A Mild Hyperthermia Hollow Carbon Nanozyme as Pyroptosis Inducer for Boosted Antitumor Immunity.

The immune checkpoint blockade (ICB) antibody immunotherapy has demonstrated clinical benefits for multiple cancers. However, the efficacy of immunotherapy in tumors is suppressed by deficient tumor immunogenicity and immunosuppressive tumor microenvironments. Pyroptosis, a form of programmed cell death, can release tumor antigens, activate effective tumor immunogenicity, and improve the efficiency of ICB, but efficient pyroptosis for tumor treatment is currently limited. Herein, we show a mild hyperthermia-enhanced pyroptosis-mediated immunotherapy based on hollow carbon nanozyme, which can specifically amplify oxidative stress-triggered pyroptosis and synchronously magnify pyroptosis-mediated anticancer responses in the tumor microenvironment. The hollow carbon sphere modified with iron and copper atoms (HCS-FeCu) with multiple enzyme-mimicking activities has been engineered to induce cell pyroptosis via the radical oxygen species (ROS)-Tom20-Bax-Caspase 3-gasdermin E (GSDME) signaling pathway under light activation. Both in vitro and in vivo antineoplastic results confirm the superiority of HCS-FeCu nanozyme-induced pyroptosis. Moreover, the mild photothermal-activated pyroptosis combining anti-PD-1 can enhance antitumor immunotherapy. Theoretical calculations further indicate that the mild photothermal stimulation generates high-energy electrons and enhances the interaction between the HCS-FeCu surface and adsorbed oxygen, facilitating molecular oxygen activation, which improves the ROS production efficiency. This work presents an approach that effectively transforms immunologically "cold" tumors into "hot" ones, with significant implications for clinical immunotherapy.

Effects of species mixtures on soil water storage in the semiarid hilly gully region.

Mixed-species plantations are promoted to restore degraded ecosystems and improve soil quality worldwide. However, differences of soil water conditions between pure and mixed plantations are still controversial and how species mixtures affect soil water storage (SWS) was not well quantified. In this study, vegetation characteristics, soil properties and SWS were continuously monitored and quantified in three pure plantations (Armeniaca sibirica (AS), Robinia pseudoacacia (RP) and Hippophae rhamnoides (HR)) and their corresponding mixed plantations (Pinus tabuliformis-Armeniaca sibirica (PT-AS), Robinia pseudoacacia-Pinus tabuliformis-Armeniaca sibirica (RP-PT-AS), Platycladus orientalis-Hippophae rhamnoides plantation (PO-HR), Populus simonii-Hippophae rhamnoides (PS-HR)). The results found that SWS of 0-500 cm in RP (333.60 ± 75.91 mm) and AS (479.52 ± 37.50 mm) pure plantations were higher than those in their corresponding mixed plantations (p > 0.05). SWS in the HR pure plantation (375.81 ± 81.64 mm) was lower than that in its mixed plantation (p > 0.05). It is suggested that the effect of species mixing on SWS was species specific. Additionally, soil properties exerted more contributions (38.05-67.24 %) to SWS than vegetation characteristics (26.80-35.36 %) and slope topography (5.96-29.91 %) at different soil depths and the whole 0-500 cm soil profile. Furthermore, by excluding the effects of soil properties and topographic factors, plant density and height were particularly important to SWS (with standard coefficients 0.787 and 0.690 respectively). The results implied that not all the mixed plantations exhibits the better soil water conditions than the compared pure plantations, which was tightly related to species selected for mixing. Our study provides scientific support for revegetation technique improvement (structural adjustment and species optimization) in this region.

[Ecological and physiological mechanisms of growth decline of Robinia pseudoacacia plantations in the Loess Plateau of China: A review.] / 黄土高原人工刺槐林生长衰退的生态生理机制.

