Promotion of Hair Regrowth in Androgenetic Alopecia with Supplemented Erzhi Wan: Exploring Its Mechanism Using Network Pharmacology and Molecular Docking.

Purpose: Supplemented Erzhi Wan (SEZW) is a Traditional Chinese Medicine commonly used in the treatment of androgenetic alopecia (AGA). This study aims to verify the effectiveness of SEZW for the treatment of AGA in mice and explore the potential molecular mechanisms underlying its function using network pharmacology and molecular docking. Methods: Forty mice were divided into five groups: Control, AGA-model, AGA-Positive, SEZW Low Dose, and SEZW High Dose. Hair regrowth in mice was evaluated by scoring hair on days 0, 14, and 28 post-treatment and weighing mouse hair on day 28 post-treatment. The targets of the active compounds of SEZW were obtained using the Traditional Chinese Medicine Database. AGA-related targets were downloaded from five databases. Then, the overlapping genes were identified. A protein-protein interaction network was constructed using the STRING database. Hub targets were determined through analysis. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed. Finally, molecular docking of active compounds and hub targets was performed. Results: Hair regrowth in mice in the SEZW treatment groups was significantly enhanced relative to that in the AGA-model mice. A total of 59 potential drug-disease targets were identified. Based on the GO/KEGG analysis results, oxidative stress and gland development were identified as potential mechanisms of action of SEZW in AGA treatment. The PI3K-Akt and AGE-RAGE signaling pathways and seven hub targets were identified as the potential underlying mechanism of SEZW function. Molecular docking results showed that the most active SEZW compounds bind stably to several of the candidate disease targets. Conclusion: SEZW is effective in the treatment of AGA in a mouse model. Combined with network pharmacological analysis, the potential mechanisms, signaling pathways, and hub targets of SEZW in the treatment of AGA were identified, providing new ideas for further studies.

Nidogen-2 (NID2) is a Key Factor in Collagen Causing Poor Response to Immunotherapy in Melanoma.

Background: The incidence of cutaneous melanoma continues to rise rapidly and has an extremely poor prognosis. Immunotherapy strategies are the most effective approach for patients who have developed metastases, but not all cases have been successful due to the complex and variable mechanisms of melanoma response to immune checkpoint inhibition. Methods: We synthesized collagen-coding gene expression data (second-generation and single-cell sequencing) from public Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) databases. Bioinformatics analysis was performed using R software and several database resources such as Metascape database, Gene Set Cancer Analysis (GSCA) database, and Cytoscape software, etc., to investigate the biological mechanisms that may be related with collagens. Immunofluorescence and immunohistochemical staining were used to validate the expression and localization of Nidogen-2 (NID2). Results: Melanoma patients can be divided into two collagen clusters. Patients with high collagen levels (C1) had a shorter survival than those with low collagen levels (C2) and were less likely to benefit from immunotherapy. We demonstrated that NID2 is a potential key factor in the collagen phenotype, is involved in fibroblast activation in melanoma, and forms a barrier to limit the proximity of CD8+ T cells to tumor cells. Conclusion: We clarified the adverse effects of collagen on melanoma patients and identified NID2 as a potential therapeutic target.

Exosomal circ_0048856 derived from non-small cell lung cancer contributes to aggressive cancer progression through downregulation of miR-1287-5p.

AIM: Present study was aimed to explore the diagnostic value and mechanism of exosome derived circular RNA (circ)_0048856 in non-small cell lung cancer (NSCLC) development. METHODS: Exosomes protein markers, CD63 and CD9, were examined by Western blot. Real time quantitative reverse transcriptase - polymerase chain reaction (qRT-PCR) was used to detect the relative expression of circ_0048856 and miR-1287-5p. Proliferation of NSCLC cell was detected by MTT assay. Transwell assay was utilized to evaluated the migration and invasion of NSCLC cells. Apoptosis rate was examined by Flow cytometry. Double luciferase report assay was used to assess the targeting association between circ_0048856 and miR-1287-5p. Diagnostic value of circ_0048856 for NSCLC was estimated by receiver operating characteristic (ROC) curve. Animal models were constructed to explore the function of exosomal circ_0048856 in NSCLC. RESULTS: Diameter of exosomes in NSCLC serum was ranged between 75 and 150 nm. Exosomes circ_0048856 was enhanced in NSCLC serum and cell lines (P < 0.001). Exosome derived circ_0048856 had high diagnostic value for NSCLC. Area under the curve (AUC) was 0.943 (95%CI=0.904-0.982, P < 0.001). at the optimal cutoff value of 1.800, the sensitivity was 0.880, and specificity was 0.800. Exosome circ_0048856 facilitated proliferation, migration and invasion, inhibited apoptosis of NSCLC cells. MiR-1287-5p could be effaced by circ_0048856. MiR-1287-5p reversed the biological behavior changes of NSCLC cells which induced by circ_0048856. In vivo, exosomal circ_0048856 could significantly promote tumor growth (P < 0.001). CONCLUSION: Exosomes derived circ_0048856 was elevated in NSCLC serum and cell lines. Exosome circ_0048856 promoted the NSCLC development by targeting miR-1287-5p. Exosome circ_0048856 had high diagnostic value for NSCLC.

