The objective of this study was to observe the effect of Astragalus membranaceus on high sugar-induced Caenorhabditis elegans, and to explore its mechanism of action. UPLC-MS method was used to identify the components of Astragalus membranaceus. A high glucose model was established by using Caenorhabditis elegans as a model organism, and the effects of Astragalus membranaceus on body length, body bending, swallowing frequency, and reactive oxygen species (ROS) of the nematode were determined; the effects of Astragalus membranaceus on the expression of mRNA of genes related to the protein skinhead-1 (SKN-1) signaling pathway were examined by using the real-time fluorescence quantitative polymerase chain reaction (PCR). The results showed that compared with the normal group, the nematode body length, body bending, and swallowing frequency expression were significantly reduced and the ROS content in the body was significantly increased in the high glucose state; after the administration of Astragalus membranaceus, the body length, body bending, and swallowing frequency expression were significantly increased, and the ROS content was significantly reduced (P < 0.01). Compared with the normal group, SKN-1, superoxide dismutas-3 (SOD-3), glutathione S-transferase 4 (GST-4), and glutathione S-transferase 7 (GST-7) expression were significantly decreased in Caenorhabditis elegans in the high glucose condition; SKN-1, SOD-3, GST-4, and GST-7 expression were significantly increased after administration of Astragalus membranaceus (P < 0.01). In the present study, we demonstrated that Astragalus membranaceus has an effect on high glucose-induced Caenorhabditis elegans nematodes, and its mechanism of action may be through the modulation of the SKN-1 signaling pathwaym in order to ameliorate the oxidative stress response induced by high glucose.

This study aimed to further explore the relevant mechanism of action by network pharmacology integrated with animal experimental verification based on previous proven effective treatment of vertebral artery type of cervical spondylosis(CSA) by Panlongqi Tablets. Bionetwork analysis was performed to establish drug-disease interaction network, and it was found that the key candidate targets of Panlongqi Tablets were enriched in multiple signaling pathways related to CSA pathological links, among which phosphatidylinositol 3-kinase(PI3 K)/serine-threonine kinase(AKT/PKB) signaling pathway was the most significant. Further, mixed modeling method was used to build the CSA rat model, and the rats were divided into normal, model, Panlongqi Tablets low-, medium-and high-dose(0.16, 0.32, 0.64 g·kg~(-1)) and Jingfukang Granules(positive drug, 1.35 g·kg~(-1)) groups. After successful modeling, the rats were administered for 8 consecutive weeks. Pathological changes of rat cervical muscle tissues were detected by hematoxylin-eosin(HE) staining, and the content of interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), vascular endothelial cell growth factor(VEGF) and chemokine(C-C motif) ligand 2(CCL2) in rat serum and/or cervical tissues was determined by enzyme-linked immunosorbent assay(ELISA). Western blot was employed to detect the protein expression levels of chemokine(C-C motif) receptor 2(CCR2), PI3 K, AKT, phosphorylated AKT(p-AKT), I-kappa-B-kinase beta(IKK-beta/IKKβ), nuclear factor kappa B(NF-κB P65) and phosphorylated nuclear factor kappa B(NF-κB p-P65) in rat cervical tissues, and positive expression of p-NF-κB P65 in rat cervical muscle tissues was detected by immunofluorescence. The results showed that Panlongqi Tablets at different doses improved the degree of muscle fibrosis and inflammation in cervical muscle tissues of CSA rats, and reduced the content of inflammatory factors IL-1β, TNF-α, VEGF, CCL2 and CCR2 in serum and/or cervical tissues. The protein expression levels of PI3 K, p-AKT, IKKβ and p-NF-κB P65 as well as the nuclear entry of p-NF-κB P65 in cervical tissues were down-regulated. These findings suggest that Panlongqi Tablets can significantly inhibit the inflammatory response of CSA rats, and the mechanism of action may be related to the down-regulation of the activation of PI3 K/AKT signaling pathway.
Animals ; Drugs, Chinese Herbal ; I-kappa B Kinase/pharmacology* ; NF-kappa B/metabolism* ; Network Pharmacology ; Phosphatidylinositol 3-Kinases/metabolism* ; Protein Serine-Threonine Kinases ; Proto-Oncogene Proteins c-akt/metabolism* ; Rats ; Signal Transduction ; Spondylosis/drug therapy* ; Tumor Necrosis Factor-alpha/metabolism* ; Vascular Endothelial Growth Factor A/genetics* ; Vertebral Artery/metabolism*

