Effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm.

OBJECTIVE: To observe the effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm. METHODS: Micro-dilution method was used to determine the minimal inhibitory concentrations (MICs) of sansanmycin, gentamycin, carbenicillin, polymyxin B, roxithromycin, piperacillin, and tazobactam. PA1 and PA27853 biofilms were observed under optical microscope after staining and under SEM after treatment with sansanmycin at different dosages and combined treatment with sansanmycin and roxithromycin. Viable bacteria in PA1 and PA27853 biofilms were counted after treatment with sansanmycin at different dosages or combined treatment with sansanmycin and roxithromycin. RESULTS: The MIC of sansanmycin was lower than that of gentamycin and polymyxin B, but was higher than that of carbenicillin. Roxithromycin enhanced the penetration of sansanmycin to PA1 and PA27853 strains through biofilms. PA1 and PA27853 biofilms were gradually cleared with the increased dosages of sansanmycin or with the combined sansanmycin and roxithromycin. CONCLUSION: Sub-MIC levels of roxithromycin and sansanmycin substantially inhibit the generation of biofilms and proliferation of bacteria. Therefore, combined antibiotics can be used in treatment of intractable bacterial infection.

[Comparison of prognosis of small hepatocellular carcinoma patients with hepatitis B virus infection versus hepatitis C virus infection].

OBJECTIVE: To compare the prognosis of small hepatocellular carcinoma patients with hepatitis B virus infection versus hepatitis C virus infection. METHODS: 413 patients receiving curative resections at Tianjin Cancer Hospital for small HCC (< or = 3 cm) from January 1997 to December 2003 were divided into four groups: HCV only (n = 75), HBV only (n = 251), HBV and HCV (n = 33), and neither HBV nor HCV (NBNC, n = 54). The preoperative status and postoperative recurrence were recorded. Survival analysis were used to assess the impact of HBV/HCV status on HCC recurrence. RESULTS: Patients with HCV were associated with older age, lower mean preoperative platelet counts and albumin levels, higher mean prothrombin time, alanine aminotransferase and total bilirubin levels. Tumors in patients with HCV are multinodular and less differentiated, and were associated with a higher incidence of vascular invasion and cirrhosis. During the follow-up, the HCV group showed a higher incidence of intrahepatic recurrence and multiple recurrent lesions than the other patients. CONCLUSIONS: HCC patients with HCV infection tended to be older, and were characterized by more severe cirrhosis and higher incidence of tumor multinodular. The statistically significant determinants of reoccurrence in patients with small HCC were HCV infection, presence of vascular invasion and multiple tumors.

[Effects of Qiulirunfei extracta on immunological function and influenza virus in mice].

OBJECTIVE: To investigate the effect of Qiulirunfei extracta on immunological function and Influenza virus type A (FM1 strain) in ICR mice. METHODS: After given Qiulirunfei extracta, mice's body weight, spleen weight and lymphocyte stimulation index (SI) were tested; its antiviral effect was also observed in mice and chick embryo. RESULTS: Qiulirunfei extracta inhibited effectively the growth of influenza virus type A (FM1 strain) in chick embryo, and reduced the mortality rate of mice infected by influenza virus type A (FM1 strain). Compared with control group, body weight, spleen index in experiment group increased greatly, and SI of lymphocyte increased significantly too. CONCLUSION: Qiulirunfei extracta shows antiviral effects, and it is a good immune system enhancer.

[Infection and latency of HSV-1 on rat bone marrow mesenchymal stem cells in vitro].

To culture bone marrow mesenchymal stem cells (BMSCs) of rat in vitro and observe HSV-1 infection on BMSCs, BMSCs were separated from the bone marrow and identified by alizarin red staining and detection of ALP. The morphology of HSV-1 infected BMSCs and the CPE were observed. The total DNA was extracted from HSV-1 infected BMSCs and the desired specific gene fragment of 477bp of HSV-1 was amplified by PCR. Results showed that after BMSCs were induced by mineral-fluid for 14 days, the ALP level was increased and the nodule calcification was formed. The induced BMSCs were manifested to have the characteristics of osteoblasts. CPE couldn't be found in HSV-1 latently infected BMSCs but the 477bp gene fragment was still detectable. HSV-1 could establish latent infection in BMSCs after 7 passages. This study indicated that rat BMSCs could be induced to differentiate into osteoblasts in vitro, therefore they can be used as the seed cells for the tissue engineering. HSV-1 can infect rat BMSCs and develop the latent infection in vitro.

