OBJECTIVE To investigate the positive rate of ?-lactamase,oprD2,aminoglycosides modifying enzyme and qacE△1 gene among clinical isolated strains of multi-resistant Pseudomonas aeruginosa in our hospital.METHODS P.aeruginosa was determined by VITEK,and MIC was determined by agar dilution method.TEM,IMP,VIM,oprD2,aac(3)-Ⅱ,aac(6′)-Ⅰ,aac(6″)-Ⅱ,ant(3″)-Ⅰ,ant(2″)Ⅰ and qacE△1 in strains were detected by polymerase chain reaction(PCR).RESULTS The positive rate of TEM,aac(3)Ⅱ,aac(6′)-Ⅱ,ant(3″)-Ⅰ and ant(2″)-Ⅰ in multi-resistant P.aeruginosa were 51.4%,48.6%,40.0%,54.3%,45.7% and 60.0%,respectively.No IMP and VIM genes were detected.CONCLUSIONS Positive rate of ?-lactamase, aminoglycosides modifying enzyme is high in 35 tested strains,and that should be paid more attention.

BACKGROUND: It is necessary to establish a high throughput screening system for anti -inflammatory drugs for rheumatoid arthritis.OBJECTIVE: To construct an eukaryotic expression vector p4CCL20-ZsGreen1-DR with the NF-kB cis-acting element 4×CCL20motif as an enhancer, SV40 as a promoter, and ZsGreen1-DR as a reporter gene.METHODS: The target fragment SV40 was PCR amplified using PGL2-control plasmid as a template. KpnⅠ/Bam HⅠ restriction sites were introduced into the flank of the target fragment. Then, pSV40-ZsGreen1-DR vector was constructed by cloning the target fragment into pZsGreen1-DR plasmid. Finally, p4CCL20-ZsGreen1-DR plasmid was constructed by cloning the double strand DNA of 4×CCL20 motif (with BglⅡ and EcoRⅠ sticky ends at the 5’ and 3’ terminus, respectively) into the corresponding restriction sites of pSV40-ZsGreen1-DR vector (upstream of SV40 promoter).RESULTS AND CONCLUSION: DNA sequencing demonstrated successful construction of p4CCL20-ZsGreen1-DR plasmid.The construction of p4CCL20-ZsGreeR plasmid might be useful to establish a high throughput screening system for anti -inflammatory drugs.

Tomography, X-Ray Computed ; trends

General regulations for the processing are the important part of processing procedures of prepared slices of Chinese crude drugs. It has an important significance on enhancing the operability of actual production, regulating production of prepared slices of Chinese crude drugs, improving quality and establishing drug safety. The article could provides suggestions and reference for future compilation work on "National processing procedures of prepared slices of Chinese crude drugs" by comparative analysis and summary on general regulations for the processing of different processing procedures of prepared slices of Chinese crude drugs in China.
China ; Drugs, Chinese Herbal ; chemistry ; standards ; Humans ; Medicine, Chinese Traditional ; standards ; Quality Control ; Safety ; Technology, Pharmaceutical ; standards

OBJECTIVETo develop an effective method for screening recombinant hairpin RNA expression plasmids using single restriction endonuclease analysis.

METHODSThe double-strand DNA fragment containing a ClaI site (the flanking sequences of which were not complementary) was annealed and ligated into small hairpin RNA (shRNA) expression vector pSilencer-4.1 that did not contain ClaI site to construct the circular pSilencer-4.1-ClaI vector. With BamHI and HindIII, the pSilencer-4.1-ClaIwas digested and ligated with the DNA template of green fluorescence protein (GFP) shRNA that did not include a ClaI site. The plasmid DNA of the positive clones was extracted and digested with ClaI, and the inserted DNA sequence of the non-linearized plasmid was identified by sequence analysis.

RESULT AND CONCLUSIONDNA sequencing showed that pSilencer-4.1-ClaI was correctly constructed and the plasmids resistant to ClaI digestion were all recombinant vectors encoding GFP shRNA. The constructed pSilencer-4.1-ClaI can be used as a universal vector to construct the shRNA expression plasmid, and the incorporated ClaI sites may allow efficient screening of recombinant shRNA expression vectors.