Production of biofuels such as ethanol from non-grain crops may contribute to alleviating the global energy crisis and reducing the potential threat to food security. Tobacco (Nicotiana tabacum) is a commercial crop with high biomass yield. Breeding of starch-rich tobacco plants may provide alternative raw materials for the production of fuel ethanol. We cloned the small subunit gene NtSSU of ADP-glucose pyrophosphorylase (NtAGPase), which controls starch biosynthesis in tobacco, and constructed a plant expression vector pCAMBIA1303-NtSSU. The NtSSU gene was overexpressed in tobacco upon Agrobacterium-mediated leaf disc transformation. Phenotypic analysis showed that overexpression of NtSSU gene promoted the accumulation of starch in tobacco leaves, and the content of starch in tobacco leaves increased from 17.5% to 41.7%. The growth rate and biomass yield of the transgenic tobacco with NtSSU gene were also significantly increased. The results revealed that overexpression of NtSSU gene could effectively redirect more photosynthesis carbon flux into starch biosynthesis pathway, which led to an increased biomass yield but did not generate negative effects on other agronomic traits. Therefore, NtSSU gene can be used as an excellent target gene in plant breeding to enrich starch accumulation in vegetative organs to develop new germplasm dedicated to fuel ethanol production.
Biomass ; Gene Expression Regulation, Plant ; Plant Breeding ; Plant Leaves/genetics* ; Plants, Genetically Modified/metabolism* ; Starch ; Tobacco/metabolism*

OBJECTIVE: To evaluate the suitability of two pretreatment methods, the nitric acid digestion method and the elution method, and two measurement modes of inductively coupled plasma-mass spectrometry(ICP-MS), the No gas mode and the helium collision(He) mode, for the determination of lithium and its compounds in the workplace air. METHODS: We collected lithium and its compounds in the air of the workplace using the microporous filter membrane, and two pretreatment methods, the nitric acid digestion and elution methods were used for processing, and measured with the No gas mode and the He mode of ICP-MS. RESULTS: The good linearity range of lithium concentration in No gas mode and He mode of ICP-MS method was 0.00-500.00 μg/L, and the correlation coefficient was 0.999. The detection limit and the lower limit of quantification of lithium were 0.04 and 0.13 μg/L respectively in the No gas mode. In He gas mode: they were 0.12 and 0.39 μg/L respectively. Using the nitric acid digestion method for pre-treatment, the recovery rate of lithium addition was 96.9%-104.9%; the within-run and the between-run relative standard deviations were 3.3%-5.0% and 2.9%-5.3% respectively. Using the elution method for pre-treatment, the recovery rate of lithium addition was 97.6%-102.1%; the within-run and the between-run relative standard deviation were 3.3%-4.6% and 3.4%-4.8%, respectively. The sample could be stored at room temperature for at least 14 days. CONCLUSION: The ICP-MS method can be used as a new technology for detecting lithium and its compounds in the air of workplace. It is recommended that the elution method and the No gas mode be the first choice when measuring lithium and its compounds.

BACKGROUND: Human infections with zoonotic coronaviruses (CoVs), including severe acute respiratory syndrome (SARS)-CoV and Middle East respiratory syndrome (MERS)-CoV, have raised great public health concern globally. Here, we report a novel bat-origin CoV causing severe and fatal pneumonia in humans. METHODS: We collected clinical data and bronchoalveolar lavage (BAL) specimens from five patients with severe pneumonia from Wuhan Jinyintan Hospital, Hubei province, China. Nucleic acids of the BAL were extracted and subjected to next-generation sequencing. Virus isolation was carried out, and maximum-likelihood phylogenetic trees were constructed. RESULTS: Five patients hospitalized from December 18 to December 29, 2019 presented with fever, cough, and dyspnea accompanied by complications of acute respiratory distress syndrome. Chest radiography revealed diffuse opacities and consolidation. One of these patients died. Sequence results revealed the presence of a previously unknown β-CoV strain in all five patients, with 99.8% to 99.9% nucleotide identities among the isolates. These isolates showed 79.0% nucleotide identity with the sequence of SARS-CoV (GenBank NC_004718) and 51.8% identity with the sequence of MERS-CoV (GenBank NC_019843). The virus is phylogenetically closest to a bat SARS-like CoV (SL-ZC45, GenBank MG772933) with 87.6% to 87.7% nucleotide identity, but is in a separate clade. Moreover, these viruses have a single intact open reading frame gene 8, as a further indicator of bat-origin CoVs. However, the amino acid sequence of the tentative receptor-binding domain resembles that of SARS-CoV, indicating that these viruses might use the same receptor. CONCLUSION A novel bat-borne CoV was identified that is associated with severe and fatal respiratory disease in humans.
Adult ; Aged ; Betacoronavirus ; genetics ; isolation & purification ; Coronavirus Infections ; diagnostic imaging ; therapy ; virology ; Female ; Humans ; Male ; Middle Aged ; Pandemics ; Pneumonia, Viral ; diagnostic imaging ; therapy ; virology ; Tomography, X-Ray ; Treatment Outcome

