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Objective:To evaluate the performance of the automated digital cell morphology instrument in detecting platelet (PLT) clumps.Methods:A total of 4271 blood samples whose PLT reached the reviewing rules of thrombocytopenia were selected from inpatients having blood analysis in Xijing Hospital from January 1 st to June 30 th, 2019, including 2 200 males and 2 071 females,with a median age of (35±7.03) years old. The smears for these cases were made, stained by Wright-Giemsa, and examined to capture PLT clumps by digital cell morphology system and manual microscope separately. The digital cell analysis system (hereinafter referred to as the instrument method) as an evaluation method and the microscope method as a reference method were used to calculate the positive rate of platelet clump detection and evaluate the comparison of two methods and bias assessments. The chi-square test was used to compare counting data rates. Results:Among 4, 271 samples reaching the reviewing rule of thrombocytopenia, 128 cases with platelet clumps were detected by manual microscope(initial) with a positive detection rate of 96.24%, and a total 133 of cases with PLT clumps were detected by microscope (initial+reconfirmation) with a positive detection rate of 100 %. Meanwhile, 129 cases with platelet clumps were detected by instrument method with a positive detection rate of 96.9%. There was no significant difference in terms of positive rate of PLT clumps detection between the instrumental method and the microscope method (initial) ( χ2 =0.115, P=0.73); the positive rate of clumps detection by the instrumental method was lower than microscope method (initial+reconfirmation), and the difference was statistically significant (χ 2 =4.061, P=0.04). For instrument method, the positive rate of PLT clumps detection by simultaneous observation of RBC analysis interface+PLT aggregation interface+WBC analysis interface was higher than only observation of PLT aggregation interface, and the difference was statistically significant (χ 2 =5.090, P=0.02). The average error of the deviation of PLT counting results before and after correction of the cases with PLT plumps missed by instrument method was significantly higher than microscope method (initial), and the difference was statistically significant (χ 2 =56.26, P<0.001). Conclusion:The automated digital cell morphology system has a good consistency with manual microscope(initial) in terms of the sensitivity of platelet clumps detection and can be used as a supplementary method for detecting platelet aggregation.
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Objective To analyze the cause of platelet aggregation in blood specimens ,so as to provide basis for reducing platelet aggregation ,and avoiding false positive of platelet count ,false report ,misdiagnosis and mistreatment .Methods The blood speci-mens which platelet was below 80 × 109 /L ,below 125 × 109 /L with histogram hinted platelet aggregation ,were smeared ,stained with Wright-Giemsa ,and observed by microscope for platelet morphological changes .The data between each groups were calculated and analyzed by statistical software SPSS version 18 .0 .Results A total of 184 cases of ethylenediaminetetraacetic acid dependent pseudothrombocytopenia(EDTA-PTCP) were found ,accounted for 0 .444 ‰ totally ,including 0 .244 ‰ of out-patients (101 cases) , 0 .159 ‰ of hospitalized patients (66 cases) ,and 0 .041 ‰ of health examination personnel (17 cases) .3 cases of multi-dependent pseudothrombocytopenia and 25 cases of pseudo platelet aggregation were found ,and accounted for 0 .007 ‰ and 0 .060 ‰ respec-tively .Conclusion The discovery of platelet aggregation which caused mainly by EDTA-PTCP ,still relies on microscopy ,and pseu-do platelet aggregation comes mainly from sampling ,so it needs to strengthen the skills training .
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Objective To investigate the long-term excessive drinking semen nitric oxide (NO) content in their sperm quality and spermatogenic cell apoptosis and infertility. Methods Nitrate reductase method was used to specific reduce the NO metabolites nitrate (NO3-) to nitrite (NO2-), which was used on behalf of the total NO level. Terminal deoxynucleotidyl transferase (TdT) and mediated nick end labeling (TUNEL) method and binocular optical microscope were used to detect and observe the rate of apoptosis of spermatogenic cells and the morphological structure. The SQA-V sperm quality automatic analyzer was used to measure sperm quality. Results In not drinking semen fertility group, NO content was (54.81±11.45)μmol / L, the rate of spermatogenic cell apoptosis was (4.52±1.23)%, sperm motility was (80.24±0.17)%, energy a+b (78.32±0.12)%, deformity rate (5.30±0.13)%, and long-term excessive alcohol infertility C group was [(128.83±22.73)μmol/L,(17.34±2.53)%,(51.18±0.58)%,(21.45±0.26)%,(21.12±3.24)%] respectively. Compared to a very significant difference (t＝10.04,17.38,6.69,15.59,17.02,P＜0.01) . In long-term excessive drinking group, the levels of NO and spermatogenesis cell apoptosis rate was significantly positive correlated (r＝0.93,P＜0.01).Apoptosis of spermatogenic cell nucleus chromatin was condensed in the formation of crescent-shaped perinuclear, nuclear was cleavaged to form apoptotic bodies. Conclusions In the long-term excessive drinking semen, NO content and spermatogenic cell apoptosis rate was increased, the sperm had poor quality. The results showed that the long-term excessive drinking can cause germ cell apoptosis in male infertility and promote the body to overproduce NO, whichmay be one of the reasons for male infertility.
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Objective To investigate the relationship between clue cell in semen and male infertility. Method Semen specimens from 957 patients were examined with microscope. Gram staining was performed when clue cell were found to be present in semen. At the same time, general characteristics of the sperm were analyzed. Results There were clue cells in 23. 5% (225/957) of the total specimens, in which, 95. 1% clue cells were gram staining positive. The concentration and vitality of living sperm in semen presented with clue cell were significantly lower, but the abnormal sperm and pH value were much higher than that in normal control (P<0.01). In addition, 28 sexual partners of 30 infertility patients were identified as having bacterial vaginosis. Conclusion Gardnerella vaginalis and short coccohacillus were mostly transmitted from infected sexual partner. It could cause squamous epithelial cell forming clue cell, and lead to infertility by chan-ging pH which mainly affect reproduction of the sperm. This study suggested that clue cell has a unique vale for the diagnosis of infertility causing by Gardnerella vaginalis and short coccohacillus.
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Objective To purify Micrococcus luteus Rpf and Rpf domain fusion protein, and to in-vestigate its effects on growth of Mycobacterium tuberculosis. Methods The recombinant plasmids pPro-EXHT-Rpf and pPro-EXHT-Rpf domain were expressed in E. Coli DHSa and then purified under denaturing condition via Ni-NTA purification system and confirmed by Western blot. The biochemical property of the M. Luteus Rpf and Rpf domain was analyzed by stimulating the resuscitation of M. Tuberculosis H37Ra which were in non-culturable' condition. Results The Rpf and Rpf domain products achieved 95% and 93% pure respectively, and the molecular weight was 30 x 103 and 12 x 103, the yield of purification was about 471 mg/L and 337 mg/L of the culture. The M. Luteus Rpf and Rpf domain from the E. Coli showed activity of stimulating the resuscitation of M. Luteus and M. Tuberculosis H37Ra in non-cuhurable' condition which could be inhibited by monoclonal antibodies of M. Luteus Rpf domain remarkably. Conclusion It was dem-onstrated that the purification of Rpf and Rpf domain have high biological activity for further functional, pharmacological and clinical investigations, and M. Luteus Rpf domain protein is fully active as M. Lateus full-length Rpf.
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Objective To investigate the immunological properties of Rv1009 domain. Methods BALB/c mice were immunized with Rv1009 domain three times at 2-week interval. ELISA was used to detect the antiRv1009 domain antibody titer in the sera of immunized mice sera. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry. Levels of secreted IFN-γ, IL-10 and IL-12 upon specific antigen stimulation were detected by ELISA. The BALB/c mice immunized with Rv1009 domain were intravenously infected with MTB H37Rv. Four weeks after the final injection, the number of CFU in spleens was determined. Results The titer of the specific antibody in sera of the immunized BALB/c mice was 1:12 800. The SI of Rv1009 domain immunized group (2. 40±0. 18) was significantly higher than that of saline immunized group (0.90±0.21). The IFN-γ,IL-10 and IL-12 levels in culture supematant of spleen lymphecytes from the fusion proteins immunized mice was (1 432±30) ng/L, (503±11) ng/L and (311±11) ng/L respectively, significant different from that of saline immunized group[(256±20) ng/L, (76±6) ng/L and(56±8) ng/L,P<0.01]. Four weeks after the final injection,compared with normal saline immunized mice (6.64±0.13), dramatic reduction in MTB replication was observed in the spleen (4.86±0.14) from BALB/c mice immunized with fusion proteins following a subsequent MTB H37Rv challenge, but the protection efficacy of mice immunized with Rv1009 domain was not as good as that of BCG vaccination group (3.81±0.16). Conclusion Rv1009 domain can be used as a candidate for the new TB vaccine.