Targeted inhibition of lncRNA CTB-191K22.5 reduces the proliferation and invasion of colorectal cancer SW480 cells and its mechanism / 国际外科学杂志

Objective:To explore the effect of down-regulation of long non-coding RNA (lncRNA) CTB-191K22.5 on the proliferation and invasion of colorectal cancer SW480 cells and the molecular mechanism.Methods:The TCGA database was used to analyze the expression differences of CTB-191K22.5 in colorectal cancer tissues and normal tissues. The CTB-191K22.5 inhibitor (Anti-CTB-191K22.5) and negative inhibitor (Control) were transfected into colorectal cancer SW480 cells, denoted as Observation group and Control group, real-time quantitative polymerase chain reaction (qRT) -PCR) was used to evaluate the inhibitory effect. MTT method and Transwell chamber method were used to evaluate the proliferation and invasion of SW480 cells. Western blot was used to evaluate the protein levels of PI 3K/AKT/mTOR signaling pathway in SW480 cells. The bioinformatics software starBase v2.0 was used to predict the target genes of CTB-191K22.5. qRT-PCR was used to evaluate the expression of CTB-191K22.5 target gene in SW480 cells. Measurement data were expressed as Mean±SD, and t-test was used for comparison between two groups. Results:Compared with normal tissues, the expression of CTB-191K22.5 in colorectal cancer tissues was significantly increased ( P<0.01). The expression of CTB-191K22.5 in SW480 cells of the Control group and Observation group were 6.60±0.85 and 1.08±0.21, respectively. The expression level of CTB-191K22.5 decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the SW480 cell proliferation ability of the Observation group decreased ( P<0.01). The invasion numbers of SW480 cells in the Control group and Observation group were (135.4 ± 16.29) and (42.24±14.59), respectively. The invasion ability of SW480 cells decreased after transfection with Anti-CTB-191K22.5 ( P<0.01). Compared with the Control group, the expression levels of PI 3K/AKT/mTOR signaling pathway protein in SW480 cells in the Observation group decreased. miR-326 may be the target gene of CTB-191K22.5. Compared with the Control group, transfection with Anti-CTB-191K22.5 significantly increased the expression level of miR-326 in SW480 cells ( P<0.01). Conclusion:CTB-191K22.5 is highly expressed in colorectal cancer tissues, and down-regulation of CTB-191K22.5 may inhibit the proliferation and invasion of colorectal cancer SW480 cells by targeting miR-326.

Cutaneous Tuberculosis in HIV Patient: A Case Report

Summary@#Tuberculosis (TB) is a serious communicable disease of major concern in endemic regions. Cutaneous tuberculosis (CTB), which accounts for less than 1% of all cases, can cause severe infection in susceptible patients. The diagnosis of CTB is challenging as it can present with a multitude of clinical presentations. The diagnosis must be supported by highly sensitive and specific investigations. This paper highlights the susceptibility of immunocompromised patients to the development of CTB and the challenges in making a diagnosis.

Tuberculosis verrucosa cutis en niños: Presentación de dos casos y revisión de la literatura / Tuberculosis verrucosa cutis in children: Two cases report and review

La tuberculosis cutánea (TBC) representa el 1.5% de todas las formas de TB extrapulmonares. Es una enfermedad proteiforme y su diagnóstico constituye un reto al clínico. La TB verrucosa cutis (TBVC) es una variante clínica paucibacilar, de TBC, donde es infrecuente aislar en medios de cultivos el Mycobacterium tuberculosis. Ocurre en niños previamente sensibilizados e inmunocompetentes que se reinfectan con este bacilo de una fuente exógena, algunas veces de un esputo o secreción de un paciente con TB activa, cuando juega en la tierra. La bacteria penetra a la piel a través de pequeñas abrasiones en zonas expuestas a traumas como lo son rodillas, tobillos o glúteos. La lesión se inicia como un papulonódulo verrucoso asintomático, parecido a una verruga común, que evoluciona a una placa verrucosa . Reportamos 2 casos de TBVC, una niña de 6 años de edad y el segundo, un niño de 10 años de edad ambos de la comarca Guna Yala, un área con alta prevalencia de TB en Panamá. En el segundo caso se logró aislar el M. Tuberculosis en medios de cultivo y demostrarse además por la reacción en cadena de la polimerasa (PCR). La respuesta a la terapia antituberculosa fue exitosa, en ambos casos, con completa regresión de las lesiones a los 2 meses de tratamiento

