Objective:To evaluate the acute oral toxicity , skin irritation and skin allergy of Thuja essential oil ( TEO) , and pro-vide experimental basis for the clinical use of TEO .Methods:The acute oral toxicity was measured by Horn ’ s assay .Totally 40 KM mice were divided into four groups and intragastrically administered with TEO at different dose of 21.50, 10.00, 4.64 and 2.15 g · kg-1 .After the 14-day observation, the death number and toxic manifestations were recorded and observed , and LD50 was calculated by checking the Horn's form of LD50 .The skin irritation test was performed on healthy adult white rabbits .Totally 9 rabbits were divid-ed into 3 groups randomly , and TEO at the concentration of 100%, 50%and 25%was painted on the skin of the rabbits .Edible vege-table oil was used as the negative control .The erythema and edema of the treated skin were evaluated and scored .Delayed skin hyper-sensitivity reaction was used to investigate the allergy of TEO .Totally 30 white guinea pigs were randomly divided into 3 groups:TEO group, the negative control (edible vegetable oil) and the positive group (1%2, 4-dinitrochlorobenzene).After the intracutaneous in-duction stage and local induction stage , TEO was used to activate the hypersensitive reaction .The skin response was observed and scored after the 24-hour and 48-hour activation.Results:The mice in 21.50 g · kg-1 TEO treatment group were all dead , while only a part of the mice in 10.00 and 4.64 g · kg-1 TEO treatment groups were dead , and no mice died in 2.15 g · kg-1 TEO treatment group.According to the Horn's form of LD50 , LD50 of TEO was 9.26 g · kg -1 for male mice and 7.94 g · kg -1 for female mice.The results of skin irritation test indicated the strong irritation effects of TEO .However , the irritation of TEO was reduced after the dilution , and 25%TEO showed no irritation to the skin of rabbits .The results of delayed skin hypersensitivity reaction showed obvious erythema and edema induced by 2, 4-dinitrochlorobenzene , while no obvious erythema and edema were found in TEO treated guinea pigs , indi-cating non-allergic effect of TEO .Conclusion:TEO has strong skin irritation in rabbits , while no obvious oral toxicity in mice and skin allergy in guinea pigs .

Objective: To evaluate the genetic toxicity of Thuja essential oil by salmonella reversion test (AMES test) and mammal micronucleus test.Methods: TA97, TA98, TA100 and TA102 were used in AMES test to evaluate the mutagenesis of Thuja essential oil.Mouse bone marrow micronucleus test was conducted to assess the chromosome toxicity of the drug.Results: Both in S9 present and absent situations, the numbers of reverse mutation of Thuja essential oil at different doses for the four strains were all less than 1-fold of that of solvent control, and the difference had no statistical significance (P>0.05), suggesting negative mutation.The micronucleus test indicated that Thuja essential oil had no influence on the rate of mouse bone marrow micronucleus (P>0.05).Conclusion: Thuja essential oil shows no obvious genetic toxicity.

Objective To investigate whether recombinant human endostatin can create a time window of vascular normalization prior to vascular pruning to alleviate hypoxia in Lewis lung carcinoma in mice. Methods Kinetic changes in morphology of tumor vasculature in response to recombinant human endostatin were detected under a confocal microscope with immunofluorescent staining in Lewis lung carcinomas in mice. The hypoxic cell fraction of different time was assessed with immunohistochemical staining . Effects on tumor growth were monitored as indicated in the growth curve of tumors . Results Compared with the control group vascularity of the tumors was reduced over time by recombinant human endostatin treatment and significantly regressed for 9 days. During the treatment, pericyte coverage increased at day 3, increased markedly at day 5, and fell again at day 7. The vascular basement membrane was thin and closely associated with endothelial cells after recombinant human endostatin treatment, but appeared thickened, loosely associated with endothelial cells in control tumors. The decrease in hypoxic cell fraction at day 5 after treatment was also found. Tumor growth was not accelerated 5 days after recombinant human endostatin treatment. Conclusions Recombinant human endostatin can normalize tumor vasculature within day 3 to 7, leading to improved tumor oxygenation. The results provide important experimental basis for combining recombinant human endostatin with radiation therapy in human tumors.

The mechanism of chromium accumulation and microstructure transformation of the fused yeast were studied in this paper.The result showed that the process of Cr6+ reduction and adsorption was accom-panied by the H+ consumption.The main adsorptive groups on the strain surface included amino,hydroxyl,phosphate group and amide,among which phosphate group played vital role in the chromium accumulation.The removal rate of chromium and reduction rate of hexavalent chromium declined 70% and 46%,respec-tively,when phosphate group was masked.During the adsorption process the chromium ions complexed on the surface of fused yeast was transported into the cell wall and combined with inclusion to form steady spe-cies and this took 90 min to reach the equilibrium.The biosorption and reduction of Cr on the cell surface would alter microstructure of cell surface,reduce cell membrane potential and increase cell membrane per-meability.

