Porphyromonas gingivalis enhances the proliferation of colorectal cancer Caco-2 cells via the JAK2-STAT3 pathway / 口腔疾病防治

Objective @# To investigate the effect of pathogenic bacterium-Porphyromonas gingivalis (P.g) on the proliferation and inflammatory factor expression of human colorectal cancer Caco-2 cells, to determine whether the Janus kinase 2-signal transducers and activators of transcription 3 (JAK2-STAT3) pathway is involved in the regulation of Caco-2 cell proliferation by P.g and to provide an experimental basis for further exploring the relationship between P.g and colorectal cancer. @*Methods @# Caco-2 cells were cultured in vitro, and P.g at different multiplicities of infection (MOIs) (0, 1, 10, 25) was selected to stimulate for 12, 24 and 48 h. The effect of P.g on the proliferation of Caco-2 cells was detected by CCK8. The stimulation time was set as 12, 24 and 48 h. MOI=0 was the control group, and MOI=1, 10 and 25 comprised the experimental group. qRT-PCR and Western blot were used to detect the changes in interleukin-6 (IL-6), interleukin-10(IL-10), JAK2 and STAT3 gene and protein (phosphorylated protein) levels in each group. @*Results @# After P.g infection of Caco-2 cells, P.g had a sustained stimulatory effect on the cells for 12, 24 and 48 h at MOI=1 and MOI=10 compared with the control group. Compared with that in the control group, the expression of pro-inflammatory factor IL-6 and related proliferative pathway protein JAK2 and STAT3 in Caco-2 cells with P.g infection increased in a concentration- and time-dependent manner (P<0.05). Additionally, the expression of IL-10, an anti-inflammatory factor, in Caco-2 cells infected with P.g decreased (P<0.05). After the addition of the JAK2 inhibitor AZ960, the proliferation of Caco-2 cells infected with P.g decreased, and the mRNA expression of STAT3 and JAK2 and the protein expression of p-STAT3 and p-JAK2 decreased (P<0.05). @*Conclusion @#P.g can promote the proliferation of the colorectal cancer cell line Caco-2, and the effect of P.g on Caco-2 cells may promote cell proliferation through the JAK2-STAT3 pathway while promoting the expression of the proinflammatory factor IL-6 and inhibiting the expression of the anti-inflammatory factor IL-10, creating an inflammatory environment conducive to cell proliferation, which may be the mechanism by which P.g affects the proliferation of Caco-2 cells.

Role of biomimetic nanomaterials made from glioma cell- derived extracellular vesicles in targeted delivery of STAT3-siRNA / 中南大学学报(医学版)

OBJECTIVES@#Glioma is the most common primary intracranial tumor and there is still no ideal treatment at present. Gene therapy, as one of the new methods for treating glioma, has attracted attention in recent years. But its application in treating glioma is very limited due to lack of effective delivery vectors. This study aims to investigate the feasibility of biomimetic nanomaterials made from glioma cells-derived extracellular vesicles (EV) for targeted delivery of signal transducers and activators of transcription 3 (STAT3)-small interfering RNA (siRNA) in treating glioma.@*METHODS@#First, U251 glioma cells-derived extracellular vessel (EVU251) was extracted by ultra-centrifugal method. Nanoparticle tracking analysis was used to characterize the particle size distribution, the transmission electron microscope was used to analyze the morphology, and Western blotting was used to verify the expression of srface characteristic protein. The homing ability was verified by cell uptake assay after labeling EVU251 with membrane dye kit PKH67; the EVU251 contents were removed by a low permeability method and then EVMU251 was prepared through a microporous membrane. Finally, the biomimetic nanomaterials EVMU251@STAT3-siRNA were prepared by loading STAT3-SiRNA with electro-dyeing method. The real-time quantitative PCR was used to quantify the successful encapsulation of siRNA, and the encapsulation and drug loading rate was calculated; then Cy5-labeled siRNA was used to evaluate the ability of biomimetic nanomaterials (EVMU251@CY5-siRNA) to target U251 cells. Lysosomal escape ability of the biomimetic nanomaterial was evaluated by lysosomal dye lyso-tracker green. At last, the ability of EVMU251@STAT3-siRNA to knock down STAT3 gene and selective killing of U251 cells was detected by cell experiments in vitro.@*RESULTS@#The size of EVU251 ranged from 50 nm to 200 nm with a natural disc shape. The expression of extracellular vesicle marker proteins could be detected on the membrane of EVU251. The cell uptake assay demonstrated that it had homing ability to target U251 cells. After EVU251 was prepared as EVMU251@STAT3-siRNA, the particle size was (177.9±5.0) nm, the siRNA loading rate was (33.5±2.2)% and the drug loading rate was (3.24±0.21)%. The biomimetic nanomaterial EVMU251@STAT3-siRNA still had the ability to target U251 cells and successfully deliver siRNA to the cytoplasm without lysosomal degradation. The EVMU251@STAT3-siRNA can effectively knock down the expression of STAT3 gene and produce selective killing ability in U251 cells.@*CONCLUSIONS@#The biomimetic nanomaterials EVMU251@STAT3-siRNA made from glioma U251 cells-derived extracellular vesicles can knock down STAT3 gene of U251 cells and produce selective killing effect, which can provide a new idea for the treatment of glioma.

