New clerodane diterpenes from Tinospora sagittata var. yunnanensis.

Four new clerodane diterpenes, namely sagittatayunnanosides A-D (1-4), were isolated from the roots of Tinospora sagittata var. yunnanensis, together with two known compounds, tinospinoside C (5) and tinospinoside E (6). The structures of the four new compounds were well elucidated by extensive analyses of the MS, IR, and 1D and 2D NMR data. The cytotoxic and antifouling activities of compounds 1-6 were evaluated.

Effect of Traditional Chinese Non-Pharmacological Therapies on Knee Osteoarthritis: A Narrative Review of Clinical Application and Mechanism.

Knee osteoarthritis (KOA) stands as a degenerative ailment with a substantial and escalating prevalence. The practice of traditional Chinese non-pharmacological therapy has become a prevalent complementary and adjunctive approach. A mounting body of evidence suggests its efficacy in addressing KOA. Recent investigations have delved into its underlying mechanism, yielding some headway. Consequently, this comprehensive analysis seeks to encapsulate the clinical application and molecular mechanism of traditional Chinese non-pharmacological therapy in KOA treatment. The review reveals that various therapies, such as acupuncture, electroacupuncture, warm needle acupuncture, tuina, and acupotomy, primarily target localized knee components like cartilage, subchondral bone, and synovium. Moreover, their impact extends to the central nervous system and intestinal flora. More perfect experimental design and more comprehensive research remain a promising avenue in the future.

Cytotoxic flavanes from Uraria clarkei.

Two new flavanes, (2R)-4'-hydroxy-2',5,7-trimethoxyflavane (1) and (2R,4R)-2',4'-dihydroxy-5,7-dimethoxyflavan-4-ol (2), were isolated from Uraria clarkei, together with two known compounds 5,7-dimethoxy-4'-hydroxyflavan (3) and 5,7,4'-trimethoxyflavan (4). The structures of the new flavanes were characterized by analyses of the MS, IR, UV, CD, 1D, and 2D NMR data. Cytotoxicity test suggested that compounds 1-4 possessed slight activity against K-562 and Hela cell lines, with the IC50 values ranging from 26.6 to 56.3 µM.

Two new sphingolipids from the leaves of Piper betle L.

Two new sphingolipids, pipercerebrosides A (1) and B (2), were isolated from the leaves of Piper betle L. Their structures, including absolute configurations, were determined by spectroscopic analysis and chemical degradation. These two compounds did not show significant cytotoxic activity against the cancer cell lines K562 and HL-60 in a MTT assay.

[Acting mechanisms and research methods of long noncoding RNAs].

Long non-coding RNAs (lncRNAs) play biological roles through a variety of mechanisms, including genetic imprinting, chromatin remodeling, cell cycle control, splicing regulation, mRNA decay, and translational regulation. LncRNAs are involved in the regulation of gene expression through the above mechanisms in different levels. Establishment and application of research technologies are important in understanding of lncRNAs functions. Microarray, RNA sequencing, Northern blot, real time quantitative reverse transcription-polymerase chain reaction, fluorescence in situ hybridization, RNA interference, and RNA-binding protein immunoprecipitation are major tools of exploring biological functions of lncRNAs. Here, we highlighted three advanced methods, i.e., fast predictions of RNA and protein interactions and domains (catRAPID), chromatin isolation by RNA purification (ChIRP), and combined knockdown and localization analysis of non-coding RNAs (c-KLAN).

A novel antioxidant isobenzofuranone derivative from fungus Cephalosporium sp.AL031.

Bioassay-guided fractionation of metabolites from the fungus Cephalosporium sp.AL031 isolated from Sinarundinaria nitida led to the discovery of a new isobenzofuranone derivative, 4,6-dihydroxy-5-methoxy-7-methylphthalide (1), together with three known compounds: 4,5,6-trihydroxy-7-methyl-1,3-dihydroisobenzofuran (2), 4,6-dihydroxy-5-methoxy-7-methyl-1,3-dihydroisobenzofuran (3) and 4,5,6-trihydroxy-7-methylphthalide (4). The structure of the new compound 1 was determined based on MS, 1D and 2D NMR spectral data. Compounds 1-4 showed potent antioxidant activity with EC50 values of 10, 7, 22 and 5 µM by 1,1-diphenyl-2-picryhydrazyl (DPPH) radical-scavenging assay.

[Mercury in tree rings: Advances, problems and prospects]. / æ ‘æœ¨å¹´è½®æ±žè®°å½•: 进展、问题和展望.

As a highly biotoxic element, mercury (Hg) can be enriched by the food chain and has negative effect on ecosystems. Changes of Hg flux and reserves in forest have important effects on its biogeochemical cycle in forest ecosystem. Due to limitation of temporal and spatial monitoring, there is not comprehensive understanding on Hg distribution. Widely distributed trees can be used as effective bio-monitors and Hg records in tree rings can be used to study Hg temporal and spatial distribution. Hg accumulated by root, leaf, bark, and other tissues can be detained in bole and record environmental Hg variations. Therefore, historical Hg trends can be restructured by analyzing Hg concentration in tree rings and the biogeochemical characteristics can be understood with Hg isotope ratio. We reviewed the method of measurement of Hg concentration and isotope ratio and application of reconstruction using Hg concentration in tree ring. We suggested the great application potential of Hg isotope ratio in atmospheric Hg construction and biogeochemistry cycle and raised concerns in further studies.

Circular RNAs in liver diseases: Mechanisms and therapeutic targets.

Circular RNAs (circRNAs) are formed from the genome through diverse back splicing and feature the closed loop. circRNAs are widely available in a variety of cells and characterized by conservation, structural stability, high abundance and tissue-specific or developmental-specific expression. Recent studies have shown that circRNAs are closely related to liver diseases, such as metabolic-associated fatty liver disease, hepatitis, liver cirrhosis and hepatocellular carcinoma. circRNAs play an important role in the progression of liver diseases, are potential diagnostic and prognostic markers, and have translational value in therapy. This article reviews the research on circRNAs in liver diseases, with a view to providing a theoretical basis and new ideas for future research and treatment of liver diseases.

A novel 18-norclerodane diterpenoid from the roots of Tinospora sagittata var. yunnanensis.

A novel 18-nor-clerodane diterpenoid named sagitone (1) was isolated from the 95% ethanol extract of dry roots of Tinospora sagittata var. yunnanensis together with the five known diterpenoids columbin (2), palmatoside C (3), fibleucin (4), tinophylloloside (5) and epitinophylloloside (6). The structure of the new compound 1 was determined based on MS, IR, 1D and 2D NMR spectral data. The compounds 1-6 did not show significant cytotoxic activity against cancer cell lines K562 and HL-60.

[Determination of seven metal elements in Yunnan wild Schizophyllum commune Fr by atomic absorption spectrometry].

The seven metal elements in Yunnan wild schizophyllum commune Fr, including Zn, Co, Ni, Cu, Fe, Cr and Mg, were determined by atomic absorption spectrometry for providing a scientific basis of the development. The results show that Zn, Co, Ni, Fe, Cr and Mg are relatively rich in schizophyllum commune Fr and the content of Cu is lower. The ratio of the content of Zn to that of Cu is 7.4, which is consistent with the relative content level of Zn and Cu contained in many anticancer Chinese traditional plants, implying that the nutritive value of Yunnan wild schizophyllum commune Fr is high.

[Studies on the chemical constituents from the roots of Tinospora sagittata var. yunnanensis (II)].

OBJECTIVE: To study the chemical constituents from the roots of Tinospora sagittata var. yunnanensis. METHODS: Various chromatographic techniques were used to isolate and purify the constituents. The structures of these compounds were elucidated on the basis of spectral analysis. RESULTS: Seven compounds were isolated from the plant and their structures were identified as tinophylloloside (1), epitinophylloloside (2), 2-deoxycrustecdysone (3), polypodine B (4) , (+)-5'-methoxyisolariciresinol 3alpha-O-beta-D-glucopyranoside (5), geniposide (6), adenine (7). CONCLUSION: All compounds are isolated from the plant for the first time,and compounds 4, 5, 6 and 7 are isolated firstly from the genus.

[Mercury Transport from Glacier to Runoff in Typical Inland Glacial Area in the Tibetan Plateau].

To investigate the transport of mercury from glacier to runoff in typical inland glacial area in the Tibetan Plateau, we selected Zhadang glacier and Qugaqie river Basin located in the Nyainqentanglha Range region and collected samples from snow pit, glacier melt-water and Qugaqie river water during 15th August to 9'h September 2011. Mercury speciation and concentrations were determined and their distribution and controlling factors in different environmental compartments were analyzed. The results showed that the average THg concentrations were (3.79 +/- 5.12) ng x L(-1), (1.06 +/- 0.77) ng x L(-1) and (1.02 +/- 0.24) ng x L(-1) for glacier snow, glacier melt-water and Qugaqie river water, respectively, all of which were at the global background levels. Particulate-bound mercury accounted for large proportion of mercury in all environmental matrices, while mercury in glacial melt-water was controlled by total suspended particle, and mercury in Qugaqie river water co-varied with runoff. With the increase of temperature, glacier melted and released water as well as mercury into glacier-fed river. Total mercury concentrations in glacier melt water, upstream and downstream peaked at 14:00, 16:00 and after 20:00, respectively, reflecting the process of mercury release from glacier and its subsequent transport in the glacier fed river. The transport of riverine mercury was controlled by multiple factors. Under the context of climate change, glacier ablation and the increasing runoff will play increasingly important roles in mercury release and transport.

Detecting AFP mRNA in peripheral blood of the patients with hepatocellular carcinoma, liver cirrhosis and hepatitis.

BACKGROUND: The low frequency of disseminated carcinoma cells in the blood now makes immunomagnetic bead sorting and reverse transcriptase-polymerase chain reaction (RT-PCR) technique more popular. METHODS: Three milliliters of peripheral blood were collected from 91 patients and 18 normal donors. The circulating carcinoma cells were enriched with CD45 and Ber-EP4 immunomagnetic beads. The alpha-fetoprotein (AFP) mRNA was amplified with nested RT-PCR. RESULTS: The total positive detection rate was 72.1%, 43.8%, 25.0%, 100%, and 66.7% in patients with hepatocellular carcinoma (HCC) untreated, liver cirrhosis (LC), hepatitis, metastasis liver cancer, and postsurgery of hepatocellular carcinoma, respectively. There was a significant difference among the patients with HCC, LC and hepatitis (HCC vs. LC, P<0.05; HCC vs. hepatitis, P<0.01) and between Class A and B of the HCC patients (P<0.05). Meanwhile, AFP mRNA was markedly expressed in HCC patients compared to the patients with no HCC (LC and hepatitis). The levels of aspartate transaminase (AST) and gamma-glutamyltranspeptidase (GGT) were significantly different in AFP mRNA-positive patients with autoimmune chronic active hepatitis B (CAHB) or LC in contrast to the corresponding negative patients. CONCLUSION: Combining negative and positive immunomagnetic bead sorting and RT-PCR technique can effectively detect circulating tumor cells. AFP mRNA is a more reliable marker of metastasis compared to serum AFP.

Survivin antisense compound inhibits proliferation and promotes apoptosis in liver cancer cells.

