OBJECTIVE: To investigate serum levels of long non-coding RNA (lncRNA) TUSC7 in patients with esophageal squamous cell carcinoma (ESCC), its association with clinicopathological parameters and its role in promoting tumor metastasis and invasion. METHODS: Serum samples were collected from 60 patients with ESCC admitted between January, 2017 and May, 2019, with 60 age- and gender-matched healthy subjects as the control group. Serum level of TUSC7 in ESCC patients and its expression in 4 ESCC cell lines was detected with RT-qPCR. The association of serum TUSC7 level with the clinicopathological features of the patients was analyzed. KYSE-30 cell models with TUSC7 overexpression or knockdown were established, and the proliferation of the cells was examined with MTT assay and their migration and invasion were assessed using wound healing and Transwell assays. Western blotting was used to detect the cellular expressions of the proteins associated with epithelial-mesenchymal transition (EMT). RESULTS: The patients with ESCC had significantly lower serum TUSC7 level than the healthy control subjects ( < 0.05). The ESCC cell lines also expressed lower levels of TUSC7 than normal cells ( < 0.05). Serum TUSC7 level was negatively correlated with tumor staging, lymph node metastasis and infiltration ( < 0.05) but was not significantly correlated with other clinicopathological parameters in ESCC patients. In the cell experiment, overexpression of TUSC7 in KYSE-30 cells significantly inhibited cell migration and invasion ( < 0.05), enhanced the expression of the EMT marker protein E-cadherin and lowered the expressions of N-cadherin, Vimentin and MMP9 ( < 0.05); knocking down TUSC7 in the cells produced the opposite effects. CONCLUSIONS The down-regulation of TUSC7 expression in the serum of ESCC patients and in ESCC cell lines is associated with the metastasis of ESCC and promotes tumor cell migration and invasion by promoting EMT, indicating the potential of serum TUSC7 level as a molecular marker for diagnosis, treatment and metastasis monitoring of ESCC.
Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition ; Esophageal Neoplasms ; genetics ; Esophageal Squamous Cell Carcinoma ; genetics ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasm Invasiveness ; RNA, Long Noncoding ; genetics

There have been many reports on berberine (BBR) effect of the inhibition on gut bacteria,but more from the protein level.In view of the preference of BBR for DNA binding,we here investigated the expression of BBR from the transcriptional expression level of the gene.The results showed that BBR had a higher affinity for UP element of Escherichia coli (E.coli) gene,and the transcription initiation region of this element contained TATA base sequence.The expression of genes sulA,recA and 16S which contain the genes of the UP element regulatory elements in the upstream of the promoter could be suppressed by BBR,and the expression of lpxC,secG and mutT which did not contain the genes of the UP element regulatory elements in the upstream of the promoter could not be inhibited by BBR.It is shown that the TATA sequence is the target of BBR.This result provides a new perspective for exploring the effect of BBR's inhibition of microbiota from gene transcription.

Berberine (BBR) is known as a classic drug for intestinal infection treatment.BBR inhibits intestinal bacteria,which is the core of its role in the treatment of intestinal infection.With the survival of local intestinal bacteria and its related metabolites on the physiological and pathological functions of the body continue to recognize the impact of it,more and more literatures have presented the effect of BBR through the impact of intestinal bacteria on the body glycol-lipid metabolism,even brain function.This allows us to re-understand the pathophysiology of BBR in inhibiting gut microbiome.In this paper,the antibacterial activity of BBR was reviewed and analyzed.The possible molecular target of BBR was analyzed according to the characteristics of prokaryotes gene expression,which was helpful to the in-depth study of BBR on intestinal bacteria.Thus,a more comprehensive understanding of the pharmacological effects of BBR is given.

Brazilein is one of the active ingredients of a Chinese medicine Caesalpinia sappan L. This study was aimed to test the vasodilator effect of brazilein on vascular smooth muscle cells in vitro from the perspective of molecular biology. The results showed that brazilein mainly acted through the influence of potassium ion channels, reducing membrane depolarization in order to affect the vessel relaxation. This effect can be influenced by blocking 5-HT receptor, and, simultaneously, correlating with the regulation of intracellular calcium ion concentration, affecting calmodulin and downregulating MLCP. In addition, the alterations of cAMP, PKA and PGI2 were involved in the pharmacological process of brazilein.

OBJECTIVETo observe the effect of Euphorbia kansui (E. KS) alcohol extracts on urination and kidney-related expressions of mice injected with normal saline and to discuss its impact on kidney.

METHODMice intraperitoneally injected with normal saline were observed for urination and changes in kidney-related histiocytic factors of after intragastrical administration of E. KS and compared with normal mice.

RESULTE. KS alcohol extracts can promote urination of mice injected with normal saline and enhance peripheral serum creatinine, with no obvious pathological change showed in tissue sections. It had a certain effect on reducing AQP2 expression and enhancing TNF-alpha expression.

CONCLUSIONEuphorbia kansui in large dose has a remarkable effect on kidney but may be accompanied with pathological reactions to some extent, especially the dose of 1.2 g x kg(-1). The pathological reactions may be related with increased serum creatinine and TNF-alpha expression.

Animals ; Aquaporin 2 ; genetics ; Euphorbia ; Interleukin-1beta ; genetics ; Kidney ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Plant Extracts ; pharmacology ; RNA, Messenger ; analysis ; Tumor Necrosis Factor-alpha ; genetics ; Urination ; drug effects

OBJECTIVETo investigate the alteration of inflammasome and receptor during IgG promoter transfected to HepG2 cells.

METHODBy assay of Elisa to evaluate the secretion of IL-1 beta, IL-8, TNF-alpha and MCP-1 after puerarine and LPS administration, and by assay of real time PCR to evaluate the expression of mRNA of IL-1 beta, IL-8,TNF-alpha and MCP-1, as well as the receptors of TLR2, 4 and NOD2, MyD88.

RESULTIgG promoter did not active innate immunity and enhance the expression and secretion of inflammasome in HepG2. Puerarine did not active the inflammasome either. LPS activated the innate immunity and increased the secretion of IL-8, TNF-alpha and MCP-1.

CONCLUSIONIgG-HepG2 cells could be used specifically as the model of allergy type II for ingredients screening. It is suggested that puerarine was suite for the activator for this type of allergy as positive control.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunity, Innate ; immunology ; Immunoglobulin G ; genetics ; Inflammasomes ; immunology ; Promoter Regions, Genetic ; genetics ; Transfection

OBJECTIVETo study the allergen in key processes during the production of Fufang Kushen injection by IgG promoter-HepG2 cells in vitro.

METHODBy transfecting a IgG promoter-regulating the expression of green fluorescent protein(GFP) plasmid into HepG2 cells, this transferred cells were incubated with common allergens (like puerarin, ovalbumin, LPS or Sal typhoid vi polysaccharide vaccine), excipients using in Fufang Kushen injection (NaOH, acetic acid, Tween-80 and ethanol) and samples from the key production processes of the injection for 30 minutes . Fluorescent photographs were analyzed the fluorescence intensity of the cells by using an image analysis software.

RESULTAll of common allergens significantly increased the IgG expression. Two of the excipicents, acetic acids and Tween-80 were shown to increased the IgG expression, while others had no effect on IgG expression. In the 8 samples from the key processes in the production of Fufang Kushen injection, two of them stimulated IgG expression.

CONCLUSIONIgG promoter-HepG2 cells are highly sensitive and specific to allergens, and thus can be applied to rapid screening of allergens in components and injections in transcriptional level. It is possible to use the IgG-promoter HepG2 cells in a real-time monitoring of allergens in the production processes of Chinese medicine injections.

Allergens ; analysis ; immunology ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Gene Expression Regulation ; immunology ; Hep G2 Cells ; Humans ; Immunoglobulin G ; genetics ; Injections ; Medicine, Chinese Traditional ; standards ; Promoter Regions, Genetic ; genetics ; Quality Control

OBJECTIVETo construct an effective system screening and evaluating possible injections and components inducing allergy type II.

METHODTransfect IgG promoter-regulated green fluorescent protein expressing plasmid into RPMI-8226 cell. The number of fluorescent cells after drug treatment was calculated and statistically evaluated.

RESULTTween 80 can suppress the expression of IgG effectively, and puerarin has activity of stimulating IgG expression, and the system has no response to KCl-treatment. No effect of Yuxingcao (Herba Houttuyniae) injection on this system is observed.

CONCLUSIONIgG promoter-drove green fluorescent protein expressing cell line can be used as system screening injections and components inducing allergy type II based on stimulating IgG promoter activity. Tween 80 can suppress the expression of IgG at the transcriptional level. Four batches of Yuxingcao injection cannot induce allergy type II by activating IgG expression.

Cell Line, Tumor ; Gene Expression ; drug effects ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Immunoglobulin G ; genetics ; Injections ; Male ; Plant Extracts ; pharmacology ; Plants, Medicinal ; chemistry ; Plasmids ; genetics ; Promoter Regions, Genetic ; genetics

OBJECTIVETo screen the active component of Wuzhuyutang (WZYT, Evodiae prescription) and investigate the regulatory effects of the components in WZYT on the TPH2 promoter, and to explore the possible molecular mechanism of WZYT on migraine.

METHODBy transfecting a TPH2 promoter regulating Red Fluorescent Protein expressing plasmid into PC12 cell, the global fluorescence intensities and calculations of fluorescent cells after components treatment were statistically evaluated.

RESULTDifferent regulatory effects of different components in WZYT with different concentrations on TPH2 promoter were observed.

CONCLUSIONTPH2 promoter drove Red Fluorescent Protein expressing cell line can be used as system screening components targeting TPH2 promoter activity. The possible mechanism of WZYT on migraine may due to its stimulating effects on TPH2 promoter, and promote the synthesis and release of 5-HT in cerebral.

Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Evodia ; chemistry ; Humans ; Migraine Disorders ; drug therapy ; enzymology ; genetics ; PC12 Cells ; Promoter Regions, Genetic ; drug effects ; Rats ; Tryptophan Hydroxylase ; genetics ; metabolism