Base Sequence ; Gene Expression ; Genetic Engineering ; methods ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Inverted Repeat Sequences ; Molecular Sequence Data ; Plasmids ; genetics ; RNA, Small Interfering ; genetics ; Restriction Mapping ; methods ; Sequence Analysis, DNA ; Time Factors

Objective To compare the efficacy of transplanting bone marrow mesenchymal stem cell (BMSC) or microenvironmental induced BMSC ( iBMSC) into the ischemic myocardium of rats with myocardial infarction. Methods iBMSC was defined as BMSC co-cultured with myocardial cells for 2 weeks. The stem cells or equal volume PBS were injected into ischemic border zone 1 wk after experimental infarction. Cardiac performance was evaluated at 1, 2, and 4 wk after cell transplantation by echocardiography and analyzed histologically at 4 wk after cell transplantations. Results Compared with PBS group, both BMSC and iBMSC transplantations reduced infarct size. iBMSC enhanced the beneficial effects of BMSC on improving cardiac function (FS: 28.5% ±4.3% in PBS, 29.0% ±2.0% in BMSC and 45. 1% ±3. 1% in iBMSC group at 4 weeks post transplantation, iBMSC group vs. PBS group P <0. 05, iBMSC group vs. BMSC group P <0. 05). Immunofluorescence microscopy results revealed co-localization of SPIO-labeled transplanted cells with cardiac markers for cardiomyocytes, indicating regeneration of damaged myocardium. Conclusion Our data suggest that iBMSC implantation is more effective on improving cardiac function than BMSC implantation in this model. iBMSC might serve as a new promising therapeutic cell source for regenerating ischemic myocardium in patients with post-infarction heart failure.

OBJECTIVETo establish polymerase chain reaction (PCR) technique for the detection of specific genes related to species of genus Bartonella, and for diagnosing clinically suspected cat-scratch disease (CSD) case complicated with pneumonia on both lungs. The appearance of Bartonella infectious diseases calls for genus and species detection and tools for identification in order to make clinical diagnosis and carry on epidemiological studies.

METHODSOne pair of primer TIle.455p-TAla.885n was designed based on the fact that tRNA(Ile)-tRNA(Ala) intergenic spacer region in 16S-23S rRNA intergenic spacer (ITS) of genus Bartonella were high variable sequences flanked by completely conserved tRNA-encoding genes. 16S-23S rRNA was longer than that which had been described in other bacteria. Two published pairs of primers were used to directly detect the specific gene fragments of Bartonella species DNA extracts from human blood, followed by PCR product Sequencing and nucleotide base sequence analysis.

RESULTSAmplification products of the three pairs of primers had the same predicted size of those in Bartonella spp. According to the different length of electrophoresis bank, the sample was identified as a species of genus Bartonella other than the positive control. Sequence analysis showed that the nuleotide sequence from the PCR product of primer TIle.455p-TAla.885n was identical to the Bartonella isolated from Yunnan in China.

CONCLUSIONSPCR-based assay provided a simple and rapid means to detect pathogenic Bartonella species in humans and mammalian hosts as well as in arthropod vecters. This study suggested that this pathogenic Bartonella species existed in patients in northern and southern parts of China.

Animals ; Bartonella ; genetics ; isolation & purification ; Bartonella Infections ; diagnosis ; microbiology ; Base Sequence ; Cat-Scratch Disease ; diagnosis ; microbiology ; Cats ; China ; Diagnosis, Differential ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Polymerase Chain Reaction ; methods ; RNA, Ribosomal, 16S ; genetics ; RNA, Ribosomal, 23S ; genetics ; RNA, Transfer, Ala ; chemistry ; genetics ; RNA, Transfer, Ile ; chemistry ; genetics ; Sequence Analysis, DNA ; Sequence Homology, Nucleic Acid

OBJECTIVETo investigate the treatment of postoperative chyle leak after surgery for digestive malignancies.

METHODSFrom December 2008 to February 2012, in the Sun Yat-sen Memorial Hospital of Sun Yat-sen University, clinical data of 19 patients with chyle leak after digestive system cancer surgery were retrospective analyzed.

RESULTSNineteen cases of chyle leak were all identified between the second and the fourth postoperative day and were all initially managed with conservative treatment including early fasting, parenteral nutrition(PN), 24-hour continuous infusion of somatostatin, and low pressure suction drainage. Eight patients were treated successfully for 6 to 10 days with a significant reduction of the daily drainage volume. Ten patients had enteral nutrition(EN) and their drain tubes were repeatedly washed with 30 ml of compound meglumine diatrizoate injection every day until the drainage volume decreased to 200 ml/day. The time to resolution of chyle leak in these ten patients ranged from 12 to 24 days. One patient had no significant decrease in fluid drainage and developed abdominal distension after one week of conservative treatment. Surgical closure of chyle leak was performed on the 11th postoperative day, abdominal cavity drainage tube was removed on the 4th postoperative day. The patient was discharged home in good condition.

