Multicomponent Therapeutics of Berberine Alkaloids

Although berberine alkaloids (BAs) are reported to be with broad-spectrum antibacterial and antiviral activities, the interactions among BAs have not been elucidated. In the present study, methicillin-resistant Staphylococcus aureus (MRSA) was chosen as a model organism, and modified broth microdilution was applied for the determination of the fluorescence absorption values to calculate the anti-MRSA activity of BAs. We have initiated four steps to seek the optimal combination of BAs that are (1) determining the anti-MRSA activity of single BA, (2) investigating the two-component combination to clarify the interactions among BAs by checkerboard assay, (3) investigating the multicomponent combination to determine the optimal ratio by quadratic rotation-orthogonal combination design, and (4) in vivo and in vitro validation of the optimal combination. The results showed that the interactions among BAs are related to their concentrations. The synergetic combinations included “berberine and epiberberine,” “jatrorrhizine and palmatine” and “jatrorrhizine and coptisine”; the antagonistic combinations included “coptisine and epiberberine”. The optimal combination was berberine : coptisine : jatrorrhizine : palmatine : epiberberine = 0.702 : 0.863 : 1 : 0.491 : 0.526, and the potency of the optimal combination on cyclophosphamide-immunocompromised mouse model was better than the natural combinations of herbs containing BAs

Application of a Diatom Transfer Function to Quantitative Paleoclimatic Reconstruction — A Case Study of Yunlong Lake, Southwest China

Although diatom records from lake sediments have been used for quantitative paleoclimatic reconstruction, their validity and sensitivity have rarely been tested rigorously. At Yunlong Lake, an alpine lake in Southwest China, we studied the seasonal succession of diatom assemblages to produce a mean surface water temperature (MSWT) transfer function. In addition, based on the spatial distribution of surface diatom assemblages with water depth, we produced a diatom-water depth (WD) transfer function. Combined with the analysis of diatom assemblages in a sediment core (YL2013-A), changes in surface water temperature and water level over the last ∼100 years were quantitatively reconstructed using the diatom-based transfer functions. Comparison with records of regional meteorology and reservoir water capacity revealed that the diatom-based lake water level reconstruction is a sensitive indicator of short-term fluctuations in precipitation, and it also reflects a long-term stepwise rise in water level caused by the impounding and large-scale extension of the reservoir. In addition, the diatom-inferred MSWT is consistent with the changes in air temperature prior to large-scale human disturbance of the site. However, after the extension of the reservoir, although the regional air temperature continued to increase, the water temperature decreased substantially. This suggests that the large increase in lake water volume in the short term led to a decrease in the average water temperature, which in turn led to the occurrence of a diatom bloom in the cold season. The results demonstrate that diatom transfer functions based on modern observations of the same lake has a high environmental sensitivity and can be used for the quantitative reconstruction of regional climate change. Overall, our findings provide a foundation for the use of lake diatom records for quantitative paleoclimatic reconstruction on various timescales

Evaluation and structure-activity relationship analysis of a new series of arylnaphthalene lignans as potential anti-tumor agents.

Arylnaphthalene lignan lactones have attracted considerable interest because of their anti-tumor and anti-hyperlipidimic activities. However, to our knowledge, few studies have explored the effects of these compounds on human leukemia cell lines. In this study, five arylnaphthalene lignans including 6'-hydroxy justicidin A (HJA), 6'-hydroxy justicidin B (HJB), justicidin B (JB), chinensinaphthol methyl ether (CME) and Taiwanin E methyl ether (TEME) were isolated from Justicia procumbens and their effects on the proliferation and apoptosis of the human leukemia K562 cell line were investigated then used to assess structure-activity relationships. To achieve these aims, cytotoxicity was assayed using the MTT assay, while intracellular SOD activity was detected using the SOD Activity Assay kit. Apoptosis was measured by both the using a cycle TEST PLUS DNA reagent kit as well as the FITC Annexin V apoptosis detection kit in combination with flow cytometry. Activation of caspase-mediated apoptosis was evaluated using a FITC active Caspase-3 apoptosis kit and flow cytometry. The results indicated that HJB, HJA and JB significantly inhibited the growth of K562 cells by decreasing both proliferation and SOD activity and inducing apoptosis. The sequence of anti-proliferative activity induced by the five tested arylnaphthalenes by decreasing strength was HJB > HJA > JB > CME > TEME. HJB, HJA and JB also decreased SOD activity and induced apoptosis in a dose-dependent manner. Activation of caspase-3 further indicated that HJB, HJA and JB induced caspase-dependent intrinsic and/or extrinsic apoptosis pathways. Together, these assays suggest that arylnaphthalene lignans derived from Justicia procumbens induce apoptosis to varying degrees, through a caspase-dependent pathway in human leukemia K562 cells. Furthermore, analysis of structure-activity relationships suggest that hydroxyl substitution at C-1 and C-6' significantly increased the antiproliferative activity of arylnaphthalene lignans while a methoxyl at C-1 significantly decreased the effect

Analysis of the Fungal Community in Ziziphi Spinosae Semen through High-Throughput Sequencing

Ziziphi Spinosae Semen (ZSS) has been widely used in traditional Chinese medicine system for decades. Under proper humidity and temperature, ZSS is easily contaminated by fungi and mycotoxins during harvest, storage, and transport, thereby posing a considerable threat to consumer health. In this study, we first used the Illumina MiSeq PE250 platform and targeted the internal transcribed spacer 2 sequences to investigate the presence of fungi in moldy and normal ZSS samples collected from five producing areas in China. Results showed that all 14 samples tested were contaminated by fungi. Ascomycota was the dominant fungus at the phylum level, accounting for 64.36⁻99.74% of the fungal reads. At the genus level, Aspergillus, Candida, and Wallemia were the most predominant genera, with the relative abundances of 13.52⁻87.87%, 0.42⁻64.56%, and 0.06⁻34.31%, respectively. Meanwhile, 70 fungal taxa were identified at the species level. Among these taxa, three potential mycotoxin-producing fungi, namely, Aspergillus flavus, A. fumigatus, and Penicillium citrinum that account for 0.30⁻36.29%, 0.04⁻7.37%, and 0.01⁻0.80% of the fungal reads, respectively, were detected in all ZSS samples. Moreover, significant differences in fungal communities were observed in the moldy and normal ZSS samples. In conclusion, our results indicated that amplicon sequencing is feasible for the detection and analysis of the fungal community in the ZSS samples. This study used a new approach to survey the fungal contamination in herbal materials. This new approach can provide early warning for mycotoxin contamination in herbal materials, thereby ensuring drug efficacy and safety

6′-Hydroxy Justicidin B Triggers a Critical Imbalance in Ca2+ Homeostasis and Mitochondrion-Dependent Cell Death in Human Leukemia K562 Cells

Justicia procumbens (J. procumbens) is a traditional Chinese herbal medicine which was used for the treatment of fever, pain, and cancer. A compound 6′-hydroxy justicidin B (HJB) isolated from J. procumbens exhibits promising biological properties. However, the mechanism of action and the in vivo behavior of HJB remain to be elucidated. In this study, we investigated the mechanism of action of HJB on human leukemia K562 cells and its pharmacokinetic properties in rats. The results demonstrated that HJB significantly inhibited the proliferation of K562 cells and promoted apoptosis. Besides, HJB resulted in decreased mitochondrial membrane potential deltaPSIm, increased the level of the calcium homeostasis regulator protein TRPC6 and cytosolic calcium. The activity of caspase-8, caspase-9 and the expression of p53 were significantly increased after treatment with HJB. Additionally, HJB has rapid absorption rate and relative long elimination t1/2, indicating a longer residence time in vivo. The results indicate that HJB inhibited the proliferation of K562 cells and induced apoptosis by affecting the function of mitochondria and calcium homeostasis to activate the p53 signaling pathway. The pharmacokinetic study of HJB suggested it is absorbed well and has moderate metabolism in vivo. These results present HJB as a potential novel alternative to standard human leukemia therapies

CircRNAs and lung cancer: Insight into their roles in metastasis

Lung cancer is the leading cause of cancer-related mortality worldwide. A major contributing factor to the poor survival rates in lung cancer is the high prevalence of metastasis at the time of diagnosis. To address this critical issue, it is imperative to investigate the mechanisms underlying lung cancer metastasis. Circular RNAs (circRNAs), a distinct type of ribonucleic acid characterized by their unique circular structure, have been implicated in the progression of various diseases. Recent studies have highlighted the close association between circRNAs and the occurrence and development of lung cancer, particularly in relation to metastasis. In this review, we provide a concise overview of the expression patterns and prognostic significance of circRNAs in lung cancer. Additionally, we summarized the current understanding of the clinical relevance of circRNAs in lung cancer metastasis. Furthermore, we systematically focused on the roles of circRNAs in each step of lung cancer metastasis, reflecting the sequential progression of this process. Notably, circRNAs exhibit dual functionality in lung cancer metastasis, acting both as facilitators and inhibitors of metastatic processes. Given their potential, circRNAs hold promise as novel biomarkers and therapeutic targets for lung cancer metastasis, warranting further investigation

Effects of arylnaphthalene lignans on caspase-3 activity in K562 cells.

<p>Active caspase-3 expression was detected by flow cytometry in K562 cells after treatment with the indicated concentrations of HJA, HJB, JB, CME, TEME, ETO and TAX for 48 h. The results are representative of three independent experiments.</p

Effect of HJB, HJA, JB, CME, TEME, POD and ETO on K562 cell proliferation.

<p>Cells were exposed to the indicated concentrations of arylnaphthalene lignans and incubated for 48± SD of three independent experiments, where each sample was tested in at least triplicate.</p

Effect of arylnaphthalene lignans on apoptosis in K562 cells.

<p>(A) The apoptosis rate and changes in the cell cycle of K562 cells treated with arylnaphthalene lignans at the indicated doses for 24 h were analyzed by PI staining and subsequent flow cytometry. Representative data are shown. (B) Apoptosis was further analyzed by annexin V/PI staining and flow cytometry of K562 cells treated with the indicated concentrations of HJA, HJB, JB, CME and ETO for 48 h. Representative FACS scatter-grams are shown. All results are representative of three independent experiments.</p

Chemical structures of 6′-hydroxy justicidin B (HJB), 6′-hydroxy justicidin A (HJA), justicidin B (JB), chinensinaphthol methyl ether (CME), Taiwanin E methyl ether (TEME), etoposide (ETO), paclitaxel (TAX) and podophyllotoxin (POD).

<p>Chemical structures of 6′-hydroxy justicidin B (HJB), 6′-hydroxy justicidin A (HJA), justicidin B (JB), chinensinaphthol methyl ether (CME), Taiwanin E methyl ether (TEME), etoposide (ETO), paclitaxel (TAX) and podophyllotoxin (POD).</p