MicroRNA-124 suppresses the invasion and proliferation of breast cancer cells by targeting TFAP4.

MicroRNA (miRNA/miR)-124 is widely accepted as the suppressor of different tumors. The present study aimed to improve understanding of the potential role of miR-124 in breast cancer. The gene expression profile change derived from the overexpression of miR-124 was investigated using RNA sequencing and bioinformatics analysis of the breast cancer cell line SKBR3. The results demonstrated that the gene expression profile of SKBR3 cells significantly changed. In addition, the transcription factor activating enhancer-binding protein 4 (TFAP4) gene was identified among the top 10 differentially expressed genes, and was identified as a novel target gene of miR-124 using a dual-luciferase reporter assay. TFAP4 knockdown in notably impaired SKBR3 cell migration and proliferation, which was consistent with decreasing migration and proliferation ability following overexpression of miR-124. Taken together, these results suggest that overexpression of miR-124 can suppress the migration and proliferation of SKBR3 cells by tarsgeting TFAP4. Thus, TFAP4 may act as a novel therapeutic target of breast cancer.

CTCF-induced upregulation of LINC01207 promotes gastric cancer progression via miR-1301-3p/PODXL axis.

BACKGROUND: Long non coding RNAs (lncRNAs) have been validated to be involved in the complicated biological processes during tumor progression. LINC01207 has been identified as an oncogene in several cancer types. However, the function of LINC01207 and its underlying molecular mechanism in gastric cancer (GC) are poorly understood. METHODS: The expression level of LINC01207, miR-1301-3p and PODXL mRNA was detected in GC tissues and cells by RT-qPCR. The level of PODXL protein was examined by western blot. Colony formation assay, EdU assay, TUNEL assay, caspase-3 activity test and transwell assays were carried out to analyze the effect of LINC01207 on GC cell proliferation, apoptosis, migration and invasion. The interaction between RNAs was confirmed by luciferase reporter assay, RNA pull-down assay and RIP assay. RESULTS: LINC01207 was expressed at high level in GC tissues and cells. Silencing of LINC01207 impaired GC cell proliferation, migration and invasion but promoted cell apoptosis. Mechanistically, LINC01207 acted as a ceRNA by sponging miR-1301-3p to upregulate PODXL. Besides, miR-1301-3p silencing or PODXL overexpression could abolish the inhibitory effect of LINC01207 knockdown on GC cell growth and migration. CCCTC-binding factor (CTCF) could transcriptionally activate LINC01207 in GC cells. CONCLUSIONS: CTCF-induced activation of LINC01207 contributes to GC progression through regulating miR-1301-3p/PODXL axis.

Identification of four genes associated with cutaneous metastatic melanoma.

BACKGROUND: Cutaneous melanoma is an aggressive cancer with increasing incidence and mortality rates worldwide. Metastasis is one of the primary elements that influence the prognosis of patients with cutaneous melanoma. This study aims to clarify the potential mechanism underlying the low survival rate of metastatic melanoma and to search for novel target genes to improve the survival rate of patients with metastatic tumors. METHODS: Gene expression dataset and clinical data were downloaded from The Cancer Genome Atlas portal. Differentially expressed genes (DEGs) were identified, and their functions were studied through gene ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. Survival and multivariate Cox regression analyses were used to screen out candidate genes that could affect the prognosis of patients with metastatic melanoma. RESULTS: After a series of comprehensive statistical analysis, 464 DEGs were identified between primary tumor tissues and metastatic tissues. Survival and multivariate Cox regression analyses revealed four vital genes, namely, POU2AF1, ITGAL, CXCR2P1, and MZB1, that affect the prognosis of patients with metastatic melanoma. CONCLUSION: This study provides a new direction for studying the pathogenesis of metastatic melanoma. The genes related to cutaneous metastatic melanoma that affect the overall survival time of patients were identified.

