Development a prediction model for identifying bacterial meningitis in young infants aged 29-90 days: a retrospective analysis.

BACKGROUND: The early diagnosis and treatment of bacterial meningitis (BM) in young infants was very critical. But, it was difficult to make a definite diagnosis in the early stage due to nonspecific clinical symptoms. Our objectives were to find the risk factors associated with BM and develop a prediction model of BM especially for young infants. METHODS: We retrospectively reviewed the clinical data of young infants with meningitis between January 2011 and December 2020 in Children's Hospital of Soochow University. The independent risk factors of young infants with BM were screened using univariate and multivariate logistic regression analyses. The independent risk factors were used to construct a new scoring model and compared with Bacterial Meningitis Score (BMS) and Meningitis Score for Emergencies (MSE) models. RESULTS: Among the 102 young infants included, there were 44 cases of BM and 58 of aseptic meningitis. Group B Streptococcus (22, 50.0%) and Escherichia coli (14, 31.8%) were the main pathogens of BM in the young infants. Multivariate logistic regression analysis identified procalcitonin (PCT), cerebrospinal fluid (CSF) glucose, CSF protein as independent risk factors for young infants with BM. We assigned one point for CSF glucose ≤ 1.86 mmol/L, two points were assigned for PCT ≥ 3.80 ng/ml and CSF protein ≥ 1269 mg/L. Using the not low risk criterion (score ≥ 1) with our new prediction model, we identified the young infantile BM with 100% (95% CI 91.9%-100%) sensitivity and 60.3% (95% CI 46.4%-72.9%) specificity. Compared with BMS and MSE model, our prediction model had larger area under receiver operating characteristic curve and higher specificity, the differences were statistically significant. CONCLUSION: Our new scoring model for young infants can facilitate early identification of BM and has a better performance than BMS and MSE models.

Identification of ADA as a Biomarker for Atypical Epstein Barr Virus Infection in Children.

OBJECTIVE: This study aims to explore the ability of adenosine deaminase (ADA) to discriminate atypical Epstein Barr virus (EBV) infection in children from acute febrile illness. METHODS: All children admitted to the Children's Hospital of Soochow University between 2018 and 2019, who were acute febrile patients and subjected to the plasma EBV-DNA polymerase chain reaction (PCR) and indirect immunofluorescence (IIF) assay for EBV-specific antibodies assays. The diagnostic value of each detection index was compared by the area under the ROC curve. RESULTS: In children with atypical Epstein Barr virus infection, the sensitivity, specificity, positive predictive value, negative predictive value and Youden index were 62.87%, 100.00%,100.00%, 61.73% and 0.63 for EBV-DNA PCR assay, 80.84%, 100.00%, 100.00%, 75.76% and 0.81 for VCA-IgG avidity and 89.22%, 87.00%, 91.98%, 82.86% and 0.76 for ADA. VCA-IgG avidity (AUC=0.904, P<0.01) and ADA (AUC=0.881, P<0.01) assays had the great diagnostic efficiency. In addition, the sensitivity, specificity and AUC were 92.75%,91.43% and 0.921(95%CI: 0.856-0.985) for ADA in the course≤3 days group, respectively. CONCLUSIONS: ADA has a good diagnostic value in the early stage of atypical EBV infection, and is not affected by primary EBV infection and reactivation. SCHLüSSELWöRTER: Adenosine deaminase, Epstein -Barr virus, Biomarker, children.

Low-dose clarithromycin therapy modulates CD4(+) T-cell responses in a mouse model of chronic Pseudomonas aeruginosa lung infection.

BACKGROUND AND OBJECTIVE: Low-dose clarithromycin (CAM) is widely used for the treatment of chronic respiratory infections. However, its anti-inflammatory mechanisms have not been fully explored. As CD4(+) T cells play an important role in the initiation of immune responses to infectious microorganisms, we aimed to investigate the effects of low-dose CAM on CD4(+) T-cell responses. METHODS: Fifty-four BALB/c mice were randomly divided into three groups: a control group (inoculated with sterile agarose beads and treated with saline from day 7), a saline group (inoculated with Pseudomonas aeruginosa-loaded beads and treated with saline from day 7) and a CAM group (identical to the saline group, except that saline was replaced by CAM solution). Bronchoalveolar lavage (BAL) fluid cell counts, bacterial load, lung tissue histology and pulmonary CD3(+) CD4(+) cell numbers were assessed. Levels of T helper (Th)1/Th2/Th17 cytokines and suppressor cytokines (interleukin (IL)-10 and transforming growth factor-ß1) were analysed. Messenger RNA (mRNA) levels for transcription factors for CD4(+) T-cell subsets were determined. RESULTS: The CAM group had lower BAL fluid cell counts, pathological scores and pulmonary CD3(+) CD4(+) cell numbers compared with the saline group, whereas the bacterial load was not significantly different. Levels of Th1/Th17 cytokines and expression of a transcription factor for naturally occurring regulatory T cells (Treg) were significantly decreased in the CAM group compared with the saline group, whereas there was no significant difference in GATA-3 mRNA expression. CONCLUSIONS: This study demonstrated a downregulation of Th1/Th17/naturally occurring Treg responses after treatment with low-dose CAM in mice with chronic P. aeruginosa lung infection.

Effect of Lactobacillus plantarum LP-Onlly on gut flora and colitis in interleukin-10 knockout mice.

BACKGROUND AND AIM: Probiotics are used in the therapy of inflammatory bowel disease. This study aimed to determine the effects of probiotic Lactobacillus plantarum LP-Onlly (LP) on gut flora and colitis in interleukin-10 knockout (IL-10(-/-) ) mice, a model of spontaneous colitis. METHODS: IL-10(-/-) and wild-type mice were used at 8 weeks of age and LP by gavage was administered at a dose of 10(9) cells/day per mice for 4 weeks. Mice were maintained for another one week without LP treatment. The colonic tissues were collected for histological and ultrastructural analysis at death after 4 weeks treatment of LP, and the feces were collected at 1-week intervals throughout the experiment for the analysis of gut flora and LP using selective culture-based techniques. RESULTS: Compared with control mice, IL-10(-/-) mice developed a severe intestinal inflammation and tissue damage, and had an abnormal composition of gut microflora. LP administration attenuated colitis with the decreased inflammatory scoring and histological injury in the colon of IL-10(-/-) mice. In addition, LP administration increased the numbers of beneficial total bifidobacteria and lactobacilli, and decreased the numbers of potential pathogenic enterococci and Clostridium perfringens, although the decrease of coliforms was not significant after LP treatment in IL-10(-/-) mice. CONCLUSIONS: Oral administration of LP was effective in the treatment of colitis, with the direct modification of gut microflora in IL-10(-/-) mice. This probiotic strain could be used as a potential adjuvant in the therapy of inflammatory bowel disease, although further studies are required in human.

Lactobacillus plantarum ameliorates colonic epithelial barrier dysfunction by modulating the apical junctional complex and PepT1 in IL-10 knockout mice.

Probiotics are efficacious in the treatment of inflammatory bowel disease. However, the precise mechanisms remain unknown. To determine whether probiotic Lactobacillus plantarum (LP) ameliorates colonic epithelial barrier dysfunction present in interleukin-10 knockout (IL-10⁻(/)⁻) mice, IL-10⁻(/)⁻ and wild-type mice received LP or the vehicle for 4 wk. Colitis was assessed by histological scores and clinical manifestation, and gut paracellular permeability was measured by Ussing chamber. Oligopeptide transporter 1 (PepT1)-mediated transepithelial transport was evaluated by measuring the plasma cephalexin concentration. The expression and distribution of apical junctional complex (AJC) proteins and PepT1 were determined by Western blotting and immunofluorescence and their mRNA by reverse transcriptase-PCR. Spontaneous colitis was observed in all IL-10⁻(/)⁻ mice in which paracellular permeability was increased, in conjunction with decreased expression and redistribution of zonula occludens-1, occludin, claudin-1, and ß-catenin. PepT1 expression was increased, accompanied with an enhanced cephalexin transport. Colonic epithelial barrier dysfunction was further confirmed by increased bacterial translocation and proinflammatory cytokine production. Treatment with LP decreased colonic paracellular permeability with restoration of expression and distribution of AJC proteins and partially prevented PepT1 expression and cephalexin transport in IL-10⁻(/)⁻ mice. Moreover, treatment with LP also prevented bacterial translocation and proinflammatory cytokine production in IL-10⁻(/)⁻ mice. Results from this study indicated that treatment with LP may ameliorate colonic epithelial barrier dysfunction in IL-10⁻(/)⁻ mice, by modulating the AJC- and PepT1-mediated transepithelial transport.

