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ObjectiveTo study the genetic diversity and genetic relationship of Pinellia ternata germplasm resources and provide the basis for germplasm identification， variety breeding， and resource conservation. MethodIn this study， 27 P. ternata were used as experimental materials to determine seven phenotypic characters， such as plant height， leaf length， and leaf width. Simple sequence repeats （SSR） primers were designed based on P. ternata transcriptome data， and polymerase chain reaction （PCR） amplification was performed on 27 P. ternata samples. The genetic diversity of P. ternata germplasm was analyzed by POPGENE32， PowerMarker V3.25， and NTSYS-PC 2.10e software. ResultA total of 10 pairs of highly polymorphic primers （PIC>0.5） and four pairs of moderately polymorphic primers （0.25<PIC<0.5） were selected. The average number of alleles detected was 3.928 6， and the average Nei's diversity index （H） and Shannon's index （I） were 0.557 8 and 1.002 9， respectively， indicating a high level of genetic diversity. Cluster analysis divided the Pinellia ternata into seven categories， and P. ternata in the same province were in the same categories. The SSR molecular ID cards of 27 P. ternata germplasm were constructed with 14 pairs of primers， and the rapid identification of P. ternata in each region was realized. ConclusionThe results of this study can lay a foundation for the genetic diversity and population structure of P. ternata and provide a scientific basis for the identification of P. ternata germplasm resources， map construction， and molecular-assisted breeding.
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Bovine viral diarrhea virus (BVDV) is an important member of the family Flaviviridae and often causes immunosuppression. Previous studies have suggested that BVDV envelope protein Erns and the nonstructural autoprotease Npro can inhibit host innate immune responses. Herein, we found that BVDV NS4B, as a nonstructural protein necessary for replication, is involved in antagonizing the main RNA virus sensing pathway. Overexpression of BVDV NS4B protein significantly inhibited Sendai virus (SeV)-induced interferon-ß promoter activity, IFN-ß mRNA and IFN regulatory factor 3 (IRF3) phosphorylation levels. We also discovered that BVDV NS4B protein significantly inhibited RIG-I like receptor (RLRs)-mediated interferon-ß (IFN-ß) promoter activity and endogenous MDA5 mRNA levels. In addition, the BVDV NS4B protein directly interacts with N-terminal CARDs of MDA5, and co-localized with MDA5 or MDA5-2CARD in the cytoplasm. In summary, the results of this study indicate that the BVDV NS4B protein acts as an interferon-ß antagonist through inhibiting the MDA5-mediated signal transduction pathway. Our study provides an in-depth understanding of the molecular mechanisms of BVDV evading the host's natural immune response.
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Diarrhea Viruses, Bovine Viral , Interferon-beta , Diarrhea , Diarrhea Viruses, Bovine Viral/genetics , Humans , Immunity, Innate , Interferon Regulatory Factor-3/genetics , Interferon Regulatory Factor-3/metabolism , RNA Helicases/metabolism , RNA, MessengerABSTRACT
Medulloblastoma (MB) is a common yet highly heterogeneous childhood malignant brain tumor, however, clinically effective molecular targeted therapy is lacking. Modulation of hedgehog (HH) signaling by epigenetically targeting the transcriptional factors GLI through bromodomain-containing protein 4 (BRD4) has recently spurred new interest as potential treatment of HH-driven MB. Through screening of current clinical BRD4 inhibitors for their inhibitory potency against glioma-associated oncogene homolog (GLI) protein, the BRD4 inhibitor
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The role of point-of-care (POC) diagnostics is important in public health. With the support of smartphones, POC diagnostic technologies can be greatly improved. This opportunity has arisen from not only the large number and fast spread of cell-phones across the world but also their improved imaging/diagnostic functions. As a tool, the smartphone is regarded as part of a compact, portable, and low-cost system for real-time POC, even in areas with few resources. By combining near-infrared (NIR) imaging, measurement, and spectroscopy techniques, pathogens can be detected with high sensitivity. The whole process is rapid, accurate, and low-cost, and will set the future trend for POC diagnostics. In this review, the development of smartphone-based NIR fluorescent imaging technology was described, and the quality and potential of POC applications were discussed.