The stability of microbial flocculant (MBF) produced by Aspergillus sojae and its application to municipal thickened sludge dewatering were studied. The results showed that the MBF had high heat and acid-base endurance with high flocculating activity in a wide range of pH from 1.5 to 12. The MBF retained 96% of flocculating activity after 35 days preservation at 4℃, but in different pH the flocculating activity difference was very apparent after 35 days store at room temperature. The experimental results also demon- strated that the MBF was better than PAM and PAC in reducing specific resistance filtration. The optimal dose of MBF used for intensifying thickened sludge dewatering is 7%(volume fraction). And the more the volume of sludge is treated, the less the cost of MBF for unit volume sludge treatment.

OBJECTIVETo investigate the feasibility of monitoring therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on a rabbit VX2 liver tumor model by using phosphorous-31 magnetic resonance spectroscopy (31P MRS).

METHODSA total of 18 healthy New Zealand White rabbits were used to generate animal models by implanting VX2 tumor chips into livers through laparotomy. Tumor-bearing animals were randomly divided into three groups and were injected with AdCMVIL12-IRES-CKb, AdCMV-Empty and saline respectively via ear veins. 31P MRS scan was performed after animals were fed with creatine solution for five days. Animals were euthanized thereafter and tumors were removed for pathological examination, immunohistochemistry (IHC) staining and protein analysis (Western blot).

RESULTSThe intrahepatic and seral expressions of creatine kinase (CKb) and IL-12 were detected only in AdCMVIL12-IRES-CKb group. Tumor diameters pre- and post- treatment in three groups were 1.63+/-0.04 vs 1.62+/-0.03 in AdCMVIL12-IRES-CKb group (P = 0.229), 1.59+/-0.05 vs 1.84+/-0.11 in AdCMV-Empty group (P = 0.003) and 1.60+/-0.02 vs 2.07+/-0.12 in saline group (P = 0.001), respectively. Pcr Changes between pre- and post- treatment among the three groups were compared (F = 6.235, P value is less than 0.05). PCr increased significantly in AdCMVIL12-IRES-CKb group as compared to AdCMV-Empty (P = 0.004) and saline group (P = 0.049), whereas no change found between AdCMV-Empty and saline group (P = 0.153).

CONCLUSION31P MRS, an effective and non-invasive functional imaging method, can be used to monitor the therapeutic effect of adenovirus vector containing IL12-IRES-CKb gene on rabbit VX2 liver tumor model through detecting metabolic product of imaging reporter gene CKb (pCr).

Adenoviridae ; genetics ; Animals ; Creatine Kinase ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; Interleukin-12 ; genetics ; Liver Neoplasms, Experimental ; genetics ; pathology ; Magnetic Resonance Spectroscopy ; Rabbits

BACKGROUNDAdoptive transfer of allogeneic tumor-specific T cells often results in severe graft-versus-host disease (GVHD). Here, we sought to maximize graft-versus-tumor and minimize GVHD by using haploidentical T cells in pre-irradiated B16-melanoma bearing mice.

METHODSC57BL/6 mice bearing B16-melanoma tumors were irradiated with 0, 5, or 7 Gy total body irradiation (TBI), or 7 Gy TBI plus bone marrow transplantation. Tumor areas were measured every 3 days to assess the influence of irradiation treatment on tumor regression. B16-melanoma bearing mice were irradiated with 7 Gy TBI; sera and spleens were harvested at days 1, 3, 5, 7, 9, 11, and 13 after irradiation. White blood cell levels were measured and transforming growth factor β1 (TGF-b1) and interleukin 10 (IL-10) levels in serum were detected using enzyme-linked immunosorbent assay (ELISA) kits. Real-time reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry were performed to test TGF-b1, IL-10 and Foxp3 mRNA levels and the proportion of CD4+CD25+Foxp3+ T-regulatory cells (Tregs) in spleens. B16-melanoma bearing C57BL/6 mice were irradiated with 7 Gy TBI followed by syngeneic (Syn1/Syn2) or haploidentical (Hap1/Hap2), dendritic cell-induced cytotoxic T lymphocytes (DC-CTLs) treatment, tumor areas and system GVHD were observed every 3 days. Mice were killed 21 days after the DC-CTLs adoptive transfer; histologic analyses of eyes, skin, liver, lungs, and intestine were then performed.

RESULTSIrradiation with 7 Gy TBI on the B16-melanoma-bearing mice did not influence tumor regression compared to the control group; however, it down-regulated the proportion of Tregs in spleens and the TGF-b1 and IL-10 levels in sera and spleens, suggesting inhibition of autoimmunity and intervention of tumor microenvironment. Adoptive transfer of haploidentical DC-CTLs significantly inhibited B16-melanoma growth. GVHD assessment and histology analysis showed no significant difference among the groups.

