Effect of ANKRD49 on migration of human lung adenocarcinoma cells and its mechanism / 中国生物制品学杂志

@#Objective To study the effect of ankyrin repeat domain 49(ANKRD49)on the migration of human lung adenocarcinoma cell line NCI-H1299 and its mechanism.Methods NCI-H1299 cells were infected with lentivirus vector carrying ANKRD49 gene and shRNA targeting ANKRD49 to construct the cell models stably overexpressing and knocking down ANKRD49. Meanwhile,the control cell models infected with empty lentivirus vector and lentivirus vector with scramble sequences were constructed respectively. The expression levels of ANKRD49 mRNA and protein were detected by real-time fluorescence quantitative PCR and Western blot. The effect of ANKRD49 on cell migration was measured by scratch test. The mRNA and protein levels of matrix metalloproteinase(MMP)-2/9 and tissue inhibitor of metalloproteinase(TIMP)-1/2 were detected by real-time fluorescence quantitative PCR and Western blot. The protein expression levels of p65,p-p65,IκBα and p-IκBα were detected by Western blot.Results The levels of ANKRD49 mRNA and protein in the ANKRD49 overexpression group were significantly higher than those in the control group(t = 70. 02 and 45. 68,respectively,each P < 0. 001). Compared with the control group,the migration ability of cells in the ANKRD49 overexpression group significantly increased at 24 h and 48 h(t = 5. 343 and 3. 282,P = 0. 005 9 and 0. 030 4,respectively);The mRNA transcription levels and protein expression levels of MMP-2 and MMP-9 significantly increased(t = 9. 304 and 6. 193,P =0. 000 7 and 0. 003 5,respectively),while the mRNA and protein expression of TIMP-1 and TIMP-2 decreased significantly(t = 3. 858 and 3. 517,P = 0. 018 2 and 0. 024 5,respectively),and the values of MMP-2/TIMP-1 and MMP-9/TIMP-2 significantly increased(t = 17. 7 and 9. 682,P < 0. 001 and < 0. 01,respectively);The expression of p-p65 and pIκBα significantly increased,the total protein levels of p65 and IκBα showed no obvious change,and the values of p-p65/p65 and p-IκBα/IκBα significantly increased(t = 3. 962 and 5. 370,P = 0. 016 7 and 0. 005 8,respectively). However,knocking down of ANKRD49 presented the opposite results.Conclusion ANKRD49 promotes the migration of NCI-H1299cells by enhan-cing the expression of MMP-2/9,the values of MMP-9/TIMP-1 and MMP-2/TIMP-2 via activating NF-κB/p65 signa-ling pathway.

Research on the antioxidant activity of metabolites from a sponge-derived fungus Alternaria sp. F49 / 药学学报

To study the chemical constituents from the the deep-sea fungus Alternaria sp. F49. Seven compounds were isolated from the EtOAc extract by using silica gel, Sephadex LH-20, ODS and HPLC methods. Based on the spectroscopic analysis, their structures were identified as (8R)-5-O-methyl-orcinotriol (1), orcinotriol (2), α-acetylorcinol (3), 3'-hydroxyalternariol 5-O-methyl ether (4), altenusiol (5), altenusin (6), and 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (7). (8R)-5-O-Methyl-orcinotriol (1) is a new phenolic compound which has never been reported in the literature. Compounds 4-7 showed strong DPPH free radical scavenging activity; whereas compounds 1-7 showed strong ABTS free radical scavenging activity.

Evaluation of aversive behavior in Rattus norvegicus experimentally infected by two distinct strains of Toxoplasma gondii (ME49 and VEG): study of epigenetic markers

ABSTRACT Background: Behavioral changes in Rattus norvegicus infected with two strains of Toxoplasma gondii (ME49 and VEG) were investigated. Methods: Rats were evaluated for motor activity and aversion or attraction to cat urine 60 days after infection. After euthanasia, arginine-vasopressin gene methylation in the central nervous system was evaluated. Results: A significant difference was observed in the methylation of the arginine-vasopressin promoter gene between rats infected with the ME49 and VEG strains. Conclusions: Although differences were not observed in many parameters, significant differences were observed in the methylation of the arginine-vasopressin promoter gene in rats infected with the two studied strains.

