Transcriptomic Effects of LINC01614 on Lung Cancer A549 Cells and Relevance of Drug Resistance / 肿瘤防治研究

Objective To investigate the biological role of LINC01614 in non-small cell lung cancer A549 cells and its drug resistance-related mechanism. Methods The CRISPR/Cas9 technology was used to construct the A549 cell model with knockdown of LINC01614. Transcriptome sequencing was performed on A549 cells knocked down with LINC01614. We validated the transcriptomic differential genes MCAM and ABCC3 at the gene level and MCAM at the protein level, detected the IC50 changes of A549 cells after knockdown of LINC01614 under the effect of different concentrations of cisplatin, and detected the effect of knockdown of LINC01614 on the migration ability of A549 cells. Results Of the 2 713 DEGs after knockdown of LINC01614, a total of 1 626 genes were up-regulated and 1, 087 genes were down-regulated. GO analysis showed that DEGs were associated with intracellular signaling, cell adhesion, and so on. Meanwhile, the KEGG analysis showed that DEGs were associated with the Wnt signaling pathway, TGF-β signaling pathway, and Rap1 signaling pathway. Selection of drug resistance-associated gene ABCC3 from DEGs for validation with MCAM: qRT-PCR results showed that knockdown of LINC01614 significantly down-regulated the expression of MCAM (P<0.05) and upregulated the expression of ABCC3 on A549 cells (P<0.05). After knockdown of LINC01614, the protein expression of MCAM, was significantly decreased in A549 cells (P<0.05); the IC50 of A549 cells to cisplatin was significantly increased (P<0.05); and the scratch healing rate of A549 cells was also significantly decreased (P<0.05). Conclusion LINC01614 may be associated with the proliferation, invasion, and apoptotic pathways of A549 cells. In addition, LINC01614 may exert its migration ability through MCAM and chemoresistance to cisplatin through ABCC3.

Metabolism of New Psychoactive Substances 4F-MDMB-BUTINACA in Zebrafish / 法医学杂志

Objective To study the metabolic transformation pathways of 4F-MDMB-BUTINACA in vivo by establishing zebrafish models. Methods Six adult zebrafish were randomly divided into blank control group and experimental group, with three fish in each group. After the zebrafish in the experimental group were exposed to 1 μg/mL 4F-MDMB-BUTINACA for 24 h, they were transferred to clean water and cleaned three times, then pretreated for instrumental analysis. The zebrafish in blank control group were not exposed to 4F-MDMB-BUTINACA. Mass spectrometry and structural analysis of 4F-MDMB-BUTINACA and its metabolites were conducted by liquid chromatography-high resolution mass spectrometry and Mass Frontier software. Results A total of twenty-six metabolites of 4F-MDMB-BUTINACA were identified in zebrafish, including eighteen phase Ⅰ metabolites and eight phase Ⅱ metabolites. The main metabolic pathways of phase Ⅰ metabolites of 4F-MDMB-BUTINACA in zebrafish were ester hydrolysis, N-dealkylation, oxidative defluorination and hydroxylation, while the main metabolic pathway of phase Ⅱ metabolites was glucuronidation. Conclusion Metabolite Md24 （ester hydrolysis） and Md25 （ester hydrolysis combined with dehydrogenation） would be recommended to be potentially good biomarkers for abuse of 4F-MDMB-BUTINACA.

Analysis Methods of Common Herbicides in Biological Material and Research Progress / 法医学杂志

Herbicides are a kind of chemical or biological agents that can effectively destroy or inhibit weed growth. Because of the widespread and frequent use of herbicides, herbicide poisonings have often been reported. At present, the main species reported to have caused poisoning are paraquat, diquat, glyphosate, and glufosinate. The main instrumental analysis method is LC-MS. This paper reviews the research progress on analysis methods of common herbicides in biological material and their application, summarizes the sample pretreatment and instrumental analysis situation of qualitative and quantitative analysis of herbicides in biological material, and collects test data of actual poisoning cases, to provide reference for clinical diagnosis and treatment and forensic identification of herbicide poisoning.