Structure-based design of peptides against HER2 with cytotoxicity on colon cancer.

In this study, we found that four novel peptides designed by molecular modeling techniques were successfully applicated with cytotoxicity on colon cancer cells sw620. First, the interactions between the Herstatin and the HER2 were explored by ational-designed approaches, which were combined with homology modeling, protein/protein docking, and structural superimposition analysis. Then, based on the results derived from theoretical analysis, four novel peptides were designed, synthesized, and experimentally evaluated for biological function; it was found that they showed a remarkable enhancement on Herceptin to inhibit the genesis and development of colon cancers, and no significant side effects on normal colon cells NCM460 were observed but Doxorubicin had. These results indicated that it is a feasible way to use the well-designed peptides derived from Herstatin to enhance the efficacy of clinical drugs Herceptin and to kill colon cancer cells selectively without harming normal colon cells. We believe that our research might provide a new way to develop the potential therapies for colon cancers.

Study on Hydrogen Fluoride at High Temperature Detection Method with Temperature Correction Based on Laser Technology.

Hydrogen fluoride(HF) is one of the important character gases for fault diagnosis of gas insulation switch (GIS) in the system of substation equipment. The high-accuracy, fast- response and real-time detection method of HF is a focus in industrial and environmental fields. In this research, the HF detection experiment system was set up at first based on laser absorption spectroscopy technology combined with anti-corrosion multiple reflection cell made by monel steel. Moreover, the laser absorption spectral characteristics of HF at different temperature were analyzed, then the coefficient partition function curve and absorption linestrength curve according to the distribution function coefficient in HITRAN database were studied. As the most important work, the concentration inversion algorithm was designed here with HF character spectrum analysis and temperature parameter correction method for accurate concentration inversion after the basic study. At last, the continuous experimental results were obtained by HF sample gases of different concentration considerating the temperature characteristic of the multiple reflection cell. When the multiple reflection cell was heat and stay stably, the biggest detection error of concentration inversion was 5.33% and 5.87% at 313 and 323 K respectively without temperature correction, and that was 1.20% and 1.47% respectively after temperature correction. By continuous detection and culculation, the detection limit is 8.7×10-5 mmol·mol-1 at 323 K which is a little higher than 6.3×10-5 mmol·mol-1 at 290 K(20 m optical length). Although the detection error with temperature correction at high temperature was higher than it at room temperature, the results show that it was lower than that without correction at the same temperature. It was verified that the this spectrum detection method and concentration inversion algorithm works stably and reliably, so this technology could realize HF real-time monitoring demand in chemical production field and it will provide the effective technical support in gas emission regulation in safety and environment protection for our country.

Enhancing the treatment effect on melanoma by heat shock protein 70-peptide complexes purified from human melanoma cell lines.

Dendritic cell (DC) vaccines are currently one of the most effective approaches to treat melanoma. The immunogenicity of antigens loaded into DCs determines the treatment effects. Patients treated with autologous antigen-loaded DC vaccines achieve the best therapeutic effects. In China, most melanoma patients cannot access their autologous antigens because of formalin treatment of tumor tissue after surgery. In the present study, we purified heat shock protein 70 (HSP70)-peptide complexes (PCs) from human melanoma cell lines A375, A875, M21, M14, WM­35, and SK­HEL­1. We named the purified product as M­HSP70­PCs, and determined its immunological activities. Autologous HSP70­PCs purified from primary tumor cells of melanoma patients (nine cases) were used as controls. These two kinds of tumor antigenic complexes loaded into DCs were used to stimulate an antitumor response against tumor cells in the corresponding patients. Mature DCs pulsed with M­HSP70­PCs stimulated autologous T cells to secrete the same levels of type I cytokines compared with the autologous HSP70­PCs. Moreover, DCs pulsed with M­HSP70­PCs induced CD8+ T cells with an equal ability to kill melanoma cells from patients compared with autologous HSP70­PCs. Next, we used these PC­pulsed autologous DCs and induced autologous specific CD8+ T cells to treat one patient with melanoma of the nasal skin and lung metastasis. The treatment achieved a good effect after six cycles. These findings provide a new direction for DC-based immunotherapy for melanoma patients who cannot access autologous antigens.

[Study on Ammonia Emission Rules in a Dairy Feedlot Based on Laser Spectroscopy Detection Method].