Cutaneous tuberculosis (CTB) comprises 1.5% of all extrapulmonary TB forms. It is a proteiform disease and its diagnosis represents a challenge to the clinician. TB verrucosa cutis (TBVC) a paucibacillary clinical manifestation of CTB, where routine cultures for Mycobacteriun tuberculosis, are commonly negative. Occurs in children previously sentitized with a moderate to high degree of immunity. It represents an exogenous reinfection of M. tuberculosis, sometimes from a contaminated sputum of a patient with active TB, at sites prone to minor abrasions, such as, knees, ankles or buttocks. The lesion begins as a solitary asymptomatic verrucous papulonodule, similar to a common wart, that evolves to a verrucous plaque. The author report 2 cases of TBVC, a 6 year old girl and the second one, a 10 year old boy, both from Guna Yala, an indigenous region, o the coast of Panama, an area with a high prevalence of TB. In the second case, M. tuberculosis was isolated in routine cultures and reported positive in Polimerase chain reaction (PCR). The antituberculous therapy was successfull in both patients, with complete regression of the lesions after 2 months of treatment

Effects of Peripheral Neuropathy on CTB-labeled Motor Neurons Following Ligation of the Tibial Nerve / 체질인류학회지

Changes in CTB labeled motor neurons of the spinal cord were observed after the induction of peripheral neuropathy by ligation of the tibial nerve. Rats were anesthetized and the tibial nerve was ligated with 3-0 silk. The rats were separated into three groups based on the length of time the tibial nerve was ligated (1, 2, or 4 weeks). After the ligation procedures were complete, the tibial nerve stumps were soaked in CTB solution. Tibial nerve segments and the spinal cord were then observed. In the control and experimental groups, CTB-labeled neurons formed a discrete population that was concentrated primarily at the L5 level, while the contributions from L4 and L6 were minor. According to the distributions, CTB-labeled neurons were divided into rostral and caudal groups. A selective decrease of CTB-labeled neurons was observed only in the caudal group, extending from the rostral L5 to one-half of the rostral L6. The total numbers of CTB-labeled motor neurons were 2,160+/-169.3, 1,002+/-245.1, 587.5+/-346.5, and 1,728+/-402.6 in the control group, 1 week group, 2 week group, and 4 week group, respectively. The selective decrease of CTB-labeled neurons in the caudal division was responsible for the decrease in the total number of labeled neurons in all groups. Following peripheral neuropathy caused by ligation of the tibial nerve, CTB-labeled neurons in the spinal cord decreased selectively. These results may provide important neuroanatomical data regarding the effects of peripheral neuropathy by ligation of the tibial nerve.

Exploring the binding affinities of p300 enzyme activators CTPB and CTB using docking method.

CREB binding protein (CBP) and E1A binding protein p300, also known as p300 are functionally related transcriptional co-activators (CoAs) and histone acetyltransferases (HATs). Some small molecules, which target HATs can activate or inhibit the p300 enzyme potently. Here, we report the binding affinities of two small molecules CTPB [N-(4-chloro- 3-trifluoromethyl-phenyl)-2-ethoxy-6-pentadecyl-benzamide] and CTB [N-(4-chloro-3-trifluoromethyl-phenyl)-2-ethoxy-benzamide] with p300 using docking method to obtain the insight of their interaction with p300. These small molecules bind to the enzyme, subsequently causing a structural change in the enzyme, which is responsible for the HAT activation. CTB exhibits higher binding affinity than CTPB, and their lowest docked energies are -7.72, -1.18 kcal/mol, respectively. In CTPB molecule, phenolic hydroxyl of Tyr1397 interacts with the non-polar atoms C(5E) and C(5F), and forms polar-non polar interactions. Similar interactions have also been observed in CTB. The residues Tyr1446 and Cys1438 interact with the non-pentadecyl atoms. Further, the docking study predicts a N-HO hydrogen bonding interaction between CTB and Leu1398, in which the HO contact distance is 2.06 Å. The long pentadecyl chain of CTPB reduces the formation of hydrogen bond with the p300. The H-bond interaction could be the key factor for the better activation of CTB.

