OBJECTIVE To study the isolation and resistance tendency of Acinetobacter calcoaceticus-baumannii to antimicrobial agents from 1998 to 2004 to provide valuable data for infection prevention and therapy. METHODS We reviewed the isolation rates,distribution in clinical specimens and wards,and the resistance rates of(A.calcoaceticus-baumannii)to 14 kinds of antimicrobial agents from 1998 to 2004. RESULTS There was an increasing tendency of isolation rates of A.calcoaceticus-baumannii every year,which was 0.18% in 1998 but 1.48% in 2004.In the seven years,there was the highest isolation rate of 70.58% in specimens from respiratory tract,the next was from the urine(9.42%),and blood(4.63%).Concerning the wards distribution,ICU had the highest rate of 47.28%.In 1998,A.calcoaceticus-baumannii had resistance rates more than 50% only to one kind of antimicrobial agents(aztreonam),but in 2004,it had increased to thirteen kinds(except cefoperazone/sulbactam).About the fourteen kinds of antimicrobial agents we inspected,that were increased in their resistance rate.The highest increasing of resistance rate was ceftazidime from 11.1% in 1998 to 88.9% in 2004,the imipenem was second for 0.0% to 64.8%,and the third was sulfamethoxazole/trimethoprim form 0.0% to 64.0%,while there still was an increasing resistance tendency to them. CONCLUSIONS The clinical isolation rate of A.calcoaceticus-baumannii is increasing,and it has higher resistance rates to many antimicrobial agents as well as an increasing resistance tendency to relatively susceptive antimicrobial agents every year.So physicians should prescribe on the basis of antimicrobial agents susceptibility tests in vitro.

To investigate biological characteristics of the IVpi-189 progeny virus derived from the culture of influenza A virus as a live-attenuated vaccine candidate. Persistent infection of a cultured cell line with influenza A virus (MDCK-IVpi) was established by incubating continuously influenza virus-infected cells at a lower temperature. The infectious progeny virus derived from MDCK-IVpi cells at the 189rd subculture was designated as the IVpi-189 strain of influenza virus. The cytopathic effect induced by IVpi-189 virus was observed under different temperature conditions. The production of infectious progeny virus was examined at 38 and 32 degrees C by plaque titration of cell-associated and released virus. IVpi-189 virus showed cytopathic effect as strong as that of IVwt in infected cell line of MDCK at 32 degrees C. However, when culture temperature was raised to 38 degrees C, the cytopathic effect induced by IVpi-189 virus was delayed and less pronounced. Virus growth in IVpi-189 virus-infected cells at 38 degrees C was significantly reduced as compared with that of IVwt virus, although both viruses yielded nearly equivalent high titers of cell-associated and released virus at 32 degrees C. The reasons of the decreased proliferative ability of IVpi-189 virus at high culture temperature were unrelated with virus inactivation or the release of progeny virus, but associated with the decreased replication of infectious progeny virus in the infected cells. IVpi-189 virus derived from MDCK cells infected persistently with influenza A virus showed biological characteristics as a potential live-attenuated vaccine candidate.
Animals ; Cell Line ; Cytopathogenic Effect, Viral ; Dogs ; Humans ; Influenza A virus ; genetics ; physiology ; Temperature ; Virus Cultivation ; methods ; Virus Replication

OBJECTIVETo identify novel multi-drug resistance-related genes, and to explore the mechanisms of multi-drug resistance.

METHODSMulti-drug resistant cell line Lovo/5-FU was established by incubation with increasing dose of 5-FU. The sensitivity to 5-FU and cis-diaminodichloroplatinum (CDDP) was measured by MTT assay. Two dimensional electrophoresis plus mass spectrum(2-DE/MS) was used to identify the differentially expressed protein between Lovo and Lovo/5-FU. The identified protein was then verified by Western blot analysis.

RESULTSThe IC50 concentrations of Lovo/5-FU to 5-FU and CDDP were increased by 31 and 3 times, compared with Lovo (both P<0.01). 2DE-MS showed that CAP-G and RhoGDI2 were up-regulated, whereas 6-PGL, DCI, Prdx-6 and Maspin were down-regulated in Lovo/5-FU. Western blot analysis confirmed that the expression levels of RhoGDI2 and CAP-G in Lovo/5-FU were increased by 6.14 and 2.98 fold respectively (both P<0.01), whereas Maspin was decreased to 5.2% of Lovo(P<0.01).

CONCLUSIONSMulti-gene and multi-pathway are involved in the development of multi-drug resistance of colorectal cancer cells. CAP-G, RhoGDI2 and Maspin are potential multi-drug resistant genes.

Cell Line, Tumor ; Colonic Neoplasms ; genetics ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; Microfilament Proteins ; genetics ; Nuclear Proteins ; genetics ; Serpins ; genetics ; rho Guanine Nucleotide Dissociation Inhibitor beta ; genetics

BACKGROUNDNontuberculous mycobacterium (NTM) had been reported to cause cutaneous infections which are difficult to interpret due to the variability of the clinical manifestations. Among NTM infections, Mycobacterium marinum (M. marinum) are mostly seen to cause skin infection. It is therefore important to establish a rapid approach for detection and identification of M. marinum from lesions of patients with suspected M. marinum infections.

METHODSSpecimens were obtained from 5 patients with swimming pool granuloma. DNA was extracted and polymerase chain reaction (PCR) was performed. PCR products were digested with Hae III and BstE II, then analysed by pattern restriction analysis to detect heat shock protein (hsp) 65 kD gene.

RESULTSThe 65 kD hsp gene was found in all specimens from patients with swimming pool granuloma. PCR restriction analysis (PRA) identified all 5 samples to be M. marinum infections, and the result was consistent with that of routine bacteriological identification. The lesions subsided or markedly improved upon treatment.

CONCLUSIONSPRA is a sensitive, specific and rapid method in identification of mycobacteria. Application of this method will be helpful for early diagnosis of mycobacterial skin infections.

Adolescent ; Adult ; Bacterial Proteins ; genetics ; Chaperonin 60 ; Chaperonins ; genetics ; Female ; Granuloma ; microbiology ; Humans ; Male ; Mycobacterium marinum ; genetics ; isolation & purification ; Polymerase Chain Reaction ; methods ; Skin Diseases, Bacterial ; diagnosis ; Staining and Labeling ; Swimming Pools

Either cetuximab or bevacizumab can improve the survival of patients with metastastic colorectal cancer (mCRC) if administered combided with cytotoxic agents. However, the effect of two or more target agents in combination is uncertain in these patients. Here, we reported a patient with mCRC successfully treated by a combination of target agents after the failure of chemotherapy. The patient received palliative resection of primary tumor followed by 9 cycles of postoperative XELOX regimen, cytokine-induced killer cell (CIK)-based biotherapy, traditional Chinese medicine, particle implantation in the lung metastatic lesions. The tumor progressed 20 months after the standard treatments. Then, the regimen cetuximab, bevacizumab and cefitinib was applied. During the treatment with targeted agents, grade IV acne-like rash and relatively severe parionychia of the toes occurred. Both of them recovered smoothly. The PET-CT reexamination at 40 days after the target treatment showed that the metabolism of mediastinal lymph nodes basically recovered to a normal level. The combination of multiple targeted agents obtained a progression-free survival(PFS) of 11 months and the patient with a good quality of life during this period.
Adenocarcinoma ; diagnostic imaging ; drug therapy ; pathology ; secondary ; Angiogenesis Inhibitors ; therapeutic use ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; therapeutic use ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bevacizumab ; Catheter Ablation ; Cetuximab ; Cytokine-Induced Killer Cells ; immunology ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Drug Delivery Systems ; Fluorouracil ; analogs & derivatives ; therapeutic use ; Humans ; Immunotherapy, Adoptive ; Liver Neoplasms ; secondary ; surgery ; Lung Neoplasms ; secondary ; surgery ; Lymphatic Metastasis ; Male ; Middle Aged ; Multimodal Imaging ; Neoplasm Staging ; Positron-Emission Tomography ; Quality of Life ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; Sigmoid Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; Tomography, X-Ray Computed

OBJECTIVETo explore the method of fabricating oriental scaffolds and investigate the biocompatibility of the scaffolds as well as cells distribution within the scaffolds in vitro.