Impacto de enfermedades orales asociadas a estrés sobre la calidad de vida / Impact on Quality of Life of Oral Diseases Related to Stress

Resumen: Se ha sugerido que enfermedades de la mucosa oral asociadas a estrés impactan negativamente la calidad de vida. Sin embargo, la información no es concluyente. El objetivo fue comparar la calidad de vida asociada a salud bucal en dos grupos: Casos (21 pacientes de una clínica de enseñanza de medicina bucal con diagnóstico de liquen plano oral, estomatitis aftosa recurrente y síndrome de boca dolorosa; Control (42 sujetos sanos) pareados por edad y género. Se aplicó el cuestionario perfil de impacto de salud oral. Resultados: El análisis estadístico mostró que el grupo casos tiene peor calidad de vida (p 0.03) comparado con el de los controles, específicamente en las dimensiones incomodidad psicológica (p 0.027); inhabilidad física (p 0.004); e incapacidad (p 0.002; RM 5.63 IC 1.58-20.80). Se concluye que los sujetos que padecen enfermedades de la mucosa oral relacionada a estrés tienen mala calidad de vida.

Abstract: It has been suggested that oral mucosa diseases related to stress have a negative impact on the quality of life. However, the information regarding which aspects are the most affected is inconclusive. The objective was to compare the quality of life associated with oral health in two groups: Cases formed by 21 patients coming from a teaching clinic, suffering oral lichen planus, recurrent aphthous stomatitis, and burning mouth syndrome; Control formed by 42 healthy subjects matched for age and gender. Oral Health Impact Profile questionnaire was applied. Statistical analysis showed that group Cases has worse quality of life (p 0.03) than the one of controls, specifically on psychological discomfort (p 0.027), physical disability (p 0.004); and handicap dimensions (p 0.002; RM 5.63 IC1.58-20.80). It was concluded that patients suffering oral mucosa disease related to stress showed poor quality of life.

Malformación genital: disparador del diagnóstico de variantes severas de síndrome de Klinefelter / Genital malformation: trigger of the diagnosis of severe variants of Klinefelter syndrome

Resumen: Introducción: El síndrome de Klinefelter y sus variantes, como alteración en el número de cromosomas sexuales, se encuentra entre los trastornos del desarrollo sexual. Sus portadores manifiestan hipogonadismo hipergonadotrófico en la pubertad; las variantes severas presentan además problemas neurocognitivos y del lenguaje desde edades tempranas. Objetivo: Describir dos pacientes portadores de mal formación genital con diagnóstico genético de variantes severas de síndrome de Klinefelter; y revisar aspectos clínicos y terapéuticos. Casos Clínicos: Caso 1: Diagnóstico de genitales atípicos al nacer: Falo pequeño y corvo con meato uretral a nivel escrotal y escroto bífido. Sin otra anomalía somática, excepto sutil clinodactilia del 5 dedo. Cariotipo: 49,XXXXY. Al año de vida se reconstruyeron los genitales. Evolucionó con retraso global del desarrollo, principalmente del lenguaje, manejado con estimulación temprana kinésica y fonoaudiológica desde los 2 meses, logró integrarse en un jardín de infantes. Caso 2: Al mes de vida se constató falo pequeño y corvo severo (más de 70°), testículos en bolsa. Cariotipo: 48,XXYY. Al año de vida se corrigió malformación del pene. Evolucionó con retraso global del desarrollo, fundamentalmente en el lenguaje expresivo, y fue manejado con el equipo de estimulación temprana desde los 4 meses, logrando adaptación en un jardín de infantes. Conclusión: Las malformaciones genitales condujeron al diagnóstico de variantes severas de síndrome de Klin efelter, y fueron corregidas alrededor del año de vida. La identificación temprana de estas variantes permitió la intervención del equipo de neuroestimulación, favoreciendo el desarrollo neurocognitivo y la integración social de estos niños.

Abstract: Introduction: Among the disorders of sexual development, Klinefelter syndrome and its variants are classified as an alteration in the number of sex chromosomes. These patients show signs of hypergonadotropic hypogonadism at puberty, however cases of severe variants also present neurocognitive and language problems from an early age. Objective: To describe two patients with genital malformation with genetic diagnosis of severe variants of Klinefelter syndrome, and to review clinical and therapeutic aspects. Clinical Cases: Case 1: Diagnosis of atypical genitalia at birth: Small and curved phallus with the urethral meatus at scrotal level, and bifid scrotum. No other somatic abnormality was observed, except for subtle clinodactyly of the fifth finger. Karyotype: 49, XXXXY. At one year of life, genitalia were reconstructed. The patient presented a global developmental delay, mainly in language, which was managed with early stimulation and speech and language therapy since he was two months old. Finally, he was able to attend kindergarten. Case 2: At one month of life, a small and severe curved phallus (more than 70°) was observed, and testicles were in the scrotum. Karyotype: 48, XXYY. At one year of life, the penile malformation was corrected. The patient presented global developmental delay, mainly in expressive language which was managed with early stimulation since the age of four months, achieving kindergarten attendance. Conclusion: Genital malformations led to the diagno sis of severe variants of Klinefelter syndrome, and were corrected around the year of life. The early identification of these variants allowed the intervention of the neurostimulation team, favoring the neurocognitive development and social integration of these children.