Study on chemical constituents from seeds of Trigonella foenum-graecum / 中草药

Objective: To research the chemical constituents from Trigonella foenum-graecum. Methods: The chemical constituents were separated and purified by sephadex LH-20, silica gel, semi-prepared HPLC and other chromatography techniques. Their structures were elucidated by their physicochemical properties and NMR data. Results: Six steroidal saponins and flavonoids (1-6) were isolated from the ethanol extracts of T. foenum-graecum and identified as 22-methoxy-trigoneoside IIb (1), gitogenin (2), diosgenin (3), luteolin (4), cynaroside (5), and luteolin-7-O-rutinoside (6). Conclusion: Compound 1 is a new compound, and compounds 5 and 6 are obtained for the first time from T. foenum-graecum.

[Identification of metabolites of Chenxiang Huaqi pill in rats based on UPLC-Q-TOF-MS].

To analyze the metabolites of Chenxiang Huaqi pill in rats by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). The separation was performed on Phenomenex Kinetex C18 column, with the acetonitrile -0.1% formic acid as the mobile phase for gradient elution at a flow rate of 0.8 mL·min⁻¹. The data were collected by the positive ion mode of ESI source. The plasma and urine total ion chromatograms of the rats in blank group and treatment group were used to analyze the targeted ion chromatograms. The results showed that 24 compounds were detected in the plasma and urine, including 5 prototype components and 19 metabolites. The major metabolic pathways included hydration, glucuronidation, demethylation, hydrolysis, hydroxylation and sulfation. The method was rapid, simple and sensitive, and can be used to rapidly identify the metabolites of Chenxiang Huaqi pill that can be absorbed in rats, providing a reference for the study of the absorption and metabolism mechanism of Chenxiang Huaqi pill in vitro.

Identification of metabolites of Chenxiang Huaqi pill in rats based on UPLC-Q-TOF-MS / 中国中药杂志

To analyze the metabolites of Chenxiang Huaqi pill in rats by using ultra performance liquid chromatography-quadrupole-time of flight-mass spectrometry (UPLC-Q-TOF-MS). The separation was performed on Phenomenex Kinetex C₁₈ column, with the acetonitrile -0.1% formic acid as the mobile phase for gradient elution at a flow rate of 0.8 mL·min⁻¹. The data were collected by the positive ion mode of ESI source. The plasma and urine total ion chromatograms of the rats in blank group and treatment group were used to analyze the targeted ion chromatograms. The results showed that 24 compounds were detected in the plasma and urine, including 5 prototype components and 19 metabolites. The major metabolic pathways included hydration, glucuronidation, demethylation, hydrolysis, hydroxylation and sulfation. The method was rapid, simple and sensitive, and can be used to rapidly identify the metabolites of Chenxiang Huaqi pill that can be absorbed in rats, providing a reference for the study of the absorption and metabolism mechanism of Chenxiang Huaqi pill .

Expression of HIF-2αin non-small cell lung cancer tissue and its relationships with MVD,Ki67,and GST-π / 吉林大学学报(医学版)

Objective:To investigate the expression of hypoxia-inducible factor-2α (HIF-2α)in non-small cell lung cancer (NSCLC)tissue,and to analyze its relationships with angiogenesis,cell proliferation and chemotherapy resistance. Methods: Total 112 cases of NSCLC and 20 cases of normal lung tissues were selected, immunohistochemical method was used to detect the expressions of HIF-2α,CD31,Ki67 and GST-π in 112 cases of cancer tissue and 20 cases of normal lung tissue,and the correlations of HIF-2α expression with microvessel density (MVD),Ki67, and GST-π were analyzed.Results:The positive expression rate of HIF-2α in NSCLC tissue was significantly higher than that in normal lung tissue (P < 0.05 ), the expression rate of HIF-2α in 112 cases of NSCLC was 47.3% (53/112).The MVD in HIF-2α protein high expression NSCLC group (31.1 ± 14.7)was higher than that in HIF-2αprotein low expression NSCLC group (24.3±15.8)(P <0.05).The cases of high expression of Ki67 in HIF-2αhigh expression group occupied 54.7% (29/53),and it was higher than that in HIF-2αlow expression group (16/59,27.1%);there was significant difference (r = 0.281,P = 0.003).The high expression of HIF-2α had no obvious correlation with the expression of GST-π (r = 0.122,P = 0.202). Conclusion:HIF-2αmay play an important role in the carcinogenesis and development of NSCLC by promoting the angiogenesis and enhancing the cell proliferation of NSCLC,but it may have no correlation with chemotherapy resistance.

