ramal surface angle (IRSA), the maximum inner ramal surface angle (MxIRSA), and the outer ramal surface angle (ORSA) to the-mid sagittal plane were MEASURED: The inner ramal surface angle in the ligular tip level (IRSA-L) and the outer ramal surface angle in the ligular tip level (ORSA-L), the ramal length (RL), and the anterior ramal length (ARL) were also measured in the lingular tip level. RESULTS: In the lingular tip level, the mean IRSA-L and ORSA-L were 28.6+/-6.3 degrees and 17.9+/-4.9 degrees respectively. The larger was the IRSA, the larger was the ORSA. In the lingular tip level, the mean ramal length was 35.8+/-3.4 mm. The larger was the IRSA-L, the shorter was the ramal length. On the lingular tip level, the mean anterior ramal length from anterior ramus to lingular tip was 19.6+/-3.3 mm. when the ramal length was longer, the anterior ramal length was also longer. On the lingular tip level, there was positive correlation vetween the IRSA and the ORSA, negative correlation between the IRSA and the ramal length, and positive correlation between the ramal length and the lingular tip level to the anterior ramus. There was no statistical meaning of data between sex and age. CONCLUSION: In the clinical view of the results so far achieved, if the direction of needle is closer to posterior it is able to contact bone on lingular tip when the internal surface of ramus is wided outer.

Linfomas no-Hodgkin asociados a SIDA: 1. Evolución durante los 20 años de la pandemia. 2. Experiencia en el INCMNSZ / Acquired inmunodeficiency syndrome-related lymphoma: 1.Evolution during the 20 years of the epidemic. 2. The experience at the Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán: 1986-2003

El objetivo de esta revisión es situar la evolución epidemiológica y la historia natural de los linfomas no-Hodgkin (LNH) asociados a síndrome de inmunodeficiencia humana adquirida (SIDA) dentro del contexto de la evolución de la pandemia originada por la infección del virus de la inmunodeficiencia humana (VIH). Inicialmente realizamos una descripción del panorama mundial desde la aparición del primer caso de infección por VIH en 1981, el pico de la epidemia en 1993 y el evento que cambió la historia natural de la enfermedad: la terapia antirretroviral altamente efectiva (TARAE), introducida en 1995 en el mundo y en 1997 en nuestro país. Presentamos evidencia clara de la disminución en la mortalidad de pacientes con infección por VIH/SIDA y su relación paralela con la reducción en la frecuencia de las tres infecciones oportunistas (neumonía por Pneumocystis carinii, enfermedad por el complejo Mycobacterium avium y retinitis por citomegalovirus) más frecuentes en esta enfermedad. Describimos los factores de riesgo para padecer LNH en pacientes con VIH/SIDA y los factores pronósticos de supervivencia y remisión en estos pacientes. Señalamos también que se ha incrementado proporcionalmente el diagnóstico de SIDA definido por la presencia de LNH a partir del uso de TARAE. No está claramente definido en la literatura que la supervivencia de los pacientes con LNH asociados a SIDA haya cambiado significativamente a partir del uso de TARAE, pero existen evidencias que sugieren que la cuenta basal de linfocitos CD4 se ha visto incrementada con TARAE, redundando esto en una mejoría en la tasa de remisiones completas y supervivencia de los pacientes con LNH asociados a SIDA. La falta de congruencia en la literatura a este respecto posiblemente esté matizada por factores como apego a terapia antirretroviral, surgimiento de resistencia a la misma y heterogeneidad en los tratamientos de quimioterapia que han recibido estos pacientes. Existen muchas controversias en cuanto al tipo de quimioterapia que deben recibir los pacientes con LNH asociados a SIDA de reciente diagnóstico, que van desde la reducción o no de las dosis estándar de quimioterapia, la combinación temporal de ésta con TARAE, el uso de inmunoterapia conjuntamente con TARAEy quimioterapia. Finalmente, presentamos los resultados preliminares del análisis de la experiencia de nuestra Institución en LNH asociados a SIDA desde 1986 hasta diciembre del 2003.

The goal of this presentation is the description of the epidemiologic evolution and changes in natural history of the human immunodeficiency virus infection (HIV) epidemic itself and its relation with the acquired immunodeficiency syndrome-related lymphoma (ARL). We have started with the description of the world's state of the HIV epidemic, its features since the first case report in the United States of America in 1981, through the peak of new diagnoses in 1993 until the event that changed the natural history of the disease: the era of the widespread use of the highly active antiretroviral therapy (HAART), introduced in 1995 in the world and in 1997 in our country. The widespread introduction of HAART led to dramatic reductions in AIDS related mortality and morbidity throughout the developed world with a marked fall in the incidence of the major opportunistic infections in AIDS. We describe the main risk factors for the development of ARL, and the prognostic factors for survival and response to treatment. There is no clear definition in the literature of the roll that has played the use of HAART in relation to survival and response to treatment in ARL, but there is evidence that the basal count of CD4 cells has increased with HAART, leading to a better survival and response in ARL. The debate regarding this issue is surely affected by factors such as degree of antiretroviral treatment compliance, antiretroviral therapy resistance and chemotherapy heterogeneity. Finally we present the preliminary results of the analysis of our experience in ARL from 1986 to 2003.