Evaluation of current disinfection practice of transvaginal ultrasound probes in a Philippine Tertiary Referral Hospital: A comparative study on the performance of manual reprocessing methods

Objectives@#There are no standard infection control regulations in transvaginal ultrasound probe disinfection followed in the most prominent local public tertiary referral hospital. Likewise, no studies have evaluated the efficacy of the current method that uses an inexpensive multipurpose antiseptic spray solution. This study aims to evaluate the efficacy of the current practice of manual disinfection of TVS probes and compare it with the performance of an acceptable manual reprocessing method. @*Methods@#A prospective, randomized, controlled study was carried out using a crossover, quasi-experimental design, collecting 119 total samples from the ultrasound transducers before (35 samples) and after disinfection with two manual reprocessing methods, either a locally manufactured multipurpose antiseptic spray (A-Septic® Multipurpose Antiseptic Spray) that is currently used for disinfection or Mikrozid Sensitive®, a ready to use impregnated wipes (42 samples each arm). Disinfection efficacy was evaluated based on microbial culture results. @*Results@#Before disinfection, bacterial growth was observed in 77.1% (27/35) of the probes. After disinfection, 80.95% (34/42) remained contaminated with the antiseptic spray and 21.43% (9/42) with the wipes. The cultures revealed many environmental and pathogenic bacterial isolates, including Burkholderia, Staphylococcus, Acinetobacter, Diphtheroids, and Pseudomonas. @*Conclusions@#The currently used method for disinfecting transvaginal transducers in the division is not adequate for decontamination and decreasing the risk of cross contamination among patients. The results call for aggressive disinfection measures and highlight the need to update local standards and formulate and institutionalize these recommendations.

Da Chaihutang Inhibits Hepatocellular Carcinoma by Regulating p38 MAPK/IL-6/STAT3 Signaling Pathway / 中国实验方剂学杂志

ObjectiveThis study aims to investigate the efficacy and underlying mechanism of Da Chaihutang （DCHT） in treating hepatocellular carcinoma （HCC） in vitro and in vivo. MethodWe employed methyl thiazolyl tetrazolium （MTT） assay and crystal violet staining to observe the proliferation of Hepa1-6 liver cancer cells treated with DCHT at different doses （0， 125， 250， 500， 1 000 mg·L-1） for different time periods （1， 2， 4， 8 days）. The orthotopic liver cancer model was established by injection of 1×106 Hepa1-6 cells into mouse， and then the model mice were randomly assigned into six groups： blank， model， DCHT （0.21， 0.625， 1.875 g·kg-1， ig， qd）， and positive control （5-fluorouracil， 25 mg·kg-1， ip， qod）. After 14 days of administration， the mice were sacrificed， and the liver samples were collected and fixed in 4% paraformaldehyde for hematoxylin-eosin （HE） staining. The Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， Cytoscape 3.7.2， STRING， and DAVID were used for the searching of the key targets of DCHT in treating HCC， the construction of protein-protein interaction （PPI） network， and the Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment. Quantitative real-time PCR was performed to determine the mRNA level of interleukin-6 （IL-6） in Hepa1-6 cells and liver tissue. Western blotting was employed to measure the protein levels of the proteins involved in the mitogen-activated protein kinase （MAPK） and signal transducers and activators of transcription 3 （STAT3） signaling pathways. ResultDCHT （500， 1 000 mg·L-1） treatment for 4 and 8 days inhibited the proliferation of Hepa1-6 cells in a dose- and time-dependent manner （P<0.05）. The in vivo assay showed that DCHT （high dose， 1.875 g·kg-1） treatment for 14 days led to high differentiation and unobvious heterogeneity of HCC cells and small necrotic area compared with the model group. Network pharmacology analysis predicted that the potential targets of DCHT in the treatment of HCC were mainly the inflammation cytokines such as IL-6， interleukin-1β （IL-1β）， and tumor necrosis factor-alpha （TNF-α） in HCC microenvironment. The potential signaling pathways involved in the treatment were mainly associated with HCC growth and differentiation， including MAPK and STAT3 signaling pathways. Compared with the blank group， DCHT （1 000 mg·L-1） treatment for 1， 2， 4， and 8 days down-regulated the mRNA level of IL-6 in Hepa1-6 cells （P<0.05）. Similar results were observed in the livers of mice treated with DCHT （0.625， 1.875 g·kg-1）. The in vitro assay demonstrated that DCHT （1 000 mg·L-1） treatment for 4 and 8 days and DCHT （500， 1 000 mg·L-1） treatment inhibited the phosphorylation of extracellular signal-regulated kinases 1/2 （ERK1/2）， c-Jun NH2-terminal kinase/stress-activated protein kinase （JNK）， p38 MAPK， and STAT3 in a dose- and time-dependent manner （P<0.05）. The in vivo assay showed that DCHT （0.625 and 1.875 g·kg-1） treatment only inhibited the phosphorylation of p38 MAPK and STAT3 （P<0.05）. ConclusionThe present study indicates that DCHT can inhibit liver cancer cell proliferation by regulating p38 MAPK/IL-6/STAT3 signaling pathway.