Background:The clinical outcome of transtrochanteric rotational osteotomy (TRO) for osteonecrosis of the femoral head (ONFH) remains controversial,and the promising clinical results of several Japanese studies could not be reproduced in American and European studies.Trying to solve controversies on TRO for ONFH rising from apparently conflicting studies,a meta-analysis was conducted to assess the 5-and 10-year hip survival rates (with conversion to artificial joint replacement and radiographic failure as endpoints) after TRO.Methods:All eligible studies were searched in seven comprehensive databases including PubMed,Web of Science,Embase,Cochrane Library,VIP Database,China Knowledge Resource Integrated Database,and Wan Fang Database prior to June 2019.The outcomes evaluated were 5-and 10-year hip survival rates after TRO.The odds ratio and risk difference for the non-comparative binary data with the 95％ confidence intervals (CIs) were calculated for each outcome.The included studies were assessed for methodologic bias and potential reasons for heterogeneity were explored.Results:Nineteen studies of TRO for ONFH were eligible for this meta-analysis according to inclusion criteria.Based on the previous report,two calculation methods (Methods 1 and 2) were adopted in this meta-analysis.Furthermore,we performed a subgroup analysis of the 5-and 10-year hip survival rates (Method 1) after TRO for ONFH:Asian sub-population and non-Asian subpopulation.Taking conversion to artificial joint replacement as the endpoint,5-and 10-year hip survival rates (Method 1) after TRO for ONFH in the Asian population were 0.86 (95％ CI =0.82-0.89) and 0.72 (95％ CI =0.65-0.78),respectively,and 5-and 10-year hip survival rates after TRO for ONFH in the non-Asian population were 0.55 (95％ CI =0.43-0.67) and 0.42 (95％ CI =0.28-0.55),respectively.The 5-and 10-year hip survival rates (Method 2) after TRO for ONFH were 0.90 (95％ CI =0.79-0.95) and 0.89 (95％ CI =0.81-0.94),respectively.Taking radiographic failure as the endpoint,5-and 10-year hip survival rates after TRO for ONFH were 0.70 (95％ CI =0.64-0.76) and 0.53 (95％ CI =0.46-0.61),respectively.Conclusions:The 5-and 10-year hip survival rates after TRO for ONFH were satisfactory in the Asian population,and were acceptable in the non-Asian population despite high early failure rates.

BACKGROUND: Repair of bone defects is not only a clinical problem, but also a hot topic in the field of orthopedics. Although autologous bone grafting is considered as the "gold standard" for bone repair, its use is limited due to the limited source of autogenous bone, bone infections and pains that are easy to occur in the donor region. Allograft bones are always associated with immune rejection, slow healing, and infection. Therefore, it is imminent to develop new materials for bone repair. OBJECTIVE: To explore the effect of rabbit adipose-derived stem cells (rADSCs) as seed cells and hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) composite as a carrier on the repair of rabbit vertebral defects. METHODS:Thirty-eight 3-month-old New Zealand white rabbits were selected,and two of them were used to culture rADSCs in vitro. Passage 3 rADSCc were inoculated on HA/β-TCP scaffolds and then cultured in vitro for 2 weeks.A 5 mm×5 mm×3 mm bone defect was prepared at the anterior edge of L4/5vertebral body in the remaining 36 rabbits. These model rabbits were then randomized into cell-scaffold composite group, scaffold group and control group with no intervention, with 12 rabbits in each group. rADSCs/HA/β-TCP composite and HA/β-TCP scaffold were implanted into the cell-scaffold and HA/β-TCP groups, respectively. Anteroposterior and lateral DR of the spine and Lane-Sandhu X-ray were performed at 4, 8, 12 postoperative weeks. All rabbits were sacrificed at 12 postoperative weeks and specimens were collected for gross and histopathological observations. RESULTS AND CONCLUSION: Under the gross observation, bone defects in the cell-scaffold group were essentially replaced by new bone tissues, which was significantly better than that in the scaffold group and control group. At 12 postoperative weeks, the material implanted was basically absorbed in the cell-scaffold group, partially absorbed in the scaffold group and poorly absorbed in the control group in which there was a clear boundary with the surrounding tissues and patchy calcified shadows were visible. X-ray results showed that the repair effect in the cell-scaffold group was better than that in the scaffold group and control group (P < 0.05). Histopathological findings showed the marked absorption of the implant in the cell-scaffold group, partial residual in the scaffold group with some fibrous calluses and osteoid tissues, and a large amount of fiber tissues and a small amount of calluses in the control group. Overall, the rADSCs/HA/β-TCA has a good ability to repair bone defects.

AIDS-Related Opportunistic Infections ; diagnosis ; microbiology ; Acquired Immunodeficiency Syndrome ; diagnosis ; microbiology ; Bronchoalveolar Lavage Fluid ; microbiology ; Female ; Humans ; Immunohistochemistry ; Male ; Methenamine ; Pneumocystis carinii ; isolation & purification ; Pneumonia, Pneumocystis ; diagnosis ; microbiology ; Polymerase Chain Reaction ; Silver Staining ; methods

The development of thrombus on the tricuspid valve is very rare. This report describes a case of acute pulmonary embolism (PE) with a mass on the tricuspid valve in a normal heart, detected by bedside transthoracic echocardiography (TTE). After successful surgical management, the histopathological examination revealed the mass from the tricuspid valve to be mixed thrombus. The early use of bedside TTE can facilitate the prompt diagnosis and aggressive therapy when PE is suspected.
Female ; Humans ; Middle Aged ; Pulmonary Embolism ; diagnosis ; Thrombosis ; diagnosis ; Tricuspid Valve ; pathology

This study was aimed to explore the expression of B cell-activating factor of TNF family (BAFF) and B cell lymphoma/leukemia-2 (BCL-2) in bone marrow mononuclear cells (BMMNC) of multiple myeloma (MM) and the significance of BAFF and BCL-2 for occurrence, development and prognosis of MM. The bone marrow of 40 cases of MM and 10 healthy persons was collected, the mononuclear cells (MNC) were isolated, the expression of BAFF and BCL-2 mRNA in BMMNC was detected by real-time PCR; the plasma was simultaneously collected and the β2-MG level was determined; the clinical staging of MM patients was performed according to Durie-Salmon (D-S) staging criterion. The results indicated that the expression level of BAFF and BCL-2 mRNA in MM patients increased, as compared with normal controls, the difference was statistical significant (p < 0.05); the expression level of BAFF and BCL-2 mRNA in plateau stage after treatment obviously decreased. The expression level of BAFF and BCL-2 mRNA in relapsed/refractory MM patients was significantly higher than that in normal controls and patients reached plateau stage (p < 0.05), there was no statistically significant difference between newly diagnosed and relapsed/refractory MM patients (p > 0.05). The expression of BAFF and BCL-2 mRNA related with D-S staging and β2-MG level. It is concluded that the expression levels of BAFF and BCL-2 mRNA increase, moreover the expression levels of BAFF and BCL-2 mRNA in newly diagnosed and relapsed/refractory MM patients are higher than those in patients reached plateau stage, which suggest the BAFF and BCL-2 may be involved in occurrence and development of MM; the relation of expression level of BAFF and BCL-2 mRNA to MM load is positive, which indicates the expression level of BAFF and BCL-2 mRNA may be a new indicator for evaluating the prognosis of MM patients.
Adult ; Aged ; B-Cell Activating Factor ; genetics ; metabolism ; Case-Control Studies ; Female ; Humans ; Lymphoma, B-Cell ; genetics ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; genetics ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics