The history and present status of phytoplasma classification are introduced briefly in this paper.The newly classification methods and rules for the description of Candidatus species are reviewed.The key problems and direction on the classification and identification of phytoplasmas in China are discussed.

Objective: To learn anxiety, depression, sleep quality and quality of life among patients with primary pruritus ani, and to analyze their correlation with anal itching symptom, so as to provide insights into the prevention and treatment of primary pruritus ani. Methods: Patients with primary pruritus ani were selected from anorectal department of the First Affiliated Hospital of Zhejiang Chinese Medical University. The Visual Analogue Scale, pruritus frequency score, Hospital Anxiety and Depression Scale, Pittsburgh Sleep Quality Index and Dermatology Life Quality Index (Chinese version) were used in the survey. Spearman rank correlation analysis was employed to analyze the correlation between the degree and frequency of pruritus ani with, anxiety, depression, sleep quality and quality of life. Results: Sixty patients with primary pruritus ani were investigated, including of 7 males and 53 females, with an average age of (34.37±10.30) years. The patients with mild, moderate and severe pruritus accounted for 28.33%, 48.33% and 23.34%, respectively. The patients with pruritus frequency of grade 1, 2 and 3 accounted for 51.67%, 38.33% and 10.00%, respectively. The patients with anxiety, depression, and coexistence of anxiety and depression accounted for 18.33%, 13.33% and 10.00%, respectively. The patients with very good, good, average and poor sleep quality accounted for 15.00%, 46.66%, 30.00% and 8.34%, respectively. No, mild, moderate and severe impacts on quality of life among the patients accounted for 10.00%, 46.66%, 33.34% and 10.00%, respectively. The degree of pruritus was positively correlated with anxiety (rs=0.405, P=0.001), depression (rs=0.343, P=0.007), sleep quality (rs=0.293, P=0.037) and quality of life (rs=0.338, P=0.008). However, there was no significant correlation between the frequency of pruritus with the above factors (all P>0.05). Conclusion The degree of pruritus in patients with primary pruritus ani is related to anxiety, depression, sleep quality and quality of life.

"Omics" and bioinformatics have brought new ideas to the study of traditional Chinese medicine. This study used metabonomics and network pharmacology to investigate the pharmacodynamic basis and regulation of Qishen Yiqi dropping pill (QDP) improving cardiac energy metabolism in rats with heart failure (HF). ¹H NMR metabonomics analysis showed that eight metabolites, including carnitine, glutamine, creatine, proline, homocitrulline, lactic acid, taurine and alanine appeared significant callback after QDP treatment for HF. The results indicate that QDP regulates the metabolism of carbohydrate, lipid, ATP and protein. The animal experiment was conducted in accordance with the regulations of the Ethics Committee for Experimental Animal Management and Animal Welfare of Institute of Materia Medica, Chinese Academy of Medical Sciences. A "drug-component-target-disease" network was established using network pharmacology, and the "component-target" sub-network related to the above energy metabolism processes was extracted by combining metabonomics results. Results revealed 79 chemical compounds and 47 potential targets of QDP involved in the regulation of energy metabolism, and identified key chemical components including ursolic acid, notoginsenoside G, ginsenoside-Rh1, and core targets such as INS, PPARG, and AKT1. The results also demonstrated the complex multi-target and multi-component relationship between QDP and HF from the perspective of energy metabolism. The molecular docking technique verified a strong interaction between some targets and chemical compounds, with affinities less than -5 kcal·mol^-1. The results of this study provide useful information for the clinical application, development, and utilization of QDP.

Objective To study the etiology of nosocomial infection and the risk factors in senile patients with acute myelogenous leukemia who received chemotherapy, then to explore the preventive measure. Methods 9lcases of senile patients with AML received 304 times of chemotherapy, then to analyze the rate of nosocomial infection, the infection site, the result of etiological examination and the relationship between the rate of nosocomial infection and the absolute of neutrophil count in peripheral blood and the duration of agranulocytosis. And then the results were compared with the non-senile patients with AML. Results The statistical indicators of nosocomial infection in senile patients with AML was higher than those younger ones. The rate of infection in lower respiratory tract, oral, alimentary canal, skin, anal area and the rate of sepsis in senile people was higher than those younger ones.Positive rate of fungal infection of senile patients was higher. The nosocomial infection in the patients whose absolute neutrophil count in peripheral blood was no more than 0.5×l09/L,and prolonged over 7 days was specific higher. Conclusions Senile patients with AML is the high risk group for nosocomial infection. Age, chemotherapy, the neutrocytopenia level and prolonged time are related to the nosocomial infection in senile patients with AML. It is necessary to take positive measure to control nosocominal infection and advance life quality of senile patients.