L. plantarum prevents enteroinvasive Escherichia coli-induced tight junction proteins changes in intestinal epithelial cells.

BACKGROUND: It is increasingly recognized that Lactobacillus plantarum (L. plantarum) has the ability to protect against Enteropathogenic Escherichia coli (EPEC)-induced damage of the epithelial monolayer barrier function by preventing changes in host cell morphology, attaching/effacing (A/E) lesion formation, monolayer resistance, and macromolecular permeability. However, the cellular mechanism involved in this protective effect still remained to be clarified. METHODS: This study was to investigate the effect of L. plantarum on the changes of Caco-2 cells responding to Enteroinvasive Escherichia coli (EIEC), the permeability of cell monolayer and the transmissivity of dextran, and the distribution and expression of the tight junction (TJ) proteins, such as Claudin-1, Occludin, JAM-1 and ZO-1 were examined when infected with EIEC or adhesived of L. plantarum after infection by confocal laser scanning microscopy (CLSM), immunohistochemistry and Western blotting, the cytoskeleton protein F-actin were observed with FITC-phalloidin. RESULTS: This study demonstrated that the transepithelial electrical resistance (TER) step down and dextran integrated intensity (DII) step up with time after infected with EIEC, but after treating with L. plantarum, the changes of TER and DII were improved as compared with EIEC group. L. plantarum prevented the damage of expression and rearrangement of Claudin-1, Occludin, JAM-1 and ZO-1 proteins induced by EIEC, and could ameliorate the injury of cytoskeleton protein F-actin infected with EIEC. CONCLUSION: L. plantarum exerted a protective effect against the damage to integrity of Caco-2 monolayer cells and the structure and distribution of TJ proteins by EIEC infection.

The relationship between air quality and respiratory pathogens among children in Suzhou City.

OBJECTIVE: We studied the short-term effects of air pollutant concentrations in Suzhou City on respiratory infections in children of different age groups. METHODS: We employed clinical data from children hospitalized with respiratory infections at the Children's Hospital of Soochow University during 2014-2016, and air quality for Suzhou City covering the same period.We investigated the relationships between the air pollutant concentrations and respiratory tract infections in children by causative pathogen using time series models with lagged effects. RESULTS: The results of single-pollutant models showed that PM2.5, PM10, NO2, SO2 and CO had statistically significant associations with respiratory tract infections in children under 3 years, with the largest effect sizes at a lag of 3 weeks. Notably, the multi-pollutant model found PM2.5 was significantly associated with viral respiratory in children under 7 months, and bacterial respiratory infections in other age groups, while PM10 concentrations were associated with viral infections in preschool children. CONCLUSION: PM2.5, PM10 and NO2 are the main atmospheric pollutants in Suzhou. The associations between pollutant concentrations and viral and bacterial respiratory infections were stronger among children under 3 years than for older age group.s PM2.5 had the strongest influence on viral and Mycoplasma pneumoniae respiratory infections when multiple pollutants were tested together.

Prevalence of plasmid-mediated AmpC beta-lactamases in a Chinese university hospital from 2003 to 2005: first report of CMY-2-Type AmpC beta-lactamase resistance in China.