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AIM: Our aim in this study was to isolate potentially novel strains of fowl adenovirus serotype-4 (FAdV-4) that is currently circulating in broiler chicken flocks in Guangdong Province, China, and to compare nucleotide and amino acid (AA) sequences of their respective hexon genes. MATERIALS AND METHODS: The experiment was carried out on poultry farms experiencing outbreaks of FAdV-4-associated hydropericardium syndrome (HPS). Tissue samples from the hearts and livers of deceased chickens were screened for FAdV-4 infection using hexon gene-specific polymerase chain reaction (PCR). RESULTS: New virus isolates were used to infect 7-day-old chicks, which went onto reproduce typical HPS signs. The hypervariable region of the FAdV-4 hexon gene was PCR-amplified and sequenced. The hexon nucleotide and deduced AA sequence identities were 99.8-99.9% and 99.5-99.8%, respectively, among the four novel isolates. In addition, the new isolates were 97-100% and 96.4-99.9% identical to the nucleotide and deduced AA sequences, respectively, of FAdV-4 hexon genes available in the National Center for Biotechnology Information GenBank database. Phylogenetic analyses, based on the hexon gene sequence, revealed that the new isolates, clustered with FAdV-C; the FAdV-A, FAdV-B, FAdV-D, and FAdV-E viruses, were more distantly related. CONCLUSION: New FAdV-4 isolates from Guangdong Province are similar to those identified in other regions of the world. This information provides critical insight into HPS epidemiology and provides a perspective for monitoring outbreaks and developing strategies for disease prevention.
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Objective:To identify the current situation of contract signing for family physician services in Luohu District, Shenzhen, and to explore the associated factors with contract signing and access to services.Methods:Using the WeChat official account of"People′s Hospital of Luohu", an online survey was conducted to investigate the status of family physician′s contract services for residents in Luohu District from May 19th to 25th, 2019. A total of 14 487 valid responses were received. Chi-square test and Logistic regression models were used to analyze the contract signing of family physician services and the related factors.Results:8 560 (59.09%) of the participants had signed with a family physician, and 12 696 of them had learnt about the service before. The awareness rate of family doctor services was 87.64%. The contract signing rate of those unemployed, who living or working in Luohu less than 5 years, who living alone, having no Shenzhen medical insurance, or having not heard of family physicians signing services were less than 50%. The signing rate was positively correlated with the awareness of family physician contract service, as well as the possibility of accepting signed medical examination, traditional Chinese medicine services and health information. Only 9.70% of the contractors received health services information.Conclusions:The contract Signing rates and awareness rate of family physicians among young and middle-aged residents in Luohu District are relatively high. However, access to contract services varies considerably. The signing rate and accepted services are mainly affected by job, length of residence in Shenzhen, type of medical insurance, and understanding of contract.
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The Centers for Medicare & Medicaid Services (CMS) contracted with the RAND Corporation to identify and/or develop standardized items to include in the post-acute care patient assessment instruments. RAND was tasked by CMS with developing and testing items to measure seven areas of health status for Medicare beneficiaries: (1) vision and hearing; (2) cognitive status; (3) depressed mood; (4) pain; (5) care preferences; (6) medication reconciliation; and (7) bladder and bowel continence. This article presents results of the first Alpha 1 feasibility test of a proposed set of items for measuring each of these health status areas. Conducted between August and October 2016, the test is one of two Alpha tests that will be completed by mid-2017 to assess the feasibility of proposed items. The results of these small-scale feasibility tests will inform a national Beta test designed to determine how well the measures perform when implemented in post-acute care settings. The Alpha 1 testing phase was successfully completed, in that all items were pilot tested among 133 patients. Items from all content areas were assessed on interrater reliability and feasibility; items from some content areas were assessed on other metrics. Items have now been revised, when necessary, based on the findings of the Alpha 1 test. Alpha 2 testing is under way with the updated, revised items.