Objective To investigate the levels of trace elements such as fluorine(F), and aluminium (Al)etc. of osteomalacia malformation children and to make etiological diagnosis in reference with clinical manifestations.Methods Urine and occipitalia hairs of 14 diseased children(patient group) from endemic fluorosis area and 13 healthy children(control group) from non-endemic area were included in the study on November, 2008, and contents of 10 elements of fluorine(F), aluminum(Al), chromium(Cr), manganese(Mn), ferrum(Fe), cuprum(Cu), zinc(Zn), arsenic (As), selenium(Se), strontium(Sr), and barium(Ba) were tested. The data were analyzed with medical soft package PEMS 3.1. Results Urinary contents of F, Al, Mn, Cu, Sr, and Se(1.18 mg/L, 112.6 μg/L,6.62,29.86 mg/L, 177.5,4.23 ng/L) in patient group were significantly different from those in control group (0.48,47.1,2.04,16.61 mg/L, 55.17,15.52 ng/L, t = 4.592,2.486,4.850,2.210 2.078,2.912, all P＜ 0.05); Hair contents of Al, Mn, As, Sr, Ba, Fe, and Se in patient group(59.27,5.26,0.96,1.50,1.29,297.13,0.45 mg/kg)were significantly different from those of control group( 18.69,0.72,1.09,0.62,0.68,69.02,1.323 mg/kg, t = 4.583,6.318,3.309,2.704,5.606,6.294, all P ＜ 0.05); in patient group, the correlation coefficients of urinary Fe to Al,Zn, As, and Se were all bigger tan 0.662(all P＜ 0.05), those of urinary Se to Mn, Ba, Cu, Zn, Sr, and As were all bigger than 0.694(all P＜ 0.05), those among urinary Mn, Sr, As, and Ba were bigger than 0.550(all P＜0.05), those of hair Al to Mn, Cr, Fe, and Cu were bigger than 0.732(all P＜ 0.05), those of hair Ba to Mn,Cr, Fe, and Sr, and of hair Mn to Cr and Fe, and those between Cr and As, between Cu and Sr were all bigger than 0.686 (all P ＜ 0.05). In control group, the correlation coefficients of urinary Cu to Zn, Se, and Ba, those of Zn to Se and Ba, and those of Cr to Mn and Ba were all bigger than 0.516(all P ＜ 0.05), those of hair Al to Mn,Fe, Cu, As, and Se, and those of hair Se to Fe, Cu, and As, those of hair Fe to Mn, Cu, and As, those of hair Cu to Zn and As, and that between Zn and As were bigger than 0.739(all P ＜ 0.05). The correlation coefficient of urinary F to Se in patient group(0.762) was significantly different from that in control group( - 0.469, u = 2.079,P ＜ 0.05). Conclusions The burden of F and Al of osteomalacia malformation children in endemic fluorosis area of Shuicheng county is too high. The contents of multi-elements in urine and hairs and their correlation are coincident with high levels of Al and F and they cause network increase of multi-element content changes and their correlation. According to bone X-ray features combining with the living environment, the diagnosis of endemic Al-F fluorosis can be made. The biological significance of reducing urinary and hair Se levels and the correlations of F and Al need to be further studied.

Objective To construct a recombinant Bacillus Calmette-Guerin ( BCG ) vaccine strain, rBCG::Rv3478-pMV261, expressing the Rv3478 protein of Mycobacterium tuberculosis and to inves-tigate its immunogenicity.Methods The gene fragments encoding Rv3478 antigen were amplified by PCR and then respectively cloned into pMV261 and pET-28a vectors to construct the recombinant expression plas-mids (Rv3478-pMV261 and Rv3478-pET-28a).The Rv3478-pMV261 plasmids were transformed into the BCG cells to construct the rBCG vaccine strains, while the Rv3478-pET-28a plasmids were transformed into Escherichia coli BL21 strains for the expression of Rv3478 protein.Polyclonal antibodies were induced in mice upon the immunization with Rv3478 protein.The rBCG vaccine strains overexpressing Rv3478 protein were screened out with Western blot assay.The C57BL/6 mice were divided into four groups including the PBS treated group, BCG treated group, rBCG::pMV261 ( R0) treated group and rBCG::Rv3478-pMV261 ( R3) treated group.All mice were sacrificed in 4 or 12 weeks after immunization.Enzyme-linked immunos-pot assay ( ELISPOT) , ELISA and flow cytometry analysis were performed to evaluate the induced humoral and cell-mediated immune responses in mice.Results The Rv3478 protein was successfully expressed and could induce polyclonal antibodies in mice.High levels of IFN-γand TNF-αwere detected in mice treated with R3, indicating that the immunization with R3 enhanced the cellular immunity.Moreover, the ratios of CD4+to CD8+T cells and the percentages of CD44+CD62L+T cells were increased in mice upon the immuni-zation with R3.Conclusion The recombinant BCG vaccine strain overexpressing Rv3478 protein could in-duce stronger cell-mediated immune responses in mice.It might be have a great significance as a new tuber-culosis( TB) vaccine strain against TB infection in the future.

