Prediction of ocular irritancy of 26 chemicals and 26 cosmetic products with isolated rabbit eye (IRE) test / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This study aims to establish and evaluate the methodology of isolated rabbit eye (IRE) test.</p><p><b>METHODS</b>IRE test was performed according to modifications of the in vitro toxicology (INVITTOX) Protocol No.85: Rabbit enucleated eye test by European Centre for the Validation of Alternative Methods (ECVAM), and then 26 chemicals and 26 cosmetic products were tested in both in vitro IRE and in vivo Draize tests. A statistical analysis was conducted to determine the relevance of the IRE test to the data generated in the Draize test.</p><p><b>RESULTS</b>IRE test was established successfully in our laboratory. It was shown that ranking correlation and class concordance were fairly well between the IRE test and the Draize test for 26 reference chemicals (Fisher's Exact Test χ(2)=51.314, P<0.001; McNemar P=0.261; Gamma=0.960, P<0.001; Kappa=0.843, P<0.001) and 26 cosmetic products (Fisher's Exact Test χ(2)=15.522, P<0.001; McNemar P=0.311; Gamma=0.967, P<0.001; Kappa=0.611, P<0.001).</p><p><b>CONCLUSION</b>IRE test was established successfully for in vitro testing of eye irritation as an alternative to Draize test.</p>

Influence of joint exposure to diazinon, propoxur and bisphenol A on phagocytosis of RAW264.7 cell / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the toxicity of joint exposure to diazinon, propoxur and bisphenol A on phagocytosis.</p><p><b>METHODS</b>Flow cytometer was employed to detect the influence of diazinon and bisphenol A, propoxur and bisphenol A in mixture (mixed according to ratio of IC(50)) on mouse macrophage RAW264.7 cells' function to phagocyte fluorescent microspheres, adopting the percentage of phagocytic cells (PP) and the phagocytic index (PI) as measurement indicators. The final concentrations of mixture of diazinon and bisphenol A were (0.4 + 0.1), (3.6 + 0.7), (36.2 + 7.2), (43.4 + 8.7), (52.1 + 10.4), (62.5 + 12.5), (75.0 + 15.0) µg/ml; while those of mixture of propoxur and bisphenol A were (0.2 + 2.0 × 10(-2)), (2.4 + 0.2), (23.7 + 2.0), (35.6 + 3.0), (53.3 + 4.4), (80.0 + 6.7), (120.0 + 10.0) µg/ml. Then based on the dose-response relationship, a 2 × 2 factorial design was then carried out among different doses of mixture with statistical significance to statistically evaluate the interaction between diazinon and bisphenol A, propoxur and bisphenol A.</p><p><b>RESULTS</b>After the joint exposure, compared to the control group (PP = (23.6 ± 2.2)%; PI = 0.36 ± 0.03), any dose of the mixture of diazinon and bisphenol A ((52.1 + 10.4), (62.5 + 12.5), (75.0 + 15.0) µg/ml) could significantly increase the levels of PP ((29.0 ± 1.4)%, t = 3.89, P < 0.05; (30.2 ± 2.3)%, t = 4.74, P < 0.05; (35.0 ± 3.4)%, t = 8.21, P < 0.05) and PI (0.43 ± 0.03, t = 3.86, P < 0.05; 0.41 ± 0.02, t = 2.95, P < 0.05; 0.46 ± 0.03, t = 5.34, P < 0.05); while that of propoxur and bisphenol A ((35.6 + 3.0), (53.3 + 4.4), (80.0 + 6.7), (120.0 + 10.0) µg/ml) reduced the levels of PP ((20.6 ± 1.1)%, t = -3.00, P < 0.05; (20.2 ± 1.0)%, t = -3.42, P < 0.05; (19.4 ± 1.3)%, t = -4.23, P < 0.05; (18.8 ± 2.1)%, t = -4.81, P < 0.05) and PI (0.31 ± 0.01, t = -4.75, P < 0.05; 0.31 ± 0.01, t = -4.58, P < 0.05; 0.30 ± 0.01, t = -4.92, P < 0.05; 0.27 ± 0.02, t = -7.80, P < 0.05) on the contrary. The 2 × 2 factorial design was carried out between the mixture of diazinon (60.0 µg/ml; PP = (28.5 ± 3.4)%; PI = 0.49 ± 0.07) and bisphenol A (12.0 µg/ml; PP = (35.7 ± 2.7)%; PI = 0.67 ± 0.07), and the mixture of propoxur (48.0 µg/ml ; PP = (28.1 ± 2.2)%; PI = 0.48 ± 0.04) and bisphenol A (4.0 µg/ml; PP = (34.4 ± 2.7)%; PI = 0.59 ± 0.07). The mixture of diazinon and bisphenol A (PP = (30.4 ± 1.4)%, F(interaction) = 6.22, P < 0.05; PI = 0.53 ± 0.03, F(interaction) = 7.35, P < 0.05) and the mixture of propoxur and bisphenol A (PP = (27.5 ± 4.1)%, F(interaction) = 4.56, P < 0.05; PI = 0.46 ± 0.08, F(interaction) = 11.13, P < 0.05) both showed a significant antagonistic interaction on phagocytosis of RAW264.7 cell.</p><p><b>CONCLUSION</b>It is suggested that the interactions between diazinon & bisphenol A and propoxur & bisphenol A both played the antagonistic role on phagocytic function of macrophages in vitro.</p>