Expression of P53, Fas, TNF-αand Cyclin E with carcinoma papillary thyroid cancer patients in serum and its clinical significance / 中国免疫学杂志

Objective:To observe the expression of the tumor suppressor gene (P53),apoptosis signal receptor (Fas),tumor necrosis factor alpha ( TNF-α) and Cyclin E ( Cyclin E) in serum and cancer tissues with papillary thyroid cancer patients ,and explore their relationship with the clinical pathology characteristics of thyroid papillary carcinoma .Methods:The puncture diagnosis in patients with papillary thyroid carcinoma as the experimental group (n=74),physical examination of healthy people as the normal control group (n=26).The two groups were fasting venous blood samples ,the experimental group in postoperative specimens from cancer tissue , adjacent normal tissue and 7 days after the fasting venous blood was sampled again.Protein content of P53,Fas,TNF-αand Cyclin E was detected by ELISA in serum , cancer adjacent normal tissue and cancer tissue;using real-time fluorescent quantitative assay to observe the gene expression of P53,Fas,TNF-αand Cyclin E in thyroid papillary carcinoma and adjacent normal tissues ; protein expression by immunohistochemical methods in papillary thyroid carcinoma and adjacent normal tissues P 53,Fas,TNF-αand Cyclin E analysis;the clinical expression with papillary thyroid cancer staging , pathological Type and has no relationship to lymph node metastasis.Results:The protein concentration in serum of patients with papillary thyroid carcinoma P 53 , Fas and TNF-αwere significantly lower than that of the normal control group ,Cyclin E protein content was significantly higher than that of normal control group ,the differences were statistically significant (P<0.01);thyroid papillary carcinoma P53,Fas and TNF-αprotein content,protein expression strength and gene expression levels were significantly lower than the normal tissues adjacent to cancer ,protein content ,Cyclin E protein expression and gene expression intensity was significantly higher than that in normal tissues ,the difference was statistically significant (P<0.05).Conclusion:The expression of p53,Fas and TNF-αin papillary thyroid carcinoma lower and expression level of Cyclin E increase ,may play an important role in papillary thyroid cancer invasion and metastasis.Combined detection of the four can be used as markers for early diagnosis of papillary thyroid cancer ,enhance the rate of early diagnosis of thyroid papillary carcinoma.

Cell budding from pre-invasive tumors: Intrinsic precursor of invasive breast lesions?

Our previous studies showed that in patients with ductal carcinoma in situ (DCIS) of the breast, the tumor cells that overlie focal myoepithelial cell layer disruptions (FMCLDs) are generally arranged as finger-like projections that bud into the stroma. These budding cells have significantly more genetic instability and invasion-related gene expression, and less estrogen receptor (ER) expression, than their epithelial cell counterparts. This study aimed to assess these cells for potential molecular markers that are uniquely associated with cell adhesion and motility. Seventeen ER-positive DCIS cases were screened by immunostaining for ER, and 7 cases which harbored FMCLD lesions were used to examine the expression of the potential markers. Two cases with both DCIS and invasive lesions were selected for comparing the differences in molecular expression between these lesion types. The results showed that expression levels of talin, E-cadherin and focal adhesion kinase (FAK) in tumor cells overlying FMCLDs were higher than those within the corresponding duct. Integrin ß1 staining was detected only in a small number of the tumor cells overlying the FMCLDs. Vinculin staining was weak (18%) or not detected (82%), and no expression was found in the tumor cells within the corresponding duct or in the pure isolated DCIS. By contrast, the expression levels of talin, vinculin and integrin ß1 in the invasive tumors were distinctly higher than those in DCIS, and the expression of FAK and E-cadherin was lower. Using electron microscopy, we found that the tight junctions between tumor cells overlying the FMCLDs were reduced compared to the adjacent tumor cells in the lumen. These results indicate that the tumor cells overlying FMCLDs are likely to represent the specific precursors of invasive breast lesions. Our findings may also facilitate the identification of specific targets for further molecular profiling, which will more completely characterize this important cell population.

