The viable but noncuturable (VBNC) state is a survival strategy when bacteria are exposed to en- vironment stress. The VBNC state is part of the life cycle of non-differentiating bacteria, and it has a far-reaching impact on traditional bacteriology. Cells in the VBNC state fail to grow on the routine bacterio- logical media, here its significance in human health and environment science are detailed. Cells entering the VBNC state exhibit dwarfing and a number of metabolic changes in respiration rates and macromolecular synthesis. This paper summarized the variations in DNA and protein comparing to the culturable cells. We also discussed the ability of cells to resuscitate from the VBNC state and return to an actively metabolizing and culturable form. Some new methods for monitoring the VBNC state were listed. Finally the future was suggested.

Objective To observe the effect of low dosage of uhrashortwave(USW) on infarction volume, B cell lymphocytoma-xl (Bcl-xl) and tumor necrosis factor alpha (TNF-?) after cerebral ischemia-reperfusion in rats and discuss its acting mechanisms. Methods Focal ischemia-reperfusion model was established in 25 rats by re- versible right middle cerebral artery occlusion with filament. The right side cerebral ischemia was lasted for 2 hours and then followed with 24 hours of reperfusion. The content of neurological deficits were evaluated by the Zea-Longa 5-degree scoring system to select rats. After surgery, the rats were divided into 3 groups: blank control group, control group and USW treatment group. The brain of all rats was taken at 24 hours after reperfusion. The cerebral infarction volume, the expression of Bcl-xl and TNF-?were measured and analyzed. Results Twenty-five rats were used in the analysis of results. When compared with the control group, the infarction volume and rate in total cerebral volume of USW group significantly decreased (t = 2.54, 2.33, P

By reverse transcription-polymerase chain reaction (RT-PCR), an open reading frame of pathogenesis-related protein 1 (PR1) was isolated from Panax notoginseng and named as PnPR1. Molecular and bioinformatic analyses of PnPR1 revealed that an open reading frame of 501 bp was predicted to encode a 166-amino acid protein with a deduced molecular mass of 18.1 kD. Homology analysis showed that the deduced amino acid sequence of PR1 protein of Panax notoginseng had a high similarity with other higher plants had the same conservative structure domain of cysteine-rich secretory protein (CAP). The recombinant expressed plasmid pET28a(+)-PnPR1 was expressed in Escherichia coli BL21. The expression conditions were optimized by induction at different times, different temperatures, different IPTG concentrations and different giving times. The optimum expression condition was 0.4 mmol.L-1 IPTG at 28 degrees C for 20 h. The successful expression of PnPR1 provides some basis for protein purification and preparation of the monoclonal antibody.
Amino Acid Sequence ; Cloning, Molecular ; Escherichia coli ; metabolism ; Molecular Weight ; Open Reading Frames ; genetics ; Panax notoginseng ; chemistry ; Phylogeny ; Plant Proteins ; genetics ; metabolism ; Plants, Medicinal ; chemistry ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Alignment

In this study the chemical constituents of the higher polar sustances from Desmodium caudatum were investigated.The compounds were isolated by using column chromatographies over silicagel, polyamide, ODS, Sephadex LH-20, and preparative HPLC. The structures of these compounds were identified on the basis of NMR and MS spectra. Thirteen compounds were obtained and their structures were identified as vanillin(1), loliolide(2), indole-3-carboxaldehyde(3), salicylic acid(4), swertisin(5), saccharumoside C(6), isosinensin (7), kaempferol 3-O-β-D-glucopyranoside-7-O-α-L-rhamnopyranoside (8), isovitexin (9), vitexin (10), nothofagin(11), resveratroloside (12), and 2"-α-rhamnopyranosyl-7-O-methylvitexin (13). Except for compound 5, the remaining compounds were isolated from D. caudatum for the first time. Compounds 2, 3, 6-8, 11-13 were separated from the genus Desmodium for the first time.
Apigenin ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Fabaceae ; chemistry ; Molecular Structure ; Saponins ; chemistry ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization

To construct the Bac-to-Bac expression system of Bombyx mori nucleopolyhedrovirus (BmNPV), a transfer vector was constructed which contained an Escherichia coli (E. coli) mini-F replicon and a lacZ: attTN7: lacZ cassette within the upstream and downstream regions of the BmNPV polyhedrin gene. B. mori larvae were cotransfected with wild-type BmNPV genomic DNA and the transfer vector through subcutaneous injection to generate recombinant viruses by homologous recombination in vivo. The genomic DNA of budded viruses extracted from the hemolymph of the transfected larvae was used to transform E. coli DH10B. Recombinant bacmids were screened by kanamycin resistance, PCR and restriction enzyme (REN) digestion. One of the bacmid colonies, BmBacJS13, which had similar REN profiles to that of wild-type BmNPV, was selected for further research. To investigate the infectivity of BmBacJS13, the polyhedrin gene was introduced into the bacmid and the resultant recombinant (BmBacJS13-ph) was transfected to BmN cells. The budded viruses were collected from the supernatant of the transfected cells and used for infecting BmN cells. Growth curve analysis indicated that BmBacJS13-ph had a similar growth curve to that of wild-type BmNPV. Bio-assays indicated that BmBacJS13-ph was also infectious to B. mori larvae.

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

OBJECTIVETo explore the feasibility of evaluating complete ischemia-reperfusion injury (IRI) of the testis by contrast-enhanced ultrasonography with microbubbles (MB) targeted to P-selectin (MBp) in rabbits.

