Objective To evaluate the capabilities of disc diffusion and Vitek2-compact GN13 methods for testing antimicrobial susceptibility of screening ESBLs ( extended-spectrumβ-lactamase) in En-terobacteriaceae clinical isolates.Methods A total of 93 Enterobacteriaceae strains were isolated from pa-tients with intra-abdominal infections in 21 hospitals during 2011 to 2012.The in vitro minimum inhibition concentration ( MIC ) values of ampicillin-sulbactam, piperacillin-tazobactam, ertapenem, ceftazidime, ceftriaxone, cefepime, imipenem, amikacin, ciprofloxacin and levofloxacin were determined by disc diffu-sion, Vitek2-compact GN13 and broth microdilution methods, respectively.Categorical agreement ( CA ) rates of disc diffusion and Vitek2-compact GN13 methods were determined by using broth microdilution meth-od as the reference method.The genes encoding ESBLs were screened in Escherichia coli (E.coli), Kleb-siella pneumoniae (K.pneumonia), Klebsiella oxytoca (K.oxytoca) and Proteus mirabilis (P.mirabilis) strains by using PCR analysis and gene sequencing.Disc diffusion and Vitek2-compact GN13 methods were used for the phenotypic confirmatory test of ESBLs and the sensitivity, specificity, positive predictive value and negative predictive value of the two tests were evaluated.Results The CA values of disc diffusion and Vitek2-compact GN13 methods for the 10 antibiotics were all >90% as compared with broth microdilution method.The major error (ME) rate for ertapenem was 3.2%and the very major error (VME) rates for am- picillin-sulbactam, ceftazidime and cefepime tests were all 2.2% by using Vitek2-compact GN13 method. The sensitivity, specificity, positive predictive value and negative predictive value of disc diffusion and Vitek2-compact GN13 methods in the phenotypic confirmatory test of ESBLs were 96.7%(29/30), 100%(20/20), 100%(30/30) and 95%(19/20), respectively.Conclusion Both disc diffusion and Vitek2-compact GN13 methods could be used for testing the antimicrobial susceptibility and the detection of ESBLs in Enterobacteriaceae clinical isolates with the advantage of accuarcy.Attention should be paid to the posibil-lity of oaurance of ME and VME when testing ertapenem, ampicillin-sulbactam, ceftazidime and cefepime by using Vitek2-compact GN13 method.

AIM: To discuss the effect of wearing customized curvature soft corneal contact lens to dry eye degree of flight attendants.
METHODS:Eighty cases (160 eyes) of flight attendants from China Southern were divided into two groups:control group 40 cases ( 80 eyes ) wearing ready-made Bausch soft corneal contact lens ( curvature 8. 4 ); the experiment group 40 cases ( 80 eyes ) , wearing Bausch soft corneal contact lens with customized curvature. Tear break-up time ( BUT ) , Schirmer Ⅰ test ( SⅠt ) and fluorescein ( FL ) staining were as dry eye evaluation index. The results was statistically analyzed.
RESULTS: BUT, SⅠt average shortening value of the experimental group were less than that of the control group, there was statistical significance (P<0. 01). FL staining positive increase, the number of experimental group was fewer than that of control group, with statistical significance (P<0. 05).
CONCLUSION: Wearing customized curvature soft corneal contact lens can prevent the flight dry eye more effectively than fixed curvature product.

Objective To investigate the correlation between the formation of pigmented biliary calculus and biliary H.pylori infection.Methods Bile from 35 patients with pigmented biliary calculus and 10 healthy controls was cultured for aerobic,anaerobic and H.pylori.The expression of H.pylori- DNA in bile,bile duct mucosa and pigmented calculus were determined by PCR.The expression of H. pylori associated protein in bile duct mucosa was determined by Western-blot and Warthin-Starry staining.Results H.pylori culture was negative in all bile samples.In 35 patients with biliary pigmen- ted calculus,H.pylori was detected by PCR in the center of calculus,bile and bile duct mucosa of 14.29%,31.43% and 56.67% patients,respectively.Among H.pylori-DNA positive bile samples,7 contained anti-CagA antibodies,and 6 contained Vac A.in addition to Vacuolating cytotoxin(35000), glycoprotein(30000),Urase Band Urase A.Bacteria resembling H.pylori by Warthin-Starry stainning were found in 7 of 30(23.33%)bile duct mueosal samples from patients with biliary pigmented calculus. H.pylori-DNA and its associated protein were not detected in all bile and bile duct mucosae samples from the healthy controls.Conclusions The evidence of H.pylori-DNA and its associated protein in biliary system might indicate the role of H.pylori in the formation of biliary pigmented calculus.

Objective To study the protective effect of polyene phosphatidylcholine on hepatic injury induced by oxaliplatin and 5?fluorouracil. Methods A subcutaneous tumor model was established by transplanting colocarcinoma HCT116 cells into 30 nude mice,which were random?ized into three groups. The polyene phosphatidylcholine group was injected with polyene phosphatidylcholine(85 mg · kg-1 · d-1)and 5?fluorouracil (20 mg·kg-1·d-1)for 7 days,and then injected with oxaliplatin(6 mg·kg-1·d-1)for 1 day. The hepatic injury group was injected with 5?fluoroura?cil(20 mg·kg-1·d-1)for 7 days and oxaliplatin(6 mg·kg-1·d-1)for 1 day. The tumor?bearing blank group was injected with normal saline. Hepat?ic injury was observed with ultrathin pathological sections. Liver homogenates were prepared to detect superoxide dismutase(SOD)and catalase (CAT)activity. Results In the hepatic injury group,pathological sections revealed dissolved cellular cytoplasm,mitochondrial membrane dam?age,cell membrane edema,and fuzzy,sinusoidal cell expansion . There were no obvious hepatic injuries observed in the polyene phosphatidylcho?line group. The expression of SOD and CAT were lower in the hepatic injury and polyene phosphatidylcholine groups compared to the tumor?bear?ing blank group(P<0.05). The expression of SOD and CAT were higher in the polyene phosphatidylcholine group compared to the hepatic injury group(P<0.05). Conclusion Polyene phosphatidylcholine has a protective effect on hepatic injury induced by oxaliplatin and 5?fluorouracil, which may be related to its effect on membrane repair and inhibition of oxidative stress.

Objective To evaluate the performance of domestic matrix-assisted laser desorption/ionization time-of-flight mass spectrometry system Clin-TOF-Ⅱ MS with BioExplorer V2.3 database ( Clin-TOF MS system) on gram-negative bacteria identification.Methods This was a methodological comparison study.A total of 1 025 gram-negative strains of 32 genus, 56 species or species complex were included in this study from 1999 to 2000 and 2014 to 2016 in Peking Union Medical College Hospital.The Bruker Biotyper MS system ( Bruker MS system ) , Bruker Autoflex Speed with Biotyper v 3.1 database were used as control.Identification by both MALDI-TOF MS systems were parallel conducted by direct smear method.The 16S rDNA sequencing based identification was performed when either MALDI-TOF MS system gave“unbelievable result” or results from two systems were not consistent.Results Amongst the isolates studied, 98.05% (1 005/1 025) was correctly identified to species or species complex level by Clin-TOF MS system.Comparatively, 99.22%(1 017/1 025) was correctly identified by Bruker MS system.There were 17 isolates just identified to genus level and 2 isolates were “no identification” by Clin-TOF MS system, meanwhile 1 Pseudomonas monteilii misidentified as P.putida.There were only two 2 isolates identified to genus level and 3 isolates were“no identification” by Bruker MS system.But it misidentified all 3 Aeromonas hydrophila (2 isolates as A.caviae and 1 isolate as A.media).It′s noted that both MS systems identified 1 Chryseobacterium gleum and 1 C. bernardetii to genus level.Conclusion The identification capability of domestic Clin-TOF MS system was good on gram-negative bacteria.