A influência do tipo de transdutor acústico na audiometria tonal liminar / Influence of the type of acoustic transducer in pure-tone audiometry

RESUMO Objetivo Comparar os limiares auditivos por via aérea obtidos em diferentes transdutores acústicos e verificar a preferência do usuário. Método Trata-se de um estudo observacional, analítico, transversal, realizado em 26 participantes de 18 a 30 anos, com audição dentro dos padrões de normalidade, sem histórico de exposição a altos níveis de pressão sonora ou queixa de zumbido no momento da avaliação. Realizou-se anamnese, meatoscopia, audiometria tonal liminar, logoaudiometria, e imitanciometria. Os limiares auditivos foram pesquisados duas vezes, cada uma com um tipo de transdutor acústico diferente: fone de inserção (E-A-R Tone) e fone circum-aural (HDA200). A ordem de realização foi aleatória, com intervalos de cinco minutos. Ao final, o participante foi questionado quanto ao conforto dos fones durante os testes. Os dados foram submetidos à análise estatística não paramétrica. Resultados Na pesquisa dos limiares auditivos, ao avaliar as medianas, o fone circum-aural apresentou resultados melhores em 250, 500, 2000 e 6000 Hz e o fone de inserção foi melhor em 3000 e 4000 Hz, sem diferença estatística para as frequências de 1000 e 8000 Hz. O fone circum-aural foi eleito o mais confortável. Conclusão O fone circum-aural apresentou melhores limiares auditivos em 250, 500, 2000 e 6000Hz quando comparado ao fone de inserção, além de ser o tipo de transdutor mais confortável relatado pelos pacientes.

ABSTRACT Purpose To compare the air-conduction hearing thresholds obtained with different acoustic transducers and verify the users' preferences regarding them. Methods This is a cross-sectional, analytical, observational study with 26 participants aged 18 to 30 years, with normal hearing and no history of exposure to high sound pressure levels or complaints of tinnitus at the time of the assessment. We surveyed their medical history and performed meatoscopy, pure-tone threshold audiometry, speech audiometry, and acoustic immittance. The auditory thresholds were surveyed twice, each time with a different type of acoustic transducer: insert (E-A-RTONE) and circumaural earphones (HDA200). The assessments were performed in a random order, with 5-minute intervals. In the end, we asked the participants which earphones they found more comfortable in the tests. The data were submitted to nonparametric statistical analysis. Results Assessing the medians in the auditory threshold survey, the circumaural earphones obtained better results at 250, 500, 2000, and 6000 Hz, while the insert earphones were better at 3000 and 4000 Hz; there were no statistical differences at 1000 and 8000 Hz. The circumaural was elected the most comfortable earphone. Conclusion The circumaural earphones had better auditory thresholds at 250, 500, 2000, and 6000 Hz than the insert earphones and were reported by the patients as the most comfortable type of transducer.

Reliability of wireless insole baropodometry of normal individual's gait / Confiabilidade da baropodometria por palmilha sem fio na marcha de indivíduos normais

ABSTRACT Objective: The plantar pressure distribution can be assessed quantitatively by computerized baropodometry such as carpet or insole. An insole-type system with wireless transmission was developed and plantar pressure results were previously validated by force platform. However, the reproducibility of the system had not been determined. Our objective was to evaluate the reliability of the results in different gait cycles, clinical characteristics and in different plantar anatomical sites. Methods: 41 healthy adults (age, 34 ± 13 years; body mass index, 25 ± 5 kg/m2; 26 [63%], male, 26 [63%] practicing physical activity) were evaluated. Baropodometer evaluations were performed in 3 walking cycles with 100 m each, and the reliability between the cycles was examined. Pressure points on the heel, first metatarsal, fifth metatarsal and total plantar pressure were analyzed and compared. Results: Moderate agreement was identified between the second and third cycles (ICC, 0.66; 95% CI, 0.14-0.83). Physical activity practitioners showed higher total plantar pressure (70.8 vs 68.2 Kpa; p = 0.04) and higher pressure in the heel (70.7 vs 68.1 Kpa; p = 0.036) in relation to sedentary ones. Conclusion: The insole was able to assess plant pressure with moderate reliability from the adaptation period. Level of Evidence III, Case control study - Investigating a diagnostic test.

RESUMO Objetivo: A distribuição da pressão plantar pode ser avaliada quantitativamente por baropodometria computadorizada tipo tapete ou palmilha. Um sistema tipo palmilha com transmissão sem fio foi desenvolvido, cujos resultados de pressão plantar foram previamente validados por plataforma de força. No entanto, a reprodutibilidade do sistema não havia sido determinada. Nosso objetivo foi avaliar a confiabilidade dos resultados em relação a diferentes ciclos de marcha, características clínicas e em diferentes sítios anatômicos plantares. Métodos: Foram avaliados 41 adultos saudáveis (idade, 34 ± 13 anos; índice de massa corpórea, 25 ± 5 kg/m2; 26 [63%], sexo masculino, 26 [63%] praticantes de atividade física). Avaliações com o baropodômetro foram realizadas em 3 ciclos de marcha com distância de 100 m, e avaliada a concordância entre os ciclos. Pontos de pressão no calcanhar, primeiro metatarsal, quinto metatarsal e a pressão plantar total foram analisados e comparados. Resultados: Houve moderada concordância entre o segundo e terceiro ciclos (CCI, 0,66; IC95%, 0,14-0,83). Praticantes de atividades físicas apresentaram pressão plantar total (70,8 vs 68,2 Kpa; p = 0,04) e no calcanhar (70,7 vs 68,1 Kpa; p = 0,036) aumentada em relação aos sedentários. Conclusão: A palmilha foi capaz de avaliar a pressão plantar com confiabilidade moderada a partir do período de adaptação. Nível de Evidência III, Estudo diagnóstico - Investigando um teste diagnóstico.