CONCLUSIONAdoptive transfer of haploidentical tumor-specific T cells in irradiation-pretreated B16-melanoma bearing mice preserved antitumor capacity without causing a GVHD response.

Animals ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Graft vs Host Disease ; Immunotherapy, Adoptive ; methods ; Male ; Melanoma, Experimental ; metabolism ; therapy ; Mice ; Mice, Inbred C57BL ; Real-Time Polymerase Chain Reaction ; T-Lymphocytes ; immunology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo explore the regeneration of mechanical sensory fibers after free nerve transplantation.

METHODNeuroelectrophysiological technique (single nerve fiber recording) was used to test the regeneration rate of mechanical sensory fibers, the proportion of rapidly and slowly adapting receptors, the stimulating thresholds of regenerated mechanoreceptors and conduction velocity of regenerated fibers. The regeneration pattern of the mechanoreceptors after free nerve transplantation to the rabbit reconstructed penis was also analyzed.

RESULTS9 months after operation, the number of regenerated mechanical sensory fiber was almost normal. The regenerated rapidly adapting receptors had a higher proportion with higher mature degree than the regenerated slowly adapting receptors. 9 months after nerve transplantation the stimulating thresholds of regenerated mechanoreceptors and conduction velocity of regenerated fibers remained below normal.

CONCLUSIONAfter free nerve transplantation to the rabbit reconstructed penis, the function of both rapidly and slowly adapting sensory nerve fiber partially recovered, but in different extent.

Animals ; Ganglia, Sensory ; physiology ; Male ; Mechanoreceptors ; physiology ; Nerve Fibers ; transplantation ; Nerve Regeneration ; Penis ; surgery ; Rabbits

OBJECTIVETo investigate the mechanism of sensory nerve regeneration of the reconstructed penis with sensory nerve implantation and to explore a new surgical technique to improve the postoperative sensory function in phallic reconstruction.

METHODSAdult male New Zealand rabbits were randomly divided into experimental group (n = 20, with sensory nerve implantation) and control group (n = 20, without sensory nerve implantation), which were both performed phalloplasty with a superficial epigastric faciovascular pedicle flap. Postoperatively, the nerve regeneration process of the reconstructed penis was observed histologically.

RESULTSIn experimental group, the amount of CGRP positive nerve fibers increased markedly with the time prolonged, whereas merely a few CGRP positive fibers scattered in deep dermis 6 months later in the other group. The cutaneous sensory nerve regeneration of the reconstructed penis in experimental group shows the procedure that the myelinated axon began to exist within 3 months, thereafter the myelinated axon and unmyelinated axon were both observed under the electron microscope.

CONCLUSIONThese findings show that the rabbit model of phalloplasty with sensory nerve implantation can acquire well sensory reinnervation, and bring a light to clinical application for restoration of sensory function in reconstructed penis.

Animals ; Male ; Nerve Regeneration ; Penis ; innervation ; surgery ; Rabbits ; Surgical Flaps ; innervation

ObjectiveThe study is to investigate the predictive values of dosimetric parameters and patient related factors in severe acute radiation pneumonitis (SARP) after concurrent chemoradiotherapy in non-small cell lung cancer (NSCLC).Methods In all,147 NSCLC patients treated with concurrent chemotherapy and 3DCRT between 2006 and 2010 was collected.Independent sample t test was used to compare parameter values between patients with SARP and those without SARP.Logistic regression was used to identify significant determined factor.Predictive value of each parameter was tested by ROC analysis.Pearson correlation was used to analyze correlations between parameters.Represent factors were identified by factor analysis.ResultsThe incidence of SARP was 9.5％ ( 14/147 ).The means lung dose (MLD),V20,V30,V40,and V50 ( x2 =4.87 -6.84,P =0.009 -0.025,respectively ) were determining factors for SARP.Our datasets shows that for SARP ＜5％,MLD,V20,V30,V40 and V50 should be ≤16.77 Gy,V20≤34.15％,.V30 ≤23.62％,.V40 ≤ 18.57％,V50 ≤ 13.02％.ROC analysis show that areas under MLD,V20,V30,V40 and V50 curves was corresponding to 0.678,0.661,0.667,0.677,and 0.651,respectively.In addition,the sensitivity and specificity of each parameter at cutoff values are:78.0％ and 48.1％ for MLD;42.9％ and 82.0％ for V2o ;78.6％ and 52.9％ for V30 ;71.4％ and 61.7％ for V40,and 57.1％ and 67.7％ for V50.Factor analysis suggest that we can choose 1 or 2 parameters from MLD,V20,or V30,and another from V40 or V50 for predicting.The incidence of SARP was greater in patients with tumorsin right lower lung than other locations ( 22.2％ vs 6.7％,x2 =6.19,P =0.0 2 3 ).Conclusions The MLD,V20,V30,V40 and V50 are determining factors for SARP.As predictive value of each parameter alone is relatively week,using two or more parameters to predict SARP is recommended.