AIM: To evaluate the effects of survivin on cell proliferation and apoptosis in liver cancer. METHODS: MTT assay was used to generate and optimize phosphorothioate antisense oligonucleotides (ODNs)-Lipofectamine2000 (LiP) compound by varying ODNs (mug):LiP (muL) ratios from 1:0.5 to 1:5. Then, liver cancer cells (HepG2) were transfected with the compound. By using RT-PCR and Western blot, the expression levels of survivin mRNA and proteins were detected in HepG2 cells treated with antisense compounds (ODNs:LiP = 1:4), and compared with those treated with sense compounds (1:4) as control. MTT assay was applied to the determination of cell proliferation in HepG2 cells. Active caspase-3 was evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscopy. Laser scanning confocal microscopy was performed to detect the subcellular localization of survivin proteins in treated and untreated cells. RESULTS: Antisense compounds (1:4) down-regulated survivin expression (mRNA and protein) in a dose-dependent manner with an IC50 of 250 nmol/L. Its maximum effect was achieved at a concentration of 500 nmol/L, at which mRNA and protein levels were down-regulated by 80%. The similar results were found in MTT assay. Antisense compound (1:4)-treated cells revealed increased caspase-3-like protease activity compared with untreated cells. Untreated cells as control were primarily negative for the presence of active-caspase-3. As shown by transmission electron microscopy, treated cells with antisense compounds (1:4) resulted in morphological changes such as blebbing and loss of microvilli, vacuolization in the cytoplasm, condensation of the cytoplasm and nuclei, and fragmented chromatin. Immunofluorescence analysis confirmed the presence of survivin protein pool inside the cytoplasm in untreated cells. Labeled-FITC immunofluorescence staining of survivin clearly showed that survivin was distributed mainly in a spotted form inside the cytoplasm. Whereas cells treated with antisense compounds were rare and weak inside the cytoplasm. CONCLUSION: Down-regulation of survivin expression induced by the antisense compounds reduces tumor growth potential, promotes apoptosis and affects the localization of survivin proteins in HepG2 cells. Furthermore, survivin protein is a key molecule associated with proliferation and apoptosis, and antisense oligonucleotides targeting survivin have a bright prospect in the therapy of liver cancer.

[Gene studies and nobel prize].

Gene is a DNA sequence which can be expressed and produces gene products (protein or RNA). By 2003, there are 51 Nobel Prize owners related to gene studies. Among them, 44 persons are in physiology or medicine (account for 24.72% of total 178), 7 persons are in chemistry (account for 5.69% of total 123). The paper reviews them in following 6 aspects: Drosophlie melanogaster is a good material for gene study; the double helix model of DNA structure provides a hard foundation in gene study; the studies on gene regulation illuminate many functions of gene; genetic central dogma researches created 11 Noble Prize laureates; gene engineering technologies make possible to modify and use genes; and the thorough studies of gene characteristic made us easier to understand many life phenomena.

Effects of daidzein on estrogen-receptor-positive and negative pancreatic cancer cells in vitro.

AIM: To study the effects of daidzein on human pancreatic cancer cells in vitro. METHODS: Human estrogen-receptor (ER)-positive pancreatic cancer cells MiaPaCa-2 and ER-negative pancreatic cancer cells PANC-1 were treated by 0.1 micromol/L, 1 micromol/L, 10 microL, 25 microL, 50 microL, 75 microL and 100 microL of daidzein, respectively. Its antiproliferative effect was studied by MTT assay. RESULTS: Daidzein inhibited the growth of MiaPaCa-2 and PANC-1 cells at the concentrations from 0.1 microL to 100 microL. A dose- and time-dependent manner was found. The IC(50) of daidzein on MiaPaCa-2 and PANC-1 cells was 45 microL and 75 micro, respectively. After MiaPaCa-2 cells were treated by daidzein for 3 d and at the concentrations more than IC(50), the inhibitory manner was identical and the inhibition appeared a saturation phenomenon, but the inhibitory manner of daidzein on PANC-1 cells was different from that of MiaPaCa-2 cells. CONCLUSION: Daidzein has antiproliferative effects on human estrogen-receptor-positive and negative pancreatic cancer cells, but their mechanisms may be different.

[Effect of survivin targeting on cell proliferation and apoptosis in pancreatic cancer cells].

OBJECTIVE: To investigate the effects of antisense survivin-Lip compound on pancreatic cancer cell proliferation and apoptosis and its mechanism. METHODS: Pancreatic cancer cells of the line PANC-1 were cultured. Survivin oligonucleotide (ODN) was transfected into the PANC-1 cells mediated by Lip reagent. The expression of survivin mRNA and that of surviving protein were detected by RT-PCR and Western blotting. MTT assay was applied to determine the proliferation of PANC-1 cells. Active caspase-3 and apoptosis rate were evaluated by flow cytometric analysis. The morphological changes were assessed by electron microscope. Lascar scanning confocal microscope immunofluorescence analysis was performed to detect the subcellular localization of survivin protein on treated cells and untreated cells. RESULTS: Antisense compound efficiently down-regulated the survivin expression (mRNA and protein) in dose-dependent manner with an IC50 of 300 nmol/L. Its maximum effect was achieved at the concentration of 500 nM, at which the expression level was down-regulated by 80%. The similar results were found in MTT assay. As revealed by gradually increased caspase-3-like protease activity and apoptosis rate in a time-dependent manner, and the morphological changes of apoptosis such as blebbing and loss of microvilli, vacualization in cytoplasm, condensation of cytoplasm and nucleus, and fragmented chromatin, treatment with antisense compound induced apoptosis and inhibited cell growth. Fluororescein isothiocyanate (FITC)-labeled immunofluorescence staining of survivin clearly showed that survivin was expressed mainly in the formation of a spotted distribution inside the cytoplasm of untreated cells. Survivin protein molecules were clearly seen in the cytoplasm of the untransfected cells and distributed like spots and almost disappeared in the transfected cells with morphological changes conforming to the changes of apoptosis. CONCLUSION: Survivin protein is a key molecular connecting proliferation with apoptosis and antisense oligonucleotides targeting survivin have a bright prospect in therapy of pancreatic cancer cells.

Non-coding RNAs and gastric cancer.

Non-coding RNAs (ncRNAs) play key roles in development, proliferation, differentiation and apoptosis. Altered ncRNA expression is associated with gastric cancer occurrence, invasion, and metastasis. Moreover, aberrant expression of microRNAs (miRNAs) is significantly related to gastric cancer tumor stage, size, differentiation and metastasis. MiRNAs interrupt cellular signaling pathways, inhibit the activity of tumor suppressor genes, and affect the cell cycle in gastric cancer cells. Some miRNAs, including miR-21, miR-106a and miR-421, could be potential markers for the diagnosis of gastric cancer. Long non-coding RNAs (lncRNAs), a new research hotspot among cancer-associated ncRNAs, play important roles in epigenetic, transcriptional and post-transcriptional regulation. Several gastric cancer-associated lncRNAs, such as CCAT1, GACAT1, H19, and SUMO1P3, have been explored. In addition, Piwi-interacting RNAs, another type of small ncRNA that is recognized by gastroenterologists, are involved in gastric carcinogenesis, and piR-651/823 represents an efficient diagnostic biomarker of gastric cancer that can be detected in the blood and gastric juice. Small interfering RNAs also function in post-transcriptional regulation in gastric cancer and might be useful in gastric cancer treatment.

Suppression of scinderin modulates epithelial­mesenchymal transition markers in highly metastatic gastric cancer cell line SGC­7901.

Scinderin is a Ca2+­dependent filamentous actin (F­actin) severing and capping protein, which has a key role in regulated secretion. However, little is known regarding the function and mechanism of scinderin in human carcinoma development and progression. In the present study, the biological function of scinderin was investigated using a cell proliferation assay, flow cytometric analysis and a Transwell assay in highly tumorigenic and the metastatic human gastric cancer cell line SGC­7901 transfected with scinderin­small hairpin RNA lentivirus. The changes in the expression of epithelial­mesenchymal transition (EMT) markers were also investigated. The results indicated that scinderin knockdown effectively suppressed proliferation, reduced migration and arrested the cell cycle of the SGC­7901 cells at G2/M phase. Furthermore, scinderin knockdown altered the expression of EMT markers; the expression of E­cadherin was significantly upregulated, along with an evident decrease in N­cadherin and ß­catenin expression. In conclusion, the present study suggested that suppression of scinderin impaired proliferation and migration of gastric cancer SGC­7901 cells and attenuates its EMT process. Scinderin may therefore be a potential target for tumor EMT and therapy against gastric cancer.