This is to report the screening, extracting and validating antitumor components and compounds from Stellera chamaejasme L. under the case of discrete distribution of active data. In this work, different components from Stellera chamaejasme L. were collected by HPD macroporous resin and polyamide resin column, and their antitumor activity on A549 were tested by MTT assay. Activity results indicate that activity of components at 30-39 min is more potent than that of Stellera chamaejasme L. extract, and the activity of components at 33.97 min is equivalent to positive drug, cis-platinum at 100 microg x mL(-1), but with totally different mode of action. Under the case of discrete activity, the weight analysis is capable of screening active components and compounds from natural products.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Drug Screening Assays, Antitumor ; Humans ; Thymelaeaceae ; chemistry

Penaeidin-2 (Pen-2) is an important antimicrobial peptide derived from the Pacific white shrimp, Penaeus vannamei, and possesses both antibacterial and antifungal activities. Recent studies suggest that recombinant penaeidins show similar activities to the native Pen-2 protein. Previous researches have shown that some antimicrobial peptides (AMPs) exhibit cytotoxic activity against cancer cells. To date, there have been no studies on the antitumor effects of Pen-2. This study evaluated the potential of recombinant pen-2 (rPen-2) in the selective killing of kidney cancer cell lines ACHN and A498, and its action mechanism. MTT assays found the maximal growth inhibition of HK-2, ACHN and A498 cells treated with 100 μg/mL rPen-2 at 48 h was 13.2%, 62.4%, and 70.4%, respectively. DNA-specific fluorescent dye staining showed a high percentage of apoptosis on cancer cells. Flow cytometry revealed that the apoptosis rate of HK-2, ACHN and A498 cells was 15.2%, 55.2%, and 61.5% at 48 h respectively, suggesting that rPen-2 induced higher apoptosis rate in cancer cells than in HK-2 cells. Laser confocal scanning microscopy demonstrated that the plasma membrane was the key site where rPen-2 interacted with and destroyed tumor cells. Scanning electron microscopy showed the morphologic changes of the cell membranes of kidney cancer cells treated with rPen-2. These results suggest that rPen-2 is a novel potential therapeutic agent that may be useful in treating kidney cancers.

Objective To investigate the effects of Sini decoction on function of hypothalamic-pituitary-adrenal axis in patients with sepsis.Methods A prospective single-blind randomized controlled trial was conducted.60 septic patients were divided into three groups with the method of random number table,20 cases in the control group,20 in the Chinese herb group,and 20 in corticoid group.All of them received routine treatment.Patients in Chinese herb group were given Sini decoction in addition (decoction of monkshood 15 g,dried ginger 15 g,honey-fried licorice 10 g) 100 mL/d orally or by nasal feeding,while patients in corticoid group were given hydrocortisone 200 mg/d intravenously instead,both for 7 days.Before the treatment,3 days and 14 days after treatment,blood was collected to determine the levels of adrenocorticotropic hormone (ACTH) and cortisol,and the result of ACTH stimulating test was observed.At the same time,acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score was recorded,and 3-day shock recovery rate and 28-day death rate were also compared among these groups.Results None of the three groups showed different result in ACTH stimulating test (x2=1.101,P=0.605).ACTH in three groups was gradually decreased.Compared with that before treatment,ACTH in Chinese herb group and corticoid groups began to decrease obviously on day 3 (ng/L:29.90 ± 3.31 vs.33.10 ±.3.31,28.20 ±.2.45 vs.33.30 ± 3.84,both P＜0.01),while in control group declined ACTH appeared later (on day 14) compared with before treatment (ng/L:29.40 ±5.63 vs.33.50 ±4.89,P＜0.05).No obvious difference in ACTH level was showed between the Chinese herb group and the cortical group (both P＞0.05).Cortisol level in both Chinese herb and cortical groups showed a raise-fall biphase trend while there was no change in the control.The cortical levels on day 3 in Chinese herb and cortical groups were much higher than that before treatment (μg/L:343.04 ± 31.20 vs.294.70 ±42.10,331.25 ±42.80 vs.280.36 ± 38.10,both P＜0.01) and that of control group (μg/L:291.61 ± 41.50,both P＜0.01),though no significant statistical difference was observed between two groups (both P＞0.05).APACHE Ⅱ score on day 14 in control,Chinese herb and cortical groups was significantly lower than that before treatment (16.8 ± 5.1 vs.20.1 ± 4.3,13.4 ± 3.2 vs.18.3 ± 3.8,15.1 ± 2.5 vs.19.5 ± 4.0,all P＜0.01),and the score was much lower in Chinese herb group comparing with that of control group (P＜0.05).No statistical difference was observed among control,Chinese herb and cortical groups in lowering 28-day death rate [35.0％ (7/20),25.0％ (5/20),20.0％ (4/20)] and improving 3-day shock recovery rate [40.0％ (8/20),70.0％ (14/20),60.0％ (12/20),all P＞0.05].Conclusions Sini decoction could elevate cortisol while lower ACTH at the early stage of sepsis.Sini decoction could also effectively improve symptoms and hypothalamic-pituitary-adrenal axis function in septic patients without affecting death rate.

Objectives To study the performance of different microbial automated inoculation systems and to evaluate the performance of the Probact microbial automated inoculation and incubation system ( Probact system) and its applications in clinical microbiology laboratory.Methods A total of 160 clinical specimens, including respiratory secretions ( n=61 ) , urine ( n=49 ) , and feces ( n=50 ) , that were submitted to the Clinical Microbiology Laboratory in Peking Union Medical College Hospital of Chinese Academy of Medical Sciences from February 2015 to April 2015 were evaluated.These specimens were processed with conventional manual method, the Probact automated inoculation system, and PREVI Isola Inoculator.The quantity of bacterial species recovery, number of effectively isolated colonies, total number of colonies recovery per plate, and time of processing the 160 specimens by the three methods were evaluated. Wilcoxon signed-rank test and Kruskal-Wallis rank sum test were used for statistical analysis.Results The Probact system had significantly higher quantity of bacterial species recovery (respiratory specimens 3.41 ±1.40, urine 1.92 ±0.86, and feces 1.16 ±0.79) than those by the Isola Inoculator (respiratory specimens 3.75 ±1.29, urine 2.24 ±0.97, and feces 1.92 ±0.72), (P=0.006, 0.011, <0.001).Compared to the manual method, Probact performed less quantity of bacterial species recovery for respiratory specimens(3.85 ±1.38), but higher in feces(0.80 ±0.81)( P<0.001).There is no significant differences for urine ( 1.84 ±1.23 ) ( P=0.266 ) .As for number of isolated colony, the Probact system ( respiratory specimens 12.16 ±7.72, urine 2.71 ±4.24, and feces 5.40 ±5.04 ) had significant smaller numbers than that of Isola Inoculator (respiratory specimens 16.56 ±5.76, urine 4.35 ± 4.89, and feces 8.40 ±3.70) (P<0.001,0.007,0.003).However, both system had larger numbers of isolated colonies than those by the manual method (respiratory specimens 11.30 ±8.42, urine 2.67 ±4.34, and feces 1.90 ±3.90) and the difference was significant for fecal specimens(P<0.001).Regarding the total number of colonies recovery, larger number was found by Isola Inoculator than that by the Probact system for fecal specimens, however, there were no significant differences for respiratory or urine specimens (P=0.524,0.738).Compared with manual method, the Probact system had significantly more numbers of colonies recovery for respiratory and fecal specimens ( P<0.001 ) . The total time for processing 160 specimens was shortest for manual method (281 min), followed by Probact system (419 min) and Isola Inoculator (495 min) .Conclusions The performance of the Probact system is better than the manual method but no superior to the Isola Inoculator.The Probact system can meet the clinical need in terms of full automation and standardization of specimen inoculation and prevention of bias of processing by laboratory staffs using manual method.