Robinia pseudoacacia has been widely planted in the Loess Plateau of China for soil and water conservation. The growth decline of R. pseudoacacia plantations has become a recently emerging challenge for the revegetation program and sustainable forest management in this region. As to the scientific definition, identified criteria and quantitative indices have not yet been comprehensively quantified, our current understanding of the ecological and physiological mechanisms for growth decline of R. pseudoacacia plantations is limited. The knowledge could enrich the basic theories of vegetation restoration and benefit the sustainable development of the afforestation project in the Loess Plateau. Through the comprehensive compilation of literatures on forest decline and tree mortality in the Loess Plateau and other regions across the world, this review summarized the mechanisms and recent research progress on growth decline for R. pseudoacacia plantations in the Loess Plateau, primarily demonstrated from ecological (e.g., climatic change, soil desiccation, the imbalance of community structure and the misconduct of forest management) and physiological (e.g., hydraulic failure, carbon starvation, genetic and molecular regulation) perspectives. Finally, we highlighted the research gap with regard to growth decline of R. pseudoacacia plantations in the Loess Plateau.

Therapeutic Observation of Abdominal Acupuncture plus Chinese Medication for Ureteral Calculus / 上海针灸杂志

Objective To observe the clinical efficacy of abdominal acupuncture plus Chinese medication in treating ureteral calculus. MethodA total of 106 patients with ureteral calculus were randomized into an abdominal acupuncture group (53 cases) and a medication group (53 cases). The abdominal acupuncture group was intervened by abdominal acupuncture plus medication, with Shuifen (CV9), Xiawan (CV10), Jianli (CV11), Zhongji (CV3), Guanyuan (CV4) and Qihai (CV6) selected as the major points. The removal of stones and the release of pain were observed in the two groups, and the efficacies were compared between the two groups.Result The two treatment methods both effectively improved the pain and promoted the removal of stones; the Visual Analogue Scale (VAS) scores were significantly changed after the intervention in both groups (P<0.05). The time taken to mitigate and release pain was significantly shorter in the abdominal acupuncture group compared to the other group (P<0.01). The total effective rate was 88.7% (47/53) in the abdominal acupuncture group, superior to 73.6% (39/53) in the medication group (P<0.05).ConclusionAbdominal acupuncture plus medication can effectively mitigate pain, promote the removal of stones and reduce patient's sufferings in treating ureteral calculus.

Therapeutic Observation of Abdominal Acupuncture plus Chinese Medication for Ureteral Calculus / 上海针灸杂志

Objective To observe the clinical efficacy of abdominal acupuncture plus Chinese medication in treating ureteral calculus. MethodA total of 106 patients with ureteral calculus were randomized into an abdominal acupuncture group (53 cases) and a medication group (53 cases). The abdominal acupuncture group was intervened by abdominal acupuncture plus medication, with Shuifen (CV9), Xiawan (CV10), Jianli (CV11), Zhongji (CV3), Guanyuan (CV4) and Qihai (CV6) selected as the major points. The removal of stones and the release of pain were observed in the two groups, and the efficacies were compared between the two groups.Result The two treatment methods both effectively improved the pain and promoted the removal of stones; the Visual Analogue Scale (VAS) scores were significantly changed after the intervention in both groups (P<0.05). The time taken to mitigate and release pain was significantly shorter in the abdominal acupuncture group compared to the other group (P<0.01). The total effective rate was 88.7% (47/53) in the abdominal acupuncture group, superior to 73.6% (39/53) in the medication group (P<0.05).ConclusionAbdominal acupuncture plus medication can effectively mitigate pain, promote the removal of stones and reduce patient's sufferings in treating ureteral calculus.

Comparison of different sequencing and assembly strategies for a repeat-rich fungal genome, Ophiocordyceps sinensis.

