Objective To explore the establishment methods of animal models of intestinal multidrug-resistant bacterial (Pseudomonas aeruginosa, PA) infection, and to provide a good model for experimental research and treatment for infection caused by drug-resistant bacteria .Methods Male BALB/c mice were divided into four groups:Normal con-trol group, MDR-PA group, MDR-PA +antibiotics group, and MDR-PA +fasting group.They were given saline, PA suspension, PA suspension after free drinking with ceftriaxone water and PA suspension after fasting , respectively , by ga-vage.Colon tissues were collected for pathological examination and inflammation scoring .The expressions of TNF-αand INF-γin the colon tissue was assessed at the end of the experiment .Results Pathology showed different degrees of inflam-matory changes in the colon tissues .Compared with the normal control group , the expressions of TNF-αand INF-γhad sig-nificant differences with that of experimental groups .Conclusions All the three methods used in this experiment can be successfully used to establish mouse models with intestinal infection caused by multidrug-resistant Pseudomonas aeruginosa, and may be chosen for experimental studies according to different research purposes .

Objective To detect and type herpes simplex virus (HSV) in genital lesions of the patients attending STD clinic. Methods Clinical data were collected and analyzed from patients with anogenital non-herpetic lesions including induration or furuncle, fissure, folliculitis, single ulcer and so on. HSV was detected and typed by culture and PCR with specimens taken from these lesions. Results One hundred and five cases were recruited in this study. Among them, 18 cases presented induration (furuncle), 15 fissure, 16 folliculitis, 7 abrasion, 12 single ulcer, 25 nonspecific erythema and 12 balanoposthitis with edema and exudation. HSV was found in 33.3%(6/18), 20%(3/15), 37.5%(6/16), 28.6%(2/7), 33.3%(4/12), 20%(5/25) and 50%(6/12) of these lesions, repectively, by PCR, while in 22.2%(4/18), 13.3%(2/15), 25%(4/16), 14.3%(1/7), 33.3%(4/12), 8%(2/25) and 41.7%(5/12), repectively, by viral culture. The positive rates of HSV from all these non-herpetic lesions were 30.5% (32/105) and 21% (22/105), respectively (? = 0.095, P = 0.114), by PCR and viral culture. The results of HSV typing were consistent between PCR and immunofluorescence with type-specific monoclonal antibodies. Among those with HSV infections, HSV-1 infection acounted for 9.4% (3/32), and HSV-2 90.6% (29/32). Conclusions The clinical manifestations of genital HSV infections vary, and HSV could be isolated from lesions of induration (furuncle), fissure, folliculitis, abrasion, single ulcer, nonspecific erythema and balanoposthitis with edema and exudation. HSV-2 is the predominant type.

Objective To establish a model of cervical spondylosis in rats. Methods Sixty SD rats (four months old) were ran-domly divided into control, muscle imbalance and posterior column instability group, ten rats in each group. Then the degeneration of X-ray film and motion function were evaluated by oblique board test. Results After analyzed the cervical films of control groups, the nature curve still existed, vertebral clearance had no abnormal. No ostensis spurs or ossifies in Luschka joint and joint processes were found. But in mus-cle imbalance and posterior column instability groups, the nature curve was disappeared or stiff, vertebral clearance were stenosis, osteosis spur formed, ossify or sclerosis emerged in Lnschka joint and joint process. Compared with control group in oblique board test, the muscle force decreased in muscle imbalance and posterior column instability group. There were no statistic significant difference between 2 and 4 months of control group, and also no difference in muscle imbalance and posterior column instability group, but there were significant differ-ence in 2 and 4 months of muscle imbalance and posterior column group. Conclusion Not only cervical posterior column instability but also muscle imbalance could result in cervical spendylosis, both muscle force balance and posterior column were important factors in maintaining spine stability.

