[Mechanism of n-butanol fraction of Wenxia Formula combining with gefitinib in treating non-small cell lung cancer based on network pharmacology and in vitro experiment].

This study combined network pharmacology, molecular docking, and in vitro experiments to explore the potential mechanism of the active components of the n-butanol fraction of Wenxia Formula(NWXF) combined with gefitinib(GEF) in treating non-small cell lung cancer(NSCLC). Ultra-performance liquid chromatography-quadrupole Orbitrap mass spectrometry(UPLC-Q-Orbitrap MS) was employed to detect the main chemical components of NWXF. The active components of NWXF were retrieved from SwissADME, and the candidate targets of these active components were retrieved from SwissTargetPrediction. Online Mendelian Inheritance in Man(OMIM) and GeneCards were searched for the targets of NSCLC. Cytoscape 3.9.0 and STRING were employed to build the protein-protein interaction(PPI) network with the common targets shared by NWXF and NSCLC. Gene Ontology(GO) annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG) enrichment were performed in DAVID to predict the potential mechanisms. Finally, molecular docking between the main active ingredients and key targets was conducted in SYBYL-X 2.0. The methyl thiazolyl tetrazolium(MTT) assay was employed to evaluate the inhibitory effects of NWXF and/or GEF on the proliferation of human non-small cell lung cancer cells(A549 and PC-9). Additionally, the impact of NWXF on human embryonic lung fibroblast cells(MRC-5) was assessed. The effectiveness of the drug combination was evaluated based on the Q value. The terminal-deoxynucleoitidyl transferase mediated nick-end labeling(TUNEL) assay was employed to examine the apoptosis of A549 and PC-9 cells treated with NWXF and/or GEF. Quantitative real-time PCR(qRT-PCR) was employed to measure the mRNA levels of epidermal growth factor receptor(EGFR), c-Jun N-terminal kinase(JNK), and Bcl2-associated X protein(Bax) in the A549 and PC-9 cells treated with NWXF and/or GEF. Western blot was employed to determine the protein levels of EGFR, p-EGFR, JNK, p-JNK, and Bax in the A549 and PC-9 cells treated with NWXF and/or GEF. A total of 77 active components, 488 potential targets, and 49 key targets involved in the treatment of NSCLC with NWXF were predicted. The results of GO annotation showed that NWXF may treat NSCLC by regulating the biological processes such as cell proliferation, apoptosis, and protein phosphorylation. KEGG enrichment revealed that the key targets of NWXF in treating NSCLC were enriched in the mitogen-activated protein kinase(MAPK), phosphatidylinositol 3-kinase(PI3K)-protein kinase B(AKT), hypoxia-inducible factor-1(HIF-1), and microRNA-related signaling pathways. Molecular docking results showed that 91.9% of the docking scores were greater than 5, indicating the strong binding capability between main active components and key targets. The cell experiments demonstrated that NWXF combined with GEF synergistically inhibited the proliferation, promoted the apoptosis, decreased p-EGFR/EGFR and p-JNK/JNK values, down-regulated the mRNA levels of EGFR and JNK, and up-regulated the mRNA and protein levels of Bax in A549 and PC-9 cells. In conclusion, NWXF combined with GEF can regulate the EGFR/JNK pathway to promote the apoptosis of NSCLC cells, thus treating NSCLC.

ELK1 Enhances Pancreatic Cancer Progression Via LGMN and Correlates with Poor Prognosis.

Pancreatic cancer is one of the most lethal cancers and its prognosis is extremely poor. Clarification of molecular mechanisms and identification of prognostic biomarkers are urgently needed. Though we previously found that LGMN was involved in pancreatic carcinoma progression, the upstream regulation of LGMN remains unknown. We used reliable software to search for the potential transcription factors that may be related with LGMN transcription, we found that ELK1 could be a new regulator of LGMN transcription that binded directly to the LGMN promoter. Moreover, knocking down of ELK1 reduced pancreatic cancer cells proliferation, invasion and survival, while LGMN restored the malignancy of pancreatic cancer in vitro and in vivo. Overexpression of ELK1 further increased cancer cells proliferation, invasion and survival. Clinically, ELK1 and LGMN were positively correlated with clinical stage, degree of differentiation and Lymph node infiltration. ELK1 and LGMN were identified as independent prognostic factors for overall survival. The patients with low expression of ELK1/LGMN survived an average of 29.65 months, whereas those with high expression of ELK1/LGMN survived an average of 16.67 months. In conclusive, our results revealed a new mechanism by which ELK1 promoted the progression of pancreatic cancer via LGMN and conferred poor prognosis.

Overexpression of GDP dissociation inhibitor 1 gene associates with the invasiveness and poor outcomes of colorectal cancer.

GDP dissociation inhibitor (GDI) regulates the GDP/GTP exchange reaction of most Rab proteins by inhibiting GDP dissociation. This study evaluated the potential prognostic and predictive value of GDI1 in colorectal cancer (CRC). To address the prognostic power of GDI1, we performed individual and pooled survival analyses on six independent CRC microarray gene expression datasets. GDI1-enriched signatures were also analyzed. Kaplan-Meier and Cox proportional analyses were employed for survival analysis. An immunohistochemistry (IHC) analysis was performed to validate the clinical relevance and prognostic significance of the GDI1 protein level in CRC tissue samples. The results revealed that GDI1 mRNA level was significantly linked with the aggressiveness of CRC, which is compatible with gene set enrichment analysis. A meta-analysis and pooled analysis demonstrated that a higher mRNA GDI1 expression was dramatically correlated with a worse survival in a dose-dependent manner in CRC patients. Further IHC analysis validated that the protein expression of GDI1 in both cytoplasm and membrane also significantly impacted the outcome of CRC patients. In CRC patients with stage III, chemotherapy significantly reduced the relative risk of death in low-GDI1 subgroup (hazard ratio (HR) = 0.22; 95% confidence interval (95% CI) 0.09-0.56, p = 0.0003), but not in high-GDI1 subgroup (HR = 0.63; 95% CI 0.35-1.14, p = 0.1137). Therefore, both high mRNA and protein levels of GDI1 were significantly related to poor outcomes in CRC patients. GD11 may serve as a prognostic biomarker for CRC.

Comparing prophylactic use of cefazolin for SSI in cesarean section: a systematic review and meta-analysis.

PURPOSE: To summarize the available evidence to explore the effect of different prophylactic cefazolin regimens on postoperative surgical site infection after cesarean section. METHODS: We searched WOS, Pubmed, and EMBASE Database also traced citations in the reference sections of the retrieved studies. English search words: Cesarean section, Surgical site infection, Cefazolin. The majority of the literature are randomized controlled trials comparing varied regimens of cefazolin. RESULTS: A total of 11 randomized controlled trials and 4 non-randomized controlled trials involving 16,328 pregnant women were eligible. There was no statistically significant difference in the risk of SSI after cesarean section when cefazolin was given at a high dose compared with cefazolin at a low dose (OR 0.77, 95% CI 0.57-1.04, I2 = 0.0%). The risk of SSI after cesarean section was reduced by prophylactic use of cefazolin before skin incision compared with that after the umbilical cord clamping (OR 0.48, 95% CI 0.29-0.82, I2 = 53.4%). Because of the extreme heterogeneity of the combined drug use, no meta-analysis results were provided. The consequences of cefazolin combined with other antibiotics (either orally or intravenously) vary widely. For pregnant women with different risk factors, cefazolin alone or the type, dose and drug route of cefazolin combined with additional antibiotics need to be further studied and explored. CONCLUSIONS: All in all, this article illustrates a better use of cefazolin for the control of Surgical incision site infection in the cesarean section. For pregnant women with cesarean section without high-risk factors of infection, the use of cefazolin is effective, but for pregnant women with different high-risk factors, the specific use of prophylactic antibiotics needs to be further explored.

Simultaneous detection of human CYP2C19 polymorphisms and antibiotic resistance of Helicobacter pylori using a personalised diagnosis kit.

OBJECTIVES: A personalised diagnosis kit for Helicobacter pylori that employs visual gene chip technology for the simultaneous detection of CYP2C19 polymorphisms and clarithromycin/levofloxacin antibiotic resistance was evaluated. METHODS: Gastric antrum mucosa biopsy specimens of 394 patients were tested using the kit. DNA sequencing and antibiotic susceptibility testing of the H. pylori were also performed. RESULTS: In total, 267 (67.8%) of the 394 specimens were positive for H. pylori using the kit and DNA sequencing, and 136 (34.5%) were positive by culturing. For human CYP2C19 and the bacterial 23S rRNA and gyrA genes, the concordance rates were 92.4% (364/394), 96.6% (258/267) and 97.0% (259/267) between the kit and DNA sequencing results, respectively. For clarithromycin and levofloxacin resistance, the concordance rates were 90.4% (123/136) and 81.6% (111/136) between the kit and antibiotic susceptibility testing results. CONCLUSIONS: The personalised diagnosis kit for H. pylori provides useful information for the choice of proton pump inhibitor and antibiotic in combination therapy.

