The susceptibility to 9 kinds of antibiotics of 393 animal pathogenic E. coli isolated from clinical samples was determined from 2000 to 2003. The resistance to TC, GM, CMP, AMP, RA, KM, FT, SM and CIP were 93.89%, 57.76%,78.63%, 77.86%, 92.11% ,47.33%, 46.82%, 76.84% and 74.81%, respectively. The isolates could be classified into eight classes according to the number of drugs to which srains were resistant. The resistance spectrum of the isolates varied from 2to 9 kinds of the above drugs. The strains that were resistant to seven kinds of drugs were more than 80 percent. The resistance rates of swine origin strains to GM,AMP and KM were higher than those of poultry origin strains, while the resistance rates of poultry origin strains to TC, CMP, SM and CIP were higher remarkably than those of swine origins. The frequency of resistance increased from 2000 to 2003,which was from 67.33% to 90.58% for CMP, 71.29% to 84.81% for AMP, 73.76%to 80.10% for SM and 61.88% to 88.48% for CIP. At the same time, the resistance rate of GM decreased from 61.39% to 53.92%. Thirty-seven strains (95 %) could make all the Kunming mice die within 72 h injected intraperitoneally with the culmice three times in vivo. The pathogenicity of wild strains with drug resistance acquired naturaly to mice did not decline.

Purpose To investigate the diagnostic value of dual energy CT spectrum curve combined with CT morphology in central lymph node metastasis of papillary thyroid carcinoma (PTC).Materials and Methods Thirty-one PTC patients who accepted dual energy CT double-phase enhanced scan before surgery were analyzed.Central lymph nodes with short diameter ＞ 5 mm were labelled using axial plus 3D positioning.Preoperative labelled lymph nodes were collected and marked during operation.The CT morphology of metastatic lymph nodes was analyzed.The spectrum curve slope (K) difference between metastatic and non-metastatic lymph nodes of arterial-phase and venous-phase primary lesion was compared.The critical K value and the diagnostic efficacy of K value combined with morphology were obtained according to receiver operating characteristic (ROC) curve.Results A total of 73 central lymph nodes were obtained from 31 patients,among which 51 were metastatic and 22 were non-metastatic.There was no significant difference in K value between metastatic and non-metastatic lymph nodes of arterial phase group (P＞0.05).While,there was a significant difference in K value between metastatic and non-metastatic lymph nodes of venous phase group (P＜0.05).For venous phase,the sensitivity and specificity of the K value in diagnosing central lymph node metastasis were 62.7％ and 59.1％,respectively,and combined with morphology,the sensitivity and specificity reached 76.5％ and 81.8％,respectively.Conclusion The K value of CT spectrum curve is of certain significance in predicting PTC central lymph node metastasis,and the K value combined with CT morphology can improve the accuracy of diagnosis.

Objective To research the effects of drought stress on activity of key enzyme in saikosaponin biosynthesis and saponins content in Bupleurum chinense; To reveal response of Bupleurum chinense to drought stress from protein level. Methods Bupleurum chinense was cultivated in potted water control experiment with 70%–80%, 60%–70%, 40%–50% and 20%–30% saturated soil water contents. The enzyme activities of 3-hydroxy-3-methyl-glutaryl-coenzyme A reductase (HMGR), isoprenyl-based coke (IPPI), farnesyl pyrophosphate synthase (FPS) and β-carotenoid synthase (β-AS) of six months and one year old Bupleurum chinense were measured. The contents of saikosaponins a and d in samples of different soil water contents were determined by HPLC. Results When the saturated moisture was 40%–50%, the enzymatic activities of HMGR, IPPI, FPS and β-AS were the highest, followed by saturated moisture 60%–70%、70%–80% and 20%–30%. The trend of saikosaponin was similar to that of the activity of key enzyme. Conclusion 40%–50% soil saturated water content can significantly increase the activity of the key enzymes in the saikosaponin synthesis pathway. The saponin content and enzyme activity show a significant positive correlation, indicating that under drought stress, Bupleurum chinense regulates the synthesis of saikosaponin by regulating the key enzyme activity of saikosaponin synthesis pathway to respond drought stress.

