Objective Study the production of the?-lactamases in eight pan-resistant citrobact- er freandi and their epidemic characteristics.Methods Modified Hodge test and EDTA-disk syner- gy test were used to screen Metallo-?-lactarnases,and isoelectric focusing to estimate the production of ESBLs and AmpC enzymes.ESBLs,AmpC,MBL genes were amplified and analyzed by PCR and DNA sequencing.Molecular epidemics were investigated by PFGE.Results All the 8 strains pro- duced TEM-1,CTX-M-14,4 of them at the same time producing CTX-M-3.No Plasmid mediated AmpC was found.Furthermore,a new subtype of MBL with one amino acid different from IMP-8 was de- tected.All the 8 strains belong to the same PFGE type.Conclusions Pan-drug-resistant citrobacter freundi produced more than one kind of?-lactamases.All these isolates were clonally related.

Objective To investigate the features and significance of pathologic changes in apoptosis of small bowel allograft during acute rejection in rats. Methods All 24 recipients were equally divided into four groups ; group A: nonoperative control; group B: allograft ; group C: isograft, group D: treatment control. The graft samples were harvested on day 3, 5, 7, 10 after transplantation, and subjected to histologic examination . Mucosal thickness, villous height and crypt depth were measured, and apoptotic cells of intestinal mucosa of grafts on day 3,5 and 7 after transplantation were examined. Results The mucosal structure was normal in group A; The degree of the inflammatory infiltrated cells ,intestinal mucosa cell apoptosis and structural injury of mucosa in group B were significantly severe compared with groups C and D. As the post-transplanted time increased, the number of musocal apoptotic cells and the degree of mucosal structural injury were significantly increased. The degree of mucosal structural injury in group C was milder than in group B. A few infiltrated cells and mild edema of mucosa occurred in group D , but no mucosal structural injury was found. Conclusions Inflammatory cell infiltration, mucosal epithelial cell apoptosis and mucosal structural damage are the main pathologic features of small bowel allograft during acute rejection. Dynamic observation of the pathologic changes and cell apoptosis of small bowel graft is of certain value in the diagnosis of acute rejection of small bowel graft and in assessment of the degree of small bowel injury.

In this study, nattokinase gene was amplified by PCR using bacillus subtilis chromosomal DNA as template and cloned into expressed vector pBV220. After transforming recombinant plasmid into E.coli HB101, the recombinant strain was yielded. It was proved that expression products was secretive and expression protein was 12% of total cell protein by SDS-PAGE. Optimum culture time and inducing time was determined as 6h and 5h respectively. The plasmid stability studies showed that recombinant plasmid has excellent segregational stability but the structural stability was not good in the host cell.

OBJECTIVETo observe the effects of local co-transfection vascular endothelial growth factor165 (VEGF165) and tissue-type plasminogen activator genes on inhibiting intimal hyperplasia and restenosis in rabbits artery after operation injury and possible mechanisms.

METHODSMicrology operation injury was used to establish the model of intimal injury of right external iliac artery in rabbits. To select 120 male New Zealand rabbits and were randomly divided into 3 groups (n = 40, in each group): Group A (physiological brine control group), Group B (pBudCE4.1 group), Group C (pBudCE4.1/VEGF165-tPA group). The vas-wall of micrology operation injury were infused respectively physiological brine, pBudCE4.1 and pBudCE4.1/VEGF165-tPA transfection solution by micro-injector. Each group were divided into 5 subgroups (n = 8, in each subgroup) randomly according to the sacrifice times (2 d, 1 week, 2 week, 4 week and 8 week after operation). The injured vascular specimen were harvested for pathology test, electric microscopy study, reverse transcription-PCR examining and immunochemistry detecting.