Ophiocordyceps sinensis is one of the most expensive medicinal fungi world-wide, and has been used as a traditional Chinese medicine for centuries. In a recent report, the genome of this fungus was found to be expanded by extensive repetitive elements after assembly of Roche 454 (223Mb) and Illumina HiSeq (10.6Gb) sequencing data, producing a genome of 87.7Mb with an N50 scaffold length of 12kb and 6972 predicted genes. To test whether the assembly could be improved by deeper sequencing and to assess the amount of data needed for optimal assembly, genomic sequencing was run several times on genomic DNA extractions of a single ascospore isolate (strain 1229) on an Illumina HiSeq platform (25Gb total data). Assemblies were produced using different data types (raw vs. trimmed) and data amounts, and using three freely available assembly programs (ABySS, SOAP and Velvet). In nearly all cases, trimming the data for low quality base calls did not provide assemblies with higher N50 values compared to the non-trimmed data, and increasing the amount of input data (i.e. sequence reads) did not always lead to higher N50 values. Depending on the assembly program and data type, the maximal N50 was reached with between 50% to 90% of the total read data, equivalent to 100× to 200× coverage. The draft genome assembly was improved over the previously published version resulting in a 114Mb assembly, scaffold N50 of 70kb and 9610 predicted genes. Among the predicted genes, 9213 were validated by RNA-Seq analysis in this study, of which 8896 were found to be singletons. Evidence from genome and transcriptome analyses indicated that species assemblies could be improved with defined input material (e.g. haploid mono-ascospore isolate) without the requirement of multiple sequencing technologies, multiple library sizes or data trimming for low quality base calls, and with genome coverages between 100× and 200×.

Non-concerted ITS evolution in fungi, as revealed from the important medicinal fungus Ophiocordyceps sinensis.

The internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA) has been widely used as a molecular marker in phylogenetic studies and has been selected as a DNA barcode for fungi. It is generally believed that nrDNA conforms to concerted evolution in most eukaryotes; however, intraindividual-intraspecific polymorphisms of this region were reported in various organisms, suggesting a non-concerted evolutionary process. In Ophiocordyceps sinensis, one of the most valuable medicinal fungi, a remarkable variation of the ITS region has been revealed. Some highly divergent sequences were thought to represent cryptic species, different species or genotypes in previous studies. To clarify the unusual ITS polymorphisms observed in O. sinensis, specific primers were designed to amplify ITS paralogs from pure cultures of both single-ascospore and tissue isolates in this study. All of the available ITS sequences, including those generated by this group and those in GenBank, were analyzed. Several AT-biased ITS paralogs were classified as pseudogenes based on their nucleotide compositions, secondary structures and minimum free energies of their 5.8S rRNAs, substitution rates, phylogenetic positions and gene expression analyses. Furthermore, ITS pseudogenes were amplified with specific primers from 10 of the 28 strains tested, including eight single-ascospore and two tissue isolates. Divergent ITS paralogs were proved to coexist in individual genomes, suggesting a non-concerted mechanism of evolution in the ITS region of O. sinensis. The hypotheses that divergent ITS paralogs represent cryptic or other species or different genotypes were thus rejected.

Effects and interaction of icariin, curculigoside, and berberine in er-xian decoction, a traditional chinese medicinal formula, on osteoclastic bone resorption.

Er-Xian decoction (EXD), a traditional Chinese medicine, has been reported to have a protective effect against bone loss in ovariectomized osteoporotic rats, and the inclusion of icariin (I), curculigoside (C), and berberine (B) in EXD displays inhibitory effects on osteoclastic bone resorption. In the present paper, we investigated the interaction and effects of I, C, B, and their combination on bone resorption activity in vitro on osteoclasts derived from rat bone marrow cells. ICB synergistically decreased the formation of bone resorption pits, the number of multinucleated osteoclasts, and the activity of tartrate-resistant acid phosphatase (TRAP) and showed antagonistic or additive effects on cathepsin K activity in the coculture system of osteoblasts and bone marrow cells in the presence of 1, 25-dihydroxyvitamin D(3) and dexamethasone. The combination of ICB also enhanced the inhibitory effects on the formation of F-actin ring, a cytoskeleton structure of osteoclasts induced from bone marrow cells with macrophage colony stimulation factor (M-CSF) and receptor activator of NF-κB ligand (RANKL). In addition, ICB synergistically improved the ratio of protein expression of osteoprotegerin (OPG) and RANKL in osteoblasts and interfered with the mitogen-activated protein kinases (MAPKs) pathway in osteoclast. These results clearly show that I, C, B, and their combination in EXD exert effects of mutual reinforcement. However, IBC does not show an intensified adverse effect in the ovariectomized murine model, as revealed by change in body and uterine weight, confirming the safety of EXD. These observations are in agreement with the rationality of the formula used in this paper.

A survey of the geographic distribution of Ophiocordyceps sinensis.

Ophiocordyceps sinensis is one of the best known fungi in Traditional Chinese Medicine. Many efforts have been devoted to locating the production areas of this species resulting in various reports; however, its geographic distribution remains incompletely understood. Distribution of O. sinensis at the county level is clarified in this work based on both a literature search and fieldwork. More than 3600 publications related to O. sinensis were investigated, including scientific papers, books, and online information. Herbarium specimens of O. sinensis and field collections made by this research group during the years 2000-2010 were examined to verify the distribution sites. A total of 203 localities for O. sinensis have been found, of which 106 are considered as confirmed distribution sites, 65 as possible distribution sites, 29 as excluded distribution sites and three as suspicious distribution sites. The results show that O. sinensis is confined to the Tibetan Plateau and its surrounding regions, including Tibet, Gansu, Qinghai, Sichuan, and Yunnan provinces in China and in certain areas of the southern flank of the Himalayas, in the countries of Bhutan, India and Nepal, with 3,000 m as the lowest altitude for the distribution. The fungus is distributed from the southernmost site in Yulong Naxi Autonomous County in northwestern Yunnan Province to the northernmost site in the Qilian Mountains in Qilian County, Qinghai Province, and from the east edge of the Tibetan Plateau in Wudu County, Gansu Province to the westernmost site in Uttarakhand, India. The clarification of the geographic distribution of O. sinensis will lay the foundation for conservation and sustainable use of the species.

Antiosteoporotic effects and proteomic characterization of the target and mechanism of an Er-Xian Decoction on osteoblastic UMR-106 and osteoclasts induced from RAW264.7.

Er-Xian Decoction (EXD) has been used for the treatment of osteoporosis disorders, menopausal syndrome, and other aging diseases in the Chinese traditional healthcare system. However, the targets and mechanism of action have not been clarified. This study was designed to investigate the effects and possible target proteins of EXD on osteoblastic UMR-106 cells and osteoclasts induced from RAW264.7 cells using the proteomic analysis technique. We found that EXD at a concentration of 50-200 microg/mL significantly enhanced osteoblastic UMR-106 cell proliferation, alkaline phosphatase (ALP) activity and formation of bone nodules, and decreased tartrate-resistant acid phosphatase (TRAP) activity and the bone resorption action of osteoclasts induced from RAW 264.7 cells. In EXD-treated osteoblasts, there were increases in the expression of heat-shock protein 1, high mobility group protein (Hmgb1), acidic ribosomal phosphoprotein P0, histone 2, carbonyl reductase 1, ATP synthase, aldolase A, and Rho GDP dissociation inhibitor (GDI)-alpha; and reduction in the expression of carbonic anhydrase 3, prohibitin, hemiferrin, far upstream element (FUSE)-binding protein. In EXD-treated osteoclasts, there were increases in the expression of vimentin, protein disulfide isomerase associated 3 and alpha-fetoprotein; and reduction in the expression of calnexin. These results indicated that EXD modulates bone metabolism through regulation of osteoblastic proliferation, apoptosis, and cell activation, and osteoclastic protein folding and aggregation.

Effects of berberine on differentiation and bone resorption of osteoclasts derived from rat bone marrow cells.