Air ; analysis ; Chromatography, Gas ; methods ; Ethylene Oxide ; analysis ; Furans ; analysis ; Methylene Chloride ; analysis ; Workplace

BACKGROUND: Xiaohuoluo pill can expel pathogenic wind, remove dampness and activate collaterals. It is used for treatment of Bi-syndrome due to wind-cold-dampness, pain and numbness in limbs.OBJECTIVE: To observe the pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity (including cellular immunity and humoral immunity), non-specific immunity [including complement 3(C3), mononuclear phagocyte system (MPS) and red blood cell (RBC)adhesion function] and free radical injury as well as pain and many other inflammations in mice.DESIGN: A randomized controlled stratified trial.SETTING: Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Hospital of Traditional Chinese Medicine.MATERIALS: Totally 628 NIH and ICR mice of 6 to 8 weeks were involved in this trial. Xiaohuoluo pills (components: Dannanxing, Zhichuanwu, Zhicaowu, Dilong, Ruxiang and so on; Chenli Pharmaceutical Factory,Guangzhou; Brach No. 19980612) were used in this trial. Rabbit antimouse immunoglobulin G (IgG) and C3 antiserum reagent kit (Guangzhou Institute of Medicine and Health) and reagent kit for measuring the antioxidizing activity of superoxide dismutase (SOD) and the level of malondialdehyde (MDA) (Jiancheng Institute of Bioengineering, Nanjing) were used.METHODS: This trial was carried out in the Guangzhou Institute of Traditional Chinese Medicine and Chinese Materia Medica; Department of Pharmacy, Guangzhou Institute of Medicine and Health during September 1998 to December 1999. ① To observe the suppressive effect of Xiaohuoluo pills on cock red blood cell (CRBC)-induced secondary immune response: Eight-four ICR mice, male and female in half, were selected.Twenty of 84 mice served as blank controls; The other 64 mice were intraperitoneally injected with cyclophos-phamide (CY) of 0.2 g/kgonce. On the 4th and 12th days, CRBC was intraperitoneally injected into the mice twice to induce immunoenhancing pathological models to form secondary immune response. Mice served as blank controls were intraperitoneally injected with the same volume of normal saline; The immunoenhanced mice were assigned into 3 groups by a lot: CY group (n=20, CY, 40 mg/kg, intragastric administration, I.g.), Xiaohuoluo pills (n=21, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (n=20, distilled water, the same volume as other groups, I.g.); once a day within 7 successive days. 19 days later, the levels of serum IgG and C3 were measured with single immunodiffusion method, and the level of circulating immune compound (CIC) was measured with polyethylene glycol precipitation method. ② To observe the suppressive effect of Xiaohuoluo pills on delayed type hypersensitivity (DTH): Fifty-four ICR mice, male and female in half, were selected. On the 1st day, the mice were sensitized by subcutaneous injection of 10 g/L 2,4-dinitrofluorobenzen e (DNFB) of 50 μL for each. On the 4th day, the sensitized mice were assigned into 3 groups by a lot: Prednisone group (n=18, prednisone, 0.01 g/kg, I.g.), Xiaohuoluo pills (n=18, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.), model group (n=18, distilled water, the same volume as other groups, I.g.), all once a day within 7 successive days. 11 days later, 10 g/L DNFB of 25μL was spread on the right ear of each mouse in each group. The swelling degree was calculated 24 hours later (The mass difference between right ear and left ear). ③ To observe the suppressive effect of Xiaohuoluo pills on immune adhesion function of RBC of mouse: Thirty-six NIH mice, male and female in half, were selected and assigned into 3 groups by a lot: CY group (n=12, CY, 20 mg/kg,I.g.), Xiaohuoluo pills (n=12, Xiaohuoluo pills suspension, 5.54 g/kg, I.g.)and blank control group (n=12, distilled water, the same volume as other groups, I.g.), once a day within 7 successive days. 7 days later, blood was taken from the orbit of mice for calculating the rosette rate of RBC-C3b receptor and the rosette rate of RBC immune compound. ④ To observe the suppressive effect o20 Mg/kg, I.g.),Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) , once a day within 7 successive days; IgM-type hemolytic concentration (HC50)was measured at 2 hours after the last administration on the 7th day [ (Sample absorption / Absorption at HC50 of CRBC) ×diluted time]. The levels of serum C3 and MDA and the activity of SOD were measured according to the method from corresponding reagent kit. ⑥ To observe the suppressive effect of Xiaohuoluo pills on agar granulation tissue hyperplasia in mice:Fifty-nine NIH mice were selected and given subcutaneous injection of 20 g/L agar of 0.5 mL for each. 24 hours later, the mice were assigned into 3 groups by a lot: diclofenac group (diclofenac, 10 mg/kg,I.g..), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume as other groups, I.g.), once a day within 7 successive days; On the 8th day, the mice were sacrificed. The hyperplasiainhibiting effect was presented in the form of the mass of agar granulation tissue in one kilogram body mass ⑦ To observe the suppressive effect of Xiaohuoluo pills on acetic distortion reaction: Sixty-three NIH mice were se lected and assigned into 3 groups by a lot: diclofenac group (diclofenac,50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg,I.g.) and model group (distilled water, the same volume of other groups, I.g.),once a day within 2 successive days. At 2 hours after the last administration, the mice were given intraperitoneal injection of 0.1 mol/L acetic acid of 0.2 mL for each one. The times of distortion of mice within 20 minutes were counted. ⑧ To observe the effect of Xiaohuoluo pills on the acute exudative inflammation evoked by dimethylbenzene, croton oil and carrageenan, and the level of prostaglandin E in the inflammatory exudates:Totally 219 NIH mice were selected and assigned into 3 groups by a lot:diclofenac group (diclofenac, 50 mg/kg, I.g.), Xiaohuoluo pills group (Xiaohuoluo pills suspension, 5.54 g/kg, I.g.) and model group (distilled water, the same volume of other groups, I.g.) once a day within 2 successive days. At 2 hours after the last administration, dimethylbenzene of 25 μL was spread on the right ear for 20 minutes, or croton oil of 25 μL was also spread on the right ear, 4 hours later, the swelling of right ear was calculated (mass of right ear-mass of left ear). 10 g/L carrageenan of 20 μL was subcutaneously injected into the right foot, 3 hours later, the swelling degree was calculated (The difference of right foot and left foot); and the level of prostaglandin E in the inflammatory exudates was measured. ⑨ t test(t' test for heteroscedasticity) was used for comparing the difference in measurement data among groups.MAIN OUTCOME MEASURES: Pharmacological effect of Xiaohuoluo pills on secondary immune response, specific immunity, non-specific immunity and free radical injury as well as pain and many other inflammations in mice.RESULTS: Totally 628 NIH and ICR mice were involved in result analysis. ① The level of IgG and CIC of mice in the model group was significantly higher than that in the other 3 groups respectively (P ＜ 0.01),while the level of C3 was significantly lower than that in the other 3 groups (P ＜ 0.05 to 0.01). ② The swelling degree of mice in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group respectively ( both P ＜ 0.01). ③ The rosette rate of RBC-C3b receptor and RBC immune compound in the blank control group was significantly higher than that in the other 2 gro ups respectively (P ＜ 0.01). ④ The phagocytic index (K value )in the diclofenac group and Xiaohuoluo pills group was significantly lower than that in the blank control group, respectively (both P ＜ 0.01).⑤ IgM-type HC50 and the level of serum MDA of CY group and Xiaohuoluo pills group were obviously lower than those in the immune control group (P ＜ 0.01),while the level of C3 was higher than that of immune control group, there was no significant difference in the activity of serum SOD between CY group or Xiaohuoluo pills group and immune control group (P ＞ 0.05). ⑥The ratio of agar granulation tissue mass to body mass in the diclofenac group or Xiaohuoluo pills group was significantly lower than that in the model group(P ＜ 0.01).⑦ The times of distortion of mice within 20 minutes in the diclofenac group or Xiaohuoluo pills group were signifi cantly less than those of model group(P ＜ 0.01,0.05).⑧The ear swelling degree of dimethylbenzene-induced inflammatory models and croton oil-induced inflammatory models,and foot swelling degree of carrageenan-induced acute inflammatory models as well as the level of prostaglandin E in the inflammatory exudates in the diclofenac group were significantly milder or lower than those in the model group(P ＜ 0.05 to 0.01),and the level of prostaglandin E in the inflammatory exudates in the Xiaohuoluo pills group was significantly lower than that in the model group (P ＜ 0.01).CONCLUSION: Xiaohuoluo pills possess pharmacological effects of immunosuppression, anti-proliferative inflammation, analgesia and antioxidation.