RESULTSThe intimal area and narrow ratio of vases in Group C at every time point after operation were significantly lessened than that in Group A and Group B (P < 0.01). The narrow ratio of vases in Group C at 8 week after operation were decreased respectively by 57.9% and 59.0% than that in Group A and B. The expression of VEGF165 mRNA in Group C were increased significantly than that in Group A and B at every time point after operation (P < 0.01), the expression reached the peak at 1 week and continued to 4 week after operation. Immunohistochemical identified that tPA positive cell in Group C were significantly increased than that in Group A and B (P < 0.01) at every time point after operation.

CONCLUSIONLocal co-transfection VEGF165 and tPA genes could restrain intimal hyperplasia and restenosis of vas, which lay a foundation for future multi-gene therapy of vascular intimal hyperplasia.

Animals ; Arteries ; pathology ; Endothelial Cells ; cytology ; Hyperplasia ; prevention & control ; In Vitro Techniques ; Male ; Myocytes, Smooth Muscle ; cytology ; Plasmids ; Rabbits ; Random Allocation ; Tissue Plasminogen Activator ; biosynthesis ; genetics ; Transfection ; Tunica Intima ; pathology ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVEThe determination of antigenicity and immunogenicity of Leptospira interrogans genus-specific outer envelope proteins (OEPs) will offer evidence for developing universal leptospiral genetic engineering vaccine and detection kit.

METHODSIn this study, Ni-NTA affinity chromatography is used to purify the recombinant products rLipL21, rOmpL1/1, rOmpL1/2, rLipL32/1, rLipL32/2, rLipL41/1 and rLipL41/2 expressed by the major genotypes of four leptospiral OEPs of 15 serogroups. SDS-PAGE is applied to examine the expression and purity of the recombinant proteins. Rabbits are intracutaneously immunized with the recombinant proteins to obtain antisera. Microscope agglutination test (MAT) is used to measure the cross inmmunoagglutination titers of antisera. The OMPs of the reference standard strains belonging to 15 serogroups of L. interrogans in China and L. biflexa strain Patoc I are prepared using salt-denature method. By each of the antisera as the first antibody, Western blot assay is established to detect the natural expressions and immunoreactivity of the four OEPs.

RESULTSThe outputs of rLipL21, rLipL32/1, rLipL32/2, rLipL41/1l, rLipL41/2, rOmpL1/1 and rOmpL1/2 are 10%, 40%, 35%, 15%, 10%, 30% and 15%, respectively. Each the purified recombinant proteins shows a single fragment after SDS-PAGE. Each the rabbit antisera displays extensive cross immunoreactivity between the products expressed by different genotypes of the same gene and the MAT titers ranging from 1:2-1:128. All the four OEPs can be detectable in the OEPs preparations. However, LipL21 is found to exist only in L. interrrogans.

CONCLUSIONThe results of this study indicate that all the four OEPs are superficial genus-specific antigens of Leptospira which can be used as the candidate antigens of leptospiral universal vaccine and detection kit.

Animals ; Antibody Formation ; Antigens, Bacterial ; immunology ; Electrophoresis, Polyacrylamide Gel ; Genetic Engineering ; Immunization ; Leptospira interrogans ; classification ; immunology ; Membrane Proteins ; Rabbits ; Recombinant Proteins ; immunology ; Serotyping

BackgroundWith the increasing prevalence of dry eye and the continuous improvement of living standards,the problem of dry eye more and more get the attention of people.At present,China still lacks the large population-based epidemiological data of dry eye. Objective To investigate the prevalence and possible risk factors of dry eye in a community of Huidong County of population aged 14 and over.Methods From September 2010 to January 2011,using questionnaires and examination of dry eye related,2800 people were selected randomly for cross-sectional survey.Those suspected as dry eye were examed by the SchirmerⅠtest ( S Ⅱ T),tear-film breakup time(BUT),corneal fluorescein staining(F1).Results In the 2475 questionnaire effectively,154 persons were diagnosed as dry eye,and the prevalence rate of dry eye was 6.22％,8.06％in females,4.14％in males.The prevalence rate increases with age.The S Ⅰ T and BUT decreased with increasing age.S Ⅰ T and BUT in females are less than males.Foreign body sensation is the primary complaints of patients.Logistic analysis showed that the most common risk factors in dry eye are age and gender.The system disease and eye diseases,eye fatigue and long exposure to dust are also main determinants.ConclusionsThe population prevalence rate of dry eye increased with age,the prevalence rate of dry eye in females is higher than that in males.The key factors associated with dry eye are age,gender,systemic disease and eye diseases,occupation,working environment.

