Combinatorial Chemistry & High Throughput Screening - Volume 25, Issue 6, 2022

Background: The efficiency of herbal medicinal products depends on the quantity of active ingredients in them, which can vary considerably in different contents. Hence, the quality control of herbal medicines is a critical concern. Objective: This paper aims to provide a succinct review of recent chemometrics applications in solving the uncertainty of the authentication of herbal medicines over the last two decades. Methods: Studies involving chemometrics applications in conjunction with various analytical methods have been categorized according to the type of research used in the quality evaluation of different samples, including chromatographic (HPLC, GC-MS) and spectroscopic analysis (UVVis, FTIR, NMR, and MS). Results: This review consists of over 90 studies illustrating the relevance of chemometrics methods in the discrimination based on the key bioactive components and phytochemical diversity of several herbs from closely related species. In addition to the prediction of the active components, the distinction between varieties and hybrids was accomplished through quantitative analysis techniques. Conclusion: Methods of chemometrics have provided an important and potent tool for the quality control and authentication of various herbs.

Background: In recent biomedical research, the area of cancer and infectious diseases occupies a leading position in the utilization of medicinal plants as a source of drug discovery. Malaysia has a diversity and a large number of underutilized fruits that are rich in phenolic compounds. Artoarpus altilis is considered an underutilized fruit that is rich in phenolic compounds. Methanol extracts of A. altilis have been previously found to contain a high content of antioxidant phytochemicals. Objective: The purpose of the study was to evaluate the cytotoxicity and toxicological effect of methanol fruit extracts against MCF-7 cells. To determine the least concentration that might kill or suppress the growth of the cancer cells was in a concentration-dependent manner. Methods: The variation in the cytotoxic activity among the extracts was indicated by determining the IC50 of each extract against cells at 72 h. The IC50 of the samples was measured using a trypan blue exclusion assay. The methanol extract of the pulp part showed the least inhibition concentration of 15.40±0.91 μg/mL on MCF-7 cells. In the study, the molecular mechanism of methanol extracts-induced apoptosis and cell cycle arrested in human cancer cells were investigated in a time-dependent-manner by using flow cytometry. The treated cells were stained with nexin to detect early and late apoptosis and with propidium iodide (PI) for cell cycle arreste associated with the DNA fragmentation; various cell arrests occurred at G1/S, S, and G2/M phases. Lastly, the gene expression analysis by RT-qPCR method was carried out by analyzing the expression of the gene of interest for the quantification of mRNA levels. Results: Results after cells were treated with IC50 were revealed by upregulating anti-apoptotic genes/downregulated of pro-apoptotic BCL-2 gene expressions triggered the treated cells into CASPASE- 3, intrinsic and extrinsic pathways. Conclusion: These findings suggest that the methanol extracts of three parts of A. altilis fruit have potential anticancer activity against MCF-7 cells mainly the pulp part of the fruit.

Background: Erchen Decoction (ECD) is a complex herbal formulation widely used for treating lipid metabolism disorder (LMD) in China. This study aims to explore the microRNA (miRNA)-related molecular targets of ECD against LMD using a network pharmacology approach (NPA) Methods: We randomly divided 20 male Sprague Dawley rats into two groups; 10 rats were normal controls, and the other 10 rats were fed a high-fat diet (HFD) for 12 weeks to establish an LMD model. Differentially expressed miRNAs (DE-miRs, HFD vs. Control) in the rats’ liver tissues were identified by miRNA sequencing and validated with qRT-PCR. Finally, the miRNArelated molecular targets for ECD activity against LMD were identified using a standard NPA by finding the intersection between identified DE-miRs-related targets and ECD-related targets. Result: We identified 8 DE-miRs and 968 targets and compared them to 262 ECD-related targets. A final list of 22 candidate targets was identified. Using a confidence score of >0.4, the network of (protein-protein interaction) PPI relationships exhibited 22 nodes and 67 edges. The GO and KEGG enrichment analyses revealed 171 molecular targets and 59 pathways, which were associated with ECD against LMD. Conclusion: The identified molecular targets and pathways suggest that complex mechanisms are involved in ECD’s mechanism of action, and immune-inflammation-related mechanisms are closely associated with the effects of ECD. The targets obtained in this study will guide future studies on the pharmacologic effects of ECD.

