[Objective]This paper aimed at discussing the application of pulse in the clinical through the analysis of medical cases by Wang Mengying. [Methods]Revisited the theory and practice of pulse in the medical cases of Wang Mengying and discussed Wang's experience in the clinical application of pulse in the diagnosis and treatment of diseases such as protration syndrome and cholera.[Results]The pulse is better than the other three in the diagnosis of patients with etiology and pathogenesis.Pulse can reflect the situation of patients ’etiology and pathogenesis.The process of disease can be plumbed through pulse diagnosis. [Conclusion] Ancient doctor attached great importance to the pulse,it can guide our clinical practice better through the study of pulse in modern clinical work.

OBJECTIVE:To improve the quality standard of Shuangshen capsule. METHODS:TLC was conducted for the qual-itative identification of Curcumnae radix and Rehmannia glutinosa in the preparation. HPLC was used for the content determination of salvianolic acid B in the preparation:the column was Diamonsil C18(2)with mobile phase of acetonitrile-0.1% H3PO4(22:78, V/V) at a flow rate of 0.5 ml/min,detection wavelength was 286 nm,column temperature was 30 ℃,and the injection volume was 2 μl. RESULTS:The TLC spots of C. radix and R. glutinosa were clear and well separated;the linear range of salvianolic ac-id B was 0.0792-0.792 μg(r=0.9999);RSDs of precision,stability and reproducibility tests were lower than 1%;recovery was 100.71%-101.82%(RSD=0.50%,n=6). CONCLUSIONS:The established standard can be used for the quality control of Shuang-shen capsule.

The weakness of the boiler high pressure steam sterilizer is put forward.The development process and actual application value of the hot steam electrical generator is mainly introduced.

Objective To find an ideal method of DNA isolation from blood and especially from clotted blood and to minimize the volume of blood collected for laboratory and clinical tests.Methods DNAs were isolated from antiagglutinated and agglutinated blood samples from auricular veins of 30 healthy subjects. The DNAs of these samples were obtained by a nonenzymatic, nontoxic procedure optimized by us and determinated by agarose gel electrophoesis and PCR. Results The yields of DNA isolated from clotted blood and antiagglutinated blood were (40.2?8.86)mg DNA/L and (39.1?10.2)mg DNA/L, and purities were 1.87?0.11 and 1.92? 0.12. The DNAs that we isolated from all samples had high molecular weight and by PCR the dimorphism of ALU alleles of the 8th intron of t-PA was easy to be obtained, so they were complete and reliable. Conclusion This method is rapid, easy, efficient and nontoxic for isolation of DNA from clotted and fresh blood and meets requirements for clinical testing and molecular biology study.

OBJECTIVE:To establish a RP-HPLC method for the determination of chlorogenic acid in Qingyan granules.METHODS:Waters Breeze System was used with a Hypersil BDS C18 column(5?m,4.6mm?150mm).The mobile phase was composed of methanol and 0.4%(V/V)H3PO4 water solution(20∶80,V/V),flow rate:1.0ml/min;wavelength of UV detector:327nm,column temperature 20℃.RESULTS:The calibration curve of chlorogenic acid was linear within the range of (174～1 392)ng(r=0.9 999).The average recovery was 100.10%,RSD=1.60%(n=5).CONCLUSION:This method is simple,sensitive,as well as reliable,and is available to quality control.

Aged, 80 and over ; Atrial Fibrillation ; complications ; Embolism ; complications ; Female ; Heart Atria ; pathology ; Heart Diseases ; complications ; Humans ; Myocardial Infarction ; etiology ; Thrombosis ; complications

OBJECTIVE:To establish a high performenc capillary zone electrophoresis method for the determination of the water-soluble active ingredients in different parts of the Salvia miltiorrhiza.METHODS:Capillary electrophoresis was conducted using uncoated capillary column(75 ?m?50.2 cm,effective length=40 cm)with 5 mmol?L-1 phosphate-borax(ph7.4)as running buffer by pressure injection(0.5 psi/4 s)and constant voltage separation(20 kV)with a detection length of 210 nm and a column temperature of 25 ℃.RESULTS:A complete baseline separation of PAH,DSS and PA was achieved within 8 min under the optimized conditions.The good linear range was from 2.5 ?g?mL-1 to 200.0 ?g?mL-1 for all the three ingredients.The average recoveries of the 3 ingredients were 100.04%,99.99%,and 100.01%,respectively,with RSD at 1.75%,1.73%,and 1.74%,respectively.CONCLUSION:The method is satisfactory in precision,recovery and linearity,and it can be used to determine three water-soluble active ingredients in different parts of the Salvia miltiorrhiza.