In this study, the rat type 2 diabetes mellitus (T2DM) model was established through tail vein injection with low dose of streptozotocin (STZ) and high fat diet for 8 weeks, and then treated with Jiaotai Pill. The oral glucose tolerance test (OGTT), fasting serum insulin (FINS), free fatty acid(FFA) levels and blood lipid were assayed. HOMA-IR was calculated. Pancreatic pathology was performed. And pancreatic triglyceride (TG) content was examined by the lipid extraction method. Pancreatic islet cell apoptosis were detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL). According to the results, the model group showed abnormal OGTT, increased FINS, HOMA-IR, FFA, lipid disorder, obvious fat accumulation and significantly increased TG content in pancreatic tissues, and enhanced pancreatic islet cell apoptosis. Compared with the model group, the Jiaotai Pill group displayed improved OGTT, reduced FINS, HOMA-IR, FFA, recovered lipid disorder, decreased fat accumulation and significantly declined TG content in pancreatic tissues, and lowered pancreatic islet cell apoptosis. In summary, Jiaotai pill could effectively treat type 2 diabetes in rats. Its mechanism may be related to the reduction in pancreatic fat accumulation and islet cell apoptosis.
Animals ; Apoptosis ; drug effects ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; administration & dosage ; Fats ; metabolism ; Glucose Tolerance Test ; Humans ; Islets of Langerhans ; cytology ; drug effects ; Male ; Pancreas ; drug effects ; metabolism ; Rats ; Rats, Wistar

Objective To investigate the clinical efficacy and related complications of bilateral minimally invasive transforminal lumbar interbody fusion via Quadrant channels for the treatment of single-level degenerative lumbar spondylolisthesis.Methods Sixty selective pa-tients with single-level mild and moderate degenerative lumbar spondylolisthesis who underwent bilateral MIS-TLIF via Quadrant channels from August 2015 to May 2017 were retrospectived investigated.Operative time,blood loss,X-Ray exposure time,postoperative drainage and related complications were recorded.Lumbar functional improvement was defined as an improvement in the JOA.The VAS was also employed at pre and postoperation(6 months),to evaluate low back and leg pain.Intervertebral fusion was assessed by Bridwell evaluation criteria and the clinical outcome was assessed by the MacNab scale at 6 months.In addition,related complications also were be analyzed.Results All patients underwent a successful MIS-TLIF surgery,of whom 58 patients were followed up for 6 months.The operation time was 170~290 minutes (mean:220.17 minutes),intraoperative blood loss was 100~260 mL(mean:168.62 mL),intraoperative radiation exposure was 15~36 times (mean:25.60 times) and postoperative drainage was 50~163 mL(mean:103.43 mL).Different significance between 6 months post-operative follow-up and pre-operation was exhibited (P<0.05) in VAS scores (t=34.97,P=0.000).At 6 months post-operative follow-up,there were significant differences(P<0.05) compared with pre-operation in JOA scores(t=36.91,P=0.000).At 6 months post-operative follow-up,21 cases of grade 1 and 37 cases of grade 2 fusion were present as determined by the Bridwell evaluation criteria(100% fusion).MacNab scale as-sessment classified 2 patients having excellent clinical outcome and 54 patients in good clinical outcome(96.55% of patients had good to excel-lent results).Amomg 60 cases,complications observed in 6 patients,direct injuries to nerve root in 1 case,dural tears occurred in 1 case,super-ficial wound infection in 1 case,pedicle screw was pulled out during the reduction of slip because of severe osteoporosis in 1 case and leg numb-ness and pain in 2 cases. Conclusion Bilateral MIS-TLIF via Quadrant channels with less complications is a safe and effective approach for the treatment of single segment degenerative lumbar spondylolisthesis,but potential complications should be prevented as well.