Objective Carbonic anhydrase 1 (CA1) not only enhances the hydration reaction but also promotes the formation of CaCO3,which is an essential step for new bone formation in vitro.However,there is no direct evidence to demonstrate the involvement of CA1 in bio-mineralization in cells and tissues.This study is aimed to evaluate the important role of CA1 in bio-mineralization and ossification in cultured cells.Methods Calcification in Saos-2 cells was induced using osteogenic medium (OM) and the calcification was determined by Alizarin Red-S staining.The expressions of ossification protein marker Runt-related transcription factor-2 (Runx2),osterix (OSX),alkaline phosphatase (ALP),osteocalcin (OCN) and bone sialoprotein (BSP) were detected in the process of bone formation by real-time PCR.The expression of CA 1 in the calcified cells were measured using real-time PCR and Western blotting.We also detected calcification in Saos-2 cells in the presence of acetazolamide,an anti-carbonic anhydrase drug to CA1,to determine the role of CA1 in biomineralization in culture cells.T test analysis was used to compare the two groups,M-ANOVA of repeated measurs was conducted for different time point.Results Following the stimulation of OM,Saos-2 cells produced a great amount of calcium-rich deposits [0.68±0.03 vs 2.76±0.13,P＜0.01].Increased transcriptions of ossification protein markers were also detected in these stimulated Saos-2 cells,indicating that the OM launched the process of bone formation in the cells.CA1 had a significantly increased expression during this process [0.25±0.03 vs 0.94±0.06,P＜0.01].Following treatment with acetazolamide,the expression of CA1 evidently declined [1.09±0.05 vs 0.55±0.07,P＜0.05],and the mineralized nodule formation was declined [2.76±0.13 vs 2.19±0.07,P＜0.01].Conclusion These findings indicate that CA1 participates in the biomineralization and ossification,and may play an important roles in bone formation.