Objective: To investigate the acquired drug-resistant genes and strains relationship in 40 strains of Pseudomonas aeruginosa isolated from burn patients. Methods: Forty strains of Pseudomonas aeruginosa isolated from burn patients hospitalized in our burn department from January 2014 to December 2015 were selected, with 20 strains from each year. Kirby-Bauer paper disk diffusion method was used to detect sensitivity of the isolated Pseudomonas aeruginosa to 9 kinds of antibiotics of cefotaxime, ceftazidime, cefepime, imipenem, meropenem, gentamicin, amikacin, ciprofloxacin, and levofloxacin. Polymerase chain reaction was applied to detect 9 kinds of acquired β-lactamase antibiotics-resistant genes, outer membrane porin protein oprD2 genes, 12 kinds of acquired aminoglycosides antibiotics-resistant genes, and 6 kinds of acquired disinfectant-resistant genes and genetic marker genes of mobile genetic elements. Among the above genes, positive expression genes were verified by DNA sequencing and comparison. Sequences of twenty-eight acquired drug-resistant genes of the above 40 Pseudomonas aeruginosa strains were analyzed by unweighted pair-group method with arithmetic means cluster analysis. Results: Forty strains of Pseudomonas aeruginosa were resistant to the above 9 kinds of antibiotics. Two kinds of acquired β-lactamase antibiotics-resistant genes of bla_TEM, bla_CARB, 5 kinds of acquired aminoglycosides antibiotics-resistant genes of aac(6′)-Ⅰb, aac(6′)-Ⅱ, ant(2″)-Ⅰ, ant(3″)-Ⅰ, and rmtB, and 3 kinds of acquired disinfectant-resistant genes and genetic marker genes of mobile genetic elements of qacE△1-sul1, merA, and intⅠ1were detected in 40 strains of Pseudomonas aeruginosa with oprD2 gene deficiency. Forty strains aggregated obviously, with a total of 7 gene modes and 3 clones. Drug-resistant gene sequences of strains of number 2 to 4, 6 to 9, 11, 14, and 17 to 39 were similar and with close relationship. Drug-resistant gene sequences of number 12 and 13 strains were similar and with close relationship. Drug-resistant sequences of number 10 and 16 strains were similar and with close relationship. Conclusions Genes of bla_TEM, bla_CARB, aac(6′)-Ⅰb, aac(6′)-Ⅱ, ant(2″)-Ⅰ, rmtB, qacE△1-sul1, merA, and intⅠ1 were prevalent in these strains of Pseudomonas aeruginosa with oprD2 gene deficiency isolated from burn patients, which may play key roles in resistance of Pseudomonas aeruginosa to β-lactamase, aminoglycoside, and quinolone antibiotics, and the drug-resistant phenotypes were in good coincidence with genotypes. Pseudomonas aeruginosa strains isolated from burn patients were with similar acquired drug-resistant genes and close relationship.

Objective: To prepare a nanoprobe, anti-human melanoma ganglioside single chain variable fragment (GD/ScFvMEL) antibody conjugated with CdTe quantum dot, and to observe its ability to specifically bind human malignant melanoma cells. Methods: The GD/ScFvMEL gene was cloned into pET32a (+), and the plasmid was then transformed into E. coli BL21 (DE3) for GD/ScFvMEL protein antibody expression. The expressed GD/ScFvMEL antibody was purified by denaturing method and further refolded by modified dialysis method. The purified GD/ScFvMEL antibody was analyzed by SDS-PAGE. The GD/ScFvMEL-QDs nanoprobe was prepared by conjugating GD/ScFvMEL antibody with CdTe quantum dot, and its specificity was observed by incubating with MGC-803 cells and melanoma A375 cells. Results: The recombinant pET32a-GD/ScFvMEL was constructed and confirmed by PCR, restriction endonuclease analysis and DNA sequencing. The proportion of expressed GD/ScFvMEL antibody in total bacteria proteins was about 40% as detected by SDS-PAGE. The purified- and refolded-GD/ScFvMEL antibody was effectively conjugated with CdTe quantum dot, and the resulting GD/ScFvMEL-QDs nanoprobe was successfully prepared. The GD/ScFvMEL-QDs nanoprobe could specifically bind melanoma A375 cells, but could not bind stomach cancer MGC-803 cells. Conclusion: We have successfully prepared an anti-human melanoma ganglioside single-chain antibody-CdTe quantum dot nanoprobe, which can specifically bind melanoma cells.