A strain of P. Aeruginosa,which was seperated from clinical environment,shows a special characteristic. It keeps normal short rod shape when cultured at 37℃, however,it forms filament without pyocyanin producing when cultured at 25℃ overnight. The filaments will divide and form short rods, simultaneously, produce pyocyanin when culture time is prolonged to over 72h or culture temperature is raised to 37℃. The preliminary study indicates that this phenomenin has nothing to do with nutritive conditions and could the inbluenced by inoculating density and irradiating with ultraviolet rays The absence of pyocyanin was not the cause of filamentous formation by the test results.

Objective To investigate the biosecurity and biocompatibility of small intestinal submucosa (SIS) as scaffold for tissue engineering and to explore the possibility to induce synovial mesenchymal stem cells (SMSCs) differentiate into cartilage with SIS as the scaffold and SMSCs as the seed cells. Methods The SMSCs were isolated and cultured from the synovial mem-brane of knee joints of rabbits by a conventional method. The SIS was harvested from pigs according to the physical method and Abraham's method. 4 rabbits are divided into the experimental group and control group. The biosecurity of SIS as scaf-folds were investigated in pyrogen test, skin sensitization test and systemic toxicity test. The SMSCs and SIS were co-cultured in vitro and induced to differentiate into cartilage to explore the biocompatibility of SMSCs and SIS, the proliferation and differ-entiation ability of SMSCs on the scaffold of SIS. The varietyies were identified by the microscope. Results The SIS isolated with the physical method and Abraham's method is a milky and translucent membrane with a smooth surface. Under the electron microscope, SIS presented a porous Stereoscopic three-dimensional network structure, which is formed by fibrous tissues' intertex-ture. Its' porosity was about 80%and its aperture was 100-200μm. Meanwhile, the biosecurity and biocompatibility of SIS were also fine. In the trial that the SMSCs and SIS were co-cultured in vitro, the SMSCs can grow, adhere to and differentiate on the sur-face of SIS and into the hollows very well. It can also secrete extracellular matrix. Through scanning microscope observation, cells contact with each other by their neuritis, or adhered to the wall of hole by cellular protrution. On the surface of SIS, the SMSCs maintain the property that it can easily differentiate into the chondrogenic lineage in the chondrogenic medium. Immunochemistry staining showed positive expression of type II collagen post 21 days of co-cultrue. Conclusion SIS can be used as the scaffold to construct tissue engineering meniscus as it has good biosecurity and biocompatibility with SMSCs without disturbing the cell form or inhibiting the growth and function of SMSCs.

Purpose: The purpose of the current study was to explore the functions and potential mechanism of miR-451a in breast cancer (BC). Methods: Quantitative reverse transcription real-time polymerase chain reaction was used to analyze the expression of miR-451a in human normal mammary cells (MCF-10A) and BC cells. Colony formation assay, terminal-deoxynucleoitidyl transferase mediated nick end labeling assay and transwell assays were conducted to validate the effect of miR-451a on proliferation, apoptosis, migration and invasion of BC cells, respectively. RNA pull-down, RNA immunoprecipitation and luciferase reporter assays were applied to investigate the upstream and downstream mechanisms of miR-451a in BC cells. Results: MiR-451a was expressed at a low level in BC cells. Overexpression of miR-451a repressed BC cells proliferation, migration and invasion. Moreover, long non-coding RNA AC092127.1 acted as a sponge of miR-451a to enhance the expression level of AE binding protein 2 (AEBP2) that was demonstrated to be the target gene of miR-451a in BC cells. Finally, rescue experiments validated that miR-451a and AEBP2 involved in AC092127.1-mediated BC cell growth, migration and invasion. Conclusion In a word, AC092127.1/miR-451a/AEBP2 axis contributes to BC cell growth, migration and invasion. Our results may help to find novel potential targets for BC treatment.

Purpose: The purpose of the current study was to explore the functions and potential mechanism of miR-451a in breast cancer (BC). Methods: Quantitative reverse transcription real-time polymerase chain reaction was used to analyze the expression of miR-451a in human normal mammary cells (MCF-10A) and BC cells. Colony formation assay, terminal-deoxynucleoitidyl transferase mediated nick end labeling assay and transwell assays were conducted to validate the effect of miR-451a on proliferation, apoptosis, migration and invasion of BC cells, respectively. RNA pull-down, RNA immunoprecipitation and luciferase reporter assays were applied to investigate the upstream and downstream mechanisms of miR-451a in BC cells. Results: MiR-451a was expressed at a low level in BC cells. Overexpression of miR-451a repressed BC cells proliferation, migration and invasion. Moreover, long non-coding RNA AC092127.1 acted as a sponge of miR-451a to enhance the expression level of AE binding protein 2 (AEBP2) that was demonstrated to be the target gene of miR-451a in BC cells. Finally, rescue experiments validated that miR-451a and AEBP2 involved in AC092127.1-mediated BC cell growth, migration and invasion. Conclusion In a word, AC092127.1/miR-451a/AEBP2 axis contributes to BC cell growth, migration and invasion. Our results may help to find novel potential targets for BC treatment.