Objective To explore the effects of microRNA(miR)-1283 on invasion,proliferation and apoptosis of HTR 8/SVneo cell line derived from human-trophoblast cells.Methods HTR-8/SVneo cells were divided into three groups,as-miR 1283 group was transfected with miR 1283 analogues,anti-miR-1283 group was transfected with miR-1283 inhibitors,and negative control group was transfected with non functional sequence.The levels of miR 1283 expression were detected by fluorescence quantitative polymerase chain reaction at 24 hours after transfection.The cell proliferation was measured by 3-(4,5)-dimethylthiazol-2-yl-(2,5)-diphenyl tetrazolium bromide assay at 24,48 and 72 hours after transfection.Apoptosis was evaluated by propidium iodide staining and flow cytometry at 48 hours after transfection.Transwell experiments were performed to evaluate cellular invasion abilities at 24 hours after transfection.Analysis of variance and Bonferroni method were applied as statistical methods.Results The level of miR 1283 in as miR 1283 group was higher than that in the negative control group with statistically significant difference (14.85±0.57 vs 7.08±0.20,P＜0.01).At 24,48 and 72 hours after transfection,the inhibitory rate of cell proliferation in as-miR-1283 group was (8.04 ± 0.27) ％,(32.47 ± 0.24) ％ and (9.23± 0.17) ％,higher than those in the negative control group [(0.72 ± 0.06) ％,(1.17 ± 0.04) ％ and (0.53 ± 0.05) ％] (P ＜ 0.01,respectively).Cell apoptosis rate was higher in as-miR 1283 group than in negative control group [(16.33 ± 0.40) ％ vs (9.39± 0.58) ％,P＜0.01].The transmembrane cell number was lower in as-miR-1283 group as comparing with the negative control group (7.25 ± 1.83 vs 16.33 ± 2.08,P＜0.01).miR-1283 expression,apoptosis and transmembrane cell number in anti-miR-1283 group had no statistical difference as compared to the negative control group (all P ＞ 0.05).Conclusions Up-regulated levels of miR-1283 could inhibit HTR-8/SVneo cell proliferation and invasion,but promote the cell apoptosis.