Characterization of SH-SY5Y neurons subject to 92kHz ultrasound stimulation / Caracterización de neuronas SH-SY5Y sujetas a estimulación por ultrasonido de 92 kHz

SUMMARY: Cellular microstructural changes due to ultrasound exposure are critical to understand and characterize in order to further the establishment of ultrasonics in cell and tissue engineering and medicine. In this study, neurite length, nuclear morphology, and cellular toxicity are assessed at varying intensities of 92 kHz ultrasound provided by a piezoceramic disk element and incident upon SH- SY5Y neurons in vitro. Findings suggest that stimulation increases neurite length up to 2.73 fold tested at α = 0.05 in an intensity dependent manner. Additionally, stimulation causes a statistically significant (α = 0.05) decrease in nuclear area and less elongated nuclei, by 1.78 fold and 1.38 fold respectively, also in an intensity dependent manner. For maximum transducer surface intensities ranging from 0 to 39.11 W/cm2, the toxicity of 92 kHz ultrasound is assessed and a nontoxic range is determined using Caspase-3 and Annexin V staining, in addition to Calcium imaging via Calcein-AM staining. Intensities of up to 1.6 W/cm2 are found to be nontoxic for the cells under the parameters used in this study.

RESUMEN: Los cambios micro estructurales celulares debidos a la exposición a los ultrasonidos son fundamentales para comprender y caracterizar el establecimiento de los ultrasonidos en la ingeniería y la medicina de células y tejidos. En este estudio, la longitud de las neuritas, la morfología nuclear y la toxicidad celular se evalúan a intensidades variables de ultrasonido de 92 kHz proporcionado por un elemento de disco piezocerámico e incidente sobre las neuronas SH-SY5Y in vitro. Los resultados sugieren que la estimulación aumenta la longitud de las neuritas hasta 2,73 veces probada a α = 0,05 de una manera dependiente de la intensidad. Además, la estimulación provoca una disminución estadísticamente significativa (α = 0,05) en el área nuclear y núcleos menos alargados, en 1,78 veces y 1,38 veces, respectivamente y también de una manera dependiente de la intensidad. Para intensidades máximas de la superficie del transductor que oscilan entre 0 y 39,11 W / cm2, se evaluó la toxicidad del ultrasonido de 92 kHz y se determinó un rango no tóxico mediante tinción con Caspasa-3 y Anexina V, además de imágenes de calcio mediante tinción con Calceína-AM. Se encontró que las intensidades de hasta 1.6 W / cm2 no son tóxicas para las células bajo los parámetros usados en este estudio.

Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis / 中国实验方剂学杂志

<b>Objective::To observe the effect of Youguiwan on the levels of cartilage transducers and activators of transcription 3 (STAT3) and interleukin-6 (IL-6) in rats with knee osteoarthritis (KOA). <b>Method::Sixty SD rats were randomly divided into six groups: sham control group, model group, glucosamine sulfate group and Youguiwan (high, middle and low-dose) groups. The modified Hulth method was used to prepare KOA models for 6 weeks. The shame control group and the model group were treated with normal saline, Youguiwan high, middle, low-dose groups received Youguiwan 4.8, 2.4, 1.2 g·kg^-1 by gavage respectively, and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g·kg^-1. The rats were administrated for 8 weeks according to the dose. After intervention, each group was put to death by femoral artery blood collection, and the keen cartilages of the rats were collected. The pathological changes were observed by htoxylin eosin (HE) staining method, and Mankin score was evaluated. The expressions of STAT3, superoxide dismutase3(SOD3) and Wnt inhibitory factor 1(WIF1) in articular cartilage were detected by immunohistochemistry. The expressions of IL-6 mRNA in articular cartilage were detected by quantitative real-time fluorescence polymerase chain reaction (Real-time PCR). The expression of WIF1 in articular cartilage was detected by Western blot. <b>Result::Compared with the sham control group, the Makin score was obviously increased in the model group, the protein expression of STAT3 was increased significantly, the mRNA of IL-6 was raised significantly, but the protein expression of WIF1 was decreased significantly (<italic>P</italic><0.01), articular cartilage was seriously damaged, and chondrocytes were arranged in disorder. Compared with the model group, the Makin score was declined obviously in the high-dose Youguiwan group, the protein expression of STAT3 was significantly reduced, the mRNA expression of IL-6 was significantly reduced in Youguiwan treatment group, while the protein expression of WIF1 was significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), the cartilage structure returned to be normal, the chondrocytes distribution was uneven, and articular cartilage surface was not smooth. <b>Conclusion::Youguiwan could significantly improve the articular cartilage degeneration of KOA rats, and inhibit the inflammation of chondrocytes, which may be related to the suppression of STAT3 and IL-6 expression.