Study on alternative methods for the acute oral toxicity in detection of chemicals / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To evaluate three alternative methods for LD50 test-Fixed Dose Procedure (FDP), the Acute Toxic Class Method (ATC) and Up and Down Procedure (UDP).</p><p><b>METHODS</b>Female SD rats (8-12 weeks of age, 160-200 g) were used. Three alternative methods from OECD were applied to assess 22 chemicals (10 cosmetic raw materials and 12 raw materials of personal and home care products). The toxicity ranking for tested chemicals was established according to Globally Harmonized System (GSH). The results LD50 test were compared for the consistency and correlation between alternative methods and traditional test.</p><p><b>RESULTS</b>For cosmetic raw materials, the concordance rate of the three alternative methods was 80% (8/10); for raw material of personal and home care products, the concordance rates of FDP, ATC and UDP was 91.7% (11/12), 75.0% (9/12) and 83.0% (10/12), respectively. The number of animals required in three alternative methods was significantly lower than that in traditional test (P < 0.05), but the time required in three alternative methods was significantly higher than that in traditional test (P < 0.05).</p><p><b>CONCLUSIONS</b>High consistency and correlation were found between each alternative method and LD50 test. FDP may be more potential when applied to assess acute oral toxicity of cosmetic raw materials.</p>

An epidemiological study on the norovirus detected in the field environment after an outbreak of norovirus gastroenteritis / 中华流行病学杂志

Objective To study the status of norovirus in environment of the patient's residence and water samples after a norovirus gastroenteritis outbreak, to provide evidences for the development of strategies for prevention and control of the disease. Methods After a norovirus gastroenteritis outbreak, anus swabs from the patient, swabs from the household environment and the water samples were collected to detect the norovirus by RT-PCR methods. Sequencing analysis was conducted on those positive specimens. Results Three specimens of the anus swabs from 9 patients and 2 samples of the 46 house environment swabs were positive to the virus. The latter were from the surface of water-closets of two families that the illness were asymptomatic. Among 5 water samples, only one was positive, which was the rivulet water that the feces of the villagers evacuated directly. Results showed that the sequences of the virus detected from the anus swabs of the patients, the swabs from the household environment and the samples of the rivulet water belonged to the same species. Conclusion It is necessary to strengthen activities as supervision and disinfection to the feces of the patients, especially on monitoring the feces that might have contaminated the water during the noroviru gastroenteritis outbreak.

