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BACKGROUND: Previous studies have demonstrated that Candida albicans possesses capsule structure. Whether capsule structure is associated with the virulence of Candida albicans?OBJECTIVE: This study investigated the pathogenic difference between the standard strains of Candida albicans and the clinically isolated strains, verified whether capsule was the virulence factor of the Candida albicans, and analyzed the association between the animal pathogenicity of different strains and capsule thickness.DESIGN: A randomized controlled animal experiment.SETTING: Department of Pathogenic Biology, Gannan Medical College.MATERIALS: This study was performed at the Scientific Research Center, Gannan Medical College between May and June 2005. A total of 120 BALB/c mice and 72 healthy adult rabbits were included. Candida albicans strains (CCCMC1a and ATCC 14053) were used. The isolated and cultured 4 strains were numbered as C1-1, C1-2, C1-3,and C1-4.METHODS: All animals were randomly divided into 6 groups with 20 mice and 12 rabbits in each group, namely,CCCMC1a, ATCC 14053, C1-1, C1-2, C1-3, and C1-4 groups. Strains smeared in sabouraud ager medium for 36 hours were diluted into the bacterial solution with physiological saline. This solution was intravenously injected into rabbit ear edge, 1.5 mL per rabbit, and intraperitoneally injected into BALB/c mice, 0.5 mL per mouse. Six hours after administration, animal response was observed, and attack time, death time, and mortality were recorded.MAIN OUTCOME MEASURES: Rabbit nephridial tissue printing slices and mouse peritoneal fluid smears were made for Hiss capsule staining microscopy. The capsule thickness of 40 randomly selected yeast cells in each strain was measured using a microscope-micrometer, and the mean capsule thickness of each strain was compared.RESULTS: Compared with C1-1, C1-3, CCCMC1a, and ATCC 14053, C1-2 and C1-4 possessed stronger animal pathogenicity. The standard strains and clinically isolated strains could form capsule in the rabbit and mouse bodies. Capsule thickness differed due to different strains and animal genera (P < 0.05-0.01). The bacterial capsule thickness was greater in the rabbit renal infection focus than in the mouse abdominal cavity. The bacterial capsule thickness of rabbit renal infection focus and mouse abdominal cavity in the C1-1, C1-2, C1-3, and C1-4 groups was greater than that of the same genus in the CCCMC1a and ATCC 14053 groups. The bacterial capsule thickness of rabbit renal infection focus and mouse abdominal cavity was the greatest in the C1-2 and C1-4 groups.CONCLUSION: Candida atbicans C1-2 and C1-4 strains have strong animal pathogenicity. C1-2 and C1-4 strains possess greater bacterial thickness than other strains. It has been primarily confirmed that capsule is possibly a virulence factor of Candida albicans, and capsule thickness is closely associated with animal pathogenicity.
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BACKGROUND: Studies have confirmed that there is secreted protein out of cell wall of Candida albicans. However, in recent researches, we found there may be a capsule-like-structure within it.OBJECTIVE: To observe the capsule-like structure of Candida albicans by means of modified quellung test.DESIGN: A controlled observation experiment.SETTING: Department of Medical Microbiology, Gannan Medical College.MATERIALS: Candida albicans from 2 clinical specimens (C1, C2) identified by preserve center of Epiphyte strains of Chinese Academy of Science (Nanjing, China) and standard strains (CCCMC1a and ATCC14053) donated by preserve center of Epiphyte strains of Peking University (Beijing, China) were taken to quellung test. Experimental rabbits (weighing 2-2.5 kg) for preparing normal serum and antiserum were donated by Experimental Animal Center of Sun Yat-sen University. Antiserum was prepared by the book with experimental strains (C1, C2, CCCMC1a, and ATCC)respectively for later quellung tests.METHODS: This experiment was carried out in the Department of Medical Microbiology, Gannan Medical College in December 2005. ① Traditional Quellung test: The culture of Candida albicans (C1, C2, CCCMC1a, and ATCC) was spread on a slide respectively. Corresponding rabbit antiserum was added onto the slide (experimental group 1) and normal serum was added onto the slide (control group1). 1% methylene blue was added into each group, and then the slides were placed in a wet box at 37 ℃ for 20 minutes; the slides were taken out and covered with a cover-slip. Under the oil immersion, the count of Candida albicans was taken and the microscopic surveying instrument was used to measure the capsule thickness of cell of Candida albicans directly. The average thickness of capsule of 40 cells was taken. ②Modified quellung test: The culture of Candida albicans (C1, C2, CCCMC1a, and ATCC) was spread on a slide respectively. Corresponding rabbit antiserum was added onto the slide (experimental group 2) and normal serum was added onto the slide (control group 2), 1% methylene blue was not added into each group, but the slides were directly placed in a wet box et 37 ℃ for 20 minutes. The slides were taken out and dried naturally, but not covered with a cover-slip during the drying process. Hiss capsule staining was applied for them.The average thickness of capsule of 40 cells was taken.MAIN OUTCOME MEASURES: The average thickness of capsule in each experimental group in traditional and modified quellung tests.RESULTS: ① Traditional quellung test of Candida albicans was positive. The thickness of culture of Candida albicans C1, C2, CCCMC1a, and ATCC of experimental group1 was larger than that of the control group1, respectively [(0.558+0.081 ) vs. (0.225+0.061) μm; (0.530+0.081) vs. (0.252+0.038) μm; (0.475+0.081) vs. (0.200+0.072) μm;(0.600+0.068) vs. (0.225+0.046) μm,P ＜ 0.01] .② The thickness of culture of Candida albicans C1, C2, CCCMC1a, and ATCC of experimental group 2 was larger than that of the control group 2, respectively[(0.541 ±0.038) vs. (0.215±0.022)μm; (0.510±0.060)vs. (0.247±0.018) μm; (0.487±0.041) vs. (0.213±0.033)μm; (0.595±0.027) vs. (0.220±0.016) μm, P ＜ 0.01]. ③ The thickness of capsule of the control group 2 was smaller than that of control group 1 (P ＜0.01); The thickness of capsule of the experimental group 2 was smaller than that of experimental group 1 (P ＜ 0.01).CONCLUSION: As a quantitative analysis test, modified quellung test is more stable and accurate than traditional quellung test.
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BACKGROUND:Airborne pollen is the most important factor to induce the seasonal allergic diseases.The production and dispersal of pollen are closely correlated with the meteorological factors.OBJECTIVE:To observe the correlation between the seven meteorological factors (air pressure,air temperature,humidity,precipitation,wind speed,evaporation capacity and sunshine) and the airborne pollen in Nanchang so as to provide theoretic evidences for preventing and treating anaphylactic disease in that region.DESIGN:Observational experiment. SETTING:Department of Allergic Reaction,Jiangxi Medical College.MATERIALS:An investigation on airbome pollen was performed for a year by Durham gravity method.Data of airborne pollen and the seven meteorological factors in urban area of Nanchang city were collected.METHODS:The top of the 4th experimental building in the southern yard of Jiangxi Medical College located in the center of Nanchang city was taken as exposed point.The pollen was surveyed by Durham gravity method from April 1st,2000 to March 31 st,2001.Meteorological data of the seven factors were collected every day and supplied by the Weather Bureau of Jiangxi Province.The correlation between meteorological factors and airborne pollen was analyzed by the multivariate Jinear stepwise regression analysis with SAS 6.12.MAIN OUTCOME MEASURES:The type,amount,seasonal distribution of airborne pollen and the correlation between the seven meteorological factors and the airborne pollen in urban area of Nanchang city. RESULTS:There were airborne pollens in Nanchang city all the year round,which included about 47 types.The main pollen season of Cupressaceae was in March,Pinaceae in March and April, and Cunninghamia R.Br in March,Chenopodium-Amaranthus and Ambrosia from August to November, Artemisia from September to November and Humulus in September.Gramineae had been scattering all seasons but mostly from June to October.In the correlation analysis between dispersal of airbome pollen and seven meteorological factors, multivariate linear stepwise regression analysis was conducted in some major polien counting in steady dispersal period and seven meteorological factors. The positive correlation was found in Pinaceae with air pressure,air temperature,wind speed and sunlight,and the negative correlation was found with precipitation and evaporation capacity.The positive correlation was found in Cupressaceae with air pressure,air temperature and sunlight,and negative correlation was found with humidity,precipitation,wind speed and evaporation capacity.The positive correlation was found in Ambrosia with air pressure,air temperature and wind speed,while negative correlation was found with humidity,precipitation,evaporation capacity and sunshine.The positive correlation was found in Artemisia with air pressure,air temperature and wind speed,while negative correlation was found with humidity,precipitation, evaporation capacity and sunshine.The positive correlation was found in Gramineae with air pressure and air temperature,while negative correlation was found with humidity,precipitation and evaporation capacity.However,if using count of total pollen in a year or half a year to analyze the relationship,the correlated factors were much fewer than that mentioned above.CONCLUSION:The dispersal of airborne pollen is related with the seven meteorological factors.It is better to use the grain amount of single pollen in its main pollen season to analyze the relationship with meteorological factors.