Cloning and Expression of Vibrio cholerae CTB Gene and the Recombinant CTB Protein Activation Assay / 中国生物工程杂志

CTB protein possessed mucosal adjuvant immunoactivity. The CTB gene was amplified by PCR method from a strain V. cholerae. The nucleotide sequence of CTB gene was 375 bp and shared 96.0%~99.2% homology with other 6 CTB genes. The recombinant plasmid pTWIN1-CTB transformed E. coli strain BL21(DE3) expressed with 0.8 mmol/L IPTG. The molecular weight of expression products was identical with expectative weight by SDS-PAGE electrophoresis. The CTB fusion proteins mainly assembled inclusion bodies and the outputs of proteins were approximately 20% of the total bacterial proteins. The CTB proteins possessed mucosal immunoactivity by GM1-ELISA assay.

Degeneration of primary afferent terminals following brachial plexus extensive avulsion injury in rats / Degeneración de los terminales aferentes primarios de rata luego de lesión extensa por avulsión del plexo braquial

Important breakthroughs in the understanding regeneration failure in an injured CNS have been made by studies of primary afferent neurons. Dorsal rhizotomy has provided an experimental model of brachial plexus (BP) avulsion. This is an injury in which the central branches of primary afferents are disrupted at their point of entry into the spinal cord, bringing motor and sensory dysfunction to the upper limbs. In the present work, the central axonal organization of primary afferents was examined in control (without lesion) adult Wistar rats and in rats subjected to a C3-T3 rhizotomy. Specific sensory axon subtypes were recognized by application of antibodies to the calcitonin gene-related peptide (CGRP), the P2X3 purinoreceptor, the low-affinity p75-neurotrophin receptor and the retrograde tracer cholera toxin subunit beta (TCbeta ). Other subtypes weres labeled with the lectin Griffonia simplicifolia IB4. Using immunohistochemistry and high resolution light microscopy, brachial plexus rhizotomy in adult rats has proven a reliable model for several neural deficits in humans. This lesion produced different degrees of terminal degeneration in the several types of primary afferents which define sub-populations of sensitive neurons. Between the C6 and C8 levels of the spinal cord,,deafferentation was partial for peptidergic GCRP-positive fibers, in contrast with elimination of non peptidergic and myelinated fibers. Dorsal rhizotomy has provided an adequate experimental model to study sensory alterations such as acute pain and allodynia as well as factors that affect regeneration into the CNS., Therefore, the differential deafferentation response must be considered inr the evaluation of therapies for nociception (pain) and regeneration for brachial plexus avulsion. The anatomical diffierences among the primary afferent subtypes also affect their roles in normal and damaged conditions.

El uso de las neuronas sensoriales primarias ha aportado avances en el entendimiento de las razones por las cuales falla la regeneración cuando el sistema nervioso central (SNC) es dañado. La rizotomía dorsal se puede usar como un modelo experimental de las lesiones por avulsión del plexo braquial, una lesión en la cual son desprendidas, en su punto de entrada en la médula espinal, las ramas centrales de los aferentes primarios causando una disfunción motora y sensorial grave e irreversible del miembro superior. En el presente trabajo, se examinó la organización central de los aferentes primarios en ratas Wistar adultas. Éstas fueron divididas en controles normales no lesionados y en animales rizotomizados entre los niveles cervical 3 y torácico 3 (C3-T3). Se estudió la deaferentación de los subtipos de axones sensoriales utilizando anticuerpos específicos contra el péptido relacionado con el gen de la calcitonina (CGRP), el receptor purinérgico (P2X3), el receptor de baja afinidad p75 para el factor de crecimiento nervioso (NGF) y contra la subunidad ®de la toxina de cólera (TCbeta ). Otro subtipo fue marcado con la lectina Griffonia simplicifolia IB4. La inmunohistoquímica y la microscopía óptica de alta resolución demostraron que el modelo animal de rizotomía completa del plexo braquial reproduce diversos déficit observados en las lesiones humanas. Esta lesión produce diferentes grados de degeneración terminal entre los diversos tipos de aferentes primarios que definen subpoblaciones de neuronas sensoriales. En los niveles de la médula espinal estudiados (entre C6 y C8), la deaferentación fue parcial para las fibras peptidérgicas GCRPpositivas, en contraste con la eliminación de las fibras no peptidérgicas y las mielinizadas. La rizotomía dorsal es un modelo experimental apropiado para estudiar las alteraciones sensoriales como el dolor agudo y la alodinia, así como los factores que podrían afectar la regeneración en el SNC. Por tanto, la respuesta de deaferentacion diferencial debe ser tenida en cuenta para la evaluación de terapias antinociceptivas y regenerativas tras la avulsión del plexo braquial. Se discute la anatomía de los subtipos de aferentes primarios y su papel en condiciones normales y después de la lesión.

Localization of Motor Neuronal Cell Bodies Innervating the Digastric Muscle of the Rat Using Cholera Toxin B Subunit (CTB) / 대한해부학회지

We used cholera toxin B subunit (CTB) as a neural tracer to localize motor neuronal cell bodies innervating the digastric muscle. After CTB injection into the left anterior belly, CTB-labelled motor neuronal cell bodies were found in caudal half of the left and right trigeminal nucleus, the left and right facial nucleus, the accessory facial nucleus and the accessory trigeminal nucleus in pons. The total number of CTB-labelled motor neuronal cell bodies were 1,179+/-119.5 in the left pons and 246+/-61.8 in the right pons after CTB injections into the left anterior belly of digastric muscle. After CTB injection into left posterior belly, CTB-labelled motor neuronal cell bodies were found only in the left ventral part of accessory facial nucleus in caudal pons and the total number of CTB-labelled motor neuronal cell bodies were 270+/-29.3.

Localization of Nerves Innervating Sublingual and Submandibular Gland in the CNS Using Cholera Toxin B Subnit and Pseudorabies Virus / 체질인류학회지

In the rat brain stem, neurons innervating the sublingual and submandibular gland were investigated by means of cholera toxin B subunit (CTB) and pseudorabies virus (PRV). Injection of CTB into the sublingual gland and PRV into the submandibular gland, neural tracer labeled neurons showed similar positions in central nervous system with PRV into the sublingual gland and CTB into the submandibular gland. CTB labeled-neurons were observed in superior salivatory nucleus, PRV labeled-neurons in superior salivatory nucleus and reticular nucleus. CTB was more fine tracer than PRV for observation of superior salivatory nucleus. The size of CTB labeled-neurons is larger in submandibular gland than in sublingual gland. The size of PRV labeled-neurons were nearly the same after injection to submandibular or sublingual gland. No neurons were labeled together with CTB and PRV. Neurons innervating sublingual and submandibular gland were localized independently in superior salivatory nucleus. These results provided a neuroanatomical data of the neurons innervating the sublingual and submandibular gland in the superior salivatory nucleus.

Protective effects of ?-crystallin on injured optic nerve fiber of LongEvens rats by CTB conjugates anterograde labeling / 第三军医大学学报

Objective To investigate in vivo the protective effects of intraocular injection of ?-crystallin after optic nerve injury.Methods Sixteen rats were divided into normal control group,and 2-week,4-week,6-week groups after optic nerve lesion.?-crystallin and bovine serum albumin(BSA) was respectively injected into the right and left corpus vitreums after the optic nerve crush injury.CTB conjugates were injected into the vitreous of all the rats at 48 h before observing optic nerve with confocal microscope(488 nm).Results It could be seen the normal stripe of the nerve fiber in the injuried optic nerve stained with CTB conjugates and some dots and lumps of fluorescence could be seen in the optic nerve.The fluorescence intensity of the ?-crystallin group was notably higher than the BSA group.Conclusion CTB conjugates anterograde labeling is a reliable and convenient method to observe the optical nerve fiber after optic nerve lesion.?-crystallin administered intraocularly at the lesion time can protect the optic nerve fiber.

The Localization of Efferent and Afferent Neurons Innervating the Rat Thymus Using the Neural Tracers / 대한해부학회지

The localizations of efferent and afferent neurons were observed following injection of neural tracers, cholera toxin B subunit (CTB) and wheat germ agglutinin-horseradish peroxidase (WGA-HRP) into the rat thymus with ages. Thirty Sprague-Dawley rats were examined at 3 weeks, 5~6 and 20 months of age. After survival times of 48~96 hours following injection of neural tracers, the rats were perfused and their brain, spinal cord, sympathetic ganglia, dorsal root ganglia and vagal ganglia were frozen sectioned (40 mm). These sections were stained by CTB immunohistochemical and HRP histochemical staining methods, and observed with polarized dark and light microscope. The results were as follows: 1. WGA-HRP and CTB labeled parasympathetic neurons were bilaterally seen in the nucleus ambiguus and medullary reticular formation of medulla with all ages. 2. WGA-HRP labeled sympathetic neurons were bilaterally labeled in superior cervical ganglia, middle cervical ganglia, stellate ganglia and T4-8 sympathetic chain ganglia. The number of labeled sympathetic neurons was increased in the thymus at 20 months of age. According to the aging, sympathetic neuronal processes were more developed, and the nerve fibers were coarse and more branched. 3. WGA-HRP labeled sensory neurons were bilaterally observed within the vagal and C1-6 dorsal root ganglia. The number of labeled sensory neurons was decreased in the thymus at 20 months of age.

Localization of Sensory Neurons Innervating the Rat Intestine Using the Cholera Toxin B Subunit(CTB) and Wheat Germ Agglutinin-Horseradish Peroxidase(WGA-HRP) / 영남의대학술지

The local arrangement of sensory nerve cell bodies and nerve fibers in the brain stem, spinal ganglia and nodose ganglia were observed following injection of cholera toxin B subunit(CTB) and wheat germ agglutinin-horseradish peroxidase(WGA-HRP) into the rat intestine. The tracers were injected in the stomach(anterior and posterior portion), duodenum, jejunum, ileum, cecum, ascending colon or descending colon. After survival times of 48-96 hours, the rats were perfused and their brain, spinal and nodose ganglia were frozen sectioned(40microM). These sectiones were stained by CTB immunohistochemical and HRP histochemical staining methods and observed by dark and light microscopy. The results were as follows: 1. WGA-HRP labeled afferent terminal fields in the brain stem were seen in the stomach and cecum, and CTB labeled afferent terminal fields in the brain stem were seen in all parts of the intestine. 2. Afferent terminal fields innervating the intestine were heavily labeled bilaterally gelalinous part of nucleus of tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS(comNTS), medial part of NTS(medNTS), wall of the fourth ventricle, ventral border of area postrema and comNTS in midline dorsal to the central canal. 3. WGA-HRP labeled sensory neurons were observed bilaterally within the spinal ganglia, and labeled sensory neurons innervating the stomach were observed in spinal ganglia T2-L1 and the most numerous in spinal ganglia T8-9. 4. Labeled sensory neurons innervating the duodenum were observed in spinal ganglia T6-L2 and labeled cell number were fewer than the other parts of the intestines. 5. Labeled sensory neurons innervating the jejunum were observed in spinal ganglia T6-L2 and the most numerous area in the spinal ganglia were T12 in left and T13 in right. 6. Labeled sensory neurons innervating the ileum were observed in spinal ganglia T6-L2 and the most numerous area in the spinal ganglia were T11 in left and L1 in right. 7. Labeled sensory neurons innervating the cecum were observed in spinal ganglia T7-L2 and the most numerous area in the spinal ganglia were T11 in left and T11-12 in right. 8. Labeled sensory neurons innervating the ascending colon were observed in spinal ganglia T7-L2 in left, and T9-L4 in right. The most numerous area in the spinal ganglia were T9 in left and T11 in right. 9. Labeled sensory neurons innervating the descending colon were observed in spinal ganglia T9-L2 in left, and T6-L2 in right. The most numerous area in the spinal ganglia were T13 in left and L1 in right. 10. WGA-HRP labeled sensory neurons were observed bilaterally within the nodose ganglia, and the most numerous labeled sensory neurons innervating the abdominal organs were observed in the stomach. 11. The number of labeled sensory neurons within the nodose ganglia innervating small and large intestines were fewer than that of labeled sensory neurons innervating stomach These results indicated that area of sensory neurons innervated all parts of intestines were bilaterally gelatinous part of nucleus tractus solitarius(gelNTS), dorsomedial part of gelNTS, commissural part of NTS(comNTS), medial part of NTS, wall of the fourth ventricle, ventral border of area postrema and com NTS in midline dorsal to the central canal within brain stem, spinal ganglia T2-L4, and nodose ganglia. Labeled sensory neurons innervating the intestines except the stomach were observed in spinal ganglia T6-L4. The most labeled sensory neurons from the small intestine to large intestine came from middle thoracic spinal ganglia to upper lumbar spinal ganglia.

ORIGINS OF THE NADPH-d POSITIVE TERMINALSIN THE MEDIAL AMYGDALOID NUCLEUS OF THE RATS / 解剖学报

Objective To estimate the effect of the NO on the medial amygdaloid nucleus(Me), we studied the origins of the NOS positive terminals in the Me. Methods Noergic afferent projection to Me was identified by a combined NADPH-d histochemical staning and retrograde CTb immunocytochemical method after microinjecting CTb into Me. Results The double labeled of neurons (NOS and CTb) were located in dorsal raphe nucleus, locus ceruleus, basolateral amygdaloid nucleus, parabrachial nucleus, ventrolateral part of periaqueductal gray.Conclution The NADPH-d positive terminals in the Me originates from the aforementioned nucleus, and may relate to the function of the Me.