The aim of this study was to investigate the prevalences of plasmid-mediated AmpC beta-lactamases (PABLs) in isolates of Escherichia coli and Klebsiella spp. from a university hospital in China. A total of 1,935 consecutive nonrepeat clinical isolates of Escherichia coli, Klebsiella pneumoniae, and Klebsiella oxytoca were collected between January 2003 and July 2005. The isolates with cefoxitin zone diameters less than 18 mm (screen positive) were selected for PCR of the bla(AmpC) genes and sequencing. Fifty-four (2.79%) isolates harbored PABLs, as demonstrated by PCR and isoelectric focusing. Sequence analysis revealed the presence of bla(DHA-1) and bla(CMY-2) genes. The Southern blot hybridization studies confirmed that bla(CMY-2) and bla(DHA-1) were located on plasmids. Based on species, PABLs were detected in 4.29% (29 isolates of DHA-1 and 1 isolate of CMY-2) of K. pneumoniae, 1.91% (11 isolates of DHA-1 and 12 isolates of CMY-2) of E. coli, and 3.03% (1 isolate of DHA-1) of K. oxytoca isolates. In contrast to our anticipation, the occurrence rate of DHA-1-producing K. pneumonia significantly decreased (P < 0.01), from 7.54% in 2003 to 2.72% in 2004. The results of random amplified polymorphic DNA analysis indicate that the prevalences of DHA-1-producing K. pneumoniae and CMY-2-producing E. coli strains were not due to epidemic strains. In conclusion, DHA-1 was the most prevalent acquired AmpC beta-lactamase in this collection of isolates from a medical center in China, and DHA-1-producing K. pneumoniae was the most prevalent bacterium harboring a PABL. To the best of our knowledge, this is the first report of CMY-2-type AmpC beta-lactamases in the Chinese mainland.

[Effects of erythromycin on the twitching motility of Pseudomonas aeruginosa].

OBJECTIVE: To observe the effects of low concentration of erythromycin on the twitching motility of Pseudomonas aeruginosa. METHODS: Pseudomonas aeruginosa 1244 (PA1244) was cultured in LB plate or LB broth with added erythromycin at different concentrations 2.5, 0.5, 0.25 mg/L, and in cultures without erythromycin as the control. The changes of PA1244's twitching motility was observed by naked eyes, immunofluorescence, Western blot, dot blot and electron microscope. RESULTS: SubMIC erythromycin inhibited the halation of twitching motility on the culture plates. The average diameters of bacterial halation after culture for 18 h were as below: group of 2.5 mg/L was (0.48 +/- 0.14) cm, group of 0.5 mg/L was (0.64 +/- 0.20) cm, group of 0.25 mg/L was (0.95 +/- 0.18) cm; the control group was (1.40 +/- 0.21) cm (F = 123.15, P < 0.01). After culture for 24 h: group of 2.5 mg/L was (0.67 +/- 0.12) cm, group of 0.5 mg/L was (0.82 +/- 0.23) cm, group of 0.25 mg/L was (1.18 +/- 0.24) cm; the control group was (1.58 +/- 0.28) cm (F = 76.37, P < 0.01). After culture for 36 h: group of 2.5 mg/L was (0.91 +/- 0.17) cm, group of 0.5 mg/L was (1.04 +/- 0.32) cm, group of 0.25 mg/L was (1.49 +/- 0.31) cm; the control group was (2.07 +/- 0.38) cm (F = 54.75, P < 0.01). Immunofluorescence showed that the key component of twitching motility was pilus VI located at the pole of the PA body. Western blot showed that the expression of pilus VI was increased with the decreasing concentration of erythromycin. Dot blot showed that pilus VI was expressed mostly at the outmost of the twitching zone and there was no significant difference between groups. Through transmission electron microscope, PA of the group of 2.5 mg/L had fewer pilus than the control group. CONCLUSION: Diverse concentrations of erythromycin have inhibitory actions on the twitching motility of Pseudomonas aeruginosa.

Identification of antibiotic resistance genes in the multidrug-resistant Acinetobacter baumannii strain, MDR-SHH02, using whole-genome sequencing.

This study aimed to investigate antibiotic resistance genes in the multidrug-resistant (MDR) Acinetobacter baumannii (A. baumanii) strain, MDR-SHH02, using whole­genome sequencing (WGS). The antibiotic resistance of MDR-SHH02 isolated from a patient with breast cancer to 19 types of antibiotics was determined using the Kirby­Bauer method. WGS of MDR-SHH02 was then performed. Following quality control and transcriptome assembly, functional annotation of genes was conducted, and the phylogenetic tree of MDR-SHH02, along with another 5 A. baumanii species and 2 Acinetobacter species, was constructed using PHYLIP 3.695 and FigTree v1.4.2. Furthermore, pathogenicity islands (PAIs) were predicted by the pathogenicity island database. Potential antibiotic resistance genes in MDR-SHH02 were predicted based on the information in the Antibiotic Resistance Genes Database (ARDB). MDR-SHH02 was found to be resistant to all of the tested antibiotics. The total draft genome length of MDR-SHH02 was 4,003,808 bp. There were 74.25% of coding sequences to be annotated into 21 of the Clusters of Orthologous Groups (COGs) of protein terms, such as 'transcription' and 'amino acid transport and metabolism'. Furthermore, there were 45 PAIs homologous to the sequence MDRSHH02000806. Additionally, a total of 12 gene sequences in MDR-SHH02 were highly similar to the sequences of antibiotic resistance genes in ARDB, including genes encoding aminoglycoside­modifying enzymes [e.g., aac(3)-Ia, ant(2'')­Ia, aph33ib and aph(3')-Ia], ß-lactamase genes (bl2b_tem and bl2b_tem1), sulfonamide-resistant dihydropteroate synthase genes (sul1 and sul2), catb3 and tetb. These results suggest that numerous genes mediate resistance to various antibiotics in MDR-SHH02, and provide a clinical guidance for the personalized therapy of A. baumannii-infected patients.

Influences of enteral nutrition combined with probiotics on gut microflora and barrier function of rats with abdominal infection.

AIM: To investigate the influences of enteral, parenteral nutrition and probiotics delivered by gut on intestinal microecology, epithelial tight junctions, immune and barrier function of rats with abdominal infection. METHODS: Rat abdominal infection models established with cecal ligation and perforation method, were divided into three groups: parenteral nutrition (PN group, n = 7), PN+enteral nutrition (EN group, n = 7) and PN + EN + probiotics (probiotics group, n = 7) via the needle jejunostomy and neck vein for five days. The total nutritional supplement of the three groups was isonitrogenic and isocaloric. Probiotics was delivered by jejunostomy 10 mL/d (1 x 10(8) cfu/mL). The rats were killed on the sixth day. The feces in the cecum were cultured for anaerobic bacterial growth and analyzed with bacterial group DNA fingerprint profile with random amplified polymorphic DNA. The transmembrane binding proteins (occludin) and IgA level in plasma cells of intestine epithelium in colon and terminal ileum were measured by an immunohistochemistry method. The ultrastructure of intestinal epithelial tight junctions in colon and small intestine was observed by electron-microscopy. Vena cava blood and the homogenated tissue of liver, lung and mesenteric lymph nodes were cultured to determine the bacterial translocations, and endotoxin in the blood from portal vein was detected. RESULTS: (1) The amount of bacteria of gut species in EN group and probiotic group was higher than that in PN group. The DNA-profiles in EN group and probiotic group were similar to that of normal rats. The number of DNA-profiles in probiotics group was much more than that in PN group and EN group. Moreover, there were strange stripes in PN group. (2) The expression of occludin and IgA in the small and large intestine in EN group (2.309 +/- 0.336, 15.440 +/- 2.383) and probiotic group (2.938 +/- 0.515, 16.230 +/- 3.183) was improved as compared with PN group (1.207 +/- 0.587, P < 0.05, 11.189 +/- 2.108, P < 0.01). The expression of occludin in probiotic group (intestine: 2.93 +/- 0.515; cecum: 3.40 +/- 0.617) was higher than that in EN group (intestine: 2.309 +/- 0.336; cecum: 2.076 +/- 0.670; P < 0.05). The expression of IgA, especially in EN group (intestine: 15.440 +/- 2.383) and probiotic EN group (large intestine: 12.516 +/- 1.542) significantly increased as compared with PN group (intestine: 11.189 +/- 2.108; cecum: 10.160 +/- 1.643; P<0.01). The intestinal epithelial tight junctions and microvilli of the probiotic group were more intact than those in the PN group. (3) The bacterial translocations in blood, liver, lung and mesenteric lymph nodes, and the levels of endotoxin were significantly reduced in probiotic (0.082 +/- 0.029) and EN (0.125 +/- 0.040) groups as compared with PN group (0.403 +/- 0.181, P < 0.05). CONCLUSION: Application of EN combined with probiotics could improve the expression of transmembrane binding proteins (occludin) and IgA, correct the intestinal flora disturbance, maintain gut barrier functions and tight junctions, and reduce the occurrence of gut bacterial translocation.

