Search

Wolfiporia cocos is a saprophytic fungus belonging to the phylum Basidiomycota. The dried sclerotium of this organism has been widely used in traditional Chinese medicine for several thousand years and it is prescribed in many formulations. The W. cocos germplasm resources are complex and diverse, and the molecular mechanisms underlying the growth and development of its sclerotia are unclear. In this study, we used genome resequencing and transcriptome analysis to evaluate the genetic diversity of W. cocos germplasm resources in China and the mechanism of sclerotium growth and development. Phylogenetic and population structure analyses revealed that all the 39 tested strains were divided into three major groups. Most of the strains were clustered into one group, and the remaining strains were clustered into the other two groups. There may be a shared origin of cultivated W. cocos in the main production areas. Transcriptome analysis and quantitative reverse transcription-polymerase chain reaction confirmed that candidate genes related to the yield of W. cocos were mainly enriched in oxidation-reduction and carbohydrate metabolism and highly expressed in the ShenChuan strain, which had the highest comprehensive cultivation score. The findings will be helpful for further understanding the evolution and population structure of W. cocos and determining the functional genes that contribute to the high yield of sclerotia.

In the mesh-connected processors, some processor elements (PEs) become ineffective due to high temperature, overload and other factors, which can affect the stability of the system. This paper deals with the problem of reconfiguring the largest possible subarray from the processor with faults under the row and column rerouting constraint. Firstly, a flexible routing scheme, based on dynamic programming, is proposed to construct the local optimal logical columns. Secondly, we discuss and revise the PEs that cannot be connected between every two logical columns under this scheme. Finally, an efficient algorithm is presented to construct the maximum subarray in polynomial time. The experimental results show that, both on the random and clustered fault scenarios, the proposed algorithm under flexible rerouting scheme is capable of constructing the larger scale logical arrays. On a 48 × 48 host array with 15% fault density, the improvement on the use of fault-free PEs is up to 6.22% for random faults. On a 256 × 256 host array, the improvement can be up to 85.60% for clustered faults. Moreover, the proposed algorithm runs faster than previous algorithms under different size arrays and fault densities, the average improvement in running time is up to 99% compared with state-of-the-art.

Chrysanthemum indicum var. aromaticum is an important resource plant for breeding excellent aromatic varieties of ornamental and medicinal chrysanthemums. However, the biosynthesis and regulation mechanisms of typically aromatic ingredients in C. indicum var. aromaticum are poorly understood, which restricted the utilization of this plant. Here, the transcriptome of C. indicum var. aromaticum was investigated and comparative analysis was conducted with the other two closely related chrysanthemum taxa C. indicum and C. nankingense. The global transcriptome profiles showed clear difference of gene expression patterns between the three chrysanthemums, of which 52 flavonoid biosynthesis-associated genes, 157 transcription factors, and 24 flavonoid Uridine diphosphate (UDP) -glucuronosyl and -glucosyltransferase genes were significantly differentially expressed. Furthermore, 22 differential expression genes (DEGs) in relation to the terpenoid backbone biosynthesis and 183 DEGs related to terpenoid diversification were identified. Of which, 14 genes involved into TS/CYP gene cluster showed higher expression in CIA than those in CI or CN. These results increased our understanding of flavonoid diversification and aromatic ingredients accumulation in plants, and will also help improve resource conservation and sustainable utilization of the important resource plant C. indicum var. aromaticum.

Ampelopsis megalophylla has been found to demonstrate anticancer activities in human cancer cells; however, the effect of total flavone extract (TFE), commonly used in Traditional Chinese Medicine, remains unclear. Furthermore, there is limited information on its effects on breast cancer cell lines. The present study aimed to investigate the inhibitory effects of TFE in different human cancer cell lines. In addition, the underlying mechanisms and the signaling pathways involved were also investigated by determining tumor cell morphological changes, and differences in the cell cycle, apoptosis, mitochondrial transmembrane potential, and related protein expression levels in a breast cancer cell line. It was found that TFE inhibited proliferation in seven cancer cell lines (HeLa, A549, MCF‑7, HepG2, A2780, SW620 and MDA‑MB‑231 and demonstrated a strong inhibitory effect on MCF‑7 cell proliferation. Cell morphological changes were also observed and arrested at the G2/M phase following treatment with TFE at different concentrations. In addition, TFE disrupted the mitochondrial membrane potential and upregulated the expression level of apoptotic proteins, including caspase‑3, ‑8 and ‑9, the Bax/Bcl‑2 ratio, and Apaf‑1 in time‑dependent manner. These results indicated that TFE induced apoptosis of the MCF‑7 cells via a mitochondrial‑mediated apoptotic pathway. In conclusion, TFE is potentially effective in treating breast cancer.