METHODSThe oriental poly (lactic-co-glycolic acid) (PLGA) scaffolds were fabricated with modified emulsion-phase separation method. The scaffolds were treated with plasma and then anchored with collagen I. Articular chondrocytes were loaded into the scaffolds. The growth status and distributing characteristic of the cells were investigated by environmental scanning electron microscope.

RESULTSThe scaffold was well compatible with the articular chondrocytes. The cells could reach to 2.5 mm depth with unilateral loading. The cells distributed evenly in the scaffold and lined along the inner pipes.

CONCLUSIONSThe oriental scaffold fabricated could significantly promote the distributing characteristics of the chondrocytes. The vertical alignment of the chondrocytes within the scaffold is closely similar to that of articular cartilage.

Cartilage, Articular ; cytology ; Cells, Cultured ; Chondrocytes ; cytology ; Glycolates ; Humans ; Lactic Acid ; Materials Testing ; Polyglycolic Acid ; Tissue Scaffolds

OBJECTIVETo evaluate the usefulness of a broad-range real-time PCR assay aimed at the 16S rRNA gene of bacteria in a clinical setting in rapid and reliable diagnosis of neonatal septicemia for improving the speed and accuracy of bacterial detection.

METHODSThe universal primer and TaqMan probe were designed based on the highly conserved sequences of the bacterial 16S rRNA gene. The chosen primers and probe did not show any likely cross hybridization with human, viral or fungal genome sequences. The TaqMan assay used the fluorescent signal on the probe, such as 6-carboxyfluorescin (6-FAM), and quenched by the standard 6-carboxytetramethylrhodamine (TAMRA) probes. The broad-range 16S rRNA gene real-time PCR array was established. Then, three common pathogenic microorganisms including Staphylococcus aureus, Staphylococcus epidermidis and Escherichia coli, which were prepared by a 10-fold dilution series respectively from 10(8) colony forming unit (CFU)/ml to 10(3) CFU/ml, as well as controls, were used for testing of both sensitivity and specificity of the real-time PCR assay. The blood samples from 830 cases of suspected septicemia, who were hospitalized in our neonatal ward and the neonatal intensive care unit (NICU) and developed clinical signs suggestive of infection, were tested with routine culture and bacterial 16S rRNA genes real-time PCR separately. In addition, 30 neonates without infection were enrolled as the negative control group.

RESULTSAll the three common pathogenic bacterial species were positive on the 16S rRNA genes real-time PCR assay. There were no cross-reaction with cytomegalovirus (CMV), Epstein-Barr virus (EBV), hepatitis B virus (HBV), fungi, human DNA and blank control, and the technique showed high specificity and sensitivity. The detection limit of the TaqMan assay was tested by amplifying serial dilutions of the three common pathogenic bacterial DNA. The minimal detection limit of the TaqMan system was equivalent to 3 CFU of bacteria, the threshold cycle (CT), which is inversely proportional to the log of the amount of target DNA initially present, was 37.90 by calculation. The real-time PCR assay was evaluated on 830 blood specimens for suspected neonatal septicemia, as compared to the results obtained from the routine bacterial cultures. The positive rate by the real-time PCR assay was 5.18% (43/830) in 830 samples, and was significantly higher than that of blood culture [2.41% (20/830) (P < 0.01)]. The real-time PCR was positive in all the 20 positive blood culture samples. Thirty non-infectious blood samples were negative by both the PCR assay and blood cultures. When blood culture was used as control, the sensitivity of the real-time PCR assay was 100%, the specificity was 97.16%, and the index of accurate diagnosis was 0.972. Moreover, three of the PCR positive amplicons were confirmed by sequencing to confirm the accuracy of the real-time PCR assay in testing clinical specimens. The sequencing showed that except for one sequence, all the others were demonstrated to be Staphylococcus aureus and Escherichia coli respectively, which was in accord with the results of the blood cultures.

CONCLUSIONSThe bacterial 16S rRNA genes real-time PCR had been established to diagnose the neonatal septicemia. The sensitivity and specificity the real-time PCR assay were higher than those of blood culture. This technique can provide a rapid way for the etiological diagnosis of neonatal septicemia, and was a convenient and accurate method in etiologic diagnosis of neonatal septicemia.