Development of a DNA microarray for detecting 8 common species of food-borne bacterial pathogens in south China / 南方医科大学学报

<p><b>OBJECTIVE</b>To prepare a DNA Microarray that can detect 8 common species of food borne bacterial pathogens in south China.</p><p><b>METHODS</b>All the 70mer oligo probes were designed on the characteristic genome loci of the 8 species of food borne bacterial pathogens. Eight subarrays corresponding to the 8 food borne bacterial pathogens were spotted onto the slide and integrated into a pan-array on the chip. A number of identified and known bacterial samples from the storage bank were selected for the validation test.</p><p><b>RESULTS</b>Based on the PPR ranking, for LM sub-array, the PPR of the 3 Listeria bacteria LM, Lin and Liv was 68.8%, 51.8% and 59.6%, respectively, while that of the non-Listeria bacterial samples was all below 43%. For VC sub-array, the PPR of VC sample was 54.1% and that of the non-VC bacterial samples was lower than 17.2%. For VP sub-array, the PPR was 66.7% for VP sample and below 24.2% for non-VP bacterial samples. For Sal sub-array, the PPR was 55.9% for Sal sample and below 50.5% for non-Sal bacterial samples. For Shi sub-array, the PPR of Shi sample and the non-Shi bacterial samples was 53.8% and below 36.6%, respectively. For SA sub-array, the PPR of SA sample and non-SA bacterial samples was 65.2% and below 22.7%, respectively. For CJ sub-array, the PPR of the 2 Campylobacter bacteria CJ and CC were 88.2% and 58.8%, respectively, and that of the non-Campylobacter bacterial samples was lower than 35.3%. For EC sub-array, the PPR of EC sample was 47.9%, and that of the non-EC bacterial samples was lower than 41.6%. Evaluation of the Biosafood-8 chip developed in this study by 18 biological samples from different origins demonstrated its good specificity and accuracy in the identification of the pathogens.</p><p><b>CONCLUSION</b>The chip we developed can clearly differentiate the target food borne pathogenic bacteria and non-target bacteria and allows specific and accurate identification of the species of the tested bacteria isolates.</p>

Molecular characters of epidemic cerebrospinal W135 Neisseria meningitidis strain firstly isolated in Guangdong province / 中华预防医学杂志

<p><b>OBJECTIVE</b>To analyze the molecular characters of the W135 Neisseria meningitidis strain firstly isolated from patients in Guangdong province.</p><p><b>METHODS</b>Biochemical profile by using the API NH system (bio-Merieux, France) was used for confirmation,and sero-grouping of the meningococcal isolates including one serogroup W135, one serogroup C and three serogroups of a Neisseria meningitidis isolated from patients in Guangdong province in recent two years were performed. The subtype was determined after amplifying porA and porB respectively from the genome DNA of Neisseria meningitidis. Multilocus sequence typing (MLST) was performed for determining the allele profiles and the sequence types (STs). The polygenetic tree was obtained by analyzing the allele profiles with program Splits tree online. The molecular characters of the serogroup W135 Neisseria meningitidis was analyzed by its evolution relationship and the variable regions in porA and porB which encoding the outer membranes proteins (OMPs).</p><p><b>RESULTS</b>The subtype determined by porA variable regions of the serogroup W135 Neisseria meningitidis was P1.5,2, which was one of the most invasive types. The types of variable regions (VRs) I, IV, V, VII with porB were 1, 1, 1, 17, and there was no VI and VIII in porB. The allele profile of the W135 strain in this study was 2, 123, 4, 3, 8, 4, 6, and its sequence type was ST-2960, which belonged to ST-11/ET-37 clone complex. The subtypes of the serogroup C and serogroup A strains were P1.20, while their types of VR IV were all 7, and they all hadn't VR VII in porB. The strain serogroup C belonged to ST-4821 clone complex, and the 3 serogroup A strains belonged to ST-5 clone complex.</p><p><b>CONCLUSION</b>The molecular character of the serogroup W135 Neisseria meningitidis should be the same with the strains isolated in foreign country, and be different from the epidemic types in the area. This serogroup W135 Neisseria meningitis isolated from patients in Guangdong for the first time was thought to be a new type appearing in the local area.</p>

MLST typing of Streptococcus suis isolated from clinical patients in Guangdong Province in 2005 / 南方医科大学学报