METHODSWe randomly divided 30 healthy adult rabbits into five groups of equal number (control, 0.5 h IRI, 1 h IRI, 2 h IRI, and 4 h IRI), prepared phospholipid MB and MBp, and performed contrast-enhanced ultrasonography of the bilateral testes with MB or MBp at an interval of 20 min at different times after IRI. When MB or MBp disappeared completely in the healthy testis at 4 to 5 min after intravenous injection, we recorded the power of the first frame (F-P) in the IRI testes followed by immunohistochemical staining of the testis tissue.

RESULTSCEU with MBp achieved a significantly higher F-P than that with MB in all the IRI groups (P < 0.05), which was (8.34 +/- 1.20) versus (1.87 +/- 0.25) 10(-5) AU at 2 hours, but there was no significant difference between MB and MBp in the control rabbits (0 AU, P > 0.05). Immunohistochemistry showed a significantly time-dependent increase in the expression of P-selectin in the vascular endothelial cells of the IRI testes, but not in those of the control.

CONCLUSIONContrast-enhanced ultrasonography with MBp can be used to evaluate the inflammatory reaction of testicular ischemia-reperfusion injury.

Animals ; Antibodies ; Disease Models, Animal ; Male ; Microbubbles ; P-Selectin ; immunology ; Rabbits ; Reperfusion Injury ; diagnostic imaging ; Testis ; blood supply ; Ultrasonography

[Objective]To discuss the microsurgical approaches of 30 patients with tentorial meningiomas and conclude their treatment experiences.[Methods]A retrospective analysis of clinical data was performed on 30 patients underwent microsurgical approaches from July 2004 to September 201 1.The pathology of these patients was confirmed after operation.The outcome and follow-up were evaluated.[Results] The meningiomas were totally removed in 19 patients subtotally in 6,and partially in 5.All the patients received 8 months to 5 years of follow-up:no mortality or death were found;no surgical related complications were noted.[Conclusion] Appropriate approaches according to the size and site of the tumors,intraoperative protection of venous sinus,facial and acoustic nerves and brain stem are the key points to improve the therapeutical outcomes and the post-operative quality of patients with infra and supra tentorial menmingiomas.

OBJECTIVETo study on growth characteristics of Curcuma wenyujin, and provide theoretical basis for the development of high-quality and high-yield medical material.

METHODThe morphological change of the plant was observed periodically, the content of volatile oil and dry matter in leaves, rhizome, root tuber was determine.

RESULTThe growth of C. wenyujin could be divided into 5 stages, i. e. seed germination, seedling, leaf growth, rhizome expansion, accumulation of dry matter, respectively. Before the stage of rhizome expansion, over 70% dry matter was accumulated in the aerial part of the plant, and during the stage of leaf growth, the maximum increase rate of dry matter in aerial part was 3.90 g/p/d. During the stage of rhizome expansion, the ratio of dry matter of rhizome increased quickly and reached above 33% , and the increase rate of dry matter of rhizome rise up to 3.83 g/p/d, in the end of the stage, the content of volatile oil in the rhizome also rose up to 1.20 mL x 100 g(-1).

CONCLUSIONDuring the whole growth stage, there are two growth centers, when the two curves of dry matter increase of aerial part and rhizome intersect, it is regarded as a signal that the growth transformed form the aerial part to rhizome. When the rate of dry matter from rhizome rise, the content of volatile oil in rhizome rises quickly with the increase of dry matter in rhizome. The optimal harvest time is in mid-December.

Curcuma ; anatomy & histology ; chemistry ; growth & development ; Germination ; Oils, Volatile ; analysis ; Plant Components, Aerial ; chemistry ; growth & development ; Plant Leaves ; chemistry ; growth & development ; Plant Roots ; chemistry ; growth & development ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development ; Rhizome ; chemistry ; growth & development ; Seasons ; Seedlings ; chemistry ; growth & development ; Seeds ; growth & development

OBJECTIVETo study the therapeutic effect of photocatalytic nano-TiO₂ on nasopharyngeal carcinoma xenograft in nude mice and underlying mechanism.

METHODSNude mice bearing human nasopharyngeal carcinoma xenograft were randomly divided into six groups: nano-TiO₂ + UV irradiation (with gradient concentration of nano-TiO₂); nano-TiO₂ alone and UV irradiation alone and blank control. The nano-TiO₂ suspension was injected into xenografts, and 24 h after UV light with the wave length of 330 - 400 nm, all the xenografts were removed and sectioned for HE staining. Ultrastructure and apoptosis of tumor cells in the xenografts were observed by transmission electron microscope (TEM). The expression of Caspase-3 was examined immunohistochemical staining and the apoptosis was detected with TUNEL.

RESULTSPathological analysis showed significant inflammatory responses (grade II and III) with local necrosis occurred in tumor tissues after nano-TiO₂ photodynamic therapy, but not in the negative control and blank control. TEM showed the nano-TiO₂ particles entered into the cytoplasm and the nucleus of tumor cells and many tumor cells had morphological changes for apoptosis. Significant positive expression of Caspase-3 and TUNEL-positive cells were found in the the xenografts with the treatments of nano-TiO₂ + UV irradiation compared to control (P < 0.01), which were enhanced with the increases in nano-TiO₂ concentration (P < 0.01).

CONCLUSIONPhotocatalytic nano-TiO₂ can inhibit the growth of nasopharyngeal carcinoma xenograft in nude mice by inducing Caspase-3 expression and apoptosis in the tumor cells.

Animals ; Carcinoma ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Mice ; Mice, Nude ; Nasopharyngeal Neoplasms ; drug therapy ; Photochemotherapy ; Titanium ; therapeutic use ; Transfection ; Xenograft Model Antitumor Assays