Phenethyl isothiocyanate inhibits metastasis potential of non-small cell lung cancer cells through FTO mediated TLE1 m6A modification.

N6-methyladenosine (m6A) modification is a prevalent RNA epigenetic modification, which plays a crucial role in tumor progression including metastasis. Isothiocyanates (ITCs) are natural compounds and inhibit the tumorigenesis of various cancers. Our previous studies show that ITCs inhibit the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells, and have synergistic effects with chemotherapy drugs. In this study, we investigated the molecular mechanisms underlying the inhibitory effects of ITCs on cancer cell metastasis. We showed that phenethyl isothiocyanate (PEITC) dose-dependently inhibited the cell viability of both NSCLC cell lines H1299 and H226 with IC50 values of 17.6 and 15.2 µM, respectively. Furthermore, PEITC dose-dependently inhibited the invasion and migration of H1299 and H226 cells. We demonstrated that PEITC treatment dose-dependently increased m6A methylation levels and inhibited the expression of the m6A demethylase fat mass and obesity-associated protein (FTO) in H1299 and H226 cells. Knockdown of FTO significantly increased m6A methylation in H1299 and H226 cells, impaired their abilities of invasion and migration in vitro, and enhanced the inhibition of PEITC on tumor growth in vivo. Overexpression of FTO promoted the migration of NSCLC cells, and also mitigated the inhibitory effect of PEITC on migration of NSCLC cells. Furthermore, we found that FTO regulated the mRNA m6A modification of a transcriptional co-repressor Transducin-Like Enhancer of split-1 (TLE1) and further affected its stability and expression. TCGA database analysis revealed TLE1 was upregulated in NSCLC tissues compared to normal tissues, which might be correlated with the metastasis status. Moreover, we showed that PEITC suppressed the migration of NSCLC cells by inhibiting TLE1 expression and downstream Akt/NF-κB pathway. This study reveals a novel mechanism underlying ITC's inhibitory effect on metastasis of lung cancer cells, and provided valuable information for developing new therapeutics for lung cancer by targeting m6A methylation.

Liver and gall bladder channel parallels in the Hippocratic Corpus and Huang Di Nei Jing with theoretical considerations.

Chinese history contains an abundance of records discussing contact with the outside world. In the ever elusive search for the origin of the meridian theory, an extended investigation into the Hippocratic Corpus reveals parallels with the Huáng Dì Nèi Jing () medical text of the Chinese tradition that provide an alternative perspective into the origins of meridian channels theory. Though this topic has been discussed in the past, this paper provides new and developed insight into theories associated with the liver and gallbladder channel systems of each tradition. Condensing the scope of the analysis between the Huáng Dì Nèi Jing and Hippocratic text traditions and focusing on the liver and gall bladder channels aids to expound on the theoretical background required to advance the comprehension and understanding regarding the origins of each tradition. This paper reveals content that further the relationship between the two traditions, including demonstrating the existence of theoretical parallels existing between the liver and gall bladder channels of these two texts.

Characteristics and clinical significance of polyploid giant cancer cells in laryngeal carcinoma.

OBJECTIVES: We aimed to construct an induction system for polyploid giant cancer cells (PGCCs), as well as to investigate PGCC features and clinical significance. METHODS: A laryngeal neoplasm-PGCC induction system was constructed using paclitaxel liposomes (PTX). We used western blots to compare expression of epithelial-mesenchymal transition-related proteins, stem cell interrelated proteins, and cyclin-associated proteins. We then measured PGCC count in tissue samples of patients with laryngeal neoplasms and analyzed its relationship with prognosis. Statistical significance was determined using t-tests. RESULTS: PTX successfully induced PGCCs. Western blotting showed that CyclinB1, CDC25C, CDK1, E-cadherin, and EIF-4A expression decreased in PGCCs compared with normal cancer cells, whereas vimentin and CD133 expression increased. Number of PGCCs in laryngeal cancer tissues and overall survival time were inversely correlated (P < .05). CONCLUSIONS: PTX successfully induces PGCC formation in laryngeal carcinoma, which may be the cause of poor prognosis in patients with laryngeal cancer.Level of Evidence: 4.

[Discussion on the body conception of the medical scholars at the early eras based on the description of meridians in bamboo slip and silk medical books].

In the paper, the basic situation and description of meridians are introduced on Zubi Shiyimai Jiujing (Moxibustion Classics of Eleven Meridians of Legs and Arms) and Yinyang Shiyimai Jiujing (Moxibustion Classic on Eleven Yin and Yang Meridians) unearthed from Mawangdui, Maishu (Book of Meridians) from Zhangjiashan, the lacquer figure from Shuangbaoshan, the lacquer figure as well as Maishu (Book of Meridians) unearthed from Laoguanshan. By analyzing the differences of the records of the aforementioned unearthed literature and the meridians described on the excavated lacquer figures in comparison with the "twelve meridians" in Huangdi Neijing (The Yellow Emperor 's Inner Classic), it is viewed that the development of meridian system, from "eleven meridians" to "twelve meridians", from "independent flowing line" to the circular motion channel as "endless cycling", is influenced by the symbolic model of image numerology of "6 for the Heaven and 5 for the Earth" and the law of the reciprocal change of yin and yang. The description of meridians by the medical scholars at the early eras are based on various kinds of practices, such as clinical diagnosis and treatment, qigong and daoyin. Hence, the construction of the knowledge system of meridians is affected by the symbol model of image numerology.

[Research on regional distribution and industrial status analysis of genuine medicinal resources of flowers in Henan province].

Flower medicinal materials usually refer to Chinese medicinal materials with a complete flower,inflorescence,or part of a flower as the different medicinal parts,they have an important share in the Chinese herbal medicine market and appeared frequently in Chinese medicine prescriptions. Firstly,the species and regional distribution of the flower medicinal materials resources in China were briefly summarized. Secondly,the characteristics,yield,producing area and origin distribution of the main flower medicinal materials in Henan province were discussed. Finally,the present situation and the main problems of the flower medicinal materials industry in Henan province were comprehensively analyzed,and the corresponding industrial development countermeasures were put forward.This research was intended to provide decision-making demonstration and scientific basis for the rational exploitation and utilization of resources,breeding of new varieties,planting division,production layout and the healthy and sustainable development of the flower medicinal materials industry in Henan province.

Benzyl isothiocyanate induces protective autophagy in human lung cancer cells through an endoplasmic reticulum stress-mediated mechanism.

Isothiocyanates, such as allyl isothiocya¬nate (AITC), benzyl isothiocyanate (BITC), phenethyl isothio¬cyanate (PEITC) and sulforaphane (SFN), are natural compounds abundant in cruciferous vegetables, which have substantial chemopreventive activities against various human malignancies. However, the mechanisms underlying the inhibition of tumor cell growth by isothiocyanates are not fully understood. Since autophagy has dual functions in cancer, in the present study we investigated the effects of BITC on autophagy induction in human lung cancer cells in vitro and in vivo. BITC (1-100 µmol/L) dose-dependently inhibited the growth of 3 different human lung cancer cell lines A549 (adenocarcinoma), H661 (large cell carcinoma) and SK-MES-1 (squamous cell carcinoma) with IC50 values of 30.7±0.14, 15.9±0.22 and 23.4±0.11 µmol/L, respectively. BITC (10-40 µmol/L) induced autophagy in the lung cancer cells, evidenced by the formation of acidic vesicular organelles (AVOs), the accumulation of LC3-II, the punctate pattern of LC3, and the expression of Atg5. Pretreatment with the autophagy inhibitor 3-MA (5 mmol/L) significantly enhanced the BITC-caused growth inhibition in the lung cancer cells. Furthermore, BITC (20-40 µmol/L) activated ER stress, as shown by the increased cytosolic Ca2+ level and the phosphorylation of the ER stress marker proteins PERK and eIF2α in the lung cancer cells. Pretreatment with the ER stress inhibitor 4-PBA (5 mmol/L) attenuated the autophagy induction and potentiated the BITC-induced cell growth inhibition. In nude mice bearing A549 xenografts, administration of BITC (100 mg·kg-1·d-1, ip) for 8 weeks markedly suppressed the lung tumor growth, and significantly enhanced both autophagy and ER stress in the tumor tissues. Our results demonstrate that BITC inhibits human lung cancer cell growth in vitro and in vivo. In addition, BITC induces autophagy in the lung cancer cells, which protects the cancer cells against the inhibitory action of BITC; the autophagy induction is mediated by the ER stress response.

[A novel immunization strategy to induce strong humoral responses against HIV-1 using combined DNA, recombinant vaccinia virus and protein vaccines].

To optimize the immunization strategy against HIV-1, a DNA vaccine was combined with a recombinant vaccinia virus (rTV) vaccine and a protein vaccine. Immune responses against HIV-1 were detected in 30 female guinea pigs divided into six groups. Three groups of guinea pigs were primed with HIV-1 DNA vaccine three times, boosted with rTV at week 14, and then boosted with gp140 protein at intervals of 4, 8 or 12 weeks. Simultaneously, the other three groups of animals were primed with rTV vaccine once, and then boosted with gp140 after 4, 8 or 12 weeks. The HIV-1 specific binding antibody and neutralizing antibody, in addition to the relative affinity of these antibodies, were detected at different time points after the final administration of vaccine in each group. The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1 gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen. In the guinea pigs that underwent the DNA-rTV-gp140 regimen, the highest V1V2-gp70 antibody was induced in the 12-week-interval group. However, the avidity of antibodies was improved in the 4-week-interval group. Using the rTV-gp140 immunization strategy, guinea pigs boosted at 8 or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4 weeks after priming. In conclusion, this study shows that the immunization strategy of HIV-1 DNA vaccine priming, followed by rTV and protein vaccine boosting, could strengthen the humoral response against HIV-1. Longer intervals were better to induce V1V2-gp70-specific antibodies, while shorter intervals were more beneficial to enhance the avidity of antibodies.

[A Study on the quotations in the Wu xing da yi (The Gist of Five Phases) cited from Nei jing (Inner Canon)].

In the Wu xing da yi (The Gist of Five Phases), written by Xiao Ji of the Sui Dynasty, there were 9 quotations cited from Huang di nei jing (Inner Canon of Huangdi). By comparison with current version of Huang di nei jing, most of the descriptions are similar, yet still with some differences, such as yin-yang nature of the five zang visceras, the liver being the root of "pi ji", the spleen being the root of "cang lin" etc. Wu xing da yi epitomizes the books about yin-yang and five phases before the Sui Dynasty, while Xiao Ji's quotations from Huang di nei jing interpreted from the views of yin-yang and five phases, are different from the annotations of later ages.

Bovine HEXIM1 inhibits bovine immunodeficiency virus replication through regulating BTat-mediated transactivation.

The bovine immunodeficiency virus (BIV) transactivator (BTat) recruits the bovine cyclin T1 (B-cyclin T1) to the LTR to facilitate the transcription of BIV. Here, we demonstrate that bovine hexamethylene bisacetamide (HMBA)-induced protein 1 (BHEXIM1) inhibits BTat-mediated BIV LTR transcription. The results of in vivo and in vitro assays show direct binding of BHEXIM1 to the B-cyclin T1. These results suggest that the repression arises from BHEXIM1-BTat competition for B-cyclin T1, which allows BHEXIM1 to displace BTat from B-cyclin T1. Furthermore, we found that the C-terminal region and the centrally located region of BHEXIM1 are required for BHEXIM1 to associate with B-cyclin T1. Knockdown of BHEXIM1 enhances BIV replication. Taken together, our study provides the first clear evidence that BHEXIM1 is involved in BIV replication through regulating BTat-mediated transactivation.

[Molecular epidemiology of viral diarrhea in Chengdu infants and young children].

OBJECTIVE: To investigated the molecular epidemiologic features of viral diarrhea in Chengdu infants and young children, and to establish baseline patterns of etiology, provides the scientific basis for the vaccine development and the epidemic situation control. METHODS: From March, 2006 to December, 2008, a total of 376 infants and young children from Chengdu area hospitalized for diarrhea in Chengdu Children's Hospital were enrolled in this study. The stool specimen collected from each patient was tested for rotavirus (RV), Calicivirus (CV), astrovirus (AstV) and adenovirus (Adv) by using enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) examination. RESULTS: Among those 376 cases,there were 142 cases (37.76%) of RV infections,which scattered predominantly in October to December. Among 234 cases RV negativity,there were 29 cases HuCV infections (15.85%), 5 cases AstV infections (1.64%), and 8 cases Adv infections (2.04%). CONCLUSION: RV appeared to be the main etiological agent of viral diarrhea in Chengdu infants and young children,the predominant serotype of RV were G3, P[8] and P[4],HuCV might be the important etiological agent besides RV.

[Li Shizhen's cognition on human life].

Through the comparison between Taoists and physicians of ancient time in our country, we make inquiry into Li Shizhen's cognition on human life and consider that Li Shizhen explicitly put forward, prior to the contemporary physicians, the original creative ideas for the first time,on human being including "the brain is the residence of original spirit", "channel is the tunnel of inner image", "vital gate is located between the two kidneys", "vital gate is the substance, triple burner, the function". In addition to these ideas, Li Shizhen also raised an idea of "spleen is the mother of vital energy", which, even though it was not created for the first time, strengthened and developed the ideas of the predecessors.