Objective To explore the whole quality control method for lactones and flavonoids in the raw material of Herb Sarcandra.Methods TLC identification of ethyl acetate extracts and spectrophotometry determination of total flavonoids had been conducted.Results Chemical compositions extracted by ethyl acetate and total flavonoids content varied greatly in the raw materials of Herb Sarcandra from different producing areas.Conclusion TLC identification combined with total flavonoids determination can be used for the quality control of the medicinal material of Herb Sarcandra.

Objective To observe the somatic embryogenesis in in-vitro cultured Dendrobium candidum wall.Ex Lindl.(DCWL),and to supply evidence for its rapid propagation and germplasm preservation.Methods Protocorm-like bodies and callus of DCWL subcultured for 30 days was used as the explants,N6 was used as the basic culture with phytohormone added,and fungal extracts as the elicitor.Protocorm-like bodies and callus of DCWL were used for induction culture.Results Somatic embryogenesis in in-vitro cultured DCWL derived from the epithelial cells or inner cells of callus of DCWL.Conclusion A large amount of buds can be obtained by the induction and culture of protocorm-like bodies and callus of DCWL.

Objective To construct a Polyphosphate kinase 1 ( ppk1) gene deletion mutant of uro-pathogenic Escherichia coli (E.coli) CFT073, and to explore the biological properties of the mutant strain . Methods The ppk1 gene deletion strain (△pk1) was constructed based on CFT073 E.coli strain by usingλRed homologous recombination technology .A comparison analysis was conducted on adhesive and invasive abilities between CFT073 wild type strain and △pk1 strain in in vitro model of human bladder cancer epithe-lial cell 5637 .Crystal violet staining method was used to evaluate the influences of ppk1 gene deletion on biofilm formation.Results The CFT073 ppk1 deletion mutant strain was successfully constructed .Com-pared with the wild type strain ,△pk1 strain showed impaired adhesive and invasive abilities to 5637 cells. Moreover , the absorbance values of crystal violet at 570 nm at each time point of the mutant strain group were also lower than those of the wild-type strain group .Conclusion The ppk1 gene deletion mutant of uro-pathogenic E.coli CFT073 could be successfully constructed by Red homologous recombination technology and its biological properties indicates that ppk1 gene plays an important role in the pathogenesis of uropatho-genic E.coli infection through regulating the abilities of adhesion , invasion and biofilm formation .

Objective To explore the role of ppk1 gene in E.coli CFT073 strain during urinary tract infection (UTI). Methods C57BL/6 mouse models of acute UTI with the wild-type(CFT073) and ppk1 mutant (△pk1) infected, were used to compare the bacteria colonization and inflammation induction abilities of bladder tissues. Results In the mouse models, the △pk1 strain showed a significantly lower infection rate (73.3% vs 93.3%) and lower adhesion frequency of bladder (0.01%vs 0.5%) than those of the CFT073 strain. The expression of IL-6 and TNF-αwere reduced in the bladder of △pk1 infected group (P<0.05). Hematoxylin-Eosin tissue staining showed that the damage degree of bladders in △pk1 infected mice were less serious than the CFT073 infected mice. Conclusion ppk1 gene plays an important role in E.coli colonization to bladder and the inflammation induction ability.

OBJECTIVE:To explore the feasibility of question-and-answer practice teaching in the pharmacy department of our hospital. METHODS:The practice teaching of pharmacy department was taken as a pilot,and the question-and-answer practice teaching was used in emergency and outpatient pharmacies,inpatient pharmacy,PIVAS,drug warehouse,clinical pharmaceutics room,preparation room and drug testing laboratory. The effects of the practice teaching on internships,teachers,pharmacy depart-ments and patients were excavated. RESULTS & CONCLUSIONS:Satisfaction of teachers,pharmacy,patients and clinics for the question-and-answer practice teaching was 100%,and satisfaction of internships was 94.5%. The question-and-answer practice teaching has improved their professional knowledge and competence,resolved difficulty of patients about drug counseling and helped the improvement of overall business level in department.

Objective To evaluate the short-term prediction of high-sensitivity cardiac troponin T (hs-cTnT) and other cardiovascular risk biomarkers in patients undergoing maintenance hemodialysis (MHD).Methods We conducted a cohort survey in 296 consecutive MHD patients whose clinical data were retrospectively analyzed.Before MHD,hs-cTnT and other relative cardiovascular biomarkers were detected.The end point (all-cause death) and time of occurring were recorded in the next 13 months.The differences between survival and all-cause death were analyzed by t-test,Mann-Whitney test and x2 test.The best two percentile cutoff point was calculated by X-tile and the survival rate was calculated by Kaplan-Meier Logistic regression analysis was applied to analyze the odd ratio between high risk and non-high risk hs-cTnT group.Non-high risk group was divided into intermediate risk and low risk group based on the 99th percentile of hs-cTnT in healthy population,to further evaluate its short-term prediction value for MHD patients.The short-term significance of hs-cTnT was proved to be independently associated with all-cause death by Logistic regression analysis.Results The mean value of serum hs-cTnT in survival group was 0.05 (0.03～0.07) ng/mL,while in the death group it was 0.07 (0.04～0.14) ng/mL,which had statistical significance (P =0.027).The best two percentile cutoff of hs-cTnT in MHD patients was 0.1 ng/mL.The survival rate in high risk group (hs-cTnT＞0.1 ng/mL) is lower than it in non-high risk group (hs-cTnT≤0.1 ng/mL) (76.67％ vs.96.62％,P ＜0.05).The odd ratios for high risk group and non-high risk group was 7.288 (P＜ 0.001).Moreover,further grouping the non-high risk group by hs-cTnT =0.014 ng/mL,intermediate risk group (hs-cTnT＞0.014 ng/mL) group has lower survival rate than low risk group (hs-cTnT≤0.014ng/mL),while there wasn't any death case occurred in the low risk group.Conclusions Hs-cTnT is an independent risk factor to all-cause death.Thus hs-cTnT can be a strong indicator of short-term prediction and prognostic evaluation.