[Characterization of macrolide-resistance mediated by transposable elements Tn1545 and Tn917 in clinical Isolates of Streptococcus pneumoniae].

OBJECTIVE: To investigate the characterization of ermB gene expression and dissemination in macrolide-resistant Streptococcus pneumoniae (Sp) in Shanghai. METHODS: Eighty-six erythromycin-resistant isolates of Sp were isolated from 3 hospitals in Shanghai. E-test or K-B disk diffusion test were used to determine the susceptibility to 12 antibiotics according to the National Committee for Clinical Laboratory Standards. Macrolide resistant genes ermB and mefE, and transposable elements Tn1545 and Tn917 were amplified by PCR. The isolates were divided into Tn1545 and Tn917 groups according to the transposable elements thereof. Double disc test with erythromycin and clindamycin discs divided the isolates into 2 macrolide resistant phenotypes: cMLS(B) (inducible) phenotype and M phenotype (resistant to erythromycin and sensitive to clindamycin). BOX-PCR was used to analyze the homology of the S. pneumoniae. RESULTS: (1) Of the 86 erythromycin-resistant isolates, the positive rates of ermB, mefE, Tn1545, and Tn917 were 94%, 46%, 87% and 7% respectively. Tn1545 and Tn917 were not detected in 5 ermB-mdfE + strains. (2) Most strains in the Tn1545 and Tn917 groups were highly resistant to erythromycin with a MIC50 of 256 microg/ml. The Tn917 group had a lower MIC to beta-lactam antibiotics and lower resistance to tetracycline, levofloxacin, and compound sulfonamide in comparison with the Tn1545 group. (3) The most common macrolide resistance phenotype of the Tn1545 group was cMLS(B) phenotype. Three strains in the Tn917 group had a 194 bp deletion in the promoter region of ermB and an insertion of TAAA motif in the N end of leader peptide, resulting in the change of the ermB gene from inducible to constitutive expression. (4) BOX-PCR showed that Tn1545 and Tn917 might spread horizontally. CONCLUSION: In Shanghai ermB-mediated cMLS(B) is the most prevalent phenotype in macrolide-resistant Streptococcus pneumoniae isolates. Primarily, the ermB gene was carried and spread horizontally by Tn1545.

Effects of wall suction/blowing on two-dimensional flow past a confined square cylinder.

A numerical simulation is conducted to study the laminar flow past a square cylinder confined in a channel (the ratio of side length of the square to channel width is fixed at 1/4) subjected to a locally uniform blowing/suction speed placed at the top and bottom channel walls. Governing equations with boundary conditions are resolved using a finite volume method in pressure-velocity formulation. The flow patterns relevant to the critical spacing values are investigated. Numerical results show that wall blowing has a stabilizing effect on the flow, and the corresponding critical Reynolds number increases monotonically with increasing blowing velocity. Remarkably, steady asymmetric solutions and hysteretic mode transitions exist in a certain range of parameters (Reynolds number and suction speed) in the case of suction.

Effect of lactobacillus on the gut microflora and barrier function of the rats with abdominal infection.

AIM: To investigate the effect of probiotics supplemented by gut on the tight junctions of epithelial cells, barrier function and the microflora of rats with abdominal infection. METHODS: After the model of cecal ligation and perforation established, SD rats were divided into two groups: parenteral nutrition (PN) group and PN+probiotics (probiotics) group, PN solution was supplemented by neck vein and probiotics was delivered via the jejunostomy tube for five days. Vena cava blood and the homogenated tissue of liver, lung and mesenteric lymph nodes were cultured to determine the bacterial translocation rate (BTR). The ultra-structure of epithelial tight junctions and microvilli of the gut were observed by electron microscopy; occluding expression was measured by indirect-immune fluorescence method; anaerobic bacterial growth by anaerobic culture and DNA fingerprint of bacterial colonies of the feces by PCR. RESULTS: The quantity of lactobacteria and bifydobacteria in probiotics group was higher than that of PN group. The profiles of DNA fingerprint expression in probiotics group were similar to that in the normal group, a new 16S rDNA sequence appeared in the profile in PN group. The occludin expression, the integrality of the gut epithelial tight junction and microvilli in probiotics group were improved as compared with PN group. The BTR and endotoxin in blood were reduced more significantly in probiotics group as compared with PN group. CONCLUSION: The probiotics could improve the gut microflora disturbance, increase occludin expression, maintain the gut epithelial tight junction and decrease the bacterial translocations rate.

[An experimental study on antimicrobial efficacy of platelet-rich plasma for bone infection prophylaxis].

OBJECTIVE: Platelet-rich plasma (PRP) contains high concentrations of platelets and leucocytes, which play a key role in antimicrobial host defense system. To evaluate the antimicrobial efficacy of autologous PRP in vitro and in vivo and to explore the mechanism of action so as to provide the experimental basis for the prevention and treatment of bone infection. METHODS: PRP was prepared with the method of two centrifugation from 15 health volunteers. Platelet-leukocyte gel (PLG) was obtained after activation of PRP with bovine thrombin. Next, PLG was incubated with Staphylococcus aureus (1 x 10(6) cfu/mL) in vitro compared with PRP, platelet-poor plasma (PPP) and PBS. Samples were taken out after 2, 4, 6, 8, 12, and 24 hours for bacterial culture and colony count. Thirty-six New Zealand adult rabbits, weighing (2.85 +/- 0.11) kg, were divided into 4 groups: PLG (n = 10), antibiotic (n = 10), infection (n = 10), and PBS (n = 6) groups. The osteomyelitis models were made by injecting 0.1 mL Staphylococcus aureus suspension (1 x 10(6) cfu/mL) into the tibial canal in PLG group, antibiotic group, and infection group; equal volumes of PBS was injected in PBS group as a control. Autologous PLG was injected immediately after operation in PLG group. Cefazolin (30 mg/kg) was injected through the auricular vein from 1 hour before operation to 72 hours after operation in antibiotic group, once per 8 hours. No treatment was given in infection and PBS groups. The efficacy of PLG for osteomyelitis prophylaxis was evaluated by microbiological, X-ray and histological observation within 28 days. RESULTS: The contents of leucocyte and platelet of PRP were 6.2 times and 5.5 times of whole blood, showing significant differences (P < 0.05); the contents of leucocyte and platelet of PPP were significantly lower than those of whole blood and PRP (P < 0.05). In vitro test showed that PLG had the most obvious bacteriostasis effect. The bacterial count reached a minimum value at 4 hours after incubation in PLG and at 6 hours after incubation in PRP. PPP had slow and no obvious bacteriostasis effect and PBS had no bacteriostasis effect. At 2, 4, 6, 8, 12, and 24 hours of incubation, the bacterial count reduced significantly when compared PLG with PRP and PPP (P < 0.05), when compared PRP with PPP (P < 0.05). In PLG group and antibiotic group, 1 rabbit died, respectively; 34 rabbits survived to the end of the experiment. There was no significant difference (P > 0.05) in temperature, body weight, erythrocyte sedimentation rate and content of leucocyte between 28 days after operation and before operation in 4 groups. After 28 days, the X-ray scores were 2.78 +/- 1.39, 1.55 +/- 1.48, 4.17 +/- 1.25, and 0 in PLG, antibiotic, infection, and PBS groups, respectively, which was significantly higher in infection group than in other 3 groups (P < 0.05). Also, the histological scores were 5.89 +/- 3.92, 3.00 +/- 2.31, 10.33 +/- 4.03, and 0, respectively, which was significantly higher in infection group than in other 3 groups (P < 0.05), and was significantly lower in antibiotic group than in PLG group (P < 0.05). The results of bacterial culture showed that the infection rates of PLG group (44.4%) and antibiotic group (20.0%) were significantly lower (P < 0.05) than that of infection group (88.9%). The quantitative analysis of bacteria showed that the number of bacteria was signficantly lower (P < 0.05) in PLG and antibiotic groups than in infection group. CONCLUSION: PRP forms into PLG after activating, it can inhibit Staphylococcus aureus reproduction in vitro and can effectively prevent bone infection in vivo.

