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 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 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.

Objective To investigate the non-medication therapy for patients withhypertension,and to provide guidance to the further non-medication therapy program.Methods 86 hypertensive patients were interviewed by self-designed questionnaire and anxiety self-assess scale (SAS),and the data was analyzed by spss 13.0 software.Results The mean score was ( 10.42 ± 3.69) ; The mean BMI was (29.96 ± 3.91 ) kg/m2,among them there were 61 patients with BMI ＞ 24 kg/m2 (70.9％ ); Hypertensive patients with blood pressure ≥ 140/90 mm Hg have worse non-medication therapy ; The average score in women was ( 11.50 ± 2.60)and (9.71 ±4.13) in men,the difference has reach the statistical significance (t =-2.466,P ＜0.05).Age,educational level and disease duration were different in non-medication therapy ( P ＜ 0.05 ).Low-fat dietary and no-smoking and no-drinking in women were better than those in man (P ＜ 0.01 ).Low-salt and lowfat dietary and regular dietary,no-smoking,weight controlling and exercise in those more than 60 years were better than those younger (P ＜ 0.05 ).Weight controlling in those with higher education lever was better than those with lower education certification (P ＜0.05 ).Exercise in those with 10 years disease duration above was better than those less than 10 years (P＜0.05).Average score of anxiety was (35.62 ±7.24),and the range was from 20 to 55.Conclusions The status of non-medication therapy for patients with hypertension was not well.We should pay more attention to develop health care education to improve patients' consciousness of nonmedication therapy,and change unhealthy life style.

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

OBJECTIVETo establish a drug-resistance cell line of human glioma mediated by MGMT.

METHODSSimulated the clinical usage of BCNU to establish a BCNU-resistant human glioma subline by cyclic exposing the U251 parent cells to a constant concentration of BCNU. The resistance index and the expression of MGMT mRNA of U251/BCNU were detected and compared the difference of in vitro proliferation between U251 and U251/BCNU.

RESULTSA subline--U251/BCNU was successfully established in about 4-month culture, which had a stable resistance to BCNU. U251/BCNU cells showed 17-fold higher resistance to BCNU than did U251 cells by MTT assay, while U251/BCNU cells expressed MGMT mRNA. The doubling time of U251 and U251/BCNU had no statistical difference.

CONCLUSIONA drug-resistance cell line of human glioma mediated by MGMT is established, which could provide experimental basis for further studies on the resistance mechanism and reversal methods of glioma chemotherapy.

ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Brain Neoplasms ; pathology ; Cell Line, Tumor ; DNA Modification Methylases ; biosynthesis ; genetics ; Drug Resistance, Neoplasm ; genetics ; Glioma ; pathology ; Humans ; O(6)-Methylguanine-DNA Methyltransferase ; metabolism ; physiology

OBJECTIVETo explicate the spinal nerve source of the rabbit penis cutaneous sensation.

METHODSTwelve adult male rabbits were randomly divided into two groups of equal number. While mechanical stimuli were given to the penis by different von Frey hairs, single fiber activities were recorded in vivo in the left (Group A) and right (Group B) S1-S4 spinal nerves, respectively. The mechanical threshold, adaptability and conduction velocity of the fibers were analyzed.

RESULTSWhen the ipsilateral penis was mechanically stimulated, discharges were detected in S2 and S3 spinal nerve fibers, but not in S1 and S4. The discharge fibers were 39.67 +/- 3.14 (S2) and 21.00 +/- 2.19 (S3) in the left spinal nerve and 40.00 +/- 3.16 (S2) and 19.67 +/- 2.58 (S3) in the right. There was no obvious difference between the numbers of the left spinal nerves and the right ones (P > 0.05).

CONCLUSIONThe rabbit penis cutaneous sensation originates from S2 and S3 spinal nerves.

Animals ; Electrophysiology ; Male ; Neurons, Afferent ; physiology ; Penis ; Rabbits ; Random Allocation ; Sensory Thresholds ; Skin ; innervation ; Spinal Nerves ; physiology

OBJECTIVETo express antibacterial peptide thanatin in the prokaryotic expression system and test its antibacterial activity.

METHODSThe DNA sequence coding for the 21 peptides of thanatin was synthesized using the preferred genetic codes of E. coli, cloned into pTYB11 plasmid, and transformed into E.coli ER2566. The expression of thanatin fused with intein was induced by IPTG in E.coli, and intein-thanatin specifically bound to the column through intein tag was cleaved overnight at 4 degrees celsius; in DTT/cysteine buffer.

RESULTSThe cleaved thanatin was eluted with a protein concentration of 245 microg/ml in the first 4 ml. The purified thanatin had showed strong antibacterial activities against G- bacteria such as Shigella flexneri, Klebsiella pneumoniae, Shigella snnei, Escherichia coli O157, toxin producing Escherichia coli, Pseudomonas aeruginosa, and fungi such as Candida albicans, with especial potency in killing drug-resistant Klebsiella pneumoniae, Pseudomonas aeruginosa, and extended-spectrum beta-lactamases (ESBL)-producing E.coli. Eighty strains of drug-resistant (ESBL-producing) and 30 strains of sensitive E. coli were used for anti-bacterial assay, and no significant differences in the antibacterial activity of thanatin were found between the sensitive and drug-resistant E. coli (P>0.05).

CONCLUSIONThe recombinant thanatin obtained shows strong antibacterial activity against drug-resistant and sensitive bacteria, and can be a potential substitute for routine antibiotics in the treatment of G- bacterial infections.

Anti-Bacterial Agents ; metabolism ; pharmacology ; Antimicrobial Cationic Peptides ; biosynthesis ; genetics ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology