The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.
Endophytes/genetics* ; High-Throughput Nucleotide Sequencing ; Plants, Medicinal ; Research Design

Using the 260 geographical distribution records of Polygonatum cyrtonema in China, combined with 53 environmental factors, the maximum entropy modeling(MaxEnt) was used to study the ecological factors affecting the suitability distribution of P. cyrtonema. The ArcGIS software was used to predict the potential distribution of the population of P. cyrtonema. The dominant factors were chosen by using the Jackknife test and the Receiver Operating Characteristic(ROC) curve was used to evaluate the simulation. The results showed that high value of area under curve(AUC) denoted good results, which significantly differed from random predictions. Based on the evaluation criterion, the accuracies of the predictions of P. cyrtonema potential distribution in the current periods were excellent. The main environmental factors affecting the suitable growth of P. cyrtonema were the monthly precipitation, the wettest monthly precipitation, the annual average temperature range and the precipitation of November, March, February, April, May and October. There are 9 environmental factors in soil type. The potential fitness of P. cyrtonema in China is high, mainly concentra-ted in Hunan, western Hubei, Guangdong, northeastern Guangxi, southeastern Guizhou, Jiangxi, southwestern Anhui, Fujian, Zhejiang, Shaanxi, southwestern Henan and Chongqing. The growth distribution of the potential distribution area of P. cyrtonema was divided, and the zoning map of the growth suitability of P. cyrtonema was formed. Through the comparative analysis of the potential distribution range based on MaxEnt and the distribution range of literature records, the understanding of the distribution range of P. cyrtonema was expanded.
China ; Ecology ; Entropy ; Polygonatum ; Research Design ; Soil

Objective:To observe serum levels of high sensitive C reactive protein(hsCRP),interleukin(IL)-6, tumor necrosis factor(TNF)-α and hepatocyte growth factor(HGF)in patients with ischemic cerebrovascular dis-ease(ICVD),and analyze their correlation with blood lipids and anticardiolipin antibody(ACA)levels.Methods:A total of 45 ICVD patients treated in our hospital from Mar 2015 to Aug 2016 were regarded as ICVD group.An-other 45 non-ICVD patients treated in our hospital during the same period were regarded as non-ICVD control group.Serum levels of hsCRP,IL-6,TNF-α,HGF,blood lipids and ACA were observed and compared between two groups,and correlation among serum levels of hsCRP,IL-6,TNF-α and HGF,blood lipids and ACA levels were analyzed in ICVD patients.Results:Compared with non-ICVD control group,there were significant rise in se-rum levels of hsCRP[(4.69 ± 1.31)mg/L vs.(8.87 ± 1.56)mg/L],IL-6[(12.17 ± 4.33)mg/L vs.(34.26 ± 5.15)mg/L],TNF-α[(28.45 ± 2.18)pg/ml vs.(48.35 ± 3.15)pg/ml],HGF[(502.34 ± 15.36)pg/ml vs. (876.25 ± 18.15)pg/ml],ACA[(4.11 ± 0.65)IU/L vs.(7.89 ± 1.02)IU/L],total cholesterol[(4.68 ± 1.12) mmol/L vs.(5.57 ± 1.21)mmol/L],low density lipoprotein cholesterol[(2.62 ± 0.49)mmol/L vs.(3.24 ± 0.87) mmol/L]and triglyceride[(1.42 ± 0.31)mmol/L vs.(1.84 ± 0.37)mmol/L],and significant reduction in serum level of high density lipoprotein cholesterol[(1.28 ± 0.25)mmol/L vs.(1.02 ± 0.22)mmol/L]in ICVD group,P=0.001 all.Pearson correlation analysis indicated that serum levels of hsCRP,IL-6,TNF-α and HGF were signifi-cant positively correlated with serum levels of ACA,TC,LDL-C and TG(r=0.468~0.632,P<0.05 or <0.01), and significant inversely correlated with HDL-C level(r= -0.571~ -0.511,P<0.05 or < 0.01)in ICVD pa-tients.Conclusion:Serum levels of hsCRP,IL-6,TNF-α and HGF significantly rise in ICVD patients,and they are closely correlated with levels of blood lipids and ACA,which can serve as important indexes monitoring ICVD.

OBJECTIVETo investigate the morphological manifestation of bone marrow cells in newly-diagnosed patients with POEMS syndrome.

METHODSThe bone marrow cells in 155 patients with POEMS syndrome were classified and counted by OLYMPUS BX51 microscope, and the abnormal morphology of bone marrow cells was observed.

RESULTSThe count of plasma cells with normal morphology was 83.9% (130/155), the count of plasma cells with abnormal morphology (< 5 percent) was 12.3% (19/155), the count of plasma cells with obvious abnormal morphology (> 10 percent) was 3.8% (6/155) in patients with POEMS syndrome.