[Survivin antisense oligodeoxynucleotides inhibits the proliferation of hepatocellular carcinoma cells and enhances 5-FU sensitivity].

OBJECTIVE: To investigate the inhibitory effect of survivin antisense oligodeoxynuleotides (ASODN) mediated by polyethylenimine (PEI) on hepatocelluar carcinoma SMMC-7721 cell proliferation and its effect on chemosensitivity to 5-FU in tumor-bearing mice. METHODS: The inhibitory effect of PEI-ASODN on SMMC-7721 cell proliferation was assayed by WST-8 test, Trypan blue exclusion test, and cell clone formation assay. In mouse models of transplanted H22 cell hepatocarcinoma and ascites tumor, the effect of 5-FU combined with PEI-ASODN on the weight and volume of the subcutaneous tumors was examined. The tumor inhibition rate in the tumor-bearing mice was calculated and the average survival time recorded. RESULTS: SMMC-7721 cells incubated with different concentrations of PEI-ASODN for 48 h showed significantly reduced cell proliferation in comparison with the control cells, while PEI or ASODN alone produced no such inhibitory effect. Incubation of SMMC-7721 cells with 0.75 micromol/L PEI-ASODN for 24, 48, 72, and 96 h resulted in significantly suppressed cell proliferation, and a 7-day incubation of the cells with PEI-ASODN at different concentrations (0.25-0.75 micromol/L) significantly inhibited the cell clone formation. In the tumor-bearing mice, the tumor weight and volume were obviously reduced with a tumor inhibition rate of 56.91% and volume inhibition rate of 57.83%, significantly different from those in saline-treated mice (P<0.01). In the mice bearing ascites tumor, the average survival time was 22.0 days in saline group and 42.7 days in 5-FU+PEI-ASODN treatment group, showing a a life-prolonging rate of 94.09% in the latter group. A synergetic effect was noted between 5-FU and PEI-ASODN. CONCLUSION: PEI-ASODN complex can significantly inhibit the proliferation of hepatocarcinoma SMMC-7721 cells and enhance 5-FU chemosensitivity of the tumor cells in vitro and transplanted H22 tumors in mice.

