OBJECTIVE To investigate systematically the existence of bactericidal/permeability increasing protein(BPI) in normal rat organs and tissues,for providing important information about BPI in clinical application.METHODS The total RNA was extracted from organs and tissues homogenates.Then the first-strand cDNA was synthesized and BPI DNA was amplified by PCR.RESULTS BPI mRNA was found in kidneys,ovary,testes,liver,small intestine,large intestine,thymus,spleen and neutrophils of 21 various organs and tissues.Among them,BPI mRNA content was the richest in testes,relatively rich in liver and large intestine.CONCLUSIONS BPI mRNA presents in many organs and tissues of rat and it indicates that the role of BPI in host defense against bacterial infection is relatively widespread.

Objective To compare sequence analysis of the yeast-like fungal isolates with traditional methods and analyze the feasibility of identification of common yeast-like fungal by sequence analysis of gene. Methods 115 yeast-like fungal isolates were collected in the clinical laboratory of Beijing Tongren Hospital. DNA of yeast-like fungal was extracted and then amplified with universal primers of part of 18S rRNA genes followed by sequencing directly. The sequences obtained were submitted to the GenBank (NCBI) to identify the fungi. At the same time, the CHROMagar Candida and Vitek 32 YBC were used to identify the fungi. The identification accuracy with three methods was compared to explore the feasibility of the identification of sequence analysis. Results 18S rRNA gene sequence analysis was compared with traditional method. There were some differences in the identification results of 13 strains. The coincidence rate between CHROMagar Candida and sequence analysis was 89. 2% (91/102) and the coincidence rate between Vitek 32 YBC and sequence analysis was 91.3% (105/115). The positivity rate of species-level identification by CHROMagar Candida , Vitek 32 YBC and the 18S rRNA gene sequence analysis were 88. 7 % ( 102/115 ), 100% ( 115/115 ), 100% ( 115/115 ). Conclusion Identification of medically important yeast-like fungal by sequence analysis of the 18S rRNA gene is reliability.

Objective To investigate the effect of propofol exposure on neuroapoptosis in pri-mary cultured cortical neurons and its mechanisms.Methods Cortical neurons were primarily cultured for seven days,then divided into two groups:control group (treated with equal volume of 20% in-tralipid),propofol-treated group (treated with 500 μmol/L propofol).The neurons were treated for 12 h.The neuron viability was determined by MTT.Neuroapoptosis was identified by Hoechest 33 258 dying.Mitochondrial membrane potential was measured by the fluorescent dye rhodamine 123 (Rh123).Western blot was performed to detect the level of cyt-c and cleaved-caspase-3.Results Neu-rons survival rate (54.4%±6.4%)in the propofol group was significantly lower than that of control group (99.8% ± 4.1%) (P < 0.05 ), the rate of neuronal apoptosis (46.5% ± 5.3%) was significantly higher than that of control group (7.2%±0.9%)(P <0.05),mitochondrial membrane potential (59.6%±4.3%)was significantly lower than that of the control group (99.9% ± 5.7%) (P <0.05 ),cyt-C protein level (0.38 ± 0.03 )was significantly higher than that of control group (0.1 5±0.02)(P < 0.05 ),level of cleaved-caspase-3 protein level (0.46 ± 0.04)was significantly higher than that of control group (0.13±0.02)(P <0.05).Conclusion Propofol induces neuroapo-tosis in primary cultured cortical neurons,which is associated with the decreased level of MPP and the increase levels of cyt-c and cleaved-caspase-3.

0.05).Left ventricular mass index and the apoptotic rate of cardiac muscle cells were decreased in low-and high-dose herbal medicine groups(P

Objective To explore the mechanisms of synergic action of commonly-used herb pair of Radix Sophorae Flavescentis (Kushen, KS) and Fructus Cnidii (Shechuangzi, SCZ) for the treatment of pruritus. Methods We predicted and analyzed the potential targets of KS and SCZ based on network pharmacology method, and then established Chinese herbs-compound skeletons-targets networks to reveal anti-pruritus targets, predicting targets, and the interaction of KS and SCZ, as well as the components of the herb pair. The RAW264.7 inflammatory cell model was established to observe the synergistic anti-inflammatory activity of KS and SCZ. Results KS and SCZ had possible synergic action on the pruritus-related targets such as histamine receptors, cannabinoid receptors and proteinase-activated receptor 2. Additionally, KS and SCZ probably had synergic regulation of inflammation-related pathway (Toll -like receptor signaling pathway , chemokine signaling and Fc epsilon RI signaling pathway) and nerve-related pathway(neurotrophin signaling pathway) during the treatment of pruritus. Flavones from KS and coumarin of SCZ had various synergistic anti-inflammatory activities(P<0.05), indicating that they played an important role in inhibiting pruritus induced by inflammation. Conclusion The method may reveal the molecular mechanism of KS and SCZ in inhibiting pruritus, which is important for the modernization of Chinese medicine and new drug development.

Objective To explore the changes of mTOR signaling in LPS/D-gal-induced acute hepatitis in mice . Methods Twenty-six healthy adult female ICR mice were divided into two groups:the control group and experimental group, 13 mice in each group .LPS/D-gal was used to induce acute hepatitis in the mice .The survival of mice was moni-tored within 24 hours after LPS/D-gal challenge .At 6 hours after challenge , samples of serum and liver tissue were collect-ed for further analysis.Results Injection of LPS/D-gal resulted in acute death of the mice within 24 hours.At 6 hours post LPS/D-gal injection , the blood levels of ALT and AST were significantly increased .The mRNA expression of inflammatory cytokines Tnfa and Il6 was up-regulated in LPS/D-gal-induced hapatitis , in which DNA fragmentation and activation of caspase-3 were subsequently observed .Immunoblot analysis showed that both mTOR pathway and NF-κB pathway were ac-tivated.Unexpectedly , inhibition of mTOR signaling could neither decrease the apoptosis in the liver nor increase the sur -vival of mice .Conclusions The results of the present study indicate that mTOR signaling may play pleiotropic roles in the pathogenesis of LPS/D-gal-induced hepatitis .

Objective To investigate the clinicopathological significance of expression of insulin like growth factor 1 receptor (IGF1R) protein in primary gastrointestinal stromal tumors (GIST) and their impacts on prognosis.Methods Between January,2005 and January,2011,84 primary GIST patients underwent surgery.Immunohistochemical analysis was performed based on tissue microarray to estimate expression pattern of IGF1R in tumor cells and nomal controls.Association of IGF1R expression with clinicopathological features and relapse-free survival (RFS) were also analyzed.Results The negative,weakly positive,positive,and strong positive expression rates of IGF1 R protein in GIST group were 20％,14％,48％ and 18％,respectively;while in the control group were 32％,40％,20％ and 8％,respectively (x2 =30.663,P ＜ 0.001).The 5-year overall survival (OS) rate was 93％.1-year,3-year and 5-year RFS rate were 99％,76％ and 60％,respectively.As shown by univariate analysis the following factors were poor prognostic indicators for RFS,non-gastric tumor location (P =0.017),large tumor size (P =0.022),high mitotic index (P ＜ 0.001),high cellularity (P =0.012),tumor rupture (P =0.013),absent or low expression of IGF1R (P =0.022).Tumor size (HR5.1-10 ≤5 cm =1.86,95％ CI:0.67-5.15;HR＞10 ≤5cm =6.71,95％ CI:0.67-5.15,overall P =0.023),and mitotic index (HR5.1-10/50HPFsvs.≤5/50HPFs =5.72,95％ CI:2.09-15.64;HR＞10/50HPF ≤5/50HPFs =3.44,95％ CI:1.13-10.45,overall P =0.002) were negative independent risk predictors by multivariate analysis.Conclusions High expression of IGF1R may be involved in the occurrence,development and poor prognostic of primary GIST.Expression of IGF1 R is correlated with high risk potential and may predict early recurrence.