β-arrestin 1 facilitates acetylation of IL-6 gene promoter to affect proliferation and apoptosis of non-small cell lung cancer cells / 肿瘤

Objective: To investigate the effects of β-arrestin 1 (ARRB1) on apoptosis and proliferation of non-small cell lung cancer (NSCLC) cells, and to explore the underlying molecular mechanism. Methods: In NSCLC cell lines A549 and H1650, the expression of ARRB1 was knocked down by transfection of siRNA or over-expressed by transfection of Flag-ARRB1 recombinant plasmids, which was verified by real-time fluorescent quantitative PCR and Western blotting. The effects of down-regulating and up-regulating ARRB1 expression on the transcription and secretion of interleukin-6 (IL-6) were detected by real-time fluorescent quantitative PCR and ELISA method, respectively. The interaction between ARRB1 and p300 was detected by protein immunocoprecipitation. The recruitment of p300 and the acetylation of histone in IL-6 promoter region after ARRB1 knock-down or overexpression were detected by chromatin immunocoprecipitation. The proliferation and apoptosis of ARRB1 silencing A549 cells were detected by CCK-8 assay and FCM, respectively. The expression and activation of IL-6/signal transducer and activator of transcription 3 (STAT3) signaling pathway-related molecules in ARRB1 silencing or overexpression NSCLC cells were investigated by Western blotting. Results: In ARRB1-silenced or overexpressed NSCLC cell lines A549 and H1650, ARRB1 enhanced the transcription and production of IL-6 (all P < 0.05). The interaction of ARRB1 and p300 was confirmed by forward and reverse immunocoprecipitation. After ARRB1 knockdown or overexpression, it was found that ARRB1 enhanced the recruitment of p300 in IL-6 promoter region (both P < 0.01) and increased the acetylating of IL-6 promoter (both P < 0.05). Moreover, ARRB1 could facilitate the growth (P < 0.01) and apoptosis inhibition of NSCLC cells. ARRB1 could promote the phosphorylation of STAT3 and the expressions of c-Myc and Bcl-2 proteins. Conclusion: In NSCLC cells, ARRB1 interacts with p300, facilitates the recruitment of p300 to IL-6 promoter, and up-regulates the acetylation of histone H3 and H4 in IL-6 promoter, leading to transcriptional activation of IL-6. So that ARRB1 positively regulates the activation of IL-6/STAT3 signaling through promoting the phosphorylation of STAT3 and the expressions of c-Myc and Bcl-2 proteins, contributing to the growth and anti-apoptosis of NSCLC cells.

Inhibition of Sijunzi Decoction extract on human breast cancer cells MDA-MB-468 / 中草药

Objective: To investigate the effect of Sijunzi Decoction extract (SDE) on the growth of human triple negative breast cancer (TNBC) cell line MDA-MB-468. Methods: MDA-MB-468 cells were treated with different concentrations of SDE. The effect of SDE on the proliferation and migration of the cells were detected by CCK-8 assay and the cell wound healing assay. The colony formation assay was performed to analyze the effect on the ability of colony formation of the cells with SDE. Hoechst 33342 staining technique and flow cytometry (FCM) were used to detect apoptosis and cell cycle of the cells. Western blotting was used to detect the expression levels of STAT3, which was related with the proliferation and apoptosis of cells. Results: Compared with the control group, SDE had a certain inhibitory effect on MDA-MB-468 cells (P < 0.05), and it was dependent on the concentration and time. Cloning formation experiments showed that SDE inhibited the clonality of the cells. The cell migration experiment showed that the wound healing ability of the cells could be weakened by the extract with the medium and high dosage (P < 0.001). The results of FCM showed that the apoptosis rate of all SDE dosage increased gradually in a dose-dependent manner. And SDE with the medium and high dosage induced apoptosis of the cells significantly (P < 0.01 and 0.001). Cell cycle was affected by SDE with the obvious reduction of the cells in G2 phase (P < 0.01). The results of Western blotting showed that the expression level of STAT3 was decreased significantly. Conclusion: SDE inhibited the proliferation and clonal formation of MDA-MB-468 cells, inhibited migration, promoted apoptosis and decreased the cells of G2 phase. which may be related to the regulation of STAT3 pathway.

Effect of Qingzao Jiufei Tang on JAK2/STAT3 Signaling Pathway and Expression of Downstream Apoptosis-related Proteins in Lung Cancer / 中国实验方剂学杂志