Objective To investigate the rule of the cerebral tissues aquaporin-4(AQP-4) expression in acute and chronic hepatic failure mice.To study the molecular biologic mechanism of the diffusion weighted imaging(DWI).Methods Sixty five male Sprague-Dawley rats were divided into 3 groups randomly,including acute(n=25),chronic hepatic failure(n=25)and control group(n=15). Thioacetamide(TAA)intraperitoneal injection produces the acute and chronic hepatic failure models.All rats in groups were examined with MR DWI.We Observed the distribution of abnormal signal on DWI.The DWI single values of top and lateral cortex of parietal lobe,peripheral region of lateral ventricle in the highest hyperintensity section of brain were measured.Blood ammonia values were examined.The pathologic and immuno-histochemistry and RT-PCR examination for brain specimen were performed.All date were analyzed with statistical methods.Results The mean values of blood ammonia were significantly different (P0.05).Conclusions Increase of the blood ammonia was the main cause for the brain energy metabolic abnormality and AQP-4 mRNA and protein expression.The hyperammonemia was the key factor in the occurrence and development of the hepatic brain edema.The abnormal findings in DWI signal could reflect the range and degree of the brain edema and AQP-4 protein expression.

Purpose To assess the feasibility of dual-energy dual-source virtual noncontrast (VNC) CT in the diagnosis of laryngeal carcinoma,in order to reduce radiation hazard.Materials and Methods Forty-nine patients with clinically identified laryngeal carcinoma underwent conventional non-contrast (CNC) scan as well as arterial and venous dual-phase and dual-energy VNC scan.The VCN images of arterial and venous phase were obtained by using the dual-energy software.The mean CT value,signal-to-noise ratio (SNR),contrast noise ratio (CNR),lesion morphology,image quality and radiation dose were compared between the VNC and CNC images.Results The mean CT value,SNR,CNR and image quality scores of CNC were higher than those of VNC images of arterial and venous phase (P＜0.05).There was no significant difference in the image quality scores of VNC between the arterial and venous phase (P＞0.05).The image quality scores of VNC were all above 3,which could meet the diagnostic requirements.There was no significant difference in the shape,size,boundary and cystic necrosis of the lesions,invasion of the surrounding space,osteolytic destruction and size of the surrounding lymph node between the CNC and VNC image.The radiation dose of dual-phase dual-energy scan was 0.42 mSv,which was 18.3％ lower than that of the conventional three-phase scan.Conclusion For laryngeal carcinoma,the VNC images from the dual-energy CT scan can be used to reduce the radiation dose without affecting image quality and diagnosis accuracy,which has good clinical value.

Abstract: Objective　To compare the efficiency of lung-microscopy and tissue homogenate in the detection of Angiostrongylus cantonensis larvae from Pomacea canaliculate, with the amin of finding a simple and rapid method suitable for different scenarios. Methods　Pomacea canaliculata was caught and collected from ponds, ditches, rivers and other environments in the confirmed epidemic areas reported by predecessors. After each snail was weighed one by one, and dissected into two parts: lung sac and muscle. Firstly, each snail lung sac was check for nodules with lung-microscopy, and the nodules were picked out with anatomical needle and press them separately. Then, the worm was found and identified for worm species under microscope, and the lung sac and snail meat then was rechecked with tissue homogenate. Results　A total of 330 snails were detected, with 19.1% (63/330) snails with Angiostrongylus cantonensis stage Ⅲ larvae were detected by tissue homogenate and 15.8% (52/330) snails with nodules were detected by lung-microscopy. Among them, 36 snails with nodules and larvae were detected by the lung-microscopy, and all of them were positive by the tissue homogenate, with a coincidence rate of 100% (36/36); 16 snails with nodules but no larvae, among which 6 snails were positive and 10 snails were negative by the tissue homogenate, The false detection rate was 19.2% (10/52). 278 snails with no nodules, but 21 of them were detected by the tissue homogenate, the missing rate was 7.6% (21/278). There was no significant difference between tissue homogenate and lung-microscopy (taking the positive determination of nodules as the standard) (χ2=1.27, P=0.26, P>0.05). There was significant difference between tissue homogenate and lung-microscopy (taking the detection of larvae as the standard)(χ2=8.66, P = 0.003, P<0.01). There was no significant difference between the two methods and tissue homogenate in the detection rate of large snails ( ≥25 g, χ2=0.08，P=0.777; χ2=2.58, P=0.108), but there was significant difference between the two methods and tissue homogenate in the detection rate of small snail (≤10 g, χ2=5.63, P=0.02). Conclusions　Compared with the tissue homogenate, lung-microscopy is simple in the detection of large snails, requires less instruments and equipment, and its detection speed is faster. It is suitable for the field investigation of the natural focus of Angiostrongylus cantonensis. The tissue homogenate has high sensitivity, can directly display the insect state and activity, has strong insect vitality and high detection rate, it's more suitable for food safety risk monitoring.