DNA ; analysis ; DNA Primers ; Escherichia coli ; genetics ; Genes, rRNA ; genetics ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Infant, Newborn ; Limit of Detection ; Nucleic Acid Hybridization ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; analysis ; Rhodamines ; Sensitivity and Specificity ; Sepsis ; diagnosis ; genetics ; Sequence Analysis, DNA ; Staphylococcus aureus ; genetics ; Staphylococcus epidermidis ; genetics

OBJECTIVETo investigate the effect of thoracic duct ligation during transthoracic esophagectomy on preventing post-operative chylothorax in different tumor locations.

METHODSBetween March 2003 and June 2007, 243 patients with thoracic esophageal carcinoma underwent esophageal resection in our hospital. All the cases were divided into five groups according to tumor localization, including cervical, upper middle, middle, lower middle and lower sections. Each was then subdivided into 2 groups: with and without intraoperative thoracic duct ligation. Statistical analysis was carried out to evaluate the relevance between ligation and non-ligation of the thoracic duct during esophagectomy and the incidence of post-operative chylothorax.

RESULTSA total of 8 cases of post-operative chylothorax was recorded and the incidence was 3.3%. Incidence with respect to tumor location was as follows: cervical section: ligation subgroup 3 cases and non-ligation subgroup 5 cases; upper middle section: no one for both ligation and non-ligation subgroups; middle section: ligation subgroup 0/26 and non-ligation subgroup 1/28 (3.6%); lower middle section: ligation subgroup 1/39 (2.6%) and non-ligation subgroup 1/35 (2.9%); lower section: ligation subgroup 1/37 (2.7%) and non-ligation subgroup 2/44 (4.5%). Logistic regression analysis revealed no significant difference between ligation and non-ligation subgroup in the prevention of post-operative chylothorax (P>0.05).

CONCLUSIONThoracic duct ligation as preventive measure can not decrease the incidence of chylothorax secondary to esophagectomy.

Aged ; Chylothorax ; etiology ; prevention & control ; surgery ; Esophageal Neoplasms ; surgery ; Esophagectomy ; adverse effects ; Female ; Humans ; Ligation ; Male ; Middle Aged ; Postoperative Complications ; prevention & control ; surgery ; Thoracic Duct ; surgery

OBJECTIVETo investigate the relationship between plasma platelet-activating factor acetylhydrolase (PAF-AH) gene 994(G--> T) mutation in exon 9 and the patients with cerebral infarction in Chinese Hans.

METHODSThe authors conducted a case-control study including 108 patients in three groups (atherosclerotic cerebral infarction group, lacunar infarction group and cerebral embolism group) and 215 normal subjects as controls. Genomic DNA was analyzed for the mutant allele by a specific polymerase chain reaction.

RESULTSThe frequency of the mutant genotype in the 102 patients with cerebral infarction was 35.19%(32.41% heterozygotes and 2.78% homozygotes), and was 38.10%(34.92% heterozygotes and 3.18% homozygotes) in the atherosclerotic cerebral infarction group, being all significantly higher than the control group's 20.46% (18.60% heterozygotes and 1.86% homozygotes)(P< 0.01); however, the frequencies of the mutant genotype in the lacunar infarction group and cerebral embolism group were 32.35% (29.41% heterozygotes and 2.94% homozygotes) and 27.27% (27.27% heterozygotes and 0 homozygotes) respectively, being not statistically different from those of the controls (P> 0.05).

CONCLUSIONThese findings show that the 994(G--> T) mutation of plasma PAF-AH gene may be an independent risk for atherosclerotic cerebral infarction, but not for lacunar infarction.

1-Alkyl-2-acetylglycerophosphocholine Esterase ; genetics ; Adult ; Aged ; Aged, 80 and over ; Cerebral Infarction ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Heterozygote ; Homozygote ; Humans ; Male ; Middle Aged ; Mutation ; Polymerase Chain Reaction

Aim To investigate the effects of Evodiamine (EVO) on proliferation and apoptosis of human leukemia cell line K562 and its potential mechanisms. Methods K562 cells were treated with EVO at different concentrations (0, 1, 2, 4, 8, 16, 32, 64 jxmol • L