OBJECTIVE To investigate the distribution of pathogens and their antibiotic resistance in lower respiratory tract infection(LRTI) from intensive care unit(ICU) in our hospital,and provide basis for rational selection of clinical drugs.METHODS Pathogens were detected from qualified sputum specimens in LRTI from ICU and identified by VITEK-AMS60 automatic microbial analyzing system.Drug susceptibility was determined by KB test.RESULTS From 320 sputum specimens 367 pathogens were detected between from Jan 2007 to Mar 2008,including 261 strains(71.1%) of Gram-negative bacilli,70 strains(19.1%) of fungi,and 36 strains(9.8%) of Gram-positive cocci.21.8% Of the isolated pathogens were Acinetobacter baumannii,with 16.7% of drug-resistant rate to cefoperazone/sulbactam and over 71% to other 13 antibiotic agents.The rate of extended spectrum ?-lactamases(ESBLs) producing Escherichia coli isolates and Klebsiella pneumoniae ones were 65.2% and 72.0%,respectively,comparing to 84.6% for meticillin-resistant Staphylococcus aureus(MRSA).CONCLUSIONS Gram-negative bacilli are the major pathogens of LRTI in ICU,in which A.baumannii shows with a high rate of drug-resistance,followed by fungi,which should attract the clinician′s more attention.

Objective To examine the antimicrobial susceptibility and prevalence of blaKPC gene in carbapenem-resistant Klebsiella pneumoniae (CRKP) strains isolated in Huashan Hospital,Fudan University.Methods The CRKP strains isolated in Huashan Hospital from January to December of 2014 were included in this study.The MICs of antibiotics were determined using CLSI broth dilution method.The blaKPC gene was amplified by polymerase chain reaction (PCR).Results A total of 205 CRKP strains were isolated,mainly from respiratory tract (76.1％,156/205) and urine specimens (18.5％,38/205).Antimicrobial susceptibility test indicated that CRKP isolates had higher resistance rates (85％-100％) to the antimicrobial agents except colistin (1.5％),tigecycline (0.5％),trimethoprim-sulfamethoxazole (51.0％) and amikacin (74.9％).Most (87.8％,180/205) of the CRKP strains were positive for blaKPC gene.Conclusions CRKP are mostly isolated from patients with lower respiratory tract infection and/or urinary tract infection in Huashan Hospital.The strains were highly resistant to the antibacterial agents tested except colistin and tigecycline.Production of KPC-type carbapenemase is the common mechanism of carbapenem resistance in these K.pneumoniae isolates.

Objective To establish a method for detection folylpolyglutamate syntbetase (FPGS),explore the change of FPGS in the drug-resistant A549 cells induced by methotrexate(MTX) enantiomer,and provide new tools to further investigate drug resistant mechanism. Methods A549 cell lines induced by L-( + )-MTX and D-( - )-MTX (25 μmol/L) were chosen to raise three cell lines as compared with MTX-sensitive cell line. Then FPGS were extracted for the CEIA-LIF and western blot was performed. After validation, FPGS antibodies were labeled by fluoreacein isothiocyanate (FITC) and produced a immune response with former-extracted FPGS. CELA-LIF can separate and detect labeled proteins according ruination time of the protein with different size and detect FPGS in drug resistant cell lines induced with L-(+)-MTX and D-(- )-MTX. The accuracy was evaluated as compared with western blot assay. Results The separation time of CEIA-LIF for labeled FPGS antibody and the immune complexes were7. 1 min and 8.9 min, and the resolving power was 4. 5. The process of protein separation and detection can be accomplished in less than 10 minutes. Western blot analysis showed there was no non-specific bands appears in the extract of these three cell lines after the freeze-thaw in liquid nitrogen. The minimum detection level in sensitive cell strains was 0. 68 mg/μl. The consent of FPGS in L-(+)-MTX and D-( - )-MTX induced cells were 46. 59% and 48. 36% compared with drug sensitive cell strains with CELA-LIF. Conclusions CELA-LIF was established in this experiment. It is efficient and sensitive for detecting of FPGS, which is similar to western blot method. The level of FPGS in L-( + )-MTX and D-( - )-MTX induced drug resistant cell lines is significantly lower, indicating the expression of FPGS is damaged.

Carbon nanotubes/Au nanoparticles ( CNT/AuNP ) composite film was fabricated on glassy carbon electrode ( GCE) by first dropping CNTs on the electrode surface and then electrodeposition of AuNPs by multi-potential step. The antibody of microcystin-( leucine-arginine ) ( anti-MCLR ) was immobilized on the modified electrode surface through adsorption on AuNPs. Subsequently, bovine serum albumin ( BSA) was used to block the non-specific adsorption to obtain the immunosensor for MCLR assay. The immunosensor could effectively capture MCLR by the specific immunoreaction between the electrode surface-confined antibody and MCLR, followed by the attachment of the anti-MCLR HRP-labeled to form a sandwich-type system. The analysis of MCLR was performed based on the catalytic reaction of HRP toward the oxidation of hydroquinone ( QH2 ) by H2 O2 . Under the optimal experimental conditions, the peak current response increased linearly with the concentration of MCLR in the range of 0 . 50-12 μg/L with a detection limit of 0. 30 μg/L (S/N=3). The developed immunosensor was used to determine MCLR in real water samples, and the recoveries of standard addition experiments were in the range of 93 . 0%-108 . 5%, with the relative standard deviation of 3 . 8%-5 . 0%.

The clinical characteristics of patients who presented in poor clinical grade due to ruptured middle cerebral artery aneurysms (MCAAs) associated with large sylvian hematomas (SylH) were analyzed and an ingenious designed prophylactic hinged craniectomy was introduced. Twenty-eight patients were graded into Hunt-Hess grades IV-V and emergency standard micro-neurosurgeries (aneurysm clipping, hematoma evacuation and prophylactic hinged craniectomy) were performed, and their clinical data were retrospectively analyzed. 46.43% of the patients reached encouraged favorable outcomes on discharge. The favorable outcome group and the poor outcome group significantly differed in terms of patients' anisocoria, Hunt-Hess grade before surgery, extent of the midline shift and time to the surgery after bleeding (P<0.05). There were no significant differences in age, sex, volume and location of the hematoma, size of aneurysm between the favorable and poor groups (P>0.05). However, ingenious designed prophylactic hinged craniectomy efficiently reduced the patients' intracranial pressure (ICP) after surgery. It was suggested that preoperative conditions such as Hunt-Hess grading, extent of the midline shift and the occurrence of cerebral hernia affect the prognosis of patients, but time to the surgery after bleeding and prophylactic hinged craniectomy are of significant importance for optimizing the prognosis of MCAA patients presenting with large SylH.

ObjectiveTo investigate the relationship between complement-3(C3) and progression of early cerebral infarction.Methods168 consecutive patients with acute cerebral infarction within the first 24 hours from onset were recruited.Fasting venous blood sample was drawn on the morning of the second or the third day after admission.C3 and other clinical chemistry variables were measured.The relationship of C3 and progressing stroke(PS) was analyzed by Logistic regression.Results68 cases(40.48%) were considered as PS.Level of serum C3 in PS patients was 1.28±0.20 g/L higher than in those with non-PS(1.16±0.25 g/L,P=(0.000)).A higher C3 accompanied a higher tendency of prevalence of PS(P=0.001).Logistic regression analysis showed that C3 was significantly correlated with PS(OR for each 1.0 g/L=7.27;95% CI=1.52～36.39; P=0.013).ConclusionElevated serum C3 may play an important role in the prevalence of PS.

Objective To compare the curative effect of open and minimally invasive treatment of acute closed achilles tendon rupture with 5 years followed-up study.Methods From September,2010 to January,2012,28 patients with acute closed Achilles tendon rupture in our hospital were followed up for 5 years.There were 21 males and 7 females.Minimally invasive percutaneous suture in 11 cases;open suture in the treatment of 17 cases.The patients were followed up at 6 months,1 year,2 years,3 years,4 years,and 5 years after AOFAS and ATRS score.Results The follow-up time ranged from 60 to 72 months.All incisions healed by first intention and no incision related complications occurred.2 groups of patients with ATRS score at 6 months after operation:the open group was 81.23±3.99,minimally invasive percutaneous group of 88.27±4.27,the difference between the two groups was statistically significantly.After 1 year,there was no significant difference in the scores between the two groups.Two groups of patients with AOFAS score at 6 months after operation:the open group was 69.00±6.23,minimally invasive percutaneous group of 79.27±4.83,the difference between the two groups was statistically significantly.At one year after operation,the open group was 85.53±3.38,and the minimally invasive group was more than 89.90±3.38.The difference between the two groups was statistically significantly.After 2 years,there was no significant difference in the scores between the two groups.Conclusion There is no significant difference between the 2 years after surgery in the treatment of acute closed Achilles tendon rupture or open surger.

OBJECTIVETo investigate the co-culture of keratinocytes with Malassezia isolates which cause the pityriasis versicolor with different color and to analyze the changes of cytokines associated with melanogenesis.

METHODSThe effects of Malassezia species with different proportions on the growth rate of keratinocytes was assessed with 5 g/L methyl thiazolyl tetrazolium (MTT). Co-culture of keratinocytes and Malassezia species were performed with isolates from hyer- and hypo-pigmentation areas of pityriasis versicolor. The supernatants were collected at different time points, and the changes of basic fibroblast growth factor (b-FGF), endothelin-1 (ET-1), nerve growth factor-beta (NGF-beta), interleukin-1alpha (IL-1alpha), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), stem cell factor (SCF) were recorded. Three control groups were established accordingly.

RESULTSWhen the ratio between keratinocytes and Malassezia species was lower than 1: 10, the growth rate of keratinocytes was not affected by Malassezia (P > 0.05). When the ratio was increased above 1:20, the growth rate of keratinocytes was significantly inhibited by Malassezia (P < 0.01). The secretions of IL-1alpha, IL-6, TNF-alpha, and ET-1 was significantly increased after the co-culture of keratinocytes and Malassezia (P < 0.01), while those of b-FGF, NGF-beta, and SCF had no significant changes (P > 0.05). Compared with the isolates from the hypo-pigmentation area, ET-1 induced by isolate from hyperpigmentation area significantly increased (P < 0.01).

CONCLUSIONWhen Malassezia isolates are co-cultured with keratinocytes, the secretions of cytokines associated with melanogenesis may differ from each other. ET-1 may play certain role in the hyper-pigmentation of pityriasis versicolor.

Cell Proliferation ; Cells, Cultured ; Cytokines ; biosynthesis ; Humans ; Keratinocytes ; cytology ; metabolism ; microbiology ; Malassezia ; isolation & purification ; physiology ; Melanins ; biosynthesis ; Tinea Versicolor ; microbiology

OBJECTIVETo explore different frequency electroacupuncture in acupuncture-drug compound anesthesia on analgesic effect and immune function in the pulmonary resection patients in order to recommend the best electroacupuncture frequency in acupuncture-drug compound anesthesia.

METHODSOne hundred and sixty-three patients scheduled for pneumonectomy were randomly divided into group A (n = 31), B (n = 34), C (n = 32), D (n = 34) and E (n = 32). Houxi (SI 3), Zhigou (TE 6), Neiguan (PC 6) and Hegu (LI 4) were selected in the five groups. Group A was treated with sham acupuncture by pasting needles without needle bodies at acupoints and electroacupuncture at the needle handle, and group B with 2 Hz electroacupuncture, and group C with 2 Hz/ 100 Hz electroacupuncture, and group D with 100 Hz electroacupuncture, and group E with 2 Hz/100 Hz transcetaneous acupoints electrical stimulation, and general anesthesia was produced in all the groups followed by 30 min electrical stimulation and the stimulation was lasted till the end of operation. The dosages of the anesthetics and the changes of surface antigen of leukomonocyte (CD3+ , CD4+ , CD8+ and CD4+/CD8+) and natural killer (NK) cell were observed at 1 day before surgery, intraoperative and 1 day after surgery.

RESULTSIn comparison with group A, the dosages of Propofol in group B and D were decreased, Fentanyl in group B, D and E were decreased. CD3+ and CD4+ in 5 groups increased at first and then decreased over time (all P < 0.01), and group E and C could inhibit the decrease of CD3+ and CD4+ in postoperation in comparison with that in the other three groups (all P < 0.05). CD8+ in 5 groups has no change in different times (all P > 0.05), and group E and D could inhibit the decrease of CD8+ in postoperation in comparison with that in the other three groups (all P < 0.05). CD4+/CD8+ in 5 groups has no change in different times (all P > 0.05), and group E and C could inhibit the decrease of CD4+/CD8+ in postoperation in comparison with that in the other three groups (all P < 0.05). NK cells of 5 groups were increased gradually in different times (all P < 0.01), with the similar regulation of NK cells (all P > 0.05).

CONCLUSIONAcupuncture-drug compound anesthesia with 2 Hz and 100 Hz electroacupuncture together with 2 Hz/100 Hz transcetaneous acupoints electrical stimulation have the best analgesic effect, and 2 Hz/100 Hz transcetaneous acupoints electrical stimulation and 2 Hz/100 Hz electroacupuncture have the best regulation of immune function. Acupuncture-drug compound anesthesia with 2 Hz/100 Hz transcetaneous acupoints electrical stimulation is recommended for that it can not only decrease the dosages of the anesthetics, but also significantly improve the immunosuppression in patients undergoing pneumonectomy.

Acupuncture Analgesia ; Adult ; Aged ; Anesthesia, General ; Anesthetics ; Electroacupuncture ; Female ; Humans ; Immune System ; immunology ; Killer Cells, Natural ; immunology ; Lung Diseases ; immunology ; surgery ; Male ; Middle Aged ; Pneumonectomy ; Young Adult