CONCLUSIONThe morphology of plasma cells in the most patients with the POEMS syndrome are normal, the minor patients of the POEMS syndrome have little abnormal plasma cell morphology, the extremely few patients showed obvious morphological abnormality in the bone marrow plasma cells. The higher proportion of plasma cells, the more easily and more abnormal plasma cells will be found.

Bone Marrow Cells ; Humans ; POEMS Syndrome ; Plasma Cells

OBJECTIVETo explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.

METHODAcute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.

RESULTThe purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.

CONCLUSIONASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.

Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Neoplastic Stem Cells ; cytology ; drug effects ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects

OBJECTIVETo investigate the incidence, pathogens, and clinical features of infection in consecutive cases from 2010 to 2012 in Peking Union Medical College Hospital.

METHODThe incidence, pathogen, treatment, and outcomes of patients with hematological diseases who had positive findings of bacterium in their samples from 2010 to 2012 were retrospectively analyzed.

RESULTSThere were 449 positive samples (5.8%) from 4 890 patients during this period, among which 388 were proved to be with pathogenic bacteria. Samples separated from patients with community-aquired infections accounted for 8.4% of all positive samples. Most community-aquired infections were caused by Gram-negative bacteria (75%), although no multidrug-resistant bacteria was observed. Samples separated from patients with nosocomial infections accounted for 91.6% of all positive samples. Respiratory tract (49.4%) and peripheral blood (32.6%) were the most common samples with positive results. Skin soft tissues (10.4%), and urine (3.7%) were less common samples. Most of the pathogenic bacteria of the nosocomial infections were Gram-negative (66.9%). The most common Gram-negative bacteria included Escherichia coli (13.8%), Pseudomonas aeruginosa (12.1%), and Klebsiella pneumonia (12.1%), while Staphylococcus aureus (10.4%), Enterococcus faecium (7.0%), and Staphylococcus epidermidis (5.1%) were the most common Gram-positive bacteria. Gram-negative bacteria consisted of most of sputum samples and peripheral blood samples. Samples from the surface of skin wound and anal swab were composed largely by Gram-positive bacteria (63.8%). The detection rates of extended-spectrum beta-lactamase-producing Klebsiella pneumonia/Klebsiella oxytoca, Escherichia coli, and Proteus mirabilis were 24.0%, 87.9% and 38.4%, respectively. The resistance to Acinetobacter baumannii was serious. Multidrug-resistant, extensive drug resistant and pan drug resistant A. baumannii acountted for 74% of all A. Baumannii infections. Stenotrophomonas maltophilia showed low resistance to sulfamethoxazole/trimethoprim, levofloxacin and minocycline. Also, 22 methicillin-resistant Staphylococcus aureus and 9 methicillin-resistant Staphylococcus Epidermidis were detected, which were only sensitive to vancomycin, teicoplanin, and linezolid. All patients were treated in the haematology wards and most of them were under agranulocytosis or immunosuppression. Finally, 22 patients reached clinical recovery through anti-infective therapy, whereas 49 patients died. Among those deaths, 42 patients attributed to severe infections and infection-associated complications. Fourteen of all the deaths might be infected with drug-resistance bacteria. There were 61 samples proved to be bacteria colonization. Nonfermenters such as Acinetobacter baumannii and Stenotrophomonas maltophilia made up for a large amount of bacteria colonization.

CONCLUSIONSThe pathogens of nosocomial infections in the hematology ward are mainly Gram-negative bacteria. The incidences and pathogens vary from different infection sites. Nosocomial infection still has a higher mortality rate. Once nonfermenters are detected positive, the pathogenic or colonial bacteria should be distinguished.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bacteria ; isolation & purification ; Bone Marrow Transplantation ; Cross Infection ; microbiology ; Female ; Hematologic Diseases ; complications ; microbiology ; Hematology ; Hospital Departments ; statistics & numerical data ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUNDVon Hippel-Lindau disease (VHL), a heritable autosomal dominant disease characterized by neoplasia in multiple organ systems, has rarely been reported in Asia. We genetically investigated a unique Chinese family with VHL disease and performed an analysis of the VHL protein stability.

METHODSGenomic deoxyribonucleic acid (DNA) extracted from peripheral blood was amplified by polymerase chain reaction (PCR) to three exons of the VHL gene in 9 members of the Chinese family with VHL disease. PCR products were directly sequenced. We estimated the effects of VHL gene mutation on the stability of pVHL, which is indicated by the free energy difference between the wild-type and the mutant protein (ΔΔG).

RESULTSThe Chinese family was classified as VHL type 1. Three family members, including two patients and a carrier, had a T to G heterozygotic missense mutation at nucleotide 515 of the VHL gene exon 1. This missense mutation resulted in the transition from leucine to arginine in amino acid 101 of the VHL protein. There was low stability of the VHL protein (the ΔΔG was 12.71 kcal/mol) caused by this missense mutation.

CONCLUSIONSWe first reported a family with this VHL gene mutation in Asia. This missense mutation is predicted to significantly reduce the stability of the VHL protein and contribute to the development of the renal cell carcinoma (RCC) phenotype displayed by this family. The genetic characterization and protein stability analysis of families with VHL disease are important for early diagnosis and prevention of the disease being passed on to their offspring.

Adult ; China ; Female ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Protein Stability ; Von Hippel-Lindau Tumor Suppressor Protein ; chemistry ; genetics ; von Hippel-Lindau Disease ; genetics

Objective To study the epidemiological characteristics on the clustering nature of pandemic (H1N1) 2009 in China.Methods Time and place distribution of pandemic (H1N1) 2009on the nature of clustering through data from Public Health Emergency Management Information System were described.Results As of August 10,2010,2773 pandemic (H1N1) 2009 clusters,a total of 77363 cases (including 20 deaths) were reported in the mainland of China.The most reported number of clusters was from schools and kindergartens with the total number of 2498 (accounted for 90.08％ of the total number).Middle schools appeared the have the most clusters (1223,accounting for 48.96％ ).The number of clusters reported in the southern provinces (cities) accounted for 77.03％ of the total,and was more than that in the northern provinces (cities).Two reported peaks in the southern provinces (cities) were in June and November,2009,respectively.There was only one reported peakin the northern provinces in September,2009.Conclusion Time and place distribution characteristics on the clusters of pandemic (H1N1) 2009 were similar to the seasonal influenza,but the beginning of winter peak was much earlier and intensity of reporting was much higher on the clusters of pandemic (H1N1 ) 2009 than that of seasonal influenza.

OBJECTIVETo investigate the diagnostic value of serum CEACAM1 in patients with pancreatic cancer.

METHODSFifty patients with pancreatic cancer and 50 with chronic pancreatitis were examine for serum levels of CEACAM1 by enzyme-linked immunosorbent assay (ELISA). The cut-off values and area under curve (AUC) of CEACAM1 was obtained by receiver operating characteristic (ROC) curve. The diagnostic efficiency of the tumor markers for pancreatic cancer was assessed by the fourfold table.

RESULTSThe serum level and positivity rate of CEACAM1 in pancreatic cancer patients were higher than those in chronic pancreatitis patients (P<0.05). Based on the ROC curve, the cut-off values and AUC of CEACAM1 were 13.835 ng/ml and 0.780, respectively (P<0.05). In pancreatic cancer patients, the diagnostic sensitivities of the tumor markers decreased in the order of CEACAM1 < CA242 < CA19-9 (P<0.05), and the specificity in the order of CA242 < CA19-9 < CEACAM1 (P<0.05).

CONCLUSIONCEACAM1 shows a higher diagnostic sensitivity than CA19-9 and CA242 for pancreatic cancer, but due to its low specificity this marker alone is not sufficient for diagnostic purposes.

Aged ; Aged, 80 and over ; Antigens, CD ; blood ; Biomarkers, Tumor ; blood ; Cell Adhesion Molecules ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Middle Aged ; Pancreatic Neoplasms ; blood ; diagnosis ; ROC Curve

Objective To elucidate the prevalence and the mutation pattern of N348I that related to the resistance seen in the AIDS patients, in China. Methods Partial pol gene of HIV-1 comprising of full protease (PR) and reverse transeriptase (RT) was obtained from plasma samples of therapy-failure individuals (n=614) and therapy-naive individuals (n=619) by using reverse transcription polymerase chain reaction(RT-PCR). 1233 sequences were then submitted to the HIV-1 drug resistance database of the Stanford University to analyze the prevalence and the emergence pattern of N348I. Results The prevalence of N348I was 6.5％ in the therapy-failure patients and 0.8％ in the naive individuals, respectively. The prevalence of N348I was more popular among those patients whose ART regimens containing zidovudine (AZT or ZDV) than those without AZT in regimens( 14.1％ vs. 4.7％, x2=10.21, P＜0.01 ). N3481 always emerged, and combined with others mutations among patients of ART, whose frequencies were: 85.0％ in combination with thymidine analog mutations (TAMs) and 52.5％ with M184V/I, respectively. Conclusion N348I was somehow prevalent in the therapy-failure patients when using the first-line antiretroviral drugs,and it emerged as unique patterns. This study laid the ground in improving the techaology of drug resistance genotypes detection and providing theoretical basis to study the mechanism of resistance and the law of molecular evolution.