Objective::To observe the effect of Qingzao Jiufei Tang on apoptosis of lung cancer, Janus protein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signaling pathway, as well as the expressions of downstream apoptosis-related proteins Bcl-2-associated X (Bax) and Cyclin D1. Method::Totally 50 male C57BL/6J mice were randomly divided into five groups: chemotherapy group (CTX), model group, high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and low-dose Qingzao Jiufei Tang group, with 10 mice in each group. The model of lung cancer was established by injecting Lewis lung cancer cells into the right axillary of mice. High-dose, middle-dose and low-dose Qingzao Jiufei Tang groups were orally given drugs (11, 5.5, 2.75 g·kg-1·d-1) two weeks before the modeling. Chemotherapy group was administered intraperitoneally at a dose of 50 mg·kg-1·(2 d)-1, while model group was administered intragastrically with the equal volume of normal saline. After inoculation, the mice in each group were continued to be administered. Two weeks later, the mice in each group were killed, and the tumors were collected. Then the JAK2 protein phosphorylation level was detected by immunohistochemistry (IHC). STAT3, Bax and Cyclin D1 protein expression levels were detected by Western blot, and apoptosis of lung cancer cells was observed by transmission electron microscopy. Result::Compared with model group, the phosphorylation levels of JAK2 and STAT3 protein in lung cancer cells were significantly decreased, the expression of Bax protein was significantly increased, and the expression of Cyclin D1 protein was significantly decreased in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group and chemotherapy group, with statistically significant differences (P<0.05, P<0.01). The results of transmission electron microscopy showed significant apoptotic phenomena in high-dose Qingzao Jiufei Tang group, middle-dose Qingzao Jiufei Tang group, low-dose Qingzao Jiufei Tang group and chemotherapy group compared with the model group. Conclusion::Qingzao Jiufei Tang had an obvious effect in promoting the apoptosis of lung cancer cells. Its mechanism may be related to the inhibition of the phosphorylation of JAK2 and STAT3 protein, the promotion of its downstream Bax protein expression and the inhibition of its downstream Cyclin D1 protein expression.

Effect of Huangqi Jianzhongtang on JAK2/STAT3 Signal Pathway in Rats with Spleen-Stomach Deficiency Cold Type Gastric Ulcer / 中国实验方剂学杂志

Objective:To investigate the effect of Huangqi Jianzhongtang on Janusprotein tyrosine kinase 2/signal transducers and transcriptional activator protein 3 (JAK2/STAT3) signal pathway in rats with spleen-stomach deficiency cold type gastric ulcer (GU). Method:A total of 60 SPF level Wistar rats were randomly divided into two groups: blank group and model group. Model rats were used to reconstruct the spleen-stomach deficiency cold type GU model by comprehensive modeling method. Model rats were divided into model group, Anweiyang group and high, medium and low-dose Huangqi Jianzhongtang groups according to the random number table, with 10 rats in each group. Rats in blank group and model group were given 10 mL·kg-1·d-1 distilled water, and 16, 8, 4 g·kg-1·d-1 Huangqi Jianzhongtang, respectively. Rats in the positive control group were given 0.14 g·kg-1·d-1 Anweiyang for 21 days. The gene expressions of JAK2 and STAT3 in the ulcer tissue were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR), the protein expressions and phosphorylation levels of JAK2 and STAT3 in the ulcer tissue were detected by Western blot, and the contents of interleukin-10（IL-10）and interleukin-17（IL-17）in the gastric tissue of each group were detected by enzyme-linked immunosorbent assay (ELISA). Result:Compared with the blank group, the general survival condition of the model group was worse, the content of IL-10 in gastric homogenate was significantly reduced, while the content of IL-17 was significantly increased (P<0.05), the protein expressions of JAK2 and STAT3 in gastric tissue was not significantly increased, whereas the gene expressions and phosphorylation levels of JAK2 and STAT3 were significantly increased (P<0.05). Compared with the model group, the content of IL-10 increased, but the content of IL-17 decreased, the gene expressions of JAK2 and STAT3 and the level of protein phosphorylation decreased in the treatment group, especially in the high-dose Huangqi Jianzhongtang group, with statistically significant differences (P<0.05). Conclusion:Huangqi Jianzhongtang can improve the survival condition of rats with spleen stomach deficiency cold type gastric ulcer, and its mechanism may be related to the intervention of gastric mucosal immune barrier dysfunction mediated by JAK2/STAT3 pathway activation.

Effect of Shenling Baizhusan on mTORC1/STAT3 Pathway in Hepatocytes of Nonalcoholic Fatty Liver Disease Rats / 中国实验方剂学杂志