Objective To explore the significance in judging the different clinical stages of relapsing polychondritis (RP) patients through examining the changes of aggrecanase and metabolic fragments of aggrecan.MethodsIn comparison with the control group (20 cases),40 patients were divided into the stable stage group (22 cases) and the active stage group (18 cases).The aggrecanase-generated neoeptitopes in cartilage matrix were analysed by immunohistochemistry and Western blot(WB) respectively.The mRNA and protein levels of aggrecanase-1,2 expressed in cartilage cells were measured by real-time reverse transcriptional polymerase chain reaction(RT-PCR) and WB respectively.The difference of these results among these three groups was analyzed accordingly.ResultsThe expression of aggrecanase-1,2 in mRNA level was measured by real-time RT-PCR.The values of aggrecanase-1,2 mRNA 2 -ΔΔC1 were 1.00 ± 0.26 and 1.00 ± 0.27 in control group,1.47 ± 0.11 and 1.57 ± 0.13 in stable stage group,2.09 ±0.12 and 2.09 ± 0.19 in active stage group respectively.By one-way ANOVA analysis,the difference between every two groups was statistically significant (F was 299.113 and 459.013,P ＜ 0.01 ).In comparison with control group,aggrecanase-1,2 increased significantly in both stable and active stage group (P ＜ 0.01 ) and aggrecanase-1,2 increased more significantly in active stage group than in stable stage group (P ＜ 0.01 ).The results from WB analysis indicated that aggrecanase-1,2 could not be detected in control group,and they were detectable in stable stage group and increased in active stage group at the relative molecular of 68 000 Da or 73 000 Da respectively.The aggrecanase-generated neoeptitopes were analyzed by WB as well.The results indicated that NITEGE and ARGSV could be detected in stable stage group and increased in active stage group at the relative molecular of 70 000 Da or 48 000 Da respectively,but there were no signals in control group.Similar with the previous WB results,no signals of NITEGE or ARGSV eptitopes were detected in normal cartilage matrix ( no red staining) by use of immunohistochemical staining.However,in stable stage group and active stage group,these eptitopes were apparently detected (obviously red staining).ConclusionWith the progression of the RP,the activity of the aggrecanase is enhanced,and the degradation of the aggrecan is increased,associated with the severity of the disease.

Objective To establish a molecular technique of internal transcribed spacer (ITS) sequencing to identify pathogenic fungi species from the fungal sinusitis tissues. Methods Total 270 sinusitis tissues samples were collected by endoscopic surgery from 2006 to 2008. The histopathology, organize spring clip culturation and ITS region (ITS region region of fungal rRNA, including ITS1-5. 8S rRNA-ITS2) sequencing were employed simultaneously. And then to evaluate the ITS sequencing as the tool for identification of pathogenic fungi directly from clinical samples. Results Of the 270 samples, histopathology positive rate was 80.0% (216/270) , organize spring clip positive rate was 80.0% (216/ 270), fungal culturation positive rate was 53.0% (143/270) , ITS region sequencing positive rate was 63. 0% [ (134 +28 +8)/270], There were 22 species and 6 genera identified by fungal culturation, and 32 species identified by ITS region sequencing. Conclusion ITS region sequencing will become a applicable tool in clinical laboratory in future.

Objective:To analyse the causes of early implantation failure and the therapeutic measures with re-implantation after the failures.Methods:6 cases of implantation failure including early infections,loosening and non-osteointegration were reviewed and trea-ted by re-implantation therapy,and the causes of failure were discussed and the effects of re-treatment were evaluated.Results:2 cases were found to be with infection of adjacent teeth after implantation and were treated by removal of the implant,socket curettage,root ca-nal therapy(RCT)and antibiotics followed by reimplantation.Implant loosening and non-osteointegration were observed in 4 cases, which were treated by the similar methods for the implant socket.Reimplantation was successful in all 6 cases followed-up for 1 -3 years.Conclusion:Preventive measure for implantation failure should include indication selection,control of infections in adjacent teeth and periodontosis,use of GBR technic and so on.Re-implantation following proper treatment of adjacent teeth and the socket of implant is effective for the treatment of implantation failure.