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Objective:To optimize the extraction technology and evaluate antioxidant activity of total flavonoids from Hedyotis dif-fusa. Methods:The content of kaempferol was detected by an HPLC method,and spectrophotometry was used to detect the content of total flavonoids. Results: The optimum extraction conditions were investigated by orthogonal design with the extraction quantities of kaempferol and total flavonoids as the evaluation indices. The antioxidant activities of the extracts were detected based on the clearance of hydroxyl radical and DPPH·model. The optimum extraction conditions were as follows:10-fold amount of 80% methanol and with ultrasonic extraction for 15 min. The clearance rate of 1ml methanol extract for scavenging DPPH· and ·OH was 73.89% and 91.27%,respectively.Conclusion:The optimum technology is stable and feasible for the extraction of Hedyotis diffuse. The extract shows good in vitro antioxidant properties, which provides powerful reference for the future development of relevant antioxidant prod-ucts.
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Euphorbia factor L2, a lathyrane diterpenoid isolated from caper euphorbia seed (the seeds ofL.), has been traditionally applied to treat cancer. This article focuses on the cytotoxic activity of Euphorbia factor L2 against lung carcinoma A549 cells and the mechanism by which apoptosis is induced. We analyzed the cytotoxicity and related mechanism of Euphorbia factor L2 with an MTT assay, an annexin V-FITC/PI test, a colorimetric assay, and immunoblotting. Euphorbia factor L2 showed potent cytotoxicity to A549 cells. Euphorbia factor L2 led to an increase in reactive oxygen species (ROS) generation, a loss of mitochondrial electrochemical potential, release of cytochromeactivation of caspase-9 and caspase-3, and cleavage of poly(ADP-ribose) polymerase, suggesting that Euphorbia factor L2 induced apoptosis through a mitochondrial pathway. The cytotoxic activity of Euphorbia factor L2 in A549 cells and the related mechanisms of apoptotic induction provide support for the further investigation of caper euphorbia seeds.
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Background and purpose: Locoregional infusion chemotherapy such as hepatic artery, or hepaticportal vein infusion is one of the most important treatments for hepatocelluar carcinoma. This study was aimed to investigate the distribution of fluorouracil(5-FU) in rat hepatoma, liver tissue and plasma after administrated by caudal vein or locoregional routes of hepatic artery, hepaticportal vein, and hepaticportal vein with ligated hepatic artery. Methods:Twenty-four tumor-bearing rats were divided into 4 groups randomly, and they were infused with 5-FU through peripheral vein(caudal vein), hepatic artery, hepaticportal vein or hepaticportal vein with ligated hepatic artery, which dose was 20 mg/kg. High performance liquid chromatography was adopted to measure the content of 5-FU in hepatoma, liver tissue and plasma, and the drug penetration rate among them were calculated. Results:The group of hepaticportal vein with ligated hepatic artery reached the highest concentrations of 5-FU in live tissue and hepatoma, which concentrations were (22.1±9.5)μg/g and (16.4±7.2)μg/g. Then was the hepatic artery group, and the concentration of the hepaticportal vein group in the hepatoma focus was much smaller than the former 2 groups, which was (8.9±3.7)μg/g. The peripheral vein group got the lowest concentrations both in the liver tissue and hepatoma, which were (9.4±3.7) and (4.3±2.2)μg/g. The concentrations of 5-FU in the plasma in the peripheral vein group, the hepatic artery group, the group of hepaticportal vein with ligated hepatic artery and the hepaticportal vein group were (26.8±12.5), (16.4±9.7), (15.9±10.1) and (14.9±8.5)μg/mL, which indicated that the drug concentrations of the latter 3 groups were much lower than the former group. The hepatoma/plasma penetration rate of 5-FU in the group of hepaticportal vein with ligated hepatic artery, the hepatic artery group, the hepaticportal vein group and the peripheral vein group were 103.47%, 92.94%, 59.58% and 16.08%. Conclusion: Compared to the peripheral venous bolus injection, locoregional infusion could significantly increase the concentrations of chemotherapy agent in hepatoma focus and liver tissue, and decrease the drug distributions in peripheral blood. And the infusion through hepaticportal vein with ligated hepatic artery and through hepatic artery reaches higher concentrations in the hepatoma focuses, which indicate that they are 2 practical and promising routes for the locoregional chemotherapy of hepatoma.
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BACKGROUND:Due to the difficulty in the control of delivery and col ection process, antibiotics are often added into the preservation fluid in order to avoid bacterial contamination but not affect cellgrowth and proliferation. OBJECTIVE:To observe the effects of meropenem on the proliferation and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS:There were two groups in this experiment:control group, preservation fluid with penicil in-streptomycin (final concentration of 100 U/mL);experimental group, preservation fluid with meropenem (final mass concentration of 1.0 mg/L). 100 umbilical cord samples were col ected in each group, and the positive rate was calculated. After isolation and culture, the passage 3 cells were used to draw a growth curve, flow cytometry analysis was used for phenotype determination, and osteogenic and adipogenic differentiation of cells were detected. RESULTS AND CONCLUSION:The contamination rates were 3%(3/100) in the experimental group and 20%(20/100) in the control group, indicating that meropenem can obviously reduce the contamination rate. In the experimental group, the morphology, differentiation potential and cellphenotype of the passage 3 cells were al normal. The proliferation ability of cells showed no difference between the two groups. Therefore, meropenem can be added to the preservation fluid.
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BACKGROUND:The growth of mesenchymal stem cells in vitro in different conditioned media is different evidently. So it is necessary to choose a more suitable medium. OBJECTIVE:To contrast and observe the proliferation of human umbilical cord mesenchymal stem cells in three kinds of media and to check the immunophenotype and differentiation ability of mesenchymal stem cells. METHODS:Human umbilical cord mesenchymal stem cells were col ected by explant method in sterile conditions. After subculturing by T75 incubation bottles, the third generation of mesenchymal stem cells were cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 medium containing 5%fetal bovine serum, Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum and Mesen PRO RSTM medium. After 1, 3, 5, 7 days of culture, the cells were counted to draw a growth curve. Immunophenotype of the third generation of umbilical cord mesenchymal stem cells were determined by flow cytometry and the ability of osteogenic and adipogenic differentiation was also detected. RESULTS AND CONCLUSION:The third generation of cells cultured highly expressed CD44, CD73, CD90, CD105, but did not express CD29, CD31, CD34, HLA-DR. The oil red O staining showed a lot of little red lipid drops after adipogenic induction;alizarin red staining showed osteoblast-like cells after osteogenic induction, indicating umbilical cord mesenchymal stem cells in vitro have the potential of multi-directional differentiation. After observation and counting, the colony and shape of cells cultured in Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum were superior to those cultured in the other two media. Therefore, it is concluded that Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing 10%fetal bovine serum is preferred for cellsubculture.