Objective::To observe the effect of Shenling Baizhusan(SBS)on the mammalian target of rapamycin complex 1 (mTORC1)/signal transducers and activators of transcription 3 (STAT3) pathway in liver hepatocyte of nonalcoholic fatty liver disease(NAFLD)rats induced by high fat diet, in order to reveal the mechanism of SBS against rat NAFLD from the perspective of inflammation. Method::Totally 80 SD rats were randomly divided into 4 groups, normal control group, model group, high-dose SBP group(30 g·kg-1), and low-dose SBS group(10 g·kg-1), with 20 rats in each group. The rats of NAFLD model were established by being fed with high-fat diets for 8 weeks, and the treatment groups were fed with high or low dose of SBS respectively. After treatment for 8 weeks, blood and liver samples of rats were collected. Alanine aminotransferase (ALT), aspartate aminotransferase(AST), total cholesterol (TC), triglyceride(TG), high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)levels in blood serum were detected with automatic biochemical analyzer. The liver tissues were observed by oil red O and hematoxylin-eosin (HE) staining. Hepatocytes were isolated by type Ⅳ collagenase perfusion in vitro. Tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-5 and IL-6 in hepatocytes were detected by enzyme-linked immunosorbent assay (ELISA), and the relevant gene and proteins expressions of mTORC1 and STAT3 in hepatocytes were detected by Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot detection respectively. Result::Compared with the normal control group, the serum levels of TG, TC, AST, ALT and LDL-C were increased significantly, the levels of TNF-α, IL-1β, IL-5 and IL-6 in hepatocytes were increased significantly, and the expression levels of mTORC1, STAT3 mRNA and proteins in hepatocytes were increased significantly(P<0.01). Compared with the model group, the hepatic lipid accumulation of the medicine intervention group was relieved significantly, the serum levels of AST, ALT, TG and LDL-C were decreased significantly, the expression levels of TNF-α, IL-1β, IL-5 and IL-6 of hepatocytes were decreased significantly, and the expressions of mTORC1, STAT3 mRNA and proteins in hepatocytes were decreased significantly(P<0.05, P<0.01). In the high-dose SBS group, the effects in improving the lipid accumulation and inhibiting the inflammatory reaction were better than those of the low-dose SBS group, and the expressions of mTORC1 and STAT3 genes and proteins in hepatocytes were significantly decreased (P<0.05, P<0.01). Conclusion::SBS can improve the fat metabolism disorder and reduce liver lipid accumulation and inflammatory reaction in NAFLD rats induced by high-fat diet. The mechanism may be correlated with the inhibition of mTORC1/STAT3 pathway relating to genes and protein expression in hepatocytes.

Variabilidad de micrófonos utilizados para análisis acústico de la voz en los últimos veinte años / Variability of microphones used for acoustic analysis of the voice in the last twenty years

Se realizó una reflexión sobre la selección, durante los últimos 20 años, de los micró-fonos utilizados en estudios científicos que consideran análisis acústico y que son he-chos por profesionales de la voz. Se revisaron los equipos escogidos en 21 artículos ­a través del motor de búsqueda de Google, principalmente Google Scholar­ en los que la metodología consideraba el análisis acústico. Solo 4 de los 21 documentos elegidos realizaron una selección acertada según los estándares actuales de microfonía para fines de muestreo. Se determinó que los procesos de estandarización deben continuar con su perfeccionamiento

A reflection was made on the selection of microphones used in scientific studies, per-formed in the last 20 years, that consider acoustic analysis and are made by voice spe-cialists. The selected equipment was reviewed in 21 articles ­found through the Goo-gle engine, mainly Google Scholar­ in which the methodology considered acoustic analysis. Only 4 of the 21 chosen articles made an accurate selection according to the most recent microphone standards for sampling purposes. It was determined that the standardization processes should continue to be refined

Quantitative research technology of tuina manipulations / 针灸推拿医学（英文版）

Tuina is a physical therapy for treatment and prevention of diseases.The predecessors had summed up the systematic tuina manipulations through experiences.In order to study the scientificity and usability of the technology,the researchers established a mathematical model of tuina manipulations,and used video technology to capture the trajectory of the manipulations.Using the mechanical sensor to sense the real manipulations,researchers developed a tuina manipulation instrument and obtained a lot of basic mechanics data about the manipulation technology.Through the summary of the research results of the predecessors,accurate,true and comprehensive mechanical parameters of technology of tuina manipulations were obtained to guide the research and development of instruments of tuina manipulations,and promote the development of the discipline of tuina science.

Research progress of janus kinase - Signal transducers and activators of transcription signal transduction pathway and neurogenesis / 解剖学报

Janus kinase-signal transducers and activators of transcription (JAK-STAT) signal transduction pathway is related to cellular biological activities and occurrence and development of many diseases. In recent years, JAK-STAT signal transduction pathway has been found in the central nervous system to regulate neurodegenerative diseases and nerve regeneration after nerve injury: however, promoting endogenous neurogenesis as a new direction of nerve regeneration research is also closely related to the positive or negative regulation of JAK-STAT signal transduction pathway. This article reviews the research progress of JAK-STAT signal transduction pathway, neurogenesis, and JAK-STAT signal transduction pathway regulating neurogenesis.

Effect of recombinant human IL-17A on growth of colon cancer cells and its mechanism / 吉林大学学报(医学版)

Objective: To explore the effect of recombinant human interleukin-17AI (IL-17A) on the growth of colon cancer cells, and to investigate the related mechanism. Methods: The colon cancer SW480 cells were divided into control group and experimental group. The SW480 cells in control group were untreated and the SW480 cells in experimental group were added with 50 fig • L_1IL-17A. The proliferation abilities of SW480 cells in two groups were detected by CKK-8 method. The levels of IL-17A in the SW480 cells in two groups were detected by ELISA, and the expression levels of signal transducers and activators of transcription 3 (STAT3) and p-signal transducers and activators of transcriptions 3 (p-STAT3) were examined by Western blotting methed. Results: Compared with control group, the proliferation ability of the SW480 cells in experimental group was increased (P < 0 . 05). The level of IL-17A in the SW480 cells in experimental group was significantly higher than that in control group (P< 0. 01). Compared with control group, the expression levels of STAT3 and p-STAT3 proteins in the SW480 cells in experimental group were significantly higher than those in control group (P < 0 . 01). Conclusion: Recombinant protein IL-17A can stimulate the growth of colon cancer SW480 cells, which may be related to the activation of STAT3 signaling pathway.

Validation and application a cell-based screening model for IL-6/JAK/STAT3 inhibitor / 药学学报

Interleukin-6 (IL-6)/janus kinase (JAK)/signal transducers and activators of transcription 3 (STAT3) is a pivotal signaling pathway in the regulation of cell proliferation, survival, differentiation and T cell activation. Aberration of this pathway is involved in multiple autoimmunity diseases and cancers, therefore the pathway is considered as a hot target for drug development. In our study, we validated a cell-based model of IL-6/JAK/STAT3 and used it in screening of its inhibitors. HEK-Blue IL-6 cells of Invivogen Inc. were used to stably express IL-6 receptor and STAT3-induced secreted embryonic alkaline phosphatase (SEAP) report gene. After stimulation by IL-6, SEAP was secreted from cells and reacted with QUANTI-Blue. The product can be detected at 655 nm. The inhibitory effect of compounds on STAT3 signaling showed as IC50 was calculated by OD value. The results shown that IL-6 specifically activated the cells, which could be applied to screen the inhibitors for IL-6/JAK/STAT3 signaling pathway. The optimized screening conditions were described as below:50 000 cells/well, 1 ng·mL-1 IL-6 incubation for 20 h and reaction with QUANTI-Blue for 1 h. Based on this condition, we screened 14 natural products based on this cell model and arctigenin, cryptotanshinone and curcumin showed potential inhibitory activities on STAT3 signaling pathway with IC50 of 1.28, 2.96 and 6.61 μmol·L-1. Our study suggests that HEK-Blue IL-6 cells were suitable for screening inhibitors for the IL-6/JAK/STAT3 signaling pathway.

The mechanism of interleukin-22 induced the secretion of vascular endothelial growth factor A in gastric cancer cell line AGS / 国际检验医学杂志

Objective To investigate the mechanism of interleukin-22(IL-22) induced the secretion of vas-cular endothelial growth factor A(VEGF-A) in gastric cancer cell line AGS .Methods Gastric cancer cell line AGS were cultured in vitro ,and recombination cytokine IL-22 were added ,or signal pathway inhibitor were pre-incubated with AGS for 1 hour and then IL-22 were added ,the level of VEGF-A were detected by enzyme-linked immunosorbent assay .Results Compared with the unstimulated group ,the secretion of VEGF-A in IL-22-stimulated group was significantly increased ,the difference was statistically significant (P＜0 .05) ,which was in a dose and time dependent manner .In addition ,IL-22-stimulated the secretion of VEGF-A by AGS was significantly decreased while pre-incubated by the signal transducers and activators of transcription 3(STAT3) inhibitor ,the difference was statistically significant (P＜0 .05) ,but such effect was not observed while AGS were pre-incubated with the nuclear factor kappa B inhibitor ,c-Jun N-terminal kinase inhibitor ,mitogen-acti-vated protein kinase kinase 1/2 inhibitor and p38 mitogen-activated protein kinase inhibitor ,there was no sta-tistical significance(P＞0 .05) .Conclusion IL-22 could induce the secretion of VEGF-A in gastric cancer cell line AGS via STAT3 signal pathway ,which may contribute to tumor progression .

Molecular mechanism of calpain in regulating macrophage polarization mediated by NF-κB/STAT3 signaling pathways / 复旦学报（医学版）

Objective To investigate the molecular mechanism of calpain in regulating macrophage polarization.Methods Macrophages (RAW264.7) were transfected with siRNA by Lipofectamine 2000 to konckdown calpain1 and calpain2,respectively.The mRNA levels of markers in M1 (IL-23,TNF-α and iNOS) and M2 (IL-10,TGF-β and Arg-1) were measured by qRT-PCR.The levels of proteins in signaling pathways (NF-κB/STAT3) were measured by Western blot.The ability of macrophage migration was detected by Transwell assay.Results The expression level of calpain1 was lower in M1 than that in M2 (P＜0.05),but the expression level of calpain2 was significantly higher in M 1 than that in M 2 (P＜0.05).In calpain1-siRNA group,the mRNA levels of M1-type macrophages markers and M2-type macrophages markers were decreased by lipopolysaccharide (LPS) stimulation;The mRNA levels of M1-type macrophages markers in calpain2-siRNA group were significantly reduced (P ＜ 0.05),while the mRNA levels of M2-type macrophages markers were significantly increased (P ＜ 0.05).In calpain2-siRNA intervention group,the total phosphorylation inhibitor of nuclear factor kappa-B kinase (p-IKK) protein level of LPS-induced macrophages was decreased;in IL-4-induced macrophages,the protein level of plasmic signal transducers and activators of transcription 3 (STAT3) was also decreased,but there was no significant difference in total level of phosphorylation-p65 (p-p65).It was also found that the ability of migration was reduced by the interventions of calpain1-siRNA and calpain2-siRNA as compared with control-siRNA intervention (P＜0.05).Conclusions Calpain2 may potentially promote M1 polarizations by NF-κB and STAT3 signaling pathways,and inhibit the ability of its migration by the interventions of calpain1-siRNA and calpain2-siRNA.