Survivin antisense oligodeoxynucleotides inhibits the proliferation of hepatocellular carcinoma cells and enhances 5-FU sensitivity / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of survivin antisense oligodeoxynuleotides (ASODN) mediated by polyethylenimine (PEI) on hepatocelluar carcinoma SMMC-7721 cell proliferation and its effect on chemosensitivity to 5-FU in tumor-bearing mice.</p><p><b>METHODS</b>The inhibitory effect of PEI-ASODN on SMMC-7721 cell proliferation was assayed by WST-8 test, Trypan blue exclusion test, and cell clone formation assay. In mouse models of transplanted H22 cell hepatocarcinoma and ascites tumor, the effect of 5-FU combined with PEI-ASODN on the weight and volume of the subcutaneous tumors was examined. The tumor inhibition rate in the tumor-bearing mice was calculated and the average survival time recorded.</p><p><b>RESULTS</b>SMMC-7721 cells incubated with different concentrations of PEI-ASODN for 48 h showed significantly reduced cell proliferation in comparison with the control cells, while PEI or ASODN alone produced no such inhibitory effect. Incubation of SMMC-7721 cells with 0.75 micromol/L PEI-ASODN for 24, 48, 72, and 96 h resulted in significantly suppressed cell proliferation, and a 7-day incubation of the cells with PEI-ASODN at different concentrations (0.25-0.75 micromol/L) significantly inhibited the cell clone formation. In the tumor-bearing mice, the tumor weight and volume were obviously reduced with a tumor inhibition rate of 56.91% and volume inhibition rate of 57.83%, significantly different from those in saline-treated mice (P<0.01). In the mice bearing ascites tumor, the average survival time was 22.0 days in saline group and 42.7 days in 5-FU+PEI-ASODN treatment group, showing a a life-prolonging rate of 94.09% in the latter group. A synergetic effect was noted between 5-FU and PEI-ASODN.</p><p><b>CONCLUSION</b>PEI-ASODN complex can significantly inhibit the proliferation of hepatocarcinoma SMMC-7721 cells and enhance 5-FU chemosensitivity of the tumor cells in vitro and transplanted H22 tumors in mice.</p>

[Survivin antisense oligodeoxynucleotides inhibits the proliferation of hepatocellular carcinoma cells and enhances the cell chemosensitivity to 5-Fu].

OBJECTIVE: To investigate the inhibitory effect of survivin antisense oligodeoxynucleotides (ASODN) mediated by polyethylenimine(PEI) on the proliferation of hepatocellular carcinoma SMMC-7721 cells, and assess its detect on the chemosensitivity of the cells to 5-FU. METHODS: The inhibitory effect of PEI-ASODN on SMMC-7721 cell proliferation was assessed using WST-8 test, trypan blue staining, and cell clone formation test. In mice bearing transplanted hepatocarcinoma and ascites tumor derived from H22 cells, 5-FU combined with PEI-ASODN was administered, and the weight and volume of the subcutaneous tumors were measured to calculate the tumor inhibition rate, and the average survival time of the mice was calculated. RESULTS: Incubation of the cells with different concentrations of PEI-ASODN for 48 h significantly inhibited the cell proliferation as compared with the control group, but PEI or ASODN alone produced no significant inhibitory effects. At 24, 48, 72, 96 h of incubation of the SMMC-7721 cells with 0.75 micromol/L PEI-ASODN, the cell proliferation was suppressed significantly, and incubation with PEI-ASODN at 0.25-0.75 micromol/L for 7 days resulted in significantly inhibited cell clone formation. No significant inhibition was detected in ASODN and PEI group. The tumor weight and volume were reduced in all the treated groups. The tumor inhibition rate was 56.91% and volume inhibition rate was 57.83% in 5-FU+PEI-ASODN group, significantly different from those in the normal saline group (P<0.01). In mice bearing ascites tumor, the average survival time was 22.0 days in saline group, and 42.7 days 5-FU+PEI-ASODN group. The life-prolongation rate of 5-FU+PEI-ASODN was 94.09% when compared with the survival time in saline group. A cooperative effect was detected between 5-FU and PEI-ASODN. CONCLUSION: PEI-ASODN complex can significantly inhibit the proliferation of hepatocarcinoma SMMC-7721 cells and enhance the chemosensitivity of the tumor cells to 5-FU.