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BACKGROUND: Clerodendron bungei steud(CBS) is a tree from genera of Verbena L. In the present pharmacological studies, CBS has showed anti-inflammatory, antineoplasic, nonspecific immunity-enhancing effects. Myoelectric effect of stimulating uterus round ligament is related with agitating adrenergic α receptor. The experiments about analgesic effect are few.OBJECTIVE: To observe the antinociceptive effect of ethanol extract of CBS in mice through hot plate and writhing tests.DESIGN: Randomized controlled experiment with animals as subjects.SETTING: Departmcent of Pharmmacology, Gannan Medical College.MATERIALS: The experiment was performed in Department of Pharmacology, Gannan Medical College from March to June 2004. A total of 120Kunming white mice, weighing(20 ± 2) g were provided by the Experiment Animal Center of Gannan Medical College.INTERVENTIONS: In the writhing test, 50 mice were randomly divided into normal saline, aminopyrine group(0. 1 g/kg), morphine group(0.01 g/kg)and extract of CBS groups(20 g/kg, 40 g/kg) . There were 10 mice in each group. Forty minutes after intraperitoneal injection. 6 mL/L acetic acid (10 mL/kg) was injected intraperitoneally, 5 minutes later, the number of writhing body and inhibitory rate of writhing body were observed for 10minutes. In hot plate test, 40 mice were randomly divided into normal saline group, morphine group(0.01 g/kg) and extract of CBS groups(20 g/kg,40 g/kg). There were 10 mice in each group. After intraperitoneal injection,the mice were put on the hot plate, and the temperature was(55 ±0.5) C. The pain threshold was recorded 15, 30 and 60 minutes after administration. And 30 white mice were randomly divided into naloxone 0. 04 g/kg + morphine 0.01 g/kg group, aloxone 0.04 g/kg + extract of CBS 40 g/kg group,naloxone 0.04 g/kg + normal saline group for hot plate test antagonized by naloxone(40 mg/kg) . The mice were injected intraperitoneally. The duration of pain reaction were recorded 15, 30, 60 and 90 minutes after administration.MAIN OUTCOME MEASURES: ① The number of writhing body and inhibitory rate of writhing body; ② The duration of pain reaction; ③ The pain threshold in the hot-plate test after antagonized by naloxone were calculated.RESULTS: All the 120 white mice entered the result analysis. ① The number of writhing body and inhibitory rate of writhing body: The number of writhing body was 2.4 ± 2.5 and 0. 6 ± 1.7 in the 20 and 40 g/kg extract of CBS injection group respectively and the inhibitory rate of the pain reaction caused by 6 mL/L acetic acid was 93.3% and 98.3% respectively. ② The pain threshold at 15, 30 and 60 minutes after administration in the hot plate test: the pain threshold was[(121.2±98.7) s, (191.2±78.6) s,(133. 1 ± 91.1 ) s] at 15, 30 and 60 minutes after injection of 20 g/kg extract of clerodendron bungei steud; it was[(233.9 ±70.4) s, (219.6 ±78.2) s,(218.3 ±92.6) s at 15, 30 and 60 minutes after 40 g/kg extract of CBS being given. The pain threshold in the extract of CBS groups was all higher than that in the control group respectively[ (13.7 ± 15.2) s, (9.7 ± 12.5) s,(22. 1 ± 15.6) s] ( P ＜ 0.01 -0. 001) . ③ In naloxone antagonized morphine test, the pain threshold at 15, 30 and 60 minutes after administration in morphine 0.01 g/kg+naloxone 0.04 g/kg group[(1.7±5.2) s (6.4 ± 8.6) s, (21.8 ± 34.0) s]was obviously smaller than that in the extract of CBS 40 g/kg +naloxone 0. 04 g/kg group[(124.9 ±79.4) s,(139.3±72.9) s, (137.9±60.8) s](P ＜0.001).CONCLUSION: Ethanol extract of CBS has strong analgesic effect and this is not manifested through the activation of opioid receptors.
ABSTRACT
Objective To determine species, counts, and seasonal variation of airborne pollen in the center of Nanchang. Methods Pollen counts were done with Durham gravity method on the top of the building in the campus of Jiangxi Medical College, Nanchang, April 1, 2000-March 31,2002. Results There were airborne pollens in all seasons of the year in atmosphere of Nanchang, which included about 47 species. The highest airborne presence (percent of total yearly pollen counts) was for Ambrosia(35.73%); followed by Pinaceae (15.94%), Artemisia (11.94%), Gramineae (5.88%), Cupressaceae (5.64%), Cunningha-mia R.Br.(5.12%), Chenopodium-Amaranthus (5.03%) , and Humulus(2.02%). The dispelling period showed a double curve: the first peak occured in Mar. and Apr. (22.56% and 11.98% respectively). The predominant pollens were tree pollen (Pinaceae, Cupressaceae, Cunninghamia R.Br., Moraceae and Salix). The second peak occured in Aug., Sep., and Oct. (9.61%, 23.77%, 19.57% respectively). The predominant pollens were grass pollen (Ambrosia, Artemisia, Gramineae, Chenopodium-Amaranthus and Humulus). Gramineae had been scattering all seasons, but it's counts were not high in every month. Conclusion There are airborne pollens dispelling in atmosphere of Nanchang in the whole year, whose dispelling period shows a double curve (the first peak occurs in Mar. and Apr. And second peak occurs in Aug. Sep. and Oct.) and the predominant pollens are Ambrosia, Pinaceae, Artemisia, Gramineae, Cupressaceae, Cunninghamia R.Br., Chenopodium-Amaranthus and Humulus. This study provides useful scientific information for production of specific allergen and for clinical prevention, specific diagnosis, specific therapy of allergic deseases.