Intensive surveillance of human S.suis infection was carried out in July and August of 2005 in Guangdong Province, which coincided with the Sichuan outbreak. Five isolated cases of human infections were identified during this period, from which 5 S. suis serotype 2 isolates were recovered. MLST analysis showed that these 5 isolates shared identical sequences of 6 MLST housekeeping genes except for one point mutation found within the thrA gene fragment, a neutral mutation (TTA to TTG) in the third nucleotide (360 nt) of the codon for leucine. MLST analysis identified 2 sequence types in the Guangdong sporadic infection. Three Guangdong isolates L-SS002, L-SS003 and L-SS005 belonged to ST7, while the other two isolates L-SS004 and L-SS006 belonged to ST1, but they all belonged to ST1 clonal complex. This finding represents a striking feature that differs from the Sichuan outbreak caused by a single ST7 SS2 clone. The 3 isolates of ST7 were probably imported from Sichuan Province, while the origin of the other 2 isolates of ST1 still remain to be clarified.

Relationship between apoptosis and activity of protein kinase B in adrenocortical cells induced by cadmium chloride / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study apoptosis induced by cadmium chloride (CdCl2) and the alteration in activity of protein kinase B (PKB/Akt) in adrenocortical cells.</p><p><b>METHODS</b>Fasciculata-glomerulosa (FG) cells of male guinea pigs were dispersed and primarily cultured in vitro. Features of apoptotic cells were observed using combined labeling with annexin-V and propidium iodide (PI) and flow cytometry. The activity of PKB/Akt was determined with immunoprecipitation and the chemiluminescence assay.</p><p><b>RESULTS</b>Apoptosis rate of FG cells increased with dose of CdCl2 two hours after treatment with 6.25-100.00 micromol/L of it, with significant difference in the groups treated with 25.00, 50.00, 100.00, micromol/L of CdCl2, as compared with the control group (P < 0.01). Regression analysis showed that occurrence of apoptosis correlated with the dose of CdCl2 in a dose-response pattern. In the meanwhile, there were obviously elevated percentages of apoptotic cells as the increase in duration of incubation ranging from 5.58% to 73.08% for incubating cells with 50.00 micromol/L of CdCl2, from 15 minutes to 4 hours. Duration of incubating cells with CdCl2 were correlated with occurrence of apoptosis in a time-effect manner. The gray scales of PKB/Akt were manifested to be decreased as the ascending of CdCl2 dosage from 6.25 to 200.00 micromol/L, with the linear correlation.</p><p><b>CONCLUSION</b>6.25 to 100.00 micromol/L CdCl2 might elicit apoptosis of adrenocortical cells. Meanwhile, PKB/Akt is decreased.</p>

The establishment and assessment on cosmetic products using 3T3 mouse fibroblast neutral red uptake phototoxicity assay / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish the 3T3 mouse fibroblast neutral red uptake (NRU-PT) phototoxicity test method, and evaluate the practicality of the method in detecting potential phototoxicity of the cosmetic products.</p><p><b>METHODS</b>Fifteen phototoxic and 9 non-phototoxic chemicals were tested in our laboratories, the phototoxic potential of the test chemicals was evaluated in a prediction model in which either the photo irritation factor (PIF) or the mean photo effect (MPE) was compared with the coherence and sensitivity of the method. 20 kinds of functional cosmetics were detected and the results were analyzed by the 3T3 NRU-PT in vitro and Guinea pig skin phototoxicity test (in vivo).</p><p><b>RESULTS</b>Both PIF and MPE of the chemicals were highly reproduced, and the correlation between in vitro and in vivo data was almost perfect. All the non-phototoxic provided a negative result, while 14 of the 15 phototoxic tested chemicals gave clear positive results. For cosmetics, the correlation between in vitro and in vivo data was consistent.</p><p><b>CONCLUSION</b>The 3T3 NRU PT test was established successfully, it should be used as a good alternative method for assessing the phototoxic potential of the chemicals and cosmetics in China.</p>