Survivin antisense oligodeoxynucleotides inhibits the proliferation of hepatocellular carcinoma cells and enhances the cell chemosensitivity to 5-Fu / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the inhibitory effect of survivin antisense oligodeoxynucleotides (ASODN) mediated by polyethylenimine(PEI) on the proliferation of hepatocellular carcinoma SMMC-7721 cells, and assess its detect on the chemosensitivity of the cells to 5-FU.</p><p><b>METHODS</b>The inhibitory effect of PEI-ASODN on SMMC-7721 cell proliferation was assessed using WST-8 test, trypan blue staining, and cell clone formation test. In mice bearing transplanted hepatocarcinoma and ascites tumor derived from H22 cells, 5-FU combined with PEI-ASODN was administered, and the weight and volume of the subcutaneous tumors were measured to calculate the tumor inhibition rate, and the average survival time of the mice was calculated.</p><p><b>RESULTS</b>Incubation of the cells with different concentrations of PEI-ASODN for 48 h significantly inhibited the cell proliferation as compared with the control group, but PEI or ASODN alone produced no significant inhibitory effects. At 24, 48, 72, 96 h of incubation of the SMMC-7721 cells with 0.75 micromol/L PEI-ASODN, the cell proliferation was suppressed significantly, and incubation with PEI-ASODN at 0.25-0.75 micromol/L for 7 days resulted in significantly inhibited cell clone formation. No significant inhibition was detected in ASODN and PEI group. The tumor weight and volume were reduced in all the treated groups. The tumor inhibition rate was 56.91% and volume inhibition rate was 57.83% in 5-FU+PEI-ASODN group, significantly different from those in the normal saline group (P<0.01). In mice bearing ascites tumor, the average survival time was 22.0 days in saline group, and 42.7 days 5-FU+PEI-ASODN group. The life-prolongation rate of 5-FU+PEI-ASODN was 94.09% when compared with the survival time in saline group. A cooperative effect was detected between 5-FU and PEI-ASODN.</p><p><b>CONCLUSION</b>PEI-ASODN complex can significantly inhibit the proliferation of hepatocarcinoma SMMC-7721 cells and enhance the chemosensitivity of the tumor cells to 5-FU.</p>

Isolation and characterization of the full coding sequence of a novel densovirus from the mosquito Culex pipiens pallens.

During an investigation of arboviruses in China, a novel densovirus (DNV) was isolated from the adult female Culex pipiens pallens. The virus, designated Culex pipiens pallens densovirus (CppDNV), caused cytopathic effect in C6/36 cells. The virus particles were icosahedral, non-enveloped and had a mean diameter of 24 nm. The complete coding region of CppDNV was found to be 3335 nt and it contained three open reading frames (ORFs). CppDNV shares 82-93 % identical nucleotides with isolates of the Aedes albopictus densovirus [isolates AalDNV-1, AalDNV-2 (C6/36 DNV) and AalDNV-3], Aedes aegypti densovirus (AaeDNV) and Haemagogus equines densovirus (HeDNV). The nucleotide sequence identity among CppDNV isolates exceeds 98 %. Phylogenetic trees based on non-structural (NS1 and NS2) and capsid (VP) genes show that CppDNV clustered with the species AaeDNV and represents a novel variant of this species within the genus Brevidensovirus.

Expression of PTEN and DNA content and invasion and metastasis of lung cancer / 中国基层医药

Objective This study was designed to determine the expression ofgeneproducts of phosphatase and tensin homoligy deleted onehromoseten(PTEN)and DNA content in non small cell lung cancer(NSCLC)tissue and investigate their association with the invasion and metastasis of NSCLC.Methods The expression of PTEN were detected in 78 cases of lung cancer tissues by immunohistochemistry SP methods,DNA content in 30 cases of lung cancer tissues was examined by flow cytometry.Results The rate of total expression depletion of PTEN was 42.3%.The rates of PTEN expression depletion in lymph node metastasis group and no lymph node metastasis group were 52.1%and 26.7%respectively,with significantant difference(P＜0.05).The patients who had high expression of PTEN had a longer survival time.The range of the DNA index(DI)distribution ranged from 1.04 to 1.93,the DNA index and aneuploid tumor had a positive correlation with the lymph node metastasis,with the increased of TNM stage,the DI and the rate of DNA aneuploid increased(P＜0.05).Conclusions Expression of PTEN is associated strongly with lymph node metastasis of lung cancer,correlations exist between DNA content and TNM stage,lymph node metastasis.They are indications of metastasis potential and prognosis of lung cancer.