ObjectiveTo investigate the expression and relationship between phosphatase and tensin homology deleted on chromosome ten(PTEN) and vascular endothelial growth factor(VEGF) in bladder transitional cell carcinoma(BTCC) and their clinical significance.MethodsExpression of PTEN and VEGF were detected in 60 specimens by immunohistochemistry test(SP method).ResultsThe positive rate of PTEN was 47%(28/60) in BTCC;VEGF was 68%(41/60).With the pathological grade and the clinical stage of tumors being higher,the lower expression level of PTEN showed(P<0.05);while the expression of VEGF increased with no statistical significance(P>0.05),but having a significant difference between specimens with lymph nodes metastasis and without lymph nodes metastasis(P<0.05).The expression of PTEN was negatively correlated with that of VEGF(r=-0.439,P<0.01).ConclusionThe expression of PTEN and VEGF in BTCC plays an important role during the progress of carcinoma and is helpful to evaluate the prognosis of patients.

Objective: To explore the potential molecular mechanism of Huanglian Jiedu Decoction in the treatment of atherosclerosis (AS) through pharmacology network. Methods: By identifying all the composition and effect targets of four herbal ingredients in Huanglian Jiedu Decoction from TCMSP platforms and literatures, and the pharmaceutical molecular-target network was constructed. The interaction network of drug-disease target by STRING platform was constructed by screening AS related targets through TTD, DrugBank and DisGeNET databases. The centre targets were analyzed by network topology. DAVID database was used to perform GO biological process analysis and KEGG pathway enrichment analysis of centre target proteins, and further construct a multi-dimensional network relationship diagram of the active component-AS target-KEGG pathway of Huanglian Jiedu Decoction. Results: A total of 71 active ingredients and 165 potential drug targets were obtained according to the screening conditions (OB ≥ 30%, DL ≥ 0.18) and accessing literatures. The main active ingredients in Huanglian Jiedu Decoction included sitosterol, quercetin, berberine, dehydrotanshinone IIA and neobaicalein, which could interfere with the formation of AS. A total of 175 disease targets were collected from the three disease databases under the search criteria of “atherosclerosis”. According to Degree, 223 centre target proteins of Huanglian Jiedu Decoction were screened, mainly invovling NOS2, NOS3, PTGS2, TNF, CYP2C9 and HMOX1, et al. GO biological process analysis identified 50 entries based on false discovery rate (FDR) ≤ 0.05, mainly including SRP-dependent targeting membrane transporters, nuclear-transcribed mRNA catabolic process, viral transcription, rRNA processing, translation and other bioanalysis process. The result of KEGG enrichment analysis showed 74 pathways were associated with AS, mainly involved in ribosome pathway, viral carcinogenesis pathway, cell cycle pathway, estrogen signaling pathway, et al. Conclusion: Huanglian Jiedu Decoction can treat AS through multi-ingredient, multi-target and multi-pathway interaction, which provides a theoretical basis for the clinical application of Huanglian Jiedu Decoction and the basic or clinical research of AS-related diseases. Meanwhile, it has a certain reference value for the research and development of new drugs and its application.

Objective To evaluate factors which maintain the stability of the ankle, and discuss indications for trans-syndesmotic fixation. Methods 12 freshly-frozen cadaver legs amputated below the knee were collected and divided into 2 groups. An ankle fracture model of unconstrained pronation-external rotation was then designed. Group A simulated the injury combined with medial malleolus fracture, and Group B the injury combined with deltoid tear. Ligaments were cut off sequentially to simulate the increasing severity of the injury. Fuji super low-pressure sensitive films and displacement transducers were used to measure the contact area of the tibiotalar articular surface and the width of the syndesmosis. The data were analyzed with SPSS to analyze the relationship of ligament injury and ankle stability. Results In Group A, the articular contact area and the syndesmotic width after section of the deltoid were significantly different from those before the section (P

Benzoates ; pharmacology ; Cells, Cultured ; Cellular Senescence ; drug effects ; radiation effects ; Gene Expression Regulation, Enzymologic ; drug effects ; radiation effects ; Humans ; Infant, Newborn ; Male ; Matrix Metalloproteinase 1 ; genetics ; Matrix Metalloproteinase 3 ; genetics ; Methoxsalen ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Retinoids ; pharmacology ; Skin ; cytology ; metabolism ; Time Factors ; Tissue Inhibitor of Metalloproteinases ; metabolism ; Ultraviolet Rays

Adult ; Fever ; etiology ; Gastrointestinal Hemorrhage ; etiology ; Humans ; Male ; Polyps ; complications ; Stomach Diseases ; complications