Effects of overexpression of human pol-beta on cellular response to DNA damage / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the biological effects of overexpression of the human DNA polymerase (pol-beta) on cellular response to DNA damage.</p><p><b>METHODS</b>The cell strain HLFbeta from the stable overexpression of the human pol-beta was contaminated with methyl methanesulfonate (MMS) for investigating the effects of the pol-beta on the cellular responses to DNA damage on the aspects such as the DNA damage, the cell cycle and the induced mutation rate.</p><p><b>RESULTS</b>The cell HLFbeta from the stable overexpression of the human pol-beta was obtained through the screening. The cellular response to DNA damage of HLFbeta induced by the MMS in the intermediate and high dosage group (ranging from 0.5 to 0.8 mmol/L) was significantly lower than that in the control group. The analysis for the cell cycle distribution showed that both the two types of cells contaminated by MMS had retardation at G(2) phase. In the HLFbeta group, the cells had the obvious G(2) phase retardation and 49.0% of the cells were retarded at G(1) phase as well when the MMS was increased to 0.5 mmol/L while in the control, only 20.1% of the cells were retarded at the G(1) phase when the same dosage of MMS was administered. Moreover, the MMS-induced mutagenesis in HLFbeta was increased from 4.5 x 10(-6) to 8.2 x 10(-6), significantly higher than that in the control group (P < 0.05).</p><p><b>CONCLUSION</b>High Pol-beta level decreases cellular DNA damage induced by MMS. Nevertheless, the overexpression of Pol-beta can also increase error-prone DNA synthesis during DNA repair process.</p>

Scanning aneugen and clastogen by micronuclei analysis using flow cytometry / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To explore a flow cytometry (FCM)-based method for discriminating aneugen- or clastogen-induced micronuclei.</p><p><b>METHODS</b>Cells were stained with anti-CD71-FITC and PI, and the PI fluorescent signal intensity of micronucleated reticulocyte (MN-RET) in the peripheral blood of NIH mouse treated with COL or CP was detected by flow cytometry.</p><p><b>RESULTS</b>The ratio of the median of the intensity of MN-RET fluorescent signals to that of nucleated cell was low in the cyclophosphamide treated mouse, while the median was high in the colchicine treated mouse.</p><p><b>CONCLUSION</b>The flow cytometry-based micronucleus assay can be used to discriminate primarily smaller MN induced by the clastogen exposure from the larger MN induced by an aneugen.</p>

The effect on apoptosis in anterior pituitary induced by cadmium chloride and its relations with caspase-9 pathway / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of cadmium chloride in anterior pituitary and the relation between apoptosis and expression of procaspase-9 mRNA.</p><p><b>METHODS</b>In vivo studies:40 SD male rats were randomly distributed into four groups which were administered with CdCl2 at different doses by gavage for 6 weeks;</p><p><b>IN VITRO STUDIES</b>the rats' anterior pituitary cells were primarily cultured for 120 hours, then treated with CdCl2 at the dose of 0, 1.56, 3.12, 6.25, 12.50, 25.00, 50.00, 100.00 micromol/L for 6 hours; The indices included: expression of procaspase-9 mRNA, detection of apoptosis with TUNEL assay.</p><p><b>RESULTS</b>The results showed the excretion of ACTH, LH seemed to be decreased dramatically and the apoptosis inclined to enhance remarkably, and further more, the expression of procaspase-9 mRNA appeared to be increased significantly as compared with those of the control. It was show that a dose-effect relationship between the CdCl2 dosing and indices above with the regression analysis and a linear correlation between the mean gray value of apoptosis cell and the relative gray value of procaspase-9 mRNA positive cell. The results indicated that damnification, for example, apoptosis could be caused by certain dose of CdCl2 in anterior pituitary cells with dose dependent manner. Caspase-9 might play a role in the occurrence of apoptosis.</p><p><b>CONCLUSION</b>It was suggested that cadmium could induce apoptosis of anterior pituitary both in vivo and in vitro in the manner of dose-dependent, and caspase-9 might play a role during above processes.</p>