OBJECTIVE: To observe the effects of berberine on osteoclastic differentiation and bone resorption action in vitro, and to investigate the cellular mechanism of its inhibitory effects on bone resorption. METHODS: The multinucleated osteoclasts (MNCs) were derived by 1,25-dihydroxyvitamin D3 and dexamethasone from bone marrow cells in the coculture system with primary osteoblastic cells. The tartrate-resistant acid phosphatase (TRAP) staining and image analysis of bone resorption pit on dental slices were used to identify osteoclast. The activity of TRAP was measured by p-nitrophenyl sodium phosphate assay. The bone resorption pit area on the bone slices formed by osteoclasts was measured by computer image processing. RESULTS: At the concentrations of 0.1, 1 and 10 micromol/L, berberine dose-dependently suppressed the formation of TRAP-positive multinucleated cells, the TRAP activity and the osteoclastic bone resorption. The strongest inhibitory effect was exhibited at the concentration of 10 micromol/L, with the inhibiting rate of 60.45%, 42.12% and 72.69% respectively. CONCLUSION: Berberine can decrease bone loss through inhibition of osteoclast formation, differentiation and bone resorption.

Synergistic hepatoprotective effect of Schisandrae lignans with Astragalus polysaccharides on chronic liver injury in rats.

The aim of this study was to investigate the synergistic hepatoprotective effect of lignans from Fructus Schisandrae chinensis (LFS) with Astragalus polysaccharides (APS) on chronic liver injury in male Sprague-Dawley rats. Subcutaneous injection of 10% CCl(4) twice a week for 3 months resulted in significantly (p<0.001) elevated serum alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP) activities compared to controls. In the liver, significantly elevated levels (p<0.001) of malondialdehyde (MDA), lowered levels of reduced glutathione (GSH) (p<0.05) and catalase (CAT) (p<0.001), superoxide dismutase (SOD) (p<0.01)were observed following CCl(4) administration. 'LFS+ASP' treatment of rats at doses of 'LFS (45mg/kg)+APS (150mg/kg)' and 'LFS (135mg/kg)+APS (450mg/kg)' displayed hepatoprotective and antioxidative effects than the administration of either LFS or APS, as evident by lower (p<0.005 or 0.001) levels of serum ALT, AST, ALP and hepatic MDA (p<0.001) concentration, as well as higher SOD (p<0.05 or 0.005), CAT activities(p<0.01 or 0.005), GSH concentration (p<0.05 or 0.005) compared to the toxin treated group. Histopathological examinations revealed severe fatty degeneration in the toxin group, and mild damage in groups treated with 'LFS+APS' were observed. The coefficients drug interaction (CDI) between each individual drug and their combination (at the same dose of their single treatment) of these foregoing parameters were all less than 1, indicating that LFS and APS display hepatoprotective and antioxidant properties and act in a synergistic manner in CCl(4) induced liver injury in rats.

Antiosteoporotic activity of phenolic compounds from Curculigo orchioides.

Six phenolic compounds isolated from Curculigo orchioides, including 2,6-dimethoxy benzoic acid (1), curculigoside A (2), curculigoside B (3), curculigine A (4), curculigine D (5) and 3,3',5,5'-tetramethoxy-7,9':7',9-diepoxylignan-4,4'-di-O-beta-D-glucopyranoside (6), together with the ethanol extract of Curculigo orchioides were evaluated for their activity on osteoblasts in neonatal rat calvaria cultures and multinucleated osteoclasts derived from rat marrow cells so as to characterize the antiosteoporotic components of this plant and explore the relationship of chemical structure with antiosteoporotic activity. The proliferation of osteoblast was assayed by MTT methods. The activity of ALP (alkaline phosphatase) and TRAP (tartrate-resistant acid phosphatase) was measured by p-nitrophenyl sodium phosphate assay. The TRAP stain was used to identify osteoclast in morphology. The resorption pit area on the bone slices formed by osteoclast was measured by computer image processing. The ethanol extract exhibited stimulatory effect on both the osteoblast proliferation and the ALP activity. Six compounds all increased the osteoblast proliferation, and compounds (1), (2) and (4) also slightly increased the osteoblastic ALP activity. Compounds (1), (2), (3), (6) and the ethanol extract decreased area of bone resorption pit, osteoclastic formation and TRAP activity. These results indicated that phenolic compounds are antiosteoporotic chemical constituents from Curculigo orchioides, and their activities are related with chemical structures.