To obtain the recombinant tumor necrosis factor-α converting enzyme (TACE) ectodomain and use it as a selective molecule for the screening of TACE peptide inhibitors, the cDNA coding catalytic domain (T800) and full-length ectodomain (T1300) of TACE were amplified by RTPCR, and the expression plasmids were constructed by inserting T800 and T1300 into plasmid pET28a and pET-28c respectively. The recombinant T800 and T1300 were induced by IPTG, and SDSPAGE and Western blotting analysis results revealed that T800 and T1300 were highly expressed in the form of inclusion body. After Ni2+-NTA resin affinity chromatography, the recombinant proteins were used in the screening of TACE-binding peptides from phage display peptide library respectively. After 4 rounds of biopanning, the positive phage clones were analyzed by ELISA, competitive inhibition assay and DNA sequencing. A common amino acid sequence (TRWLVYFSRPYLVAT) was found and synthesized. The synthetic peptide could inhibit the TNF-α release from LPS-stimulated human peripheral blood mononuclear cells (PBMC) up to 60.3 %. FACS analysis revealed that the peptide mediated the accumulation of TNF-α on the cell surface. These results demonstrate that the TACE-binding peptide is an effective antagonist of TACE.

ObjectiveTostudytheincidenceandclinicalcharacteristicsoftheco-infectedsexuallytransmittedinfections(STI)inpatientswithgenitalherpes.MethodsTheclinicaldataof287caseswithherpeticlesionsorpatientswithsuspectedherpeslesionswerecollected,andthepathogensofsexuallytransmittedinfectionsweredetected.ResultsGenitalherpeswasconfirmedin64.8%(186/287)oftherecruitedcases.HIVantibodiesweredetectedin68cases,andnoHIVantibodywasdetected.Theco-infectionssuchascondylomaacuminatum,activeorlatentsyphilis,genitalcandidiasisandotherSTIswerediscoveredin23.1%(43/186)ofpatientswithgenitalherpes.Allgenitalherpescasesco-infectedwithotherSTIswerecausedbyHSV-2.ConclusionTheco-infectionsarecommoninpatientswithgenitalherpes,andthefeaturesofthelesionsmaybechangedbytheseco-infections.

Objective Surface modification of hydrophobic nanoparticles is a key process for their application in the biological medicine fields. The aim of present study is to prepare the immunofluorescent probes by conjugation of hydrophilic surface-engineered quantum dots (QDs) and monoclonal antibody for use of fluorescence labeled cells tracing. Methods The bovine serum albumin (BSA) with excellent water-solubility and biocompatibility was employed as the emulsifying agent, and used for surface modification of hydrophobic QDs under ultrasonication. The diameter, fluorescence spectrum and cytotoxicity of BSAcoated QDs (BSA-triP.QDs) were analyzed. Then the BSA-coated QDs were conjugated with trastuzumab, which can be recognized and bound specifically to HER2. SKBR-3 breast cancer cells, with over-expression of HER-2, were labeled with hydrophilic QDstrastuzumab and imaged on a fluorescence planar imaging system. Results The experimental results revealed that BSA as the emulsifying agent, combined with other polymers, can effectively phase transfer hydrophobic QDs. The BSA functionalized QDs exhibited excellent colloidal stability with fine hydrodynamic size distribution (about 70nm) in a wide range of pH and ionic strengths values. Moreover, no significant cytotoxic effect was observed in SKBR-3 cells treated with BSA-coated QDs. After being coupled with trastuzumab, the hydrophilic QDs can be used as an immunofluorescence probe for HER2-positive breast cancer cell imaging. Conclusions The advantages of BSA-coated QDs include straight forward synthesis, high colloidal stability, and promising immunofluorescence characteristic when coupled with specific antibody. These are therefore proved to be ideal nano systems for biomedical labeling, targeting, and imaging.

Cytochrome P450 enzymes participate in the biotransformation of various organic compounds in living organisms. Cytochrome P450 enzymes are present and active in various brain regions, which mediate the synthesis and metabolism of various endogenous substances in the brain, so the enzymes play an important role in maintaining homeostasis of the central nervous system. And cytochrome P450 enzymes in brain are widely involved in metabolism of central drugs such as tramadol, so the changes of their activities or expressions may affect the drug's efficacy in brain. In addition, in view of the important roles of cytochrome P450 enzymes in the biotransformation of endogenous and exogenous substances in brain, the importance of their genetic polymorphism or activity changes in neuropsychiatric diseases is being gradually described. Therefore, in order to provide theoretical basis for the development of related targeted therapeutic drugs and explore whether cytochrome P450 enzymes in the brain can be used as therapeutic targets for central diseases, the development in the function of cytochrome P450 enzymes in brain and their recent advances in neuropsychiatric diseases by reviewing the relevant research in recent years are summarized.