[Complete genome sequence analysis of a newly isolated paramyxovirus Tianjin strain].

Paramyxovirus Tianjin strain is the high-pathogenic virus to primate and might also cause human lower respiratory tract infection. To determine the genome structure, variation features and phylogenetic position, the complete nucleotide sequence of paramyxovirus Tianjin strain was analyzed. The homology comparison and phylogenetic analysis of the nucleotide and the deduced amino acid sequences among paramyxovirus Tianjin strain and the 28 strains in seven genera and the 7 unclassified viruses of Paramyxoviridae were performed. The results suggested that Tianjin strain is a member of the Respirovirus genus in the Paramyxovirinae, Paramyxoviridae and has the closest relationship to Sendai virus. Its genome length and composition are similar to the previously published Sendai virus except one extra glutamic acid residue increasing at the C terminus of Large protein due to the genomic RNA mutation at position A15240C. 440 unique nucleotide variations of Tianjin strain lead to 110 amino acid residue changes, making it differed from any other Sendai viruses. The phylogenetic analysis reveals paramyxovirus Tianjin strain doesn't belong to any of the three known evolution lineages of Sendai viruses and locates at a separate evolution branch. The obvious distinctions of genome nucleotide sequence, host tropism and pathogenicity suggest that paramyxovirus Tianjin strain might represent a novel genotype of Sendai virus.

In vivo pharmacokinetics of hesperidin are affected by treatment with glucosidase-like BglA protein isolated from yeasts.

Hesperidin is an abundant flavanone glycoside in citrus fruits and has been reported to possess a wide range of biological activities. However, hesperidin has poor bioavailability. Here, we tested the hypothesis that hesperetin found in chenpi will have a better bioavailability than hesperidin and that treatment of hesperidin with the glucosidase-like yeast Bg1A protein will increase its bioavailability. The results indicate that hesperidin in pure or extract form is hydrolyzed by BglA protein extracted from Sporobolomyces singularis or expressed in Escherichia coli BL21 (DE3). This biotransformation affected the plasma pharmacokinetics of total hesperetin in rats, in that the plasma T max was significantly shorter after administration of BglA protein-treated hesperidin than after administration of hesperidin extract. In addition, the area under the curve values for total hesperetin after administration of Bg1A-treated hesperidin were approximately 4-fold higher by oral administration and 3-fold higher by intravenous administration, respectively. In contrast, the plasma clearance value and volume of distribution after administration of Bg1A-treated hesperidin extract or pure hesperetin were significantly smaller than after administration of untreated hesperidin extract or pure hesperidin. This is the first study that systemically determines the absolute bioavailability of hesperidin and hesperetin simultaneously, shows clearly that hesperetin is more bioavailable than hesperidin regardless of the route of administration, and shows that prior transformation of hesperidin to hesperetin via fermentation should significantly increase its bioavailability because of the action of the yeast glycosidase-like protein BglA.

[Construction of recombinant eukaryotic expression plasmid of murine chemokine MIP-1alpha and coimmunization with HSV-II DNA vaccine].

AIM: To construct the recombinant eukaryotic expression plasmid of murine macrophage inflammatory protein-1alpha (MIP-1alpha) and investigate the effect of MIP-1alpha as an adjuvant on immune response induced by herpes simplex virus type II glycoprotein D (HSV-II gD) DNA vaccine. METHODS: Using total RNA from RAW264.7 cells stimulated with LPS, the whole code sequence of murine MIP-1alpha was amplified by RT-PCR and inserted into pcDNA3 at Hind III/Xba I restriction sites. The recombinant eukaryotic expression plasmid Pm was transiently expressed in COS-7 cells and its specificity was demonstrated by RT-PCR and Boyden chemotaxis chamber assay. BALB/c mice were immunized with gD DNA vaccine and/or MIP-1alpha, and the effect of MIP-1alpha on gD DNA vaccine was evaluated by detecting anti-HSV-II antibody, antigen-specific lymphoproliferative responses, and examining survival rates after mice were challenged intravaginally with HSV-II. RESULTS: The recombinant eukaryotic expression plasmid of murine MIP-1alpha was constructed, and it was revealed that immune responses of HSV-II gD DNA vaccine were enhanced by coimmunization with MIP-1alpha. CONCLUSION: The murine MIP-1alpha can be used as an adjuvant of HSV-II gD DNA vaccine.