To investigate the influence of the thalidomide on the growth of multiple myeloma cells from untreated, relapsed or refractory patients and summarize its mechanisms, thalidomide influence on colony growth of untreated, relapsed or refractory multiple myeloma cells cultured by semisolid methylcellulose was observed. The level of interleukin-6 (IL-6) autosecreted by myeloma cells was tested by IL-6-dependent cell line when myeloma cells were treated with thalidomide at 200 microgram/ml, and in the same concentration of thalidomide the expression of IL-6 receptor were tested by flow cytometry. Results showed that colony growths of myeloma cell from untreated and relapsed or refractory patients were all colonies were inhibited when treated by thalidomide up to 75 microgram/ml or 100 microgram/ml concentration. The inhibition was concentration-dependent, higher concentration cause more inhibition. After treatment with thalidomide at 200 microgram/ml, the concentrations of IL-6 secreted by myeloma cells were (148.5 +/- 96.7) microgram/ml, and the levels of IL-6 receptor expressed on the cell surface were 16.7% and 20.2% in untreated and relapsed or refractory patients, respectively, and those were significantly lower than those levels in the cells before exposure to thalidomide. It was concluded that thalidomide can inhibit growth of both relapsed or refractory cells and untreated myeloma cells in vitro. Therefore, it can be used to treat untreated multiple myeloma patients. Inhibiting tumor cells secreting level of IL-6 and reducing the expression of IL-6 receptor on myeloma cell surface is one of the mechanisms for thalidomide to remedy multiple myeloma patients
Angiogenesis Inhibitors ; pharmacology ; Cell Division ; drug effects ; Dose-Response Relationship, Drug ; Humans ; Interleukin-6 ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Receptors, Interleukin-6 ; biosynthesis ; Thalidomide ; pharmacology ; Tumor Cells, Cultured

OBJECTIVETo screen and verify differentially expressed genes in prostate cancer.

METHODSUsing DNA microarray, we screened differentially expressed genes in prostate cancer tissue and its adjacent tissue followed by verification by PCR.

RESULTSA total of 1 444 genes were found to be differentially expressed (differentiation ≥ 1.5-fold; P≤ 0.05) in the prostate cancer tissue, of which 769 (53%) were up-regulated and 675 (47%) down-regulated. Fifty percent of the differentially expressed genes showed a 1.5- to 2-fold differentiation, including 396 up-regulated and 182 down-regulated ones. Additionally, 308 up-regulated and 334 down-regulated genes exhibited a >2- to 5-fold, 46 up-regulated and 78 down-regulated genes a > 5- to 10-fold, and 19 up-regulated and 81 down-regulated genes a > 10-fold differentiation. Verification by subjecting 15 most significantly up-regulated and another 15 most markedly down-regulated genes to quantitative real-time PCR (qRT-PCR) showed that most of the genes had a transcriptional profile similar to that in the microarray data, with a Pearson correction coefficient of 0.83 between the microarray data and qRT-PCR results. Totally, 10 significantly differentially expressed genes were identified.

CONCLUSIONDNA microarray analysis provides reliable information on differentially expressed genes in prostate cancer and benign tissues. The 10 significantly differentially expressed genes verified by qRT-PCR could possibly become new bio-markers and specific molecules for tumor identification.

Cell Differentiation ; Down-Regulation ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Oligonucleotide Array Sequence Analysis ; Polymerase Chain Reaction ; Prostatic Neoplasms ; genetics ; Transcriptional Activation ; Up-Regulation