Objective: The objective of this study is to construct a prognostic model using genetic markers of liver cancer and explore the signature genes associated with the tumor immune microenvironment. Methods: Cox proportional hazards regression analysis was carried out to screen the significant HR using the dataset of TCGA Liver Cancer (LIHC) gene expression data. Then LASSO (least absolute shrinkage and selection operator) was performed to select the minimal variables with significant HR of genes. Thus, the prognostic model was constructed by the minimal variables with their HR. Time-dependent receiver-operating characteristic (ROC) curve and area under the ROC curve (AUC) value was used to assess the prognostic performance. Then the patients were divided into high and low-risk groups by the median of the model. Survival analysis was performed on the two groups with testing and an independent dataset. Furthermore, enrichment analysis of signature mRNAs and lncRNAs and their co-expression genes was performed. Then, Spearman rank correlation was used to calculate the correlation between immune cells and genes in the prognostic model, and abundance difference of the immune cells in high and low risks groups was tested. Results: A total of 5989 genes with significant HR were identified. 6 key genes (three mRNAs: DHX37, SMIM7, and MFSD1, three lncRNAs: PIWIL4, KCNE5, and LOC100128398) screened by LASSO were used to construct the model with their HR value respectively. The AUC values of 1 and 5-year overall survival were 0.78 and 0.76 in discovery data and 0.67 and 0.68 in testing data. Survival analysis performed significantly discriminated high and low groups with testing and independent data. Furthermore, many immune cells such as nTreg found a significant correlation with the genes in the prognostic model, and many immune cells showed significantly different abundance in high and low-risk groups. Conclusion: In the study, we used Univariate Cox analyses and LASSO algorithm with TCGA gene expression data to construct the prognostic model in liver cancer patients. The prognostic model comprised of three mRNAs, including DHX37, SMIM7, MFSD1, and three lncRNAs, including PIWIL4, KCNE5, and LOC100128398. Furthermore, these gene expression levels were associated with the abundance of some immune cells, such as nTreg. Also, many immune cells have significantly different abundance in high and low-risk groups. All these results indicated that the combination with all these six genes could be the potential biomarker for the prognosis of liver cancer.

Background: Hepatocellular carcinoma (HCC) is one of the most common malignant tumors of the digestive tract in the world. Therefore, it is important to carry out studies on the molecular mechanisms of early diagnosis and treatment of HCC to reduce mortality. Methods: Bioinformatic analysis was performed to explore the significant role of GCSF on the occurrence and development of neoplasm. Differently expressed genes (DEGs) were screened, and the significant hub genes related to GCSF were identified by the multiple algorithms of Cytoscape. Functional annotation for DEGs, pathological stage, and overall survival analysis were implemented. In addition, the verification for the role of GCSF on HCC was made via the clinical samples. A total of 70 participates diagnosed as HCC were recruited from November 2014 to November 2019. The immunohistochemistry assay, qRT-PCR, receiver operating characteristic (ROC) curves, and overall survival analysis were carried out. Results: GCSF was related to the tumor size, and the expression of GCSF was up-regulated in hepatocellular carcinoma tissues. The enrichment results of GO and KEGG analysis were mainly enriched in “Inflammatory response”, “Protein binding”, “Metabolic pathways”, and “Proteasome”. The tumor diameter (P < 0.001), and survival time (P < 0.001) were significantly associated with the expression of GCSF via the verification of clinical data. The univariate and multivariate Cox proportional regression analysis manifested that high expression of GCSF in patients with HCC was related to poor OS. Conclusion: The expression level of GCSF is significantly associated with the prognostic survival of HCC, and it is expected to become a new prognostic marker of HCC, providing a novel idea for future basic research as well as targeted therapy.

Objective: We aimed to investigate the feasibility of multi-acquisition with variable resonance image combination slab selectivity inversion recovery (MAVRIC SL IR) sequence on 3.0 T MRI in patients with anterior cervical discectomy and fusion (ACDF) surgery compared to bandwidth-optimized short tau inversion recovery (STIR) sequence. Methods: Paired sagittal MR images of MAVRIC SL IR and bandwidth-optimized STIR sequences were acquired and analyzed for 21 patients after ACDF surgery with PEEK cage-plate construct. Quantitative comparisons were made on the metal artifact areas of paired mid-sagittal images. In qualitative analysis, the consistency of fat suppression and visibility of anatomic structures (bonemetal interface, surrounding soft tissues, and spinal cord) were independently assessed, based on a five-point scale by two musculoskeletal radiologists, who were blind to the images and patient details. Results: The application of the MAVRIC SL IR sequence resulted in a significant reduction of 48% in the mean area of metal artifacts (t = -7.141, p < 0.001). Based on the comments received from both the reviewers, the MAVRIC SL IR sequence showed greater visibility of the bone-metal interface (p < 0.001), considerable visibility of the surrounding soft tissues (p > 0.05) but worse visibility of the spinal cord (p < 0.001), including the consistency of fat suppression (p < 0.001) relative to the bandwidth-optimized STIR sequence. Conclusion: With significantly reduced metal artifacts, the MAVRIC SL IR sequence can be implemented in patients undergoing ACDF surgery with PEEK cage-plate construct for 3.0 T MRI, despite the poor visibility of the spinal cord.