CONCLUSIONMost postoperative chyle leak after surgery for digestive malignancies can be successfully managed with conservative treatment. Somatostatin and the drainage are the main therapeutic approaches. When chyle leak is not resolved with conservative treatment, surgical treatment should be considered to prevent serious complications.

Adult ; Aged ; Anastomotic Leak ; therapy ; Chyle ; Digestive System Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Postoperative Complications ; therapy ; Retrospective Studies

OBJECTIVETo evaluate the efficacy of cyclosporine A-nanoparticles emulsion (CsA-NP) combined with adipose tissue-derived stem cells (ASCs)transplantation therapy for acute myocardial infarction (AMI) in a miniswine model.

METHODSCsA-NP emulsion was prepared by the high-pressure homogenization method. Models were performed by coronary angioplasty for percutaneous balloon occlusion of left anterior descending artery (LAD). A total of 17 miniswines survived after AMI were divided into four groups: control group (n=5), CsA-NP group (n=4), ASCs group (n=4), and CsA-NP+ASCs group (n=4). ASCs or saline were delivered by intracoronary injection one week after AMI.Before cell transplantation and 8 weeks after cell transplantation, delayed-enhanced magnetic resonance imaging (DE-MRI) was performed to evaluate cardiac function and viability. The infarcted myocardium and implanted cells were histologically studied.

RESULTSEight weeks after treatment, the left ventricular ejection fraction (LVEF)significantly increased in the CsA-NP+ASCs group when compared with the ASCs group [(53.6 ± 2.4)% vs. (48.3 ± 1.8)%, P<0.05]; meanwhile, the infarct size significantly decreased [(6.2 ± 1.7)cm(3) vs.(7.5 ± 0.6) cm(3), P<0.05] and the thickness of the ventricular wall significantly increased (P<0.05). Histology showed that the number of surviving cells increased nearly by three times in the CsA-NP+ASCs group, and the expressions of the cardiomyocyte specific markers (cTnT and α-actin) were detected. Histological samples also showed that CsA-NP+ASCs group reduced fibrotic tissue, and down-regulated the activation of Caspase-3.

CONCLUSIONThe CsA-NP+ASCs combination therapy can enhance the viability of ASCs by improving LVEF and preventing LV expansion, which may be explained that CsA-NP has the anti-apoptotic effect and can promote the survivals and proliferation of ASCs.

Adipocytes ; cytology ; Animals ; Caspase 3 ; metabolism ; Cyclosporine ; therapeutic use ; Disease Models, Animal ; Myocardial Infarction ; therapy ; Nanoparticles ; Random Allocation ; Stem Cell Transplantation ; Swine

OBJECTIVETo discuss the diagnosis and surgical treatment of carotid body tumour.

METHODSThirty-eight cases (41 carotid body tumours) admitted from 1983 to 2002 were analyzed retrospectively.

RESULTSAll patients were examined by ultrasound and angiography routinely before operation. The diagnostic coincidence rates of ultrasound and angiography are 95% and 98% respectively. Thirty-eight tumours were resected successfully. Twelve patients underwent external carotid artery severed, internal carotid artery repaired and tumour resected operation. Eleven patients underwent external carotid artery severed and tumour resection. Eight patients underwent tumour simple resected operation. Six patients underwent external carotid artery severed and internal carotid artery reconstruction. One patient underwent tumour resection, while common, external, and internal carotid artery were partially resected.

CONCLUSIONSFor suspected carotid body tumour patients, ultrasound and angiography reconstruction of carotid artery should be applied routinely before operation. The early stage and one stage tumour resected operation as well as internal carotid artery reconstruction are the key factors of decreasing operative complication rate.

Adult ; Angiography, Digital Subtraction ; Carotid Arteries ; diagnostic imaging ; surgery ; Carotid Artery Diseases ; diagnosis ; surgery ; Carotid Body Tumor ; diagnosis ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reoperation ; Retrospective Studies ; Ultrasonography, Doppler, Color ; Vascular Surgical Procedures ; methods