Reconfiguring a high-performance subarray of a VLSI array with faults is to construct a maximum target array with the minimum number of long interconnects, which can reduce communication costs, capacitance and dynamic energy dissipation. An existing work proved that the high performance VLSI subarray can be constructed in polynomial time using network flow algorithm. However, because of the disadvantage of the previous network model and the low-performing of standard network flow algorithms for reconfiguration, the efficiency of these algorithms is poor for constructing the high performance VLSI subarray. In this paper, we present an efficient multiple shortest augmenting paths algorithm for rapidly constructing high performance VLSI array. Firstly, we proposed an efficient data structure to construct the network model of the VLSI array with faults, which can dramatically reduce the size of the model compared with previous algorithm. Secondly, a multiple shortest augmenting path algorithm based on the new data structure is proposed, which can significant reduce the running time. Finally, we conduct solid experiments to highlight the efficiency of the proposed method in terms of the running time compared to the standard network flow algorithms. The experimental results show that on a 64 × 64 host array with 0.1% faults, the size of the network model can be reduced by about 50% and the average improvements in running time is up to 85.10% compared with four standard network flow algorithms.

In this study, we isolated and identified the pathogens by fulfilling Koch's postulates. Disease samples with typical spot blight symptoms were collected and pathogens were isolated from the diseased tissues. The pathogen was identified based on its biological characteristics and molecular analysis of internal transcribed (rDNA-ITS) and large subunit (LSU) sequences. Phylogenetic tree were constructed using MEGA7 software and pathogenicity tests were performed usingBased on Koch's postulates, we identified the pathogen causing spot blight on

Chinese goldthread (Coptis chinensis Franch.), a member of the Ranunculales, represents an important early-diverging eudicot lineage with diverse medicinal applications. Here, we present a high-quality chromosome-scale genome assembly and annotation of C. chinensis. Phylogenetic and comparative genomic analyses reveal the phylogenetic placement of this species and identify a single round of ancient whole-genome duplication (WGD) shared by the Ranunculaceae. We characterize genes involved in the biosynthesis of protoberberine-type alkaloids in C. chinensis. In particular, local genomic tandem duplications contribute to member amplification of a Ranunculales clade-specific gene family of the cytochrome P450 (CYP) 719. The functional versatility of a key CYP719 gene that encodes the (S)-canadine synthase enzyme involved in the berberine biosynthesis pathway may play critical roles in the diversification of other berberine-related alkaloids in C. chinensis. Our study provides insights into the genomic landscape of early-diverging eudicots and provides a valuable model genome for genetic and applied studies of Ranunculales., Coptis chinensis represents an early-diverging eudicot lineage with diverse medicinal applications. Here, the authors report its chromosome-scale genome assembly, infer a single ancient whole-genome duplication, and characterize the function of CYP719 in diversification of protoberberine-type alkaloids.

For a rapid enrichment and separation of minor components from Malus hupehensis, the selection of suitable solvent system is the great challenge for liquid-liquid extraction with a three-phase solvent system and high-speed counter-current chromatography. According to the concept of "like dissolves like," the similarity of the average polarity between solvent system and target compounds was the significant characteristic of liquid-liquid extraction with a three-phase solvent system and high-speed counter-current chromatography separation. The polarity parameter model provides a way to calculate the polarity of unknown compounds. Under the guidance of the polarity, an efficient enrichment and separation approach was established through liquid-liquid extraction and high-speed counter-current chromatography with solvent systems composed of n-hexane-ethyl acetate-acetonitrile-water (5:3:5:7, v/v), n-hexane-ethyl acetate-methanol-water (1:2:1:2, v/v), respectively. Thus, the total content of minor compounds was increased from 2.6% to 17.2%, and two novel compounds (6´´-O-coumaroyl-2´-O-glucopyranosylphloretin and 3´´´-methoxy-6´´-O-feruloy-2´-glucopyranosylphloretin) were obtained. The discovery of the new dihydrochalcones expanded the structural diversity of compounds produced by the genus Malus. The experimental results demonstrated that compound polarity can be described by the polarity parameter model and is an important reference for investigating optimum solvent systems for liquid-liquid extraction with a three-phase solvent system and high-speed counter-current chromatography.