Outbreak of infection caused by Enterobacter cloacae producing the novel VEB-3 beta-lactamase in China.

Over a 4-month period from November 2002 to February 2003, 27 ceftazidime-resistant or cefotaxime-resistant nonrepetitive Enterobacter cloacae isolates were collected from 27 patients hospitalized at HuaShan Hospital, Shanghai, People's Republic of China. The Etest did not detect extended-spectrum beta-lactamases (ESBLs) in those 27 isolates; however, screening by the NCCLS ESBL disk test and confirmatory tests detected ESBLs in 4 of 27 isolates and PCR detected ESBLs in 23 of 27 isolates. The majority of ESBL producers exhibited the same repetitive extragenic palindromic PCR pattern but harbored different ESBL genes. CTX-M-3 was the most prevalent ESBL in our study. Interestingly, 12 clonally related E. cloacae isolates possessed a novel bla(VEB)-type beta-lactamase, bla(VEB-3). Bla(VEB-3) was encoded by the chromosome and was located in an integron. Nine of the 12 isolates harbored both the bla(VEB-3) and the bla(CTX-M-3)-like ESBLs. This is the first report of a VEB-1-like ESBL in China and the first report of the simultaneous presence of VEB-1 and CTX-M-3-like ESBLs in an isolate.

[Influences of enteral nutrition combined with probiotics on the gut microecology and barrier function of the rats with abdominal infection].

OBJECTIVE: To investigate the influences of enteral nutrition (EN), parenteral nutrition (PN) and probiotics supplement on the intestinal microecology, and barrier function of the rats with abdominal infection. METHODS: Twenty-one Sprague-Dawley (SD) rats with abdominal infection were randomly divided into three groups, and received PN (PN group, n=7), PN+ EN (PN+ EN group, n=7) or PN+ EN+ probiotics (probiotics group, n=7) respectively with isonitrogen and isocaloric nutrition. The rats were sacrificed after six days. The feces in cecum were cultured for anaerobic bacterial growth and DNA fingerprint spectrum was analyzed by randomly amplified polymorphic DNA technique. The transmembrane binding protein (occludin) and IgA levels in colon and terminal ileum were detected by immunohistochemistry method. The bacterial translocation rate and endotoxin level were also measured. RESULTS: The germ numbers of different species were both higher in PN+ EN and probiotic group than those in PN group. The bands of DNA fingerprint spectrum were significantly decreased in PN group, but the bands in both PN+ EN group and probiotic group were similar to that in the normal rats. The expression levels of occludin and IgA in the intestine and colorectum were higher in both PN+ EN group and probiotic group compared with those of PN group (P< 0.05, P< 0.01, respectively), the expression level of occludin was higher in probiotic group than that in PN+ EN group (P< 0.05). The overall bacterial translocation rates and endotoxin levels were significantly reduced in both probiotic and PN+ EN group (P< 0.05), but there was no difference between probiotic group and EN group. CONCLUSION: EN combined with probiotics can increase occluding and IgA expressions, improve the intestinal microecology,maintain the gut barrier function, and decrease the incidence of gut bacterial translocation.