Activated hepatic stellate cells directly induce pathogenic Th17 cells in chronic hepatitis B virus infection.

Th17 cells are involved in liver fibrosis by activating hepatic stellate cells (HSCs). We aimed to investigate whether HSCs are able to regulate the function of Th17 cells and to determine the relevant mechanism. Sixty-five patients diagnosed with chronic hepatitis B (CHB) were enrolled in this study. To determine the effect of HSCs on T cells, naïve CD4+T cells and Th17 cells were sorted from CHB patients and cultured with or without activated-HSCs, and cytokine expression and gene transcription were analyzed. In addition, the regulatory mechanism of HSCs was investigated. ELISA and qRT-PCR showed that Th17 cells from CHB patients were more pathogenic, on the basis of the expression of IL-17A, IL-23R, RORC, CCL20 and CCR6, and meanwhile, they could activate the primary HSCs. Co-culture experiments indicated that activated HSCs dramatically promoted proliferation of CD4+T cells in a time- and dose-dependent manner. In addition, they could induce naïve CD4+T cells to become Th17 cells which had a more pathogenic phenotype. Moreover, activated HSCs-mediated induction of Th17 cells might depend on the release of IL-1ß and IL-6 as well as on the COX-PGE2 pathway. Th17 cells cooperated with HSCs in a proinflammatory feedback loop might provide a better understanding of the pathogenic role of Th17 cells in the chronicity of HBV infection.

[Effect of estrogen receptor alpha gene polymorphism on the variations of T lymphocyte subsets and its related cell factors in female patients with primary cholestasis cirrhosis].

OBJECTIVE: To study the effects of estrogen receptor (ER) alpha gene polymorphism on the migration of T lymphocyte subsets and related cytokines in the female patients with primary biliary cirrhosis(PBC). METHODS: This study was conducted with sixty female PBC patients without treatment as the study group and fifty-two healthy people wtih sex and age met the requirements of the study as the control group. The polymorphism of restriction enzyme cutting site of Xba I and Pvu II in intron 1 of ERa gene was detected by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). CD4+, CD8+, CD4+CD25+ and CD4+CD28- T lymphocytes in peripheral blood were quantitatively detected by flow cytometry. RT-PCR method was used to detect the expression of TNFa, IL-2, IFNgamma, IL-4, IL-6 and IL-10 in peripheral mononuclear cells. RESULTS: The positive rate of Pp in ERa gene Pvu II enzyme gene subtypes of female PBC patients was significantly greater than that of the control group, and the positive rate of pp gene subtype was significantly smaller than that of the control group (X2 = 7.2880, P = 0.0261). The difference of Xba I genotype and allele frequency between the female PBC patients group and the control group was not of statistical significance (X2 = 6.5382, P = 0.5833). The proportion of CD4+ T in T lymphocytes of PBC patients was increased to 45.31%+/-5.26%, compared with 33.81%+/-3.87% in the control group; and the proportion of CD8+ T lymphocytes was decreased to 27.78%+/-1.43 % from 31.83%+/-1.73% in the control group. In comparison with the control group, the proportion of CD4+CD25+ T lymphocytes decreased significantly, while that of CD4+CD28- T lymphocytes rose significantly. The expression levels of TNFa, IL-2 and IFNgamma mRNA were 0.59+/-0.19, 0.71+/-0.29 and 0.67+/-0.21 respectively, which were significantly higher than that in the control group (0.22+/-0.13, 0.31+/-0.14, 0.27+/-0.13) (t = 6.93, 5.07, 7.01, P value less than 0.01); the expression level of IL-6 mRNA was increased to 0.45+/-0.21 from 0.34+/-0.16 in the control group (t = 1.84, P value less than 0.05); and the difference of the expression levels of IL-4 and IL-10 mRNA between two groups was not of statistical significance. CONCLUSION: Pp of gene Pvu II was a genetically susceptible genotype in female PBC patients, and the allele p was a susceptible gene. Th1 cell subsets and related cytokines were dominant in peripheral blood of PBC patients. As a background of genetic susceptibility, ERa gene polymorphism could affect the shift of T lymphocyte subsets and the expression of the related cytokines in PBC patients.