BACKGROUND: Wnt signaling pathway plays an important role in the regulation of stem cells, but its regulatory effect on limbal stem cells is not well defined. OBJECTIVE: To investigate the regulation of limbal stem cells via Wnt signaling pathway and its function in the treatment of limbal stem cell deficiency. METHODS: Rat limbal segments were digested with Dispase and trypsin/EDTA and then single limbal stem cells were seeded in 3D-Matrigel. In experimental group LiCl (500 μmol/L) was added to the culture system. Control group received no LiCl. The expression levels of p63α, CK12, CEBPδ and Ki67 in limbal stem cells were detected by qRT-PCR on the 7th day of culture. The expression level of β-catenin in limbal stem cells was detected by immunofluorescence staining. The rat model of limbal stem cell deficiency was made by alkali burn method. Rats in treatment group were treated with Wnt-activated limbal stem cells by subconjunctival injection. Rats in control group were treated with PBS. Rats were checked by slit lamp every day. On the 4th day after treatment, the immunofluorescence staining and hematoxylin-eosin staining were applied to evaluate the repair of limbus. RESULTS AND CONCLUSION: (1) The limbal stem cells aggregated in 3D-Matrigel. β-Catenin was negative in the control group. β-Catenin was found in the cytoplasm and nucleus of the limbal stem cells in the experimental group. (2) qRT-PCR results showed there was no significant difference in the levels of p63α, CK12 or CEBPδ between control and experimental groups (P > 0.05). The Ki67 level in the experimental group was significantly higher than that in the control group (P < 0.05). (3) The rat models of limbal stem cell deficiency were established. The degree of corneal opacity in the treatment group was significantly lower than that in the control group (P < 0.05). Hematoxylin-eosin staining results observed that the corneal epithelial cells in the treatment group were neatly arranged, the cell size was uniform, and repair was good. Immunofluorescence staining showed that β-catenin was found in the cytoplasm and nucleus of most corneal epithelial cells in the treatment group. While β-catenin was only weakly positive in the control group, and was invisible in the cytoplasm or nucleus. (4) These results indicate that activation of Wnt signaling pathway in limbal stem cells enhances their proliferation and keeps them in an undifferentiated self-renewal state. Wnt-activated limbal stem cells can promote the regeneration of corneal epithelium and reduce the degree of corneal opacity in the treatment of limbal stem cell deficiency. The regulation of limbal stem cells via Wnt signaling pathway is expected to provide new ideas for the treatment of limbal stem cell deficiency.

Objectives To investigate the infection and colonization of Mycoplasma genitalium and Ureaplasma urealyticum in different male populations, to explore the association of M. genitalium and U. urealyticum with nongonococcal urethritis (NGU) respectively. Methods A case-controlled, cross sectional study of four different male populations was performed, namely: NGU patients (G1), non-NGU subjects attending STD clinic (G2), men who had sex with men participating in a health education program (G3), and healthy volunteers (G4). Nested PCR and culture were used to detect U. urealyticum. Nested PCR and PCR product sequencing were applied to detect M. genitalium. Results The prevalence rates of M. genitalium in the four study populations were 25.0%(25/100), 6.4%(6/94), 5.5%(6/110) and 0% respectively. Significant difference was found between each two groups except G2～G3 with a p value of 0.80. By multivariate regression analysis, controlling for the age of first sex, new sexual partners, urethritis and condom use in the previous 3 months, M. genitalium was only associated with urethritis (P= 0.004, OR = 6.754, 95% CI 1.833～24.893). The direct sequencing of PCR products showed gene mutations, in comparison with the reference sequence in GenBank, in 3 samples. The prevalence rates of U. urealyticum by PCR in 4 groups were 40.0%, 44.7%, 22.7% and 46.9% respectively, and there was no significant difference between G1～G2, G1～G4 or G2～G4 with a p value of 0.419, 0.325, 0.868 respectively, but the prevalence rate of U. urealyticum in G3 was significantly lower than that in other groups. Conclusions M. genitalium is strongly associated with NGU and the prevalence rate is significantly higher in groups with high risk sexual behaviors than that in general population. There is no association between the colonization of U. urealyticum and NGU.

AMP-Activated Protein Kinases/metabolism* ; Adenosine Monophosphate/metabolism* ; Kidney/metabolism* ; Phosphorylation