AIM: To identify and determine compounds in supercritical CO_2 extraction from Hypericum perforatum L. METHODS: The contents of flavones and hyperforin were determined by reverse phase HPLC.The condition in supercritical fluid extraction(SFE) was: extraction temperature at 40 ?C,extraction pressure at 20 MPa,separation temperature at 45 ?C,separation presure at 8 MPa,extraction time 1 h. RESULTS: There was no flavones and the content of hyperforin was 180.6 mg/g in the sample of SFE using neat CO_2.The flavones and hyperforin were found in SFE extract using ethanol as modifier and the contents of rutin,quercetin,hyperoside,hyperforin were 1.10 mg/g、1.84 mg/g、1.33 mg/g、26.19 mg/g,respectively. CONCLUSION: The concentration of hyperforin by SFE with neat CO_2 is significantly higher than that with modified supercritical CO_2 extraction. The method of SFE with neat CO_2 can be used to prepare extracts with high concentration hyperforin.

With the rapid development of clinical laboratory technology and clinical improvement,the discordance between department of microbiology and clinical departments got more and more.Based on the analysis of interface issues existed between them,we discussed and explored how to do a wonderful job to communicate with each other.(Chin J Lab Med,2013,36:375-376)

AIM:To explore the effect on stability of hyperforin with different medicinal herbs and the changes of hyperforin content in herb combinations with Hypericum perforatum L.(HP).METHODS:Several common antidepressant medicinal herbs were selected separately to match with HP.The herbal samples consisted of HP group,HP with Curcuma wenyujin Y.H.Chen et C.Ling group,HP with Curcuma longa.L.group,HP with Epimedium brevicornum Maxim group and HP with Wuling powder(WY)group.The contents of hyperforin with each group were determined by reverse phase HPLC.RESULTS:The rates of extraction and retention of hyperforin in HP with Curcuma wenyujin Y.H.Chen et C.Ling group and HP with Curcuma longa.L.group were significantly lower than that of HP group.There were no obvious differences in the extraction rates of the HP group,HP with Epimedium brevicornum Maxim group and HP with WY group.The retention rate of hyperforin in HP group with WY powder was higher than that of HP group.The retention rate of hyperforin in HP with Curcuma wenyujin Y.H.Chen et C.Ling group,was lower than that of HP group.CONCLUSION:The results show that compatibility of HP with traditional Chinese medicine have effects on the stability of active herbal component.

Klebsiella Pneumoniae is an important pathogen for various infections clinically,in which hypervirulent Klebsiella pneumoniae (hvKP) mainly causes pyogenic liver abscess.By now,there are no standardized method to identify hvKP strains.hvKP strains are usually with hypermucoviscous phenotype,and the prevailing serotypes and clonal types are K1,K2 and CC23,CC65,respectively.Genes rmpA,magA,alls and kfu contribute to the pathogenicity of hvKP.This paper reviews the identification,clonal types and serotypes,hypermucoviscous phenotype and some other virulent genes of hvKP.

Objective Exploring different strengthening immune strategy on long-term immune memory effects for tuberculosis vaccine lays the theoretical foundation.Methods The fusion protein AMM,adjuvant DDA and BCG-PSN mixed built AMM subunit vaccine.Choose only 50 C57BL/6 mice (SPF),according to the table of random number method was divided into 5 groups and each group 10 only,the experiment 1 group injected mice to phosphate buffer solution (PBS) as control; Experiment 2 mice only injected BCG vaccine (BCG) from early; Experiment 3 mice BCG initial free,in 10 weeks with AMM subunit vaccine strengthen immunity; The four mice from early after BCG,respectively on 8 weeks,10 weeks with AMM subunit vaccine strengthen immune mice one time.Experimental 5 groups of mice after BCG initial free,respectively on 6 weeks,8 weeks,10 weeks with AMM subunit vaccine strengthen immune mice a.28 weeks to remove spleen lymphocytes in mice,add 2.5μg/ml Ag85B stimulus 72 h after collecting spleen cell culture supernatant,with ELISA test Ag85B specificity splenic lymphocyte caused IFN ppar-gamma level.Results Spleen cells by antigen Ag85B stimulation,BCG initial free,AMM subunit vaccine strengthen immune once,twice,three times group and PBS compared all had difference (P ＜0.01 orP ＜0.05) ;BCG initial free-AMM subunit vaccine strengthen immune once,twice,three times group and BCG compared all had difference (P ＜0.01 or P ＜0.05) ; BCG initial free-AMM subunit vaccine strengthen immune once and twice or three times group compared with difference (P ＜0.01 orP ＜0.05) ; Initial free-AMM subunit vaccine strengthen immune two group and three times than group difference was statistically significant (P ＞0.05).Conclusion Mice strong immune strengthened the number is not the more the better,the appropriate number of strengthen the immune to induce the most lasting immunity memory,subunit vaccine strengthen immunization strategy may affect the immune memory is one of the important factors.

OBJECTIVE:To compare clinical efficacy and safety of budesonide and beclometasone assisting with terbutaline by atomization inhalation in the treatment of acute pediatric asthma attack. METHODS:150 pediatric patients with acute asthma attack were randomly divided into group A and group B,with 75 cases in each group. Both group received symptomatic support treatment as low-flow oxygen inhalation,eliminating phlegm,relieving a cough,dilating bronchus and anti-infection,and atomization inhala-tion of budesonide,1 ml each time,bid. Based on this,group A was given atomization inhalation of budesonide,2 ml each time, bid;group B was given atomization inhalation of beclometasone,2 ml,bid. Treatment course of 2 groups lasted for 7 d. Clini-cal efficacy,asthma control,PEF% and the occurrence of ADR were observed in 2 groups;the time of symptoms and signs dis-appearance and hospitalization time were recorded,and symptoms and signs of children were scored 1 day before treatment and 7 days after treatment. RESULTS:During treatment,2 cases and 3 cases were withdraw from the test in group A and group B, respectively. The total effective rates of group A and B were 91.78% and 94.44%,and the rate of asthma control 95.89% and 94.44%,without statistical significance(P>0.05). The time of breathing difficulties disappearance,cough disappearance time, wheezing disappearance time and hospitalization time of group A were separately (2.11 ± 0.54),(3.28 ± 0.93),(4.38 ± 1.05), (5.83±1.29)d;those of B group were separately(2.07±0.52),(3.30±0.96),(4.45±1.08),(5.90±1.33)d,there was no sig-nificant difference between 2 groups(P>0.05). The clinical symptoms and signs score of group A before and after treatment were separately (4.28 ± 0.94) and (0.15 ± 0.04);those of before and after treatment of group B were separately (4.23 ± 0.91) and(0.11±0.03). There was significant difference before and after treatment(P<0.05),but there was no significant difference between 2 groups (P>0.05). The PEF% of group A before and after treatment were separately (51.4 ± 9.8)% and (67.2 ± 11.2)%;those of before and after treatment of group B were separately(52.0±7.1)%and(68.3±8.7)%. There was significant difference before and after treatment (P<0.05),but there was no significant difference between 2 groups (P>0.05). No obvi-ous ADR was found in 2 groups. CONCLUSIONS:Budesonide and beclometasone assisting with terbutaline by atomization inha-lation in the treatment for acute pediatric asthma attack possess the same effects and can efficiently relieve symptoms and signs, increase the rate of asthma control and shorten the rehabilita-tion course with good safety.

Objective To explore the changes of frequency, BAFF expression and sensitivity to apoptosis of CD19 + CD5 + B cells in endometriosis patients and investigate the relationship between CD19 +CD5 + B cells and endometriosis. Methods Flow cytometry and quantitative PCR were used to assess the frequency, BAFF expression and sensitivity to apoptosis of CD19+ CD5 + B cells in the peripheral blood,peritoneal fluids and endometrium biopsies of 39 patients with endometriosis and 31 patients with myoma of uterus ( control subjects). Results: All 39 endometriosis patients had higher frequencies ( all P ＜ 0. 01 ) of CD19 + CD5 + B cells in the peripheral blood, peritoneal fluids and endometrium biopsies than that in 31 control subjects. CD19 + CD5 + B cells from endometriosis patients expressed higher BAFF, and were more resistant to CD95L-induced apoptosis than the cells from control subjects ( P ＜ 0. 01 ). Conclusion CD19 + CD5 + B cells might play an important role in the pathogenesis of endometriosis through higher BAFF expression and more resistance to CD95L-induced apoptosis.

OBJECTIVE To evaluate the antibacterial effect of sulbactam combined with minocycline against 110 multidrug-resistant Acinetobacter baumannii strains.METHODS Checkerboard method was designed and agar dilution method was used for the MIC testing of sulbactam combined with minocycline against 110 multidrug-resistant A.baumannii strains,and the fractional inhibitory concentration(FIC) index was calculated according to MIC value.RESULTS The MIC50 of sulbactam was reduced significantly and the antimicrobial activities had been reinforced remarkably when combined with minocycline.The FIC results suggested that the main interaction be synergism(22%) and additivity(74%),with only 5% non-effect and without antagonism.CONCLUSIONS Synergism and additivity are the main effects for subactam when combined with minocycline against multidrug-resistant A.baumannii.

Objective To study the function of aminophenylboronic acid(APB)and clavulanate(CA)for detecting ESBLs in Enterobacter cloacae.Methods The phenotype of ESBLs of 61 Enterobacter cloacae isolates was detected with adding single beta-lactamase inhibitor CA to ceftazidime(CAZ)and cefotaxime(CTX),and double beta-lactamase inhibitors CA/APB to ceftazidime(CAZ)and cefotaxime(CTX)respectively.PCR was used to detect ESBLs genes of 61 Enterobacter cloacae isolates.The results of the enzymatic inhibitor potentiation test and PCR were compared and analyzed.Results With adding single enzymatic inhibitor CA to CAZ,28 isolates of Enterobacter cloacae producing ESBLs were detected,while 14 isolates were detected with adding CA to CTX.With adding double enzymatic inhibitors CA/APB to CAZ,28 isolates of Enterobacter cloacae producing ESBLs were detected,while 44 isolates were detected with adding CA/APB to CTX.By PCR positive ESBLs genes were detected in 47 isolates of Enterobacter cloacaes.Conclusions The potentiation test with double beta-lactamase inhibtion can be used to detect ESBLs in Enterobacter cloacae.

Objective To survey the bacterial species and drug resistance of bacteria isolated from blood, urine and other samples in 12 military hospitals located at different areas in China. Methods A total of 1099 non-repetitive bacterial isolates were collected from 12 military hospitals and sent to the General Hospital of PLA for re-identification and drug susceptibility test. The minimal inhibitory concentrations (MICs) of antimicrobial agents were determined by agar dilution method. The results were evaluated according to the standards of CLSI (2007) and analyzed by WHONET 5.4 software. ESBLs, AmpC ?-lactamases were detected using the confirmatory test and APB discs method, respectively. Results Gram positive cocci and gram negative bacilli constituted 39.7% and 60.3% of 1099 clinical isolates respectively. Methicillin-resistant Staphylococcus aureus (MRSA) accounted for 62%, and methicillin-resistant coagulase negative Staphylococcus (MRSCN) accounted for 92%. ESBLs-producing and AmpC-producing strains of Escherichia coli accounted for 51.1% and 11.3%, respectively, and of Klebsiella pneumoniae accounted for 45.1% and 16.2%, respectively. As to caftazidime, amikacin, cefotaxime, cefoxitin and levofloxacin, the resistance rate in Escherichia coli (E. coli) and Klebsiella pneumoniae isolated from blood was lower than that isolated from urine. However, as to meropenem, ceftazidime, polymyxin and minocyclin, the resistance rate in Pseudomonas aeruginosa and Acinetobacter spp isolated from blood was higher than that isolated from urine. Conclusion MRSA, MRSCN, and producers of ESBLs and AmpC ?-lactamases are common in military hospitals. Resistance pattern of bacteria from blood differs from that of bacteria from urine. It is necessary for military hospitals to take the bacterial distribution and resistance levels to antimicrobial agents under surveillance in order to guide the proper use of antibiotics for military doctors, and the results may serve as guidelines in the use of antimicrobial agents in war time.