Background: Developing the high-efficiency and low-risk small-molecule greennematocide is the key of effective control of the nematodes. Paeonol, is a naturally occurring phenolic compound, isolated from the root bark of Paeonia suffruticosa and the whole plant of Cynanchum paniculatum. Due to its crucial phenolic ketone skeleton, modern biological science research has indicated that paeonol has a wide range of biological activities. Methods: The structural modification of paeonol into paeonol carbonyl hydrazone derivatives is a potential approach for the development of novel nematodes, which showed more toxicity than paeonol. However, there are no reports on the nematicidal activity of paeonol carbonyl hydrazone derivatives to control Heterodera glycines. Results: We always endeavor to discover and develop biorational natural products-based pesticidal agents, 4 significant intermediates and 21 novel 3/5(3,5)-(di)nitro/chloropaeonol carbonyl hydrazone derivatives were prepared, and their structures well characterized by ¹H NMR, HRMS, MS, and mp. Due to the steric hindrance, the substituents on the C=N double bond of all hydrazine compounds adopted E configuration. Results of nematicidal activity revealed that, among all compounds, especially 5-nitropaeonol (5) and 3,5-dinitropaeonol (7) displayed the most potent nematicidal activity H. glycines in vivo with LC50 values of 0.0323 and 0.0367 mg/mL, respectively. Conclusion: It suggested that for the 3/5(3,5)-(di)nitro/chloropaeonol carbonyl hydrazone derivatives, a nitro group introduced at C5 position of 1 was necessary for obtaining the potent compound as nematicidal agents. These preliminary results will pave the way for further modification of paeonol in the development of potential new nematicides.

Objective: The goal of this study was to investigate the status of FEN1 in colorectal cancer (CRC) and determine the potential correlation between FEN1 expression level and clinicopathological parameters in CRC patients. Methods: Expression of FEN1 in CRC tissue on tissue microarray was detected using immunohistochemistry (IHC). The relationship between FEN1 expression status and clinicopathologic characteristics of CRC was analyzed by the Chi-square test. The survival data of TCGA Colon Cancer (COAD) were obtained from ucsc xena browser (https://xenabrowser.net/). Patients were separated into higher and lower expression groups by median FEN1 expression. The association with prognosis of CRC patients was determined by Kaplan-Meier survival analysis with Log-rank test. Results: FEN1expression level and cellular localization had wide variability among different individuals; we classified the staining results into four types: both positive in nucleus and cytoplasm, both negative in nucleus and cytoplasm, only positive in the nucleus, only positive in the cytoplasm. Moreover, FEN1 expression status only correlated with patient’s metastasis status, and the patients in the NLCL group showed more risk of cancer cell metastasis. Conclusion: Our results indicate that FEN1 expression level and cellular localization had wide variability in CRC and is not a promising biomarker in CRC.

Background: Suicide has grown in global prevalence as a public health problem. We aimed to evaluate the association of socioeconomic factors, biochemical and hematologic tests, and suicide ideation. Methods: In this cross-sectional study, 8267 Iranian adults aged 35 – 65 years old were enrolled. The assessment of suicide ideation was made by the completion of Beck’s depression inventory (BDI) questionnaire; according to one specific item on the questionnaire: “have you ever decided to suicide in the past week?” Results: According to our results, 6.9 % of subjects had ideation of suicide. The results showed high levels of FBG, RBC, MCHC, and hs-CRP were associated with suicide ideation. Obese, single subjects, and current-smokers had a higher risk of suicide ideation. Conclusion: Increased physical activity, obesity, and smoking are associated with a high risk of suicide ideation; whilst, a high MCHC is related to a low risk of suicide ideation in Iranian adults.

