To study the ecological distribution and diversity of endophytic fungi associated with Ferula of medicinal plants in Xinjiang. The endophytic fungi were isolated from roots, stems and leaves of Ferula by microbiology research methods and technology. The endophytic fungi were identified using ITS rDNA sequence analysis and morphology analysis. The composition, diversity and preference of endophytic fungal community were analyzed with Shannon-Wiener biodiversity index (H') and Sorensen coefficient (Cs). A total of 337 strains endophytic fungi were isolated and classified into 38 genera, Alternaria, Aureobasidium and Fusarium were the dominant genera. Among the 337 isolates, the endophytic fungi of F. sinkiangensis were the most, The Shannon-Wiener biodiversity index (H') associated with roots of F. fukanensis was the highest, reached 1.85. The highest Sorensen coefficient ( Cs) was between leaf of F. sinkiangensis and leaf of F. ovina, reached 0.75. From the result, endophytic fungi were widely distributed in six Ferula, there are some notable differences between distribution and composition of the endophytic fungi isolated from different issues and different species of Ferula, show a certain degree of species and tissue preference. The results obtained in this study will provide realistic basis and theoretical basis for further study the secondary metabolites of endophytic fungi associated with Ferula, and the relationship between endophytic fungi and their host plants.
Biodiversity ; Ecology ; Endophytes ; isolation & purification ; Ferula ; microbiology ; Fungi ; classification ; isolation & purification ; metabolism

Objective To observe the expressions of bone morphogenetic protein-2(BMP-2)and bone morphogenetie protein-7(BMP-7)in the synovial tissue of fluorosis rats and its correlation with pathogenic mechanism of fluorosis arthritis.Methods Thirty-two SD rats were randomly divided into 4 groups:the control group,low,moderate and high-dose fluoride group.The control group ate commou fodder.The low,moderate and high dose fluoride group were fed with fodder composed of 25%.35%and 68%of corn(containing fluorine of 148.00 mg/kg)in chronic endemic fluorosis region in Guizhou Province.After 140 days,the expressions of BMP-2 and BMP-7 protein were determined by immunohistochemistry and assayed the absorbanee by computer image-pattern analysis system.Light microscope was used to observe the synovial tissue by Hematoxin Eosin,and calculated the pathological integral of synovium according to pathological grade standard.Results The expressions of BMP-2 (32.50±2.73)and BMP-7(38.90±2.56)in the control group was spare.Compared with the control group,the expressions of BMP-2(59.43±5.12,79.82±6.41,101.76±7.56)and BMP-7(55.10±4.82,78.42±5.61,98.46± 6.05)in the synovial tissue was up-regulated in each experimental groups(P＜0.05),especially in the moderate dose and the high-dose groups(P＜0.05).Compared with the control group(0.54±0.21).the pathological integral of synovium increased(P＜0.05)in each experimental groups(1.04±0.98,4.69±1.28,8.60±2.07).The expressions of BMP-2 and BMP-7 in the synovial tissue was found to be positively related with the pathological integral of synovium(r=0.98,0.99,P＜0.05).Conclusion The BMP-2 and BMP-7 play an important role in the development of fluorosis arthritis,probably by affecting osteogenesis.

Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.

Objective To investigate the epidemic characteristics of mycoplasmal pneumonia outbreak in a middle school, and to provide suggestions for outbreak prevention and control.Methods Principle and method of field epidemiology was used and,speciously,questionnaire investigation was carried out for the outbreak of mycoplasmal pneumonia.Outbreak characteristics and corresponding emergency intervention were discussed.Results There was an outbreak of mycoplasmal pneumonia in 2 classes of the middle school,and the attack rate was 42.50%.The attack rate of other classes was 6.75%.Timely identification and diagnosis of the disease,and comprehensive control to prevent the spread of the epidemic,brought the disease under control.Conclusion The outbreak of mycoplasmal pneumoniae infection is associated with the close contact in daily activities.Emergency intervention can prevent the further spread of the disease.

Objective To investigate the effect of reducing blood lipid in three effective parts of Rheum palmatum L.based on variation coefficient weight.Methods First of all,the SD rat model of hyperlipidemia was given the water extract,the ethanol extract and the ethyl acetate extract of Rheum palmatum L.,then the content of total cholesterol (TC),high densitylipoprotein cholesterol (HDL-C),low density lipoprotein cholesterol (LDL-C),triglyceride (TG) in serum were determined and general survival of rats was analyzed.Then using fuzzy matter-element model on variation coefficient weight to evaluate the comprehensive reducing blood lipid effect of different extract parts of Rheum palmatum L..In the end,combining with the result of the value of Euclid approach degree,compared the effect of normal group (M1),the model group (M2),the positive control group (Xuezhikang,M3),the water extract group(M4),the ethanol extract group (M5),the ethyl acetate extract group (M6) of blood lipid.Results Compared with model group,the water extract part and the ethanol extract part of Rheum palmatum L.could reduce the congtents of TC,LDL-C,TG and rise HDL-C.Combining with the result of the value of Euclid approach degree,the water extract was 0.74,the ethanol extract was 0.54.Combining with the result of the value of Euclid approach degree,the order of its magniude is M3 ＞ M4 ＞ M1 ＞ M5 ＞ M6 ＞ M2.Conclusion The water extract part is the effective part of reducing blood lipid.The method and model can be used to objectively evaluate the comprehensive effect of reducing blood lipid of Rheum palmatum L.'s three extracted parts.