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BACKGROUND:High-quality and efficient umbilical cord-derived mesenchymal stem cells could be obtained by establishing and improving the method to avoid fungal contamination and by effectively decreasing pol ution rate of isolated culture during umbilical cord col ection. OBJECTIVE:To explore the effects of amphotericin B on growth and differentiation potential of umbilical cord-derived mesenchymal stem cells. METHODS:Umbilical cord-derived mesenchymal stem cells were separated from healthy ful-termed delivery fetus using col agenase digestion method and treated with amphotericin B. Subsequently, umbilical cord-derived mesenchymal stem cells were amplified and cultured in vitro with MesenPRO RSTM medium. The third passage of umbilical cord-derived mesenchymal stem cells in logarithmic phase was obtained to analyze their morphology, proliferation and immunophenotype, and induced to differentiate into osteoblasts and adipocytes in vitro. RESULTS AND CONCLUSION:After treatment with amphotericin B, umbilical cord-derived mesenchymal stem cells were successful y isolated and cultured in vitro. Flow cytometry results revealed that human umbilical cord-derived mesenchymal stem cells strongly expressed CD44, CD105 and CD73, CD90, and negatively expressed HLA-DR, CD29, CD31, and CD34. Amphotericin B-treated human umbilical cord-derived mesenchymal stem cells can stil differentiate into adipocytes and osteoblasts in vitro.
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Not only can esthetic clasps removable partial dentures reserve some advantages that removable partial denture itself has such as non-preparation or less preparation and cheap price, but also can bring metal-free smile to the patients, which is an new effective and affordable treatment option for partial edentulism. This article introduced the basic concepts, principle, various types of estheticdesign and related clinical application.
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Humans , Dental Clasps , Denture Design , Denture Retention , Denture, Partial, Removable , Esthetics , Esthetics, DentalABSTRACT
Aim To observe the effects of amlodipine on cell cycle,cell cycle-related genes and cyclin expression of human breast carcinoma MCF-7 cells.And to probe effects of amlodipine on cell cycle and its mechanism of human breast carcinoma MCF-7 cells.Methods In vitro growth inhibitory effects of amlodip-ine on human breast carcinoma MCF-7 cells were determined by MTT assay,cell cycle distribution was detected by flow cytometry,the Mrna expression levels of cell cycle-related genes cyclinD1 and p21 were treated by RT-PCR,and the protein expression of cell cycle protein cyclinD1 and p21 were assessed by Western blot.Results With dose and time dependently,amlodipine could inhibit the proliferation of human breast carcinoma MCF-7 cells in vitro.The IC_(50) was 14.439 μmol·L~(-1).When breast cancer cells MCF-7 were treated with 7.22 μmol·L~(-1)(1/2 IC_(50)),14.439 μmol·L~(-1)(IC_(50))and 28.88 μmol·L~(-1)(2IC_(50))amlodipine for 48 h,the ratios in the G_0/G_1 phase were significantly increased as compared with control group(P<0.05).Amlodipine inhibited the expression of Mrna and protein of cyclinD1,while increased the expression of Mrna and protein of p21.Conclusions Amlodipine exhibits obvious anti-tumor activities on human breast carcinoma MCF-7 cells and arrest cell cycle in G_1 phase.The mechanism of G_1 phase arresting may be related to modulating the Mrna and protein expression of cell cycle-related gene cyclinD1 and p21.
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OBJECTIVE Through the investigations of the pathogen monitoring and drug resistance,we find the evidence for controlling the spread of bacterial drug resistance.METHODS The medical hospital of 1180 patient cases frim 2005 to 2007 were retrospectively investigated and analyzed.RESULTS For recent two years there were isolated 170 strains of pathogenic bacteria and the top five were Pseudomonas aeruginosa,Staphylococcus epidermidis,Candida albicans,Staphylococcus aureus and Escherichia coli.P.aernuginosa was in the first place and its drug rssistance was severe.The sensitive ratio of MRSA and MRCNS to vancomycin was 100% and their resistance ratio to oxacillin,penicillin,amoxicillin/clavulanic acid and ampicilin/sulbactam was 100%;The E.coli and in ESBLs-producting Klebsiella pneumoniaehad multidvug vesistance to various kinds of antibacterials.CONCLUSIONS The resistance of the pathogenic bacteria is increasing year by year,and the multi-drug resistance is also seen,we should use the antibiotic properly in clinicsl to control the increase in bacterial drug resistance.
