Association of occupational medicamentosa-like dermatitis induced by trichloroethylene and infection with human herpesvirus 6 and cytomegalovirus / 中国职业医学

OBJECTIVE: To explore the potential association between occupational medicamentosa-like dermatitis induced by trichloroethylene( OMDT) and past infection,reactivation and recent infection of human herpesvirus 6( HHV6) and human cytomegalovirus( HCMV). METHODS: Twenty OMDT patients were recruited as case group by using judgment sampling method. Twenty healthy workers occupationally exposed to trichloroethylene for more than half a year were randomly selected as exposure group. Twenty healthy people with no exposure history to trichloroethylene were randomly selected as control group. The enzyme linked immunosorbent assay was used to qualitatively determine the titer of HHV6 and HCMV immunoglobulin( Ig) G,Ig M antibodies from serum samples of these subjects. The polymerase chain reaction was used to qualitatively detect HHV6 and HCMV DNA from whole blood DNA samples of these subjects. The differences of previous infection rate,reactivation rate and recent infection rate of HHV6 and HCMV among these three groups of patients with different clinical types of OMDT were analyzed. RESULTS: The prevalence of HHV6 and HCMV infection in the case group was higher than that in the control group,and the difference was statistically significant( 65. 5% vs 20. 0%,75. 0% vs15. 0%,P < 0. 017). The reactivation rate of HHV6 and HCMV in the case group was higher than that in the control group,but the difference was not statistically significant( P > 0. 017). The recent infection rate of HHV6 and HCMV in the case group was not significantly different from that in the control group( P > 0. 017). There was no significant difference in the past infection rate,reactivation rate and recent infection rate of HHV6 and HCMV between the exposure group and the control group( P > 0. 05),meanwhile in different clinical types of OMDT patients( P > 0. 05). CONCLUSION: OMDT may be associated with past infection of HHV6 and HCMV.

Direct determination of manganese in urine by Graphite Furnace Atomic Absorption Spectrometry / 中华劳动卫生职业病杂志

Objective@#To establish a method for the determination of manganese in urine by graphite furnace atomic absorption spectrometry (AAS) without the use of matrix modifier.@*Methods@#The urine samples were 5 times diluted with 1% nitric acid then directly determined by AAS. Zeeman was used for background correction.@*Results@#The linear range for determination of manganese in urine was 5~60 μg/L (urine) . The correlation coefficient was greater than 0.995 with the detection limit of 1.5 μg/L and with the lower limit of quantification of 5.0 μg/L. The relative standard deviations (RSDs) of within-run precision was between 1.1%~4.3%, the RSDs of between-run precision was between 3.3%~7.0%. The average recovery was 102.6%. The samples can be stored for 14 days at room temperature, 4℃, -8 ℃ and -35 ℃.@*Conclusion@#The method is feasible for determination of manganese in urine.

Correlation between HLA-B~* 13:01 allele and liver dysfunction in patients with occupational medicamentosa-like dermatitis due to trichloroethylene / 中国职业医学

OBJECTIVE: To explore the correlation between human leukocyte antigen( HLA)-B~* 13 : 01 allele and liver dysfunction in patients with occupational medicamentosa-like dermatitis due to trichloroethylene( OMDT). METHODS: Twenty patients with OMDT were chosen as study subjects by using a convenient sampling method. The sequence-based genotyping method was used for detecting HLA-B~* 13 : 01 allele in the DNA samples from peripheral blood of all study subjects. The serum levels of total protein,albumin,total bilirubin,direct bilirubin and alanine aminotransferase,aspartate aminotransferase and alkaline phosphatase activities in patients were examined. The correlation between the number of HLA-B~* 13 : 01 alleles and the liver function indices was also analyzed. RESULTS: There were 16 patients carrying HLA-B~* 13: 01 allele. The serum total protein in the HLA-B~* 13: 01 carriers was higher than that of non-carriers( P < 0. 05). The serum total protein was positively correlated with the number of patients carrying HLA-B~* 13: 01 alleles( P < 0. 05). CONCLUSION: The degree of liver function damage in OMDT patients may be related to carrying the HLA-B~* 13: 01 allele.

Detection of HLA-B*13:01 gene by dual allele-specific real-time polymerase chain reaction in patients with trichlorethylene-induced dermatitis / 中华劳动卫生职业病杂志

Objective@#To investigate the detection of a human leukocyte antigen-B (HLA-B) allele HLA-B*13:01 by dual allele-specific real-time polymerase chain reaction (PCR) in patients with trichlorethylene-induced dermatitis.@*Methods@#A total of 20 patients with trichlorethylene-induced dermatitis who were admitted and treated from January 2014 to October 2016 were enrolled as case group, and 20 persons who underwent physical examination from January to October, 2016 were enrolled as control group. Peripheral cubital venous blood samples were collected from all subjects, and dual allele-specific real-time PCR was used to detect the HLA-B*13:01 gene. The two groups were compared in terms of the proportion of subjects carrying HLA-B*13:01 gene.@*Results@#There were no significant differences between the case group and the control group in median age (25.0 years vs 27.0 years, Z=0.30, P>0.05) and the proportion of male subjects (60.0% vs 70.0%, χ2=0.44, P>0.05) . The mean time of exposure to trichloroethylene was 30.8 days in the case group, while the subjects in the control group were not exposed to trichloroethylene. The case group had a significantly higher frequency of HLA-B*13:01 gene than the control group (80.0% vs 20.0%, χ2=14.40, P<0.01) with an odds ratio of 16.00.@*Conclusion@#Dual allele-specific real-time PCR can be used for detection of the HLA-B*13:01 gene in patients with trichlorethylene-induced dermatitis.

Isolation and Identification of Platelet by Stepped Centrifugal Method / 现代检验医学杂志

Objective To establish a method for the isolation and identification of platelets.Methods 10 healthy volunteers were selected to collect the EDTA anticoagulant venous blood of 3 tubes,each tube was 2 ml,which was divided into the whole blood cell tube,platelet rich plasma (control group),and stepped centrifugal platelet extract (experiment group).Platelet was isolated by simple centrifugation method(PRP) and stepped centrifugal method.The two groups were full blood count and analyzed by microscopic morphology and platelet activity test.Leukocyte specific HGB gene and platelet mitochondrial ND1 gene content was analyzed by real time PCR.Results Platelets were extracted and detected in control group and experimental group.Platelets were found and white blood cells and red blood cells were not remained in experimental group.Platelets and sporadic white blood cells were found in control group.The platelet pick up rate of experiment group was significantly higher than control group,the difference was statistically significant.Experimental gene content HGB of experiment group was significantly lower than control group,the difference was statistically significant (t=-3.281,-2.865,P＜0.05).ND1 gene content of experiment group higher than the control group,the difference was not statistically significant.There was no significant difference for platelet activity test between experimental group and control group (t=-0.046,-0.799,P＞ 0.05).Conclusion A isolation and identification method of stepped centrifugal platelet was established.The method can be used for the study of platelet gene and the functional analysis of platelets.

Application of a modified culturing method for lymphocytes in cytogenetic research / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a modified method for microculturing whole human blood for cytogenetic analysis.</p><p><b>METHODS</b>A novel tube rack was designed to overcome the drawbacks of directly culturing the cells within centrifuge tubes. The fractions of human plasma, human serum and two commercial fetal bovine sera were analyzed with 15% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The influence of adding 0%, 5%, 10%, 15%, 20%, 25% and 30% autologous plasma to the culture on lymphocyte transformation rate and mitotic index (MI) was examined.</p><p><b>RESULTS</b>The SDS-PAGE analysis showed a significant difference between commercial fetal bovine sera, and that the components of human plasma were similar to those of fetal bovine serum. The value of MI in lymphocyte was evidently increased along with addition of autologous plasma. However, this has exerted no significant effect on the transformation rate. With the addition of 10% autologous plasma, the MI value has become much higher than the conventional method.</p><p><b>CONCLUSION</b>A modified method was established by application of a novel tube inclined rack and optimization of whole blood inoculation. This method is easier and cheaper, and is suitable for application in clinical practice.</p>

Surface display of lead-binding protein PbrR on Escherichia coli and preliminary study of intestinal ;colonization by the recombinant bacteria / 中华微生物学和免疫学杂志

Objective To construct a recombinant Escherichia coli ( E. coli) with surface-dis-played lead specific binding protein PbrR and to further study intestinal colonization by the recombinant bac-teria in mice and gastrointestinal tolerance of the bacterial surface-displayed PbrR. Methods Chimeric pro-tein Lpp-OmpA coding sequence was chemically synthesized and inserted into the expression vector pET-21a to construct the outer membrane display vector pLOA. PbrR coding sequence was also obtained by chemical-ly synthesis and inserted into pLOA to generate the outer membrane display plasmid pLOA-pbrr. E. coli BL21 (DE3)pLysS was transformed with pLOA-pbrr and induced by IPTG. The expressed recombinant proteins were analyzed by 15% SDS-PAGE and Western blot assay. Lead adsorption capacity of the cell surface-dis-played PbrR in the simulated intestinal juice and tolerance of the recombinant E. coli to simulated gastric juice were analyzed, respectively. KM mice were orally given the induced recombinant bacteria by gastric lavage for 7 consecutive days and then were continually fed until day 30. The contents of recombinant bacte-ria in stool samples were detected by dilution plate method on day 7, 15 and 30. The recombinant protein with His tag was detected by immunoblotting on day 7 and 15. Results Based on Lpp-OmpA, the PbrR outer membrane display vector was successfully constructed. The recombinant fusion protein Lpp-OmpA-PbrR-His tag was highly expressed in E. coli. The recombinant E. coli strains displaying PbrR on their outer membrane accumulated a significant level of Pb2+ in simulated intestinal juice. Moreover, those strains showed a tolerance to gastric acid in vitro and could colonize in the intestinal tracts of mice via oral infection. The surface-displayed recombinant fusion protein showed a better tolerance to the environment of digestive tract. Conclusion The recombinant E. coli strain displaying PbrR on its surface showed a stronger capabili-ty of lead accumulation from simulated intestinal environment and could colonize in the intestinal tracts of mice. The surface-displayed recombinant PbrR also showed a good tolerance to digestive juice. This study paved the way for further researches on the selective elimination of lead by biosorption based on animal mod-els.

The rabbit experimental study for toxicokinetics of chlorpyrifos impacted by hemoperfusion / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate toxicokinetic parameters impacted by hemoperfusion after oral chlorpyrifos exposure, to investigate the adsorption effect of hemoperhusion for chlorpyrifos poisoning.</p><p><b>METHODS</b>12 rabbits were divided into two groups after oral exposure with chlorpyrifos 300 mg/kg body weight. Control group: without hemoperfusion; hemoperfusion group: hemoperfusion starts 0.5 h after chlorpyrifos exposure and lasts for 2h. Blood samples were collected at different times, concentrations of chlorpyrifos were tested by GC, then, toxicokinetic parameterswere calculated and analysis by DAS3.0.</p><p><b>RESULTS</b>In hemoperfusion group, peak time was (7.19±3.74) h, peak concentrations was (1.37±0.56) mg/L, clearance rate was (13.93±10.27) L/h/kg, apparent volume of distribution was (418.18±147.15) L/kg The difference of these parameter were statistically significant compared with control group (P<0.05).</p><p><b>CONCLUSION</b>Hmoperfusion will decrease the inner exposure and load dose of rabbits with chlorpyrifos poisoning.</p>