Objective To screen the effective parts of loosening bowel and relieving constipation in mice from different extracted parts of Angelica and to investigate the utilization of fuzzy matter-element model in evaluating the effect of moisturizing intestine to relaxing the bowels of Angelica's different extracted parts.Methods Kunming mice were randomly divided into 11 groups,including normal group,model group (constipation),positive control group (Nongsuo Danggui Wan and Runchang Wan:20 μL · g-1),medicated group consisted by 7 different extracted parts of Angelica (3.4 g · kg-1 Angelica herbs).The constipation model was formed by the compound difenoxate solution 5.0 mg · kg-1 and finished after 30 min.We used the first defecation time as well as the dark stool points in 6 h,the weight of stools,the pushing rate of intestine and intestinal moisture as detection index and then using fuzzy matter-element model on variation coefficient weight to evaluate the comprehensive effect of different extract parts of Angelica.Results Time of first black stool discharge as well as the dark stool points in 6 h,the weight of stools,the pushing rate of intestine and intestinal moisture in model group were (241.90 ± 48.10) min,(9.00 ± 5.40) count,(149.10 ± 86.80) mg,(48.90 ± 16.50) ％,(1.45 ± 0.16) g,respectively.The 70％ ethanol impregnation part (part 2) on the factors were (172.60 ± 40.70) min,(17.30 ± 4.50) count,(223.70 ± 63.40) mg,(73.70 ± 13.10) ％,(1.71 ± 0.18) g,respectively.The difference between part 2 and model group was statistically significant in the factors (P ＜0.05 or P ＜0.01).Combining with the result of the value of Euclid approach degree,the maximum is 0.71 (part 2:70％ alcohol impregnation part).Conclusion The 70％ alcohol impregnation part is the effective part of loosening bowel to relieve constipation in mice.The method and model can be used to objectively evaluate the effect of moisturizing intestine to relaxing the bowels of Angelica' s different extracted parts.

Midkine is a heparin-binding growth factor, which plays important roles in the regulation of cell growth and differentiation. The non-tagged recombinant human midkine (rhMK) is therefore required to facilitate its functional studies of this important growth factor. In the present work, rhMK was expressed in Escherichia coli (E. coli) BL21 (DE3). The expression of midkine was efficiently induced by isopropyl-beta-D-thiogalactopyranoside (IPTG). After sonication, midkine was recovered in an insoluble form, and was dissolved in guanidine hydrochloride buffer. Renaturation of the denatured protein was carried out in the defined protein refolding buffer, and the refolded protein was purified using S-Sepharose ion-exchange chromatography. The final preparation of the rhMK was greater than 98% pure as measured by sodium dodecylsulfate-polyacrylamid gel electrophoresis (SDS-PAGE) and reverse phase high performance liquid chromatography (RP-HPLC). The purified rhMK enhanced the proliferation of NIH3T3 cells.
Animals ; Base Sequence ; Cell Proliferation ; drug effects ; Cytokines ; biosynthesis ; genetics ; isolation & purification ; pharmacology ; Escherichia coli ; genetics ; Humans ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Recombinant Proteins ; biosynthesis ; isolation & purification

OBJECTIVETo explore the impact of the waist circumference change on new onset diabetes (NOD) in the impaired fasting glucose (IFG) population.

METHODSA total of 12 657 subjects who took part in the health examination from 2006 to 2007 and from 2010 to 2011 from the employees of Kailuan Group and met the inclusion criteria were selected as the observation cohort.Of the 12 657 subjects, 10 697 were male, 1960 were female, with age of (49.9 ± 11.3) years old. According to the baseline waist circumference (WC) measurements and its quartile in the health examinations during 2006 to 2007, the observation population was divided into four groups (first, second, third and the fourth quartile groups) . Multiple logistic regression analysis was used to test the relation between the increasing of WC and NOD.

RESULTSThe incidences in the IFG population of NOD were 4.27% (1884/12 657) in the total population;4.25% (1581/10 697) in male and 4.44% (303/1960) in females, respectively (P < 0.05) . Along with increasing WC in the 4 quartile groups, the incidences of NOD was progressively increased, which were 2.19% (235/3083) , 3.07% (333/3114) , 4.47% (473/3037) and 7.08% (843/3423) , respectively;2.34% (213/2626) , 3.06% (282/2645) , 4.37% (393/2582), 7.00% (693/2844) in males and 1.38% (22/457) , 3.12% (51/469) , 5.05% (80/455) , 7.45% (150/579) in female (P < 0.05) . Multiple logistic regression analysis showed that compared with the first quartile group, the second, third and fourth quartile group had increased risk of NOD after adjusting for age, gender and other risk factors, the OR (95%CI) values were 1.38(1.13-1.68), 1.79 (1.47-2.09) and 3.10 (2.57-3.75), respectively.