Scolopendra mutilans L. Koch, 1878 is an important Chinese animal with thousands of years of medicinal history. However, the genomic information of this species is limited, which hinders its further application. Here, the complete mitochondrial genome (mitogenome) of S. mutilans was sequenced and assembled by next-generation sequencing. The genome is 15,011 bp in length, consisting of 13 protein-coding genes (PCGs), 14 tRNA genes, and two rRNA genes. Most PCGs start with the ATN initiation codon, and all PCGs have the conventional stop codons TAA and TAG. The S. mutilans mitogenome revealed nine simple sequence repeats (SSRs), and an obviously lower GC content compared with other seven centipede mitogenomes previously sequenced. After analysis of homologous regions between the eight centipede mitogenomes, the S. mutilans mitogenome further showed clear genomic rearrangements. The phylogenetic analysis of eight centipedes using 13 conserved PCG genes was finally performed. The phylogenetic reconstructions showed Scutigeromorpha as a separate group, and Scolopendromorpha in a sister-group relationship with Lithobiomorpha and Geophilomorpha. Collectively, the S. mutilans mitogenome provided new genomic resources, which will improve its medicinal research and applications in the future.

It remains a technical challenge to accurately identify close species of herbal medicines, especially from adulterants, because of their highly identical phenotypes and chemical compositions. Here, we report a direct, sequencing-free, high-curvature nanostructuring-based electrochemical herb sensor (nanoE-herb sensor) to identify herbal species quickly and accurately using ITS2 barcodes. We engineer a nano-roughened carbon-supported gold nanostructuring array by photolithograph-free, one-step electrodeposition. The 3D fractal nanostructures exhibit a high deflection angle that largely enhances DNA hybridization efficiency, particularly for the midcomplementary hybridization, as compared to the 2D planar surface. More importantly, such a trans-scale array biointerface (including macroscale carbon and nanoscale gold branches) can overcome the detection barrier of slow diffusion of a long genomic sequence and inaccessibility of the sequestered variations in ITS2 secondary structures through the out-protruded 3D functional nanostructures. Our nanoE-herb sensor achieves a detection limit of 0.18 fM for the 64-mer fragment of saffron ITS2 barcode with midhybridization and shows superior specificity against even single-base mismatch. The sensor also precisely differentiates saffron from six other adulterants by directly detecting unpurified asymmetric PCR amplicons (∼500 bp) with ITS2 sequences, suggesting its great potential in the field identification of herbal medicinal species and pathogenic bacteria with specific DNA barcodes.

Background/Aims: Increasing evidence showed that miR-1-3p plays a major role in malignant tumor progression. However, the specific biological function of miR-1-3p in bladder cancer is yet unknown. Methods: The expression levels of miR-1-3p in bladder cancer tissues and cell lines were examined by qRT-PCR. Bisulfite sequencing PCR was used for DNA methylation analysis. The target of miR-1-3p was validated by a dual luciferase reporter assay, and the effects of miR-1-3p on phenotypic changes in bladder cancer cells were investigated in vitro and in vivo. Results: The expression of miR-1-3p in bladder cancer cells was downregulated as compared to normal SV-HUC-1 cells. Also, the expression of miR-1-3p was significantly lower in bladder cancer tissues than the corresponding non-cancerous tissues. The methylation status of CpG islands was involved in the regulation of miR-1-3p expression. miR-1-3p inhibited the bladder cancer cell proliferation, migration, and invasion by directly targeting the 3’-UTR of glutaminase. It also exerted an anti-tumor effect by negatively regulating the glutaminase in a xenograft mouse model. Furthermore, GLS depletion resulted in the prolonged expression of γH2AX. Conclusion: Taken together, these results demonstrated that miR-1-3p acts as a tumor suppressor via regulation of glutaminase expression in bladder cancer progression, and miR-1-3p might represent a novel therapeutic target for the treatment of bladder cancer.

In this research paper, a fast, quantitative, analytical model for magnesium oxide (MgO) content in medicinal mineral talcum was explored based on near-infrared (NIR) spectroscopy. MgO content in each sample was determined by ethylenediaminetetraacetic acid (EDTA) titration and taken as reference value of NIR spectroscopy, and then a variety of processing methods of spectra data were compared to establish a good NIR spectroscopy model. To start, 50 batches of talcum samples were categorized into training set and test set using the Kennard–Stone (K-S) algorithm. In a partial least squares regression (PLSR) model, both leave-one-out cross-validation (LOOCV) and training set validation (TSV) were used to screen spectrum preprocessing methods from multiplicative scatter correction (MSC), and finally the standard normal variate transformation (SNV) was chosen as the optimal pretreatment method. The modeling spectrum bands and ranks were optimized using PLSR method, and the characteristic spectrum ranges were determined as 11995–10664, 7991–6661, and 4326–3999 cm−1, with four optimal ranks. In the support vector machine (SVM) model, the radical basis function (RBF) kernel function was used. Moreover, the full spectrum data of samples pretreated with SNV, the characteristic spectrum data screened using synergy interval partial least squares (SiPLS), and the scoring data of the first four ranks obtained by a partial least squares (PLS) dimension reduction of characteristic spectrum were taken as input variables of SVM, and the MgO content reference values of various sample were taken as output values. In addition, the SVM model internal parameters were optimized using the grid optimization method (GRID), particle swarm optimization (PSO), and genetic algorithm (GA) so that the optimal C and g-values were determined and the validation model was established. By comprehensively comparing the validation effects of different models, it can be concluded that the scoring data of the first four ranks obtained by PLS dimension reduction of characteristic spectrum were taken as input variables of SVM, and the PLS-SVM regression model established using GRID was the optimal NIR spectroscopy quantitative model of talc. This PLS-SVM regression model (rank = 4) measured that the MgO content of talcum was in the range of 17.42–33.22%, with root mean square error of cross validation (RMSECV) of 2.2127%, root mean square error of calibration (RMSEC) of 0.6057%, and root mean square error of prediction (RMSEP) of 1.2901%. This model showed high accuracy and strong prediction capacity, which can be used for rapid prediction of MgO content in talcum.

The leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) is highly expressed in many human malignancies and is correlated with poor prognosis. However, the function of LETM1 in bladder cancer still remains unknown. In the present study, we analyzed the expression levels of LETM1 in bladder cancer tissues and non-cancerous tissues as well as in four bladder cancer cell lines (T24, EJ, 5637 and J82) and a human bladder epithelial immortalized cell line SV-HUC-1. Small interfering RNA (siRNA) was employed to knockdown the expression of LETM1 in the T24 cells. The proliferation of T24 cells was significantly repressed as evaluated by CCK-8 assays. Transwell migration and invasion assays indicated that knockdown of LETM1 suppressed cell migration and invasion significantly. Flow cytometric analysis revealed that cells had accumulated at the S-phase when the expression of LETM1 was suppressed. Moreover, we found that several oncogenic proteins in the Wnt/β-catenin signaling pathway, namely β-catenin, cyclin D1 and c-Myc were significantly decreased by the LETM1 siRNA. Collectively, these results revealed that the knockdown of LETM1 exhibited tumor suppressive effects, possibly by controlling the downstream Wnt/β-catenin signaling pathway.

// Guanghui Hu 1 * , Binbin Dong 2, * , Jingwei Zhang 1, * , Wei Zhai 1, 3 , Tiancheng Xie 1 , Bisheng Huang 1 , Chi Huang 4 , Xudong Yao 1 , Junhua Zheng 1 , Jianping Che 1, 4 and Yun-Fei Xu 1 1 Department of Urology, Shanghai Tenth People’s Hospital, Tongji University, Shanghai, China 2 Department of Urology, Yancheng Third People’s Hospital, Yancheng, Jiangsu, China 3 Department of Urology, Renji Hospital, Shanghai Jiaotong University, Shanghai, China 4 Department of Urology, Shanghai Tenth People’s Hospital, Nanjing Medical University, Nanjing, Jiangsu, China * These authors have contributed equally to this work Correspondence to: Yun-Fei Xu, email: xuyunfeibb@sina.com Jianping Che, email: chejianping77@126.com Wei Zhai, email: jacky_zw2002@hotmail.com Keywords: renal cell carcinoma, HOTAIR, hippo pathway, SAV1 Received: December 31, 2016 Accepted: March 14, 2017 Published: April 25, 2017 ABSTRACT The long noncoding RNA HOTAIR promotes the development and progression of several tumors. Here, the clinical significance and role of HOTAIR in renal cell carcinoma (RCC) tumorigenesis were explored. The results showed that increased expression of HOTAIR predicted a poor prognosis of RCC after surgery. HOTAIR promoted RCC cell proliferation and growth in vitro and in vivo . The expressions of HOTAIR and Salvador homolog 1 (SAV1) were inversely correlated in clinical RCC samples. HOTAIR downregulated SAV1 by directly binding to the SAV1 protein and enhanced histone H3K27 methylation. Loss of function of SAV1 activated the Hippo pathway. HOTAIR could be a potential therapeutic target in RCC.

Scolopendra mutilans L. Koch, 1878 is an important Chinese animal with thousands of years of medicinal history. However, the genomic information of this species is limited, which hinders its further application. Here, the complete mitochondrial genome (mitogenome) of S. mutilans was sequenced and assembled by next-generation sequencing. The genome is 15,011 bp in length, consisting of 13 protein-coding genes (PCGs), 14 tRNA genes, and two rRNA genes. Most PCGs start with the ATN initiation codon, and all PCGs have the conventional stop codons TAA and TAG. The S. mutilans mitogenome revealed nine simple sequence repeats (SSRs), and an obviously lower GC content compared with other seven centipede mitogenomes previously sequenced. After analysis of homologous regions between the eight centipede mitogenomes, the S. mutilans mitogenome further showed clear genomic rearrangements. The phylogenetic analysis of eight centipedes using 13 conserved PCG genes was finally performed. The phylogenetic reconstructions showed Scutigeromorpha as a separate group, and Scolopendromorpha in a sister-group relationship with Lithobiomorpha and Geophilomorpha. Collectively, the S. mutilans mitogenome provided new genomic resources, which will improve its medicinal research and applications in the future.

Background It is an established fact that excess of glucocorticoids could cause the harmful effects, such as suppression on the male reproduction. Although glucocorticoids pharmacologically inhibit the Leydig cell function, their roles in Leydig cell development are unclear. Therefore, the present study was designed to investigate effects of synthetic glucocorticoid dexamethasone (DEX) on rat stem Leydig cell proliferation and differentiation. Methods Male Sprague-Dawley rats received a single intraperitoneal injection of 75 mg/kg EDS to eliminate Leydig cells and an in vitro culture system of the seminiferous tubules was established from Leydig cell-depleted testis. Using basal medium and Leydig cell differentiation-inducing medium (LIM) in the culture system, we examined the effects of DEX (0–100 nM) on the proliferation and differentiation of the stem Leydig cells in vitro, respectively. Results Results showed that LIM is a good agent to induce stem Leydig cell differentiation into Leydig cells that produce testosterone in vitro. DEX inhibited the differentiation of stem Leydig cells by reducing the expression levels of Cyp17a1 and Scarb1 and that NR3C1 antagonist RU38486 reversed the DEX-mediated effects. However, DEX are not involved with the proliferation of stem Leydig cells. Conclusions DEX suppressed the differentiation of rat Leydig cells in vitro and glucocorticoid-induced effects acted through NR3C1. This suppression partially targets on Cyp17a1 and Scarb1 gene expression.

The development of testicular inflammation affects the normal male reproductive function. The proteasome activator complex subunit 3 (REGγ) has been suggested to regulate experimental colitis. However, to the best of our knowledge, a potential association between REGγ and testicular inflammation has not been demonstrated. The present study successfully established inflammatory models in C57 mice, primary Leydig cells and the TM3 cell line. It was observed that the absence of REGγ conveyed a significantly protective effect toward testosterone secretion in Leydig cells. REGγ deficiency significantly decreased the expression levels of phosphorylated transcription factor p65 and inflammatory factors in testis tissues, primary Leydig cells and the TM3 cell line. Inflammation also upregulated the expression levels of REGγ. Furthermore, the degradation of the nuclear factor light‑chain‑enhancer of activated B cells (NF‑κB) inhibitor ε (IkBε) signaling pathway regulated REGγ and NF‑κB expression. Double knockdown of REGγ and IkBε restored the response in wild‑type cells to LPS‑induced inflammation. In summary, these results demonstrated that REGγ regulates NF‑κB activity by specifically degrading IkBε to regulate inflammation in testicular Leydig cells.

Mineral traditional Chinese medicines (TCMs) are natural minerals, mineral processing products, and some fossils of animals or animal bones that can be used as medicines. Mineral TCMs are a characteristic part of TCMs and play a unique role in the development of TCMs. Mineral TCMs are usually identified according to their morphological properties such as shape, color, or smell, but it is difficult to separate TCMs that are similar in appearance or smell. In this study, the feasibility of using Raman spectroscopy combined with support vector machine (SVM) for rapid identification of nine easily confused mineral TCMs, i.e., borax, gypsum fibrosum, natrii sulfas exsiccatus, natrii sulfas, alumen, sal ammoniac, quartz, calcite, and yellow croaker otolith, was investigated. Initially, two methods, characteristic intensity data extraction and principal component analysis (PCA), were performed to reduce the dimensionality of spectral data. The identification model was subsequently built by the SVM algorithm. The 3-fold cross validation (3-CV) accuracy of the SVM model established based on extracting characteristic intensity data from spectra pretreated by first derivation was 98.61%, and the prediction accuracies of the training set and validation set were 100%. As for the PCA-SVM model, when the spectra pretreated by vector normalization and the number of principal components (NPC) is 7, the 3-CV accuracy and prediction accuracies all reached 100%. Both models have good performance and strong prediction capacity. These results demonstrate that Raman spectroscopy combined with a powerful SVM algorithm has great potential for providing an effective and accurate identification method for mineral TCMs.

A new dimeric naphtho-γ-pyrone, asperpyrone F (1), along with six known ones, asperpyrones B (2) and C (3), fonsecinones A (4) and B (5), aurasperones A (6) and E (7), have been isolated from the solid culture of the edible fungus Pleurotus ostreatus. The structures of 1–7 were determined mainly by NMR and MS experiments. The absolute configuration of compound 1 was assigned via the circular dichroism (CD) data analysis. Compounds 1–7 showed modest antioxidant and immunomodulatory activities. All compounds were isolated from the fungus P. ostreatus for the first time.

In the present experiment, a green and highly efficient extraction method for flavonoids established on deep eutectic solvents (DESs) was investigated by using the response surface methodology. The DES-based high-speed countercurrent chromatography (HSCCC) solvent systems were developed for the separation of high purity compounds from the DES extract of Malus hupehensis for the first time. Under the optimal conditions (liquid-to-solid ratio of 26.3 mL/g, water content of 25.5%, and extraction temperature of 77.5°C), the yield of flavonoids was 15.3 ± 0.1%, which was superior to that of the methanol extraction method. In accordance with the physical property of DES-based HSCCC solvent systems and K values of target compounds, DES-based HSCCC solvent systems composed of choline chloride/glucose-water-ethyl acetate (ChCl/Glu-H2O-EAC, 1:1:2, v/v) was selected for the HSCCC separation. Thus, five flavonoids (two novel compounds 1-2, 6´´-O-coumaroyl-2´-O-glucopyranosylphloretin and 3´´´-methoxy-6´´-O-feruloy-2´-O-glucopyranosylphloretin; three know compounds 3-5, namely, avicularin, phloridzin, and sieboldin) were efficiently separated from Malus hupehensis. DESs are the environment friendly and highly efficient solvents as the components of extraction solvent and HSCCC solvent system, and can be re-utilized many times. However, ethyl acetate can be soluble with a few hydrogen bond donors, such as urea, carboxylic acid and polyol, through the shake flask test. It is the great difficulty for the efficient and rapid separation of target compounds from the DESs extract because of the DESs residual in the HSCCC fractions. ChCl and Glu are the great choices of DESs without this problem. In addition, K values increased with the increase of the molar ratio of ChCl/Glu and the content of water, which could effectively guide us to choose the suitable DES-based HSCCC solvent system. The twice HSCCC separation results indicated that DES was the valuable and green solvent for the HSCCC separation of pure compounds from the extract for the first time, and showed the recycle superiority of DES-based HSCCC solvent system.

For a successful high-speed countercurrent chromatography (HSCCC) separation of multiple components, the suitable solvent system selection is a critical operation. Despite the difference in separation mechanism between HSCCC and HPLC, K values of compounds in the solvent system and the retention factor of compounds in HPLC were connected with polarity, and the polarity mainly depended on the structure and chemical properties of compounds. On the basis of the concept of “like dissolves like”, the average polarity of the solvent system is equal to the average polarity of multiple components at the “sweet point” log K = 0. The result of theoretical deduction showed a negative linear correlation between the polarity of each component and the logarithm of the gradient range of the organic phase. Therefore, the average polarity of unknown multiple components could be obtained through the polarity parameter model established in the HPLC analysis. The suitable solvent system composed of n-hexane–ethyl acetate–methanol–water (8:3:8:1, v/v) was selected through the average polarity of multi-components combining K values of the enriched samples. In the context of bioassay, an efficient HSCCC separation procedure was established, and two groups of analogue benzaldehyde derivatives (four main antioxidants, namely, isodihydroauroglaucin, isoaspergin, isotetrahydro-auroglaucin, and flavoglaucin; two minor antioxidants, namely, 6′-oxo-chaetopyranin and chaetopyranin) were obtained from Hypsizygus marmoreus. The predicted polarity values of multi-components were sufficient to meet the HSCCC experimental requirements. The HPLC analysis of reference compounds and multi-components showed a significant consistency with different chromatographic columns. Therefore, the polarity parameter model established in the HPLC analysis was a simple, rapid, and helpful tool for looking an appropriate solvent system, which was a forwarding step for the HSCCC separation of multiple components.

A method to optimize a high-speed countercurrent chromatography solvent system from many possible solvent systems was guided by average polarity and K values of crude samples, which was successfully applied in separation of the naphtho-γ-pyrones. This method expanded the scope of spectrophotometer application in the solvent systems selection, and showed the merits of simplicity, rapidity, and low reagent consumption. As a consequence of utilizing the method, an efficient separation procedure was established, and two groups of naphtho-γ-pyrone isomers (fonsecinone B and asperpyrone F; asperpyrone C, fonsecinone A, and aurasperone A) were obtained from Pleurotus ostreatus by high-speed countercurrent chromatography with a solvent system composed of petroleum ether/ethyl acetate/methanol/water (5:5:5:5, v/v). The results indicate that high-speed countercurrent chromatography could be a powerful technology for separation isomers and analogues, and the method guided by average polarity and K values of crude samples is feasible to optimize solvent system.

Herbal material is both a medicine and a commodity. Accurate identification of herbal materials is necessary to ensure the safety and effectiveness of medication. With this work, we initiated an identification method to investigate the species authenticity for herbal products of Celastrus orbiculatus and Tripterygum wilfordii utilizing DNA barcoding technology. An ITS2 (internal transcribed spacer two) barcode database including 59 sequences was successfully established to estimate the reliability of species-level identification for Celastrus and Tripterygium. Our findings showed that ITS2 can effectively and clearly distinguish C. orbiculatus, T. wilfordii and its congeners. Then, we investigated the proportions and varieties of adulterant species in the herbal markets. The data from ITS2 region indicated that 13 (62%) of the 21 samples labeled as “Nan-she-teng” and eight (31%) of the 26 samples labeled as “Lei-gong-teng” were authentic; the remaining were adulterants. Of the 47 herbal products, approximately 55% of the product identity were not in accordance with the label. In summary, we support the efficacy of the ITS2 barcode for the traceability of C. orbiculatus and T. wilfordii, and the present study provides one method and reference for the identification of the herbal materials and adulterants in the medicinal markets.