Background: Due to the strong association between the chemistry of medicinal plants and their biological properties, it is important to determine their phytochemical composition to justify experimental tests. Objective: The aim of this study was to evaluate the in vitro antioxidant and the enzyme inhibitory properties and to identify the bioactive compounds present in the extracts of Ephedra nebrodensis growing in Algeria. Methods: Total phenolic and flavonoids content in these extracts were quantified by Folin- Ciocalteu and aluminum chloride methods. The antioxidant capacity was assessed using DPPH, ABTS, β-carotene/linoleic acid, CUPRAC and FRAP assays, and in vitro cholinesterase activity against acetylcholinesterase and butyrylcholinesterase were evaluated. The chemical constituents of the extracts were analyzed by high-performance liquid chromatography coupled with mass spectrometric detection and gas chromatography. For the acute toxicity study, extracts were administered to mice at single dose of 2 g/kg and 5 g/kg by gavage. Results: Plant extracts were rich in phenolic compounds. Ethyl acetate extract presented the highest phenolic (238.44 ± 1.50 μg GAE /mg of extract) and flavonoid (21.12 ± 0.00 μg QE /mg of extract) contents. Likewise, ethyl acetate extract showed potent radical scavenging and reducing properties. Ethanol-acetone extract showed inhibitory activity against acetylcholinesterase, and was a potent inhibitor of butyrylcholinesterase. In all extracts, flavonoids were the most abundant compounds. The phytochemical investigation showed the presence of alkaloids (ephedrine and pseudo-ephedrine). In the acute toxicity, the LD50 was superior to 5 g/kg body weight. There were no alterations in the histology of the liver and kidneys. Conclusion: This study demonstrated a good antioxidant potential and anticholinesterase activity of aerial parts of E. nebrodensis.

Background: Traditionally, Portulaca oleracea Linn. is used for treating abscesses, dysentery and liver diseases. In addition, recent studies have reported its efficacy as an analgesic, as neuroprotective, anti-inflammatory, bronchodilatory, and anticancer agent, besides antioxidant, wound healing and other important pharmacological actions. Aims and Objectives: For curing liver diseases, Chinese use juice from the fresh leaves of Portulaca oleracea Linn. Hence, to prove this claim, an in-vivo hepatoprotective study of extracts from Portulaca oleracea Linn. against carbon tetrachloride hepatotoxic rats was carried out. Methods: The in-vitro antioxidant activity by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and hepatoprotective activity of extracts of Portulaca oleracea Linn. was assessed against the carbon tetrachloride-induced toxicity rat model. Results: The extracts of Portulaca oleracea Linn produced concentration-dependent percentage inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical and most significantly, by methanol extract. Among the extracts, methanol extract showed a significant decrease in Serum glutamic oxaloacetic transaminase (SGOT), Serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP) levels and a significant increase in total protein (TP) levels against carbon tetrachloride-induced liver toxicity. Conclusion: From the results obtained, it was concluded that methanol extract of Portulaca oleracea Linn. possesses significant DPPH free radical scavenging and hepatoprotective potential.

Background: The gonadotrophin-releasing hormone agonist (GnRHa) has gained widespread popularity in achieving pituitary suppression before ovarian stimulation with exogenous gonadotropins in assisted reproductive technology protocols. However, a very small part of patients may develop ovarian hyper response after the sole administration of GnRHa without gonadotropins. Case Report: A 32-year-old female diagnosed with polycystic ovary syndrome presented for her second IVF cycle in our reproductive center. Twenty-eight days after 3.75mg triptorelin was administrated on day 2 of her menstrual cycle, bilateral ovaries were significantly enlarged and presented multiple cystic masses. The hormone profile was as follows: E24870pg/ml, P 13.19ng/ml, FSH 14IU/L, and LH 10.77IU/L. The patient felt symptoms of mild ovarian hyperstimulation syndrome. In the subsequent IVF treatment cycle, antagonist protocol was performed. It showed that follicles developed slowly and exogenous gonadotropins were used for 13 days. Finally, seven oocytes were obtained, and only one blastocyst graded 4BC formed. Conclusion: Ovarian hyperstimulation following the sole administration of GnRHa can occur, but the mechanism is still not yet clear. Antagonist protocol may be an alternative fertility strategy, but the risk of poor embryo quality and low pregnancy rate of transplantation should be warned.