CONCLUSIONThe incidence of NOD in the IFG population increased as the WC increased.

Adult ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; epidemiology ; Female ; Glucose Intolerance ; epidemiology ; Humans ; Incidence ; Male ; Middle Aged ; Risk Factors ; Waist Circumference

Glioblastoma is a common brain tumor and the overall survival rate of the patients is very low, so it is an effective way to develop the potential chemotherapy or adjuvant chemotherapy drugs in glioblastoma treatment. As a well-known antimalarial drug, artesunate(ARTs) has clear side effects, and recently it has been reported to have antitumor effects, but rarely reported in glioblastoma. Different concentrations of ARTs were used to treat the glioblastoma cells, and then the inhibitory effect of ARTs on glioblastoma proliferation was detected by MTT assay; Ki67 immunofluorescence assay was used to detect the proliferation of cells; Soft agar experiment was used to explain the clonal formation abilities ; Flow Cytometry was used to detect the cell cycle; and Western blot assay was used to determine the expression of key cell cycle protein. MTT assay results indicated that ARTs-treated glioblastoma cell A172, U251, U87 were significantly inhibited in a time-and-dose dependent manner as compared to the control group(DMSO treatment group). Soft agar experiment showed that ARTs could significantly reduce the clonal formation ability of glioblastoma. Furthermore, Flow cytometry analysis showed that ARTs could obviously increase the cell proportion in G₀/G₁ phase and reduce the cell proportion in S phase. Western blot results showed that the expressions of cell cycle-related proteins CDK2, CDK4, cyclin D1 and cyclin B1 were all obviously down-regulated. Above all, ARTs may inhibit the proliferation of glioblastoma cells by arresting cell cycle in G₀/G₁ phase through down-regulating the expression of CDK2, CDK4, cyclin D1, cyclin B1. These results may not only provide a novel method for rediscovering and reusing ARTs but also provide a new potential drug for treating glioblastoma.
Antineoplastic Agents ; pharmacology ; Apoptosis ; Artesunate ; pharmacology ; Cell Cycle Checkpoints ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cyclin B1 ; metabolism ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 2 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Glioblastoma ; drug therapy ; pathology ; Humans

Objective To select the effective parts of blood nourishing and to investigate the utilization of fuzzy matter-element model in evaluating the effect of blood nourishing of Angelica's different extracted parts.Methods KM mice half males and half females were randomly divided into 10 groups,including blank control group,model group (blood-deficiency),positive control group (Complex Prescription Glue Mucilage:0.02 mL· g-1),experimental group consisted by 7 different extracted parts of Angelica (3.4 g kg-1 Angelica herbs) Acetylphenyhydrazine (APH) and cyclophosphamide (CTX) were used to copy the blood-deficiency model mice by administration APH at the second day (20 mg · kg-1) and the fifth day (40 mg · kg-1) respectively,since the fifth day,ip CTX 40 mg · kg-1 everyday,once a day for four days.Periphera hemogram and organ index,including thymus index (TI),spleen index (SI),were detected at the 10th day.Then using Fuzzy Matter-element model on variation coefficient weight.Results The TI,SI,red blood cell (RBC),platelet count(PLT),hematokrit (HCT),hemoglobin (HGB),white blood cell (WBC) were (7.70 ± 1.81) × 10-2,(56.90 ± 12.23) × 10-2,8.44 ± 1.59,1453.10 ± 102.50,56.54 ± 6.69,170.00 ± 26.39,2.10 ± 0.49,respectively in model group.The 70％ ethanol impregnation part (part 2) for TI,SI,RBC,PLT,HCT,HGB,WBC were 0.10 ± 2.66 ×10-2,(33.83 ±8.97) × 10-2,11.94 ±2.07,1749.60 ±211.50,55.53 ±9.42,55.53 ±9.42,2.60 ± 0.47,respectively.The part 2 can increase TI,RBC,PLT,HCT,WBC and reduce SI,HGB.Compared with model group,the difference was statistically significant in SI,RBC,PLT,HCT,WBC (P ＜ 0.05 or P ＜ 0.001).There were no significant differences between part 1 and part 2 (P ＞ 0.05).Combining with the result of the value of Euclid approach degree,in part 1-7,the maximum is 0.50 (part 2),is greater than 0.40 (part 1).Conclusion The 70％ alcohol impregnation part is the effective part of blood nourishing.The method and model can be used to objectively evaluate the comprehensive effect of blood nourishing of Angelica's different extracted parts.