It needs on-line monitoring of ammonia concentration on dairy feedlot to disclose ammonia emissions characteristics accurately for reducing ammonia emissions and improving the ecological environment. The on-line monitoring system for ammonia concentration has been designed based on Tunable Diode Laser Absorption Spectroscopy (TDLAS) technology combining with long open-path technology, then the study has been carried out with inverse dispersion technique and the system. The ammonia concentration in-situ has been detected and ammonia emission rules have been analyzed on a dairy feedlot in Baoding in autumn and winter of 2013. The monitoring indicated that the peak of ammonia concentration was 6.11 x 10(-6) in autumn, and that was 6.56 x 10(-6) in winter. The concentration results show that the variation of ammonia concentration had an obvious diurnal periodicity, and the general characteristic of diurnal variation was that the concentration was low in the daytime and was high at night. The ammonia emissions characteristic was obtained with inverse dispersion model that the peak of ammonia emissions velocity appeared at noon. The emission velocity was from 1.48 kg/head/hr to 130.6 kg/head/hr in autumn, and it was from 0.004 5 kg/head/hr to 43.32 kg/head/hr in winter which was lower than that in autumn. The results demonstrated ammonia emissions had certain seasonal differences in dairy feedlot scale. In conclusion, the ammonia concentration was detected with optical technology, and the ammonia emissions results were acquired by inverse dispersion model analysis with large range, high sensitivity, quick response without gas sampling. Thus, it's an effective method for ammonia emissions monitoring in dairy feedlot that provides technical support for scientific breeding.

[Laboratory Investigation of DNAPL Migration Behavior and Distribution at Varying Flow Velocities Based on Light Transmission Method].

The migration and distribution of dense non-aqueous phase liquid (DNAFL) in subsurtace are attectea ny many factors. We selected PCE as the substitute contaminant, and performed several well-controlled two-dimensional sandbox experiments to investigate the effect of flow velocity on DNAPL infiltration and redistribution. Light transmission method (LTM) was used to monitor the transport process of DNAPL in the sandbox and quantitatively measure DNAPL saturation. The spatial moments based on measured DNAPL saturation were used to describe the average spatial behavior of DNAPL plume at various times. Experimental results showed a strong correlation between results obtained by LTM and the known amounts of DNAPL added into the sandbox (R2 >0.98). The LTM accurately reflected the infiltration and redistribution processes. The results of DNAPL saturation and first moment (mass center) showed that the increased velocity promoted not only lateral but also vertical migration, leading to an inclined percolation path. Also vertical migration reacted more sensitive to flow velocity. The second moment (spread variance) showed that the increased velocity promoted lateral and vertical spread, increasing the pollution scope. The histogram of DNAPL saturation showed a unimodal distribution at low flow velocity, but showed a bimodal distribution at lager flow velocity, and the distance between two peaks became higher with the increasing flow velocity.

Oncogenicity of LHX4 in colorectal cancer through Wnt/ß-catenin/TCF4 cascade.

The LHX genes play an important role in a number of developmental processes. Potential roles of LHXs have been demonstrated in various neoplastic tissues as tumor suppressors or promoters depending on tumor status and types. The aim of this study was to investigate the function role of LHXs in the human colorectal cancer (CRC). The gene expression changes of LHXs in CRC tissues compared with noncancerous colorectal tissues was detected using real-time quantitative reverse transcriptase-polymerase chain reaction (QRT-PCR) analysis and immunohositochemistry. And we identified the gene LHX4 that were significantly upregulated in CRC by QRT-PCR analysis and immunohistochemistry. Furthermore, we discovered that LHX4 promoted cancer cell proliferation in vitro, and LHX4 expression correlated with elevated ß-catenin levels in CRC and ß-catenin function was required for LHX4's oncogenic effects. Mechanistically, LHX4 facilitate TCF4 to bind to ß-catenin and form a stable LHX4/TCF4/ß-catenin complex and transactive its downstream target gene. LHX4 mutations that disrupt the LHX4-ß-catenin interaction partially prevent its function in tumor cells. All in all, LHX4 is a commonly activated tumor promoter that activate Wnt/ß-catenin signaling in cancer cells of CRC.

Discrimination of three dimensional fluorescence spectra based on wavelet analysis and independent component analysis.

Fluorescence spectroscopy is a rapid and non-destructive method for monitoring water quality. In this work, wavelet analysis, together with independent component analysis (ICA), was applied for component recognition of seriously overlapped, multi-component, three dimensional fluorescence spectra. Wavelet analysis extracts the features of the spectra and amplifies differences among phenolic homologs. ICA analysis in blind signal separation was used to separate single component before multiple linear regression (MLR). The proposed method increases the correct classification rate and enriches the spectra library. As such, it is a useful alternative to traditional techniques in component recognition.