The present study investigated the possible tumor-suppressing function of microRNA (miR)-612 and the underlying molecular mechanism of its action in bladder cancer in vitro and in vivo. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was carried out to quantify the expression levels of miR‑612 in bladder cancer tissues and cell lines. The data demonstrated that the level of miR‑612 expression was significantly reduced in bladder cancer tissues and cell lines, as compared with that in non‑cancerous tissues and cells. Reduced miR‑612 expression was associated with advanced tumor, lymph node and metastasis stages, and with distant metastasis of bladder cancer. A functional study revealed that transfection of cells with an miR‑612 mimic suppressed bladder cancer cell growth, colony formation, migration, invasion and epithelial-mesenchymal transition. Bioinformatics analysis identified that miR‑612 targeted the expression of malic enzyme 1 (ME1), and this was confirmed by western blot and luciferase reporter assay results. Furthermore, the ME1 expression levels were inversely associated with miR‑612 expression in bladder cancer tissue specimens. In addition, knockdown of ME1 expression using ME1 siRNA mimicked the effect of ectopic miR‑612 overexpression in bladder cancer cells in terms of tumor cell growth, migration and invasion. By contrast, ME1 overexpression weakened the inhibitory effect of the miR‑612 mimic in bladder cancer cells. In conclusion, the present study demonstrated that miR‑612 may function as a tumor suppressor in bladder cancer by targeting ME1 expression.

Aldosterone (ALDO) is a primary endogenous mineralocorticoid, appearing as the main hormone controlling sodium and water homeostasis. Its emerging role in the development of many organs has gained interest over the past few years. In the testis, Leydig cells contain mineralocorticoid receptors and ALDO stimulates androgen synthesis via the mineralocorticoid receptors in rat adult Leydig cells. Although ALDO pharmacologically promoted the Leydig cell function, its role in Leydig cell development was unclear. In the present study, we investigated effects of ALDO on rat stem Leydig cell (SLC) proliferation and differentiation. Using an in vitro culture system of the seminiferous tubules from Leydig cell-depleted testis and EdU incorporation into the SLC to judge its DNA synthesis and measurement of medium testosterone production, steroidogenesis-related gene and protein expression, we found that: 1) ALDO suppressed EdU incorporation into SLCs at 100 nM via mineralocorticoid receptor-mediated mechanism; 2) ALDO mainly down-regulated Lhcgr and Scarb1 expression possibly via reducing Leydig cell number. In conclusion, ALDO pharmacologically blocked rat SLC development.

Transurethral resection of bladder tumor (TURBT) using a wire loop is considered the gold standard for staging and treating non-muscle invasive bladder cancer (NMIBC). TURBT is associated with serious disadvantages that facilitate tumor recurrence. The present study evaluated the safety and efficacy of the bipolar button electrode for en bloc resection of NMIBC. From January 2013 to July 2016, 82 consecutive patients newly diagnosed with NMIBC received transurethral en bloc resection with bipolar button electrode. Operative details, pathological result, and intraoperative and postoperative complications regarded as safety outcomes were documented. Each patient was followed up for ≥ 18 months. A total of 118 neoplasms were removed en bloc from 82 patients. The mean tumor diameter was 2.42 ± 1.34 cm. The average operation time was 35 ± 14 min. No complications such as bladder bleeding, vesicle perforation, and obturator nerve reflex occurred during the treatment. Pathological evaluations showed urothelial carcinoma with stage Ta low grade in 26 patients, T1 high grade in 51 patients, and T2 high grade in 5 patients. In addition, the bladder detrusor muscle layer was provided in all cases. The 18-month recurrence-free survival was 88.5% (23/26) and 74.5% (38/51) for Ta and T1 patients, respectively. The current results demonstrated that transurethral en bloc resection with bipolar button electrode is an effective, feasible, and safe treatment for NMIBC.

The leucine zipper-EF-hand containing transmembrane protein 1 (LETM1) has been reported to serve an important role in a number of human malignancies and is correlated with poor prognosis. However, little is known about the role of LETM1 in renal cell carcinoma (RCC). In the present study, the expression levels of LETM1 were investigated in RCC cell lines (Caki-1, 786-O, OS-RC-2, A498 and ACHN) and the HK-2 normal human renal tubular epithelial cell line. Short interfering RNA (siRNA) was used to knock down the expression of LETM1 in 786-O and A498 cells. The results indicated that the constitutive expression of LETM1 was notably upregulated in RCC cell lines. Knockdown of LETM1 significantly decreased cell proliferation, migration and invasion. Mechanistically, it was revealed that the knockdown of LETM1 expression sharply downregulated the protein expression of β-Catenin, Cyclin D1 and c-Myc in 786-O and A498 cells. In conclusion, these results suggest that knockdown of LETM1 exhibits tumor suppressive effects, at least in part by controlling the downstream Wnt/β-Catenin signaling pathway. Therefore, LETM1 may act as a novel therapeutic target for the treatment of RCC.