Objective To investigate the role and the mechanism of ppk1 gene (coding for polyphosphate kinase 1) in oxidative stress resistance in uropathogenic Escherichia coli (UPEC).MethodsMutant strains with ppk1-deletion (△pk1) and complemented strains (△pk1-C) were constructed based on the UPEC strain CFT073.A comparative analysis was conducted to analyze survival rates of CFT073, △pk1 and △pk1-C strains at different time points while they were under oxidative stress.Differences in protein expression between CFT073 and △pk1 strains were analyzed using mass spectrometric analysis.Differences between CFT073 and △pk1 strains in expression of katG and katE genes were analyzed using real-time quantitative RT-PCR.Results The survival rate of △pk1 strains was lower than that of CFT073 strains at every time point, while the survival rate of △pk1-C strains was basically the same as that of CFT073 strains.Gel image analysis and mass spectrometric analysis revealed that six proteins were down-regulated and one was up-regulated in △pk1 strains as compared with those in CFT073 strains.Expression of the catalase-coding genes katG and katE in △pk1 strains were respectively (20.5±8.2)% and (20.9±6.9)% of those in CFT073 strains (P<0.05).Conclusion The ppk1 gene plays an important role in oxidative stress resistance in UPEC by modulating the expression of catalase-coding genes katG and katE.

OBJECTIVETo study the changes in biological behaviors of meningitis E. coli K1 strain E44 after deletion of polyphosphate kinase 1 (ppk1) gene and explore the role of ppk1 in the pathogenesis of E. coli K1-induced meningitis.

METHODSThe wild-type strain E. coli K1 and ppk1 deletion mutant were exposed to heat at 56 degrees celsius; for 6 min, and their survival rates were determined. The adhesion and invasion of the bacteria to human brain microvascular endothelial cells (HBMECs) were observed using electron microscopy and quantitative tests. HBMECs were co-incubated with wild-type strain or ppk1 deletion mutant, and the cytoskeleton rearrangement was observed under laser scanning confocal microscope.

RESULTSThe survival rate of the ppk1 deletion mutant was significantly lower than that of the wild-type strain after heat exposure. The ppk1 deletion mutant also showed lowered cell adhesion and invasion abilities and weakened ability to induce cytoskeleton rearrangement in HBMECs.

CONCLUSIONSppk1 gene is important for E.coli K1 for heat resistance, cell adhesion and invasion, and for inducing cytoskeletal rearrangement in HBMECs.

Brain ; cytology ; Cells, Cultured ; Cytoskeleton ; Endothelial Cells ; cytology ; microbiology ; Escherichia coli ; genetics ; physiology ; Escherichia coli Proteins ; genetics ; Gene Deletion ; Humans ; Phosphotransferases (Phosphate Group Acceptor) ; genetics

Objective:To establish an automatic planning method using volumetric-modulated arc therapy (VMAT) for primary liver cancer (PLC) radiotherapy based on predicting the feasibility dose-volume histogram (DVH) and evaluate its performance.Methods:Ten patients with PLC were randomly chosen in this retrospective study. Pinnacle Auto-Planning was used to design the VMAT automatic plan, and the feasibility DVH curve was obtained through the PlanIQ dose prediction, and the initial optimization objectives of the automatic plan were set according to the displayed feasible objectives interval. The plans were accessed according to dosimetric parameters of the planning target volume and organs at risk as well as the monitor units. All patients′ automatic plans were compared with clinically accepted manual plans by using the paired t-test. Results:There was no significant difference of the planning target volume D 2%, D 98%, D mean or homogeneity index between the automatic and manual plans ((58.55±2.81) Gy vs.(57.98±4.17) Gy, (47.15±1.58) Gy vs.(47.82±1.38) Gy, (53.14±0.95) Gy vs.(53.44±1.67) Gy and 1.15±0.05 vs. 1.14±0.07, all P>0.05). The planning target volume conformity index of the manual plan was slightly higher than that of the automatic plan (0.77±0.08 vs. 0.69±0.06, P<0.05). The mean doses of normal liver, V 30Gy, V 20Gy, V 10Gy, V 5Gy and V< 5Gy of the automatic plan were significantly better than those of the manual plan ((26.68±11.13)% vs.(28.00±10.95)%, (29.96±11.50)% vs.(31.89±11.51)%, (34.88±11.51)% vs.(38.66±11.67)%, (45.38±12.40)% vs.(50.74±13.56)%, and (628.52±191.80) cm 3vs.(563.15±188.39) cm 3, all P<0.05). The mean doses of the small intestine, the duodenum, and the heart, as well as lung V 10 of the automatic plan were significantly less than those of the manual plan ((1.83±2.17) Gy vs.(2.37±2.81) Gy, (9.15±9.36) Gy vs.(11.18±10.49) Gy, and (5.44±3.10) Gy vs.(6.25±3.26) Gy, as well as (12.70±7.08)% vs.(14.47±8.11)%, all P<0.05). Monitor units did not significantly differ between two plans ((710.67±163.72) MU vs.(707.53±155.89) MU, P>0.05). Conclusions:The automatic planning method using VMAT for PLC radiotherapy based on predicting the feasibility DVH enhances the quality for PLC plans, especially in terms of normal liver sparing. Besides, it also has advantages for the protection of the intestine, whole lung and heart.