Application of flow cytometry on functional assessment of health food / 中华预防医学杂志

<p><b>OBJECTIVE</b>To establish flow cytometry (FCM) methods and evaluate their application value for measuring the index for enhancing immune function of health food.</p><p><b>METHODS</b>In mice experiment model, the dosage groups were respectively oral fed with three test substances according to 5, 10, 30 times of the recommended dose for human body; both the negative and positive control groups were fed with equivalence purified water once a day. The positive control was fed with 25 mg/kg body weight levamisole for 3 days before finishing the administration, and the immune two percent of sheep erythrocytes were administrated at the last day. In rats experiment model, the test substance was given by mixing feed according to 25 and 50 times of the recommended dose for human body. At the end of the experiment, indices below were simultaneously detected. (1) The classical indices included: spleen lymphocyte transformation test by using ConA (MTT assay); spleen NK cell activity test (LDH assay); delayed-type hypersensitivity test by using sheep erythrocyte (foot palm thickening) method and phagocytosis activity tested by mice peritoneal macrophages. (2) FCM indices included: T and B lymphocytes quantitating in mice peripheral blood, activated antigen expression level in the surface of T lymphocytes and NK cells and phagocytosis activity for fluospheres in mice peritoneal macrophages.</p><p><b>RESULTS</b>(1) Compared with the negative control group, there were no significant differences in T and B lymphocytes proportion and the number of lymphocytes in mice peripheral blood after given 0.83, 1.67, 5.01 g/kg protein powder; (2) mice peripheral blood T lymphocyte sub-cluster CD(69)(+)/CD(3)(+) of 3.75, 7.50, 15.0 ml/kg bw Cen-Rong Cream groups were all significantly increased (P < 0.05), which were shown a good coherence with the classic test index; (3) mice peripheral blood NK cell sub-cluster CD(69)(+)/NKG2D(+) of 0.83, 1.67 g/kg protein powder groups were both significantly increased (P < 0.05), which was kept in good coherence with those of NK cell activity test (LDH assay); rats peripheral blood NK cell sub-cluster CD(161a+)/CD(25)(+) of 1.50 g/kg ganoderma lucidum and cordycepicmycelia group was significantly increased (P < 0.05); (4) the phagocytosis activity in mice peritoneal macrophages: there were no significant difference found between the controls and the dosage groups in the classic test. However, in the FCM test, the percentage of phagocytic cells of 0.15, 0.30, 0.90 g/kg ganoderma lucidum and cordycepicmycelia groups and the phagocytic index of 0.30, 0.90 g/kg were enhanced.</p><p><b>CONCLUSION</b>It suggests that was shown in detecting and assessing enhancing immune function of health food the results tested by FCM were fairly consistent with those by using traditional methods, most of them would have higher sensitivity. It should be valuable to applying FCM in the measurement and assessment of enhancing immune function of health food and worth while to further study as to enlarging its application.</p>

Effects of jing'an capsule on rat sperm quality / 中华男科学杂志

<p><b>OBJECTIVE</b>The effects of Jing'an capsule on the quality of rat sperm were studied to supply data for its safe clinical use.</p><p><b>METHODS</b>Eighty SD male rats were evenly and randomly divided into three test groups and one control group. Then the test groups were continuously given Jing'an capsule at different dosages: 557.1 mg/kg, 5,571 mg/kg or 11,420 mg/kg and the control group was given starch (20 g/L). Sixty days later, one of the epididymides, the sperm density was made and the sperm motility and morphology were investigated. The data obtained were analyzed by ANOVA, chi 2 test and Kruskal-Wallis test.</p><p><b>RESULTS</b>The sperm density, motility and morphology were variable at different dosages. Compared with the control, the sperm density increased significantly at the dosage of 557.1 mg/kg(P < 0.05), and a significant decrease was observed in the sperm density and motility (P < 0.05) at the dosage of 11,420 mg/kg. Although the rate of abnormal sperm morphology decreased at the dosage of 557.1 mg/kg and increased at the dosage of 5,571 mg/kg or 11,420 mg/kg compared with the control, there was no statistic significance(P < 0.05).</p><p><b>CONCLUSIONS</b>A low dosage of Jing'an capsule might ameliorate the quality of sperm, while a high dosage could do damage to sperm.</p>