Effect of an Active Polyphenol Compound LM49 on Polarization of RAW264.7 Macrophages / 中国药学杂志

OBJECTIVE: To investigate the effect of LM49(2,4′, 5′-trihydroxy-5,2′-dibromobenzophenone) on M1/M2 phenotype in RAW264.7 macrophages by LPS plus INF-γ and its regulation mechanism on M1/M2 polarization. METHODS MTT assay was used to determine the effect of LM49 on cell viability. Different concentrations of LM49 (5, 10, 20 μmol•L-1) were used to intervene in LPS combined with IFN-γ induced macrophages, the expression of macrophage subsets of markers and the effect on JAK/STAT of the signaling pathway were examined by flow cytometry, RT-PCR and Western-blot. RESULTS: Compared with LPS/INF-γ group, LM49 significantly inhibited the number of CD16/32+ cells and the mRNA expression of iNOS, IL-6 and TNF-α, increased the number of CD206+ cells and the mRNA expression of Arg-1 and IL-10, and decreased the ratio of M1/M2 in macrophage. It was demonstrated that LM49 significantly reduced the proteins expression levels of TLR4, Myd88, NF-κB and STAT1, while inhibiting the phosphorylation levels of p-JAK2 and p-STAT1 by Western-blot. CONCLUSION: The study demonstrates that LM49 inhibits M1 polarization and promotes M2 polarization in RAW264.7 macrophages via inhibiting TLR4-Myd88-NF-κB and JAK2-STAT1 pathway, regulating the balance of M1/M2 ratio in macrophages.

A new monoterpene glycoside compound from roots of Paeonia lactiflora / 中草药

Objective: To study the constituents from the roots of Paeonia lactiflora. Methods: The chemical constituents were isolated by various chromatographic techniques and their structures were elucidated by physicochemical properties and NMR data. Results: Eleven compounds were obtained and characterized as (4S)-perillic acid 6-O-α-L-arabinopyranosyl-(1'→6'')- β-D-glucopyranosyl (1), 4,9-dihydroxy-8,10-dehydrothymol-1-O-β-D-glucoside (2), emodin-8-O-β-D-glucoside (3), resveratrol (4), β-D-glucopyranosyl benzoate (5), ilexperphenoside A (6), catechol (7), methyl gallate (8), ethyl gallate (9), 3-methoxygallic acid (10), 1,2,3,4,6-penta-O-galloyl-β-D-glucopyranoside (11). Conclusion: Compound 1 is identified as a new compound, named perillic acid glycoside, compounds 2 and 5 are identified from the genus Paeonia for the first time.

The Prognostic Implication of CD49d Expression by Flow Cytometry and Trisomy 12 Detection by Fluorescent in situ Hybridization in Chronic Lymphocytic Leukemia

Chronic lymphocytic leukemia (CLL) is the most common chronic lympho‑proliferative disorder. This study done to detect the level of cluster of differentiation (CD)49d in CLL patients by flow cytometry and itscorrelation with the prognosis (survival) and with (trisomy12) detected by fluorescent in situ hybridization (FISH).Methods:Clinico-hematological profiles done to fourty CLL patients. CD49d tested by flow cytometry and trisomy12 was detected by FISH.Results:CLL patients classified according to modified Rai staging system into: low risk 12.5%, intermediate risk 22.5% and high risk 65%. CD49d and trisomy12 positivitywere detected in 29Original Research Article patients (72.5%) and 22 patients (55%), respectively. There was a significant positive correlation between the percentage of trisomy12 and of CD49d cells in CLL patients (P =0.034). And also, between CD49d and CD38 (P =0.034). On the other hand, there was no significant relation between both CD49d and trisomy12 expression and modified Rai staging system.As regard to overall survival (O.S) and disease free survival (DFS), both CD49d, trisomy12 positive cases were associated with shorter disease free, and overall survivals compared to the negative cases.Regarding to the relation between the use of combination of fludarabine, cyclophosphamide, and rituximab (FCR) as a standard treatment in CLL and OS and DFS of patients in our study, we found that FCR account for the better outcome associated with its use.Conclusion:CLL B-cell membrane expression of CD49d as measured by flow cytometry is a powerful prognostic parameter in patients with CLL. Its positive correlation with the trisomy12 and CD38 and the association of both CD49d and trisomy12 with short survival times indicate that they may have roles in the prognosis of CLL

Expressions of ubiquitin-specific proteinase 49 and FK506 binding protein 51 in spinal dorsal horns and their roles in rats with diabetic neuropathic pain / 中华神经医学杂志

Objective To delineate the expressions of ubiquitin-specific proteinase 49 (USP49)and FK506 binding protein 51 (FKBP51) in the spinal dorsal horns and their roles in rats with diabetic neuropathic pain (DNP).Methods Male SD rats were fed with high glucose and high fat diet with an intraperitoneal injection of 1％ streptozocin to establish DNP rat models.DNP rats were allocated into DNP group,control shRNA group (KC group) and knockdown USP49 group (KD group,n=10);saline,Ad-scrambled shRNA (3×108 PFU/mL) and Ad-USP49-shRNA (3×108 PFU/mL) were intrathecally injected into the rats of the three groups.Another 10 normal rats were chosen as control group (C group).Thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured one d before model establishment,one and 7 d after model establishment,one,three and 7 d after intrathecal injection.The mRNA and protein expressions of USP49,FKBP51 and glucocorticoid receptor α (GRα)in dorsal horn neurons were detected by real time-PCR and Western blotting.Results (1) TWL showed no significant difference among the four groups one d before model establishment (P＞0.05);three and 7 d after intrathecal injection,TWL in KD group was significantly prolonged as compared with that in the DNP group,and significantly shortened as compared with that in C group (P＜0.05).(2) MWT showed no significant difference among the four groups one d before model establishment (P＞0.05);three and 7 d after intrathecal injection,MWT in KD group was significantly higher as compared with that in the DNP group,and significantly lower as compared with that in C group (P＜0.05).(3) As compared with DNP group,KD group had significantly decreased mRNA expressions of USP49 and FKBP51,and statistically increased mRNA GRα expression (P＜0.05).(4) As compared with DNP group,KD group had significantly decreased protein expressions of USP49 and FKBP51,and statistically increased protein GRα expression (P＜0.05).Conclusion Specific knockdown of USP49 expression is associated with improvement of heat hyperalgesia and mechanical hyperalgesia in DNP rats,which may be attributed to the de-ubiquitination ofFKBP51 by USP49.

Stem cell analysis with intraductally treated mammary epithelial cells

PURPOSE: The activity of mammary stem cells (MaSCs) is essential to mammary growth, differentiation and regeneration in cycles of pregnancy, lactation, and involution. The capability to recruit the mammary gland through the cycles is attributed to stem cells. It was shown that the intraductal (i.duc) injection of pegylated liposomal doxorubicin (PLD) to multiparous FVB/N mice was associated with a significantly reduced outgrowth potential of mammary gland cells. We have explored i.duc PLD's effect on stem cell number and function in mouse mammary gland and aldehyde dehydrogenase (ALDH)'s availability as a mouse MaSC marker.METHODS: The total mammary epithelium was purified from 6 to 8-month-old FVB/N control and i.duc PLD-administered mice treated twice and analyzed by flow cytometry and limiting dilution cleared mammary fat pad transplants.RESULTS: There was no significant difference in the proportions of stem cell-enriched population (CD49(fhigh)CD24(med)) between control and i.duc PLD-treated groups. However, we found a significant reduction in the outgrowth potential of CD49(fhigh)CD24(med) and CD49(fhigh)CD24(med)ALDH(+) cells from i.duc PLD-treated mammary glands. We discovered that adding ALDH to CD49(fhigh)CD24(med) had the possibility of better marker selection for MaSC of mice.CONCLUSION: We present i.duc administration of PLD to reduce MaSC function, but not the number; and ALDH activity may add further selection of MaSCs to CD49f CD24 in mouse mammary glands. Screening of chemotherapeutic drugs or other natural products by this method of stem cell analysis may provide safe i.duc treatment in breast cancer.