Polymyxin E shows effective treatment of the infection induced by resistant gramnegative bacteria, but its nephrotoxicity severely limits the clinical application of this drug. In this work, methoxypolyethylene glycols 2000 (mPEG2K)-polymyxin E (PME) was synthesized via chemical grafting reaction and had been characterized. The antimicrobial activity and cytotoxicity of mPEG2K-PME in vitro were investigated on Escherichia coli and HK-2 cells, separately. Intra-abdominal infection model was further established in order to study the therapeutic effect and the toxic effect on kidney of mice. The results showed that mPEG2K-PME exhibited significant inhibitory effect on Escherichia coli and had a lower toxicity on HK-2 cells in vitro. At the same time, mPEG2K-PME had a good efficacy in the treatment of Escherichia coli infected mice in vivo. Moreover, nephrotoxicity caused by mPEG2K-PME was significantly reduced compared to free PME. mPEG2K-PME is promising in development of new preparations with high efficiency and low toxicity.
Animals ; Cell Line ; Colistin ; pharmacology ; toxicity ; Escherichia coli ; drug effects ; Escherichia coli Infections ; drug therapy ; Humans ; Kidney ; cytology ; drug effects ; Mice ; Polyethylene Glycols ; chemistry

OBJECTIVETo investigate effects of the variation of immune function in high humidity environment in different time, and lay a foundation for further study of the related mechanism.

METHODThirty SD rats were divided into 3 groups (n = 10): 20 day group, 40 day group in 90% relative humidity chamber and control group in normal relative humidity. Peripheral blood and spleens were collected to detect the levels of T lymphocyte subsets by Flow Cytometery.

RESULTSIn peripheral blood of the 20 day group rats, the CD3+ %, CD4+ %, CD8+ % and CD4+/CD8+ were 52.91 +/- 6.27, 37.80 +/- 4.11, 14.85 +/- 3.73 and 2.72 +/- 0.82 separately. Expect CD3+ %, they all had significant differences (P < 0.05). In addition, the data of the 40 day group rats showed no diversity in statistics. In spleen, CD8+ % of the 20 day group rats was 6.23 +/- 2.87 with significant differences (P < 0.05) and IgG, IgA and IgM did not change a lot in blood serum of the high humidity groups except C3 of the 20 days group (P < 0.05).

CONCLUSIONIn high humidity environment, the immune function of the rats increased in the initial stage. As time went on, the immune function gradually went to normal level through the self adjustment.

Objective By the method of multiple polymerase chain reaction (PCR),we intend to amplify different regions (DFR) of Yersinia pestis DNA,and to establish a multiple DFR genotyping technique for detection of Yersinia pestis.Methods According to the product size of 23 DFRs and pMT plasmid,24 primers were optimized and combined,then multiple primers in one PCR reaction system were added,and positive template DNA was amplified.Meanwhile,200 wild strain DNAs were amplified by multiple PCR and normal PCR,to verify the coincidence rate of the two methods.Results Totally 24 target segments were amplified through the positive DNA template.Through different permutation and combination,24 primers were optimized and combined into 9 groups.Totally 200 wild strain DNAs were used for verification,the coincidence rate of multiple PCR and normal PCR was 100％.Conclusions Multiple PCR is applicable and feasible for DFR genotyping of Yersinia pestis.It is an efficient,economic and high accuracy experimental method for large quantities of Yersinia pestis DFR genotyping.

Objective To investigate the significances of karyotyping analysis on umbilical cord vein blood lymphocytes in the diagnosis of abnormal karyotypes in middle to late period of pregnant fetus.Methods A volume (0.5 ～ 1 ml) of umbilical cord vein blood was extracted from pregnant women in third trimester pregnancy with prenatal detection indications,and collected in sterilized anticoagulant tube.Lymphocytes were cultured and collected for karyotyping analysis after fixed and dropped on slides.Data were analyzed statistically.Results Lymphocytes were cultured successfully in 1 211 cases out of total 1 213 cases collected.Totally 142 abnormal karyotypes were found,which includes 81 cases (detection rate 6.68 ％) of non-heteromorphic abnormal chromosomes and 61 cases (detection rate 5.03％) of heteromorphic chromosomes.Among these abnormal karyotypes,50 cases (accounting for 35.21％ in total abnormal cases) of aneuploidy include 4 cases of chimerical karyotype.Structural abnormalities were found in 31 cases (accounting for 21.83％ in total abnormal cases) samples including 11 cases of translocations,17 cases of inversion and 3 cases of deletion.Conclusions Based on our findings,karyotyping analysis on umbilical cord vein blood lymphocytes could be an effective method for detect abnormal karyotypes in middle to late period of pregnant fetus and played an important role in prenatal diagnosis.

Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.