[Measurement and analysis of lead in soil using X-ray fluorescence spectroscopy].

The present paper analyzed the characteristics of X-ray fluorescence spectroscopy (XRF) of metal element lead in soil using the NITON XLt793 portable X-ray fluorescence spectra of heavy metal analyzer under laboratory conditions. The characteristic spectral lines of L(alpha) (energy: 10. 55 keV) and L(beta) (energy: 12. 61 keV) with different matrix elements were selected respectively for lead in the experiment. By measuring the intensities of the characteristic spectral line with different Pb concentration, the results demonstrate that the relation between concentration [mass fraction 10 x 10(-6) - 1 800 x 10(-6)] of Pb element and the intensity of the characteristic spectrum is well linear. The calibration curve of Pb was plotted based on the different concentration measurement results, and the limit of detection of 7.89 x 10(-6) was obtained for Pb in soil.

A new purification method for enhancing the immunogenicity of heat shock protein 70-peptide complexes.

When purified from a tumor, certain heat shock protein 70 (HSP70)-peptide complexes (PCs) can function as effective vaccines against the tumor from which the complexes were isolated. The immunogenic mechanisms of HSP70 preparations imply that tumor-derived HSP70-PCs exhibit antigens associated with antigen-presenting cells such as dendritic cells (DCs), inducing antigen-specific cytotoxic CD8+ T cells. However, some important membrane-resident tumor-associated peptides, such as the HER-2/neu (c-erbB2) oncogenic protein, cannot be purified from HSP70 by traditional methods. In the present study, a new approach for the purification of HSP70-PCs from HER-2-overexpressing breast cancer cells was established. The detergent 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS) was used to obtain more effectual tumor peptides. The new purified product was named HSP70-HER-2-PC, and its immunological activities were determined. Traditionally purified HSP70-PCs (without CHAPS) and recombinant human HSP70-HER-2 protein complexes (recombined in vitro) were used as controls. These three HSP70-associated tumor antigenic complex pulsed dendritic cells (DCs) were used to stimulate an antitumor response. The mature DCs pulsed with HSP70-HER-2-PCs stimulated autologous T cells to secrete higher levels of type I cytokine compared to the two control groups. Moreover, DCs pulsed with HSP70-HER-2-PCs induced the most specific CD8+ T cells that specifically killed the same tumor cells. These findings provide a basis for new approaches in enhancing HSP70-based immunotherapy for HER-2-associated or other membrane antigenic peptide-related cancers.

Down-regulation of cyclin D1 by small interfering RNA inhibits cell growth and induces apoptosis of laryngeal squamous cell carcinoma.

OBJECTIVE: The objective of the study was to explore the inhibitive role of cyclin D1 gene silence in laryngeal squamous cell carcinoma by lentivirus-mediated RNA interference. MATERIALS AND METHODS: Cd1-RNAi-Lentivirus and the control lentivirus (GFP-Lentivirus) were transfected into Hep-2 cells. Reverse transcriptase polymerase chain reaction was performed to explore the cyclin D1 expression level in Cd1-RNAi-Lentivirus-transfected Hep 2 cells. The apoptosis and viability of Cd1-RNAi-Lentivirus-treated Hep-2 cells were measured with flow cytometry and methyl thiazolyl tetrazoliym assay. In an animal experiment, 10 mice bearing Hep-2 cell tumor were intratumorally injected with Cd1-RNAi-Lentivirus; and the other 10 mice were injected with GFP-Lentivirus. Terminal deoxytransferase-mediated dUTP nick end labeling stains and transmission electron microscope were used to observe the apoptosis in the xenografts. RESULTS: Cyclin D1 was knocked down after Cd1-RNAi-Lentivirus was transfected into Hep-2 cells. The proliferative ability of Hep-2 cells was significantly inhibited by Cd1-RNAi-Lentivirus, and a significant apoptosis of Hep-2 cells was also observed after Cd1-RNAi-Lentivirus transfection. The average weight and volume of tumors in the Cd1-RNAi-Lentivirus-treated group were significantly lower than those in the control group (P < .01). The significant apoptosis was detected with terminal deoxytransferase-mediated dUTP nick end labeling stain and transmission electron microscope. CONCLUSIONS: The present findings suggest that cyclin D1 gene silence by lentivirus-mediated RNA interference can inhibit growth and promote apoptosis of laryngeal squamous cell carcinoma.