Table of Content

30 September 2015, Volume 30 Issue 9

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Orginal Article

Change of red blood cell distribution width and its significance in patients with primary biliary cirrhosis

FU Haitao, YANG Min, HUANG Fenglou, LIANG Yan, ZHONG Renqian

2015, 30(9):  871-873.  DOI: 10.3969/j.issn.1673-8640.2015.09.001

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To investigate the clinical significance of red blood cell distribution width (RDW) in patients with primary biliary cirrhosis (PBC).

A total of 98 patients with PBC and 88 healthy subjects were enrolled, and their data were analyzed retrospectively. The change of RDW in PBC patients and the relationships with total bilirubin(TBil), creatinine(Cr), albumin(Alb), platelet(PLT), gamma glutamyltransferase(GGT), alkaline phosphatase(ALP) and Mayo risk score were analyzed.

RDW in PBC group (14.9%±2.8%) was higher than that in healthy control group (12.3%±0.4%, P<0.05), and there was statistical significance for TBil, Cr, PLT, Alb, GGT and ALP between the 2 groups (P<0.05). RDW was positively correlated with TBil, ALP and Mayo risk score (r=0.61, 0.46 and 0.51, P<0.05), and was negatively correlated with Alb and PLT (r=-0.69 and -0.46, P<0.05).

The increasing of RDW is a potential index for the severity of PBC.

Study on the serology identification of para-Bombay phenotype and the investigation of a family

ZHU Zehang, LU Yuanshan

2015, 30(9):  874-876.  DOI: 10.3969/j.issn.1673-8640.2015.09.002

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To study the correct serology identification methods of para-Bombay phenotype and search compatible donors for rare blood type.

A total of 4 cases of positive and reverse typing discrepancy were identified comprehensively. ABO positive and reverse typing, Lewis blood type identification, H antigen detection, red blood cell absorption-elution test, cold antibody detection and hemagglutination inhibition test for blood type substance in saliva were performed. A family investigation was made.

All the 4 cases were confirmed as para-Bombay phenotype. Proband c's sister was also proved that she was the same para-Bombay phenotype.

The cases of positive and reverse typing discrepancy should be added other essential tests for para-Bombay phenotype. The detection of cold antibody assists in not only identifying para-Bombay phenotype but also selecting compatible blood for transfusion. Family investigation makes possible observe source and inheritance of para-Bombay phenotype and provide homogeneous donors for transfusion.

Clinical significance on the determinations of serum autoantibodies and thyroid hormone in patients with vitiligo

WU Junqi, YAO Aiping, WANG Meiyan, JIANG Yixiu, LU Xiaodong

2015, 30(9):  877-880.  DOI: 10.3969/j.issn.1673-8640.2015.09.003

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To investigate the levels and clinical significance of serum autoantibodies and thyroid hormone in patients with vitiligo.

The anti-nuclear antibody (ANA), anti-nuclear antibody spectrum(ANAS), free triiodothyronine (FT3), free thyroxine (FT4), thyroid stimulating hormone (TSH), thyroglobulin antibody (TG-Ab) and thyroid peroxidase antibody (TPO-Ab) were determined in 123 patients with vitiligo and 60 healthy controls by indirect immunofluorescence, immunoblotting and chemiluminescence. The determination items were statistically compared between vitiligo group and control group.

The positive rates of ANA, TG-Ab and TPO-Ab in vitiligo group were higher than those in control group(P<0.01), while ANAS was not statistically significant between the 2 groups (P>0.05). FT3 and FT4 were not statistically significant in the 2 groups(P>0.05). TSH in vitiligo group was higher than that in control group (P<0.05).

The relationship is close between vitiligo and thyroid autoimmune diseases. ANA, TG-Ab, TPO-Ab and TSH determinations can be usually used in vitiligo patients with autoimmune diseases.

Diagnostic significance on the dynamic monitoring of serum procalcitonin level for recurrent infection

MU Yuejing, WANG Weijia, XU Shengnan, KAN Lijuan, HUANG Yanhua, OUYANG Nengliang, XU Quanzhong, YAN Haizhong, ZHANG Xiuming

2015, 30(9):  881-885.  DOI: 10.3969/j.issn.1673-8640.2015.09.004

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To investigate the clinical application significance of serum procalcitonin(PCT) level dynamic monitoring for diagnosing recurrent infection, and to investigate the PCT cut-off value for recurrent infection.

The retrospective study enrolled 150 patients with bacterial infection in Zhongshan Hospital, Sun Yat-sen University, and the change of serum PCT level was monitored dynamically. Among them,100 patients suffered from recurrent infection, while the other 50 patients acquired effective treatment. The PCT levels of recurrent infection group and effective treatment group in the same period were observed, and the difference between them was analyzed. A total of 60 blood samples from healthy subjects were also collected, and the difference between recurrent turning points of PCT and PCT levels of healthy subjects was analyzed. The diagnostic performance of PCT, white blood cell (WBC) and interleukin 6 (IL-6) for recurrent infection with receiver operating characteristic (ROC) curve was evaluated, and a primary diagnostic cut-off value of PCT was established for recurrent infection.

In recurrent infection group and effective treatment group, the serum PCT levels showed different trends. The recurrent infection group showed an obvious rebound, and the level was higher than that in effective treatment group (P<0.05). The rebound turning points of serum PCT level in recurrent infection group were still higher than those in healthy subjects(P<0.05). The diagnostic significance of PCT for recurrent infection was confirmed by ROC curve analysis, and the area under the ROC curve for PCT was 83.5%. The most appropriate cut-off value was 2.29 ng/mL, and the sensitivity was 89.0%, the specificity was 83.0%, the positive predictive value was 84.0%, and the negative predictive value was 88.3%. The areas under the ROC curves of WBC and IL-6 for recurrent infection were 55.1% and 61.0%, respectively.

It was of great clinical application significance to monitor serum PCT level dynamically for the diagnosis and differential diagnosis of recurrent infection.

Primary investigation on in vitro bacteriostatic action of Amur-corktree to Staphylococcus aureus

ZHAO Lingxu, WANG Lei

2015, 30(9):  886-889.  DOI: 10.3969/j.issn.1673-8640.2015.09.005

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To investigate in vitro bacteriostatic action of Amur-corktree to Staphylococcus aureus.

The minimum inhibitory concentrations (MIC) of Amur-corktree to 20 isolates of Staphylococcus aureus were determined by agar dilution method, normal dilution method and micro-dilution method.

The MIC for inhibiting 90% bacteria (MIC90) of agar dilution method was 12.5 mg/mL, the MIC90 of normal dilution method was 25 mg/mL, and the MIC90 of micro-dilution method was 25 mg/mL.

It exists an obvious in vitro bacteriostatic action of Amur-corktree to Staphylococcus aureus.

Evaluation on the efficiency of different media and GBS enrichment broth for GBS detection

LU Tingyan, SHEN Li, TANG Zhenhua

2015, 30(9):  890-893.  DOI: 10.3969/j.issn.1673-8640.2015.09.006

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To evaluate the reliability of Group B Streptococcus(GBS) screening medium, Colombia blood agar medium and GBS enrichment broth detection results and investigate the viability of clinical application by analyzing the positive rate of GBS and the performance of selective plates.

A total of 242 samples from distal vaginal segment and around crissum and 15 midstream urine samples were collected among 35-37-week pregnant women. The samples were inoculated directly and with one more step of GBS enrichment broth on Colombia blood agar and GBS screening media, respectively, and the growth of GBS in the 2 kinds of media was observed.

As for inoculating directly, the detection rates of GBS in Colombia blood agar medium for 24 h and 48 h were 7.39% (19/257), and the 24 h and 48 h detection rates of GBS in GBS screening medium were 5.45%(14/257) and 7.39%(19/257), respectively. As for inoculating with one more step of GBS enrichment broth, for neither 24 h nor 48 h, the detection rate of GBS in Colombia blood agar medium was 9.73%(25/257), and the detection rate in GBS screening medium was 8.95%(23/257), both of which had an evident increasing comparing with the former one. There was no statistical significance for the detection rate between the 2 kinds of media (P>0.05).

By enriching 18 h-24 h in GBS enrichment broth, followed by inoculating in GBS screening medium or Colombia blood agar medium, the growth of GBS from clinical samples in media could be observed just after 24 h, and this method has simple recognition ability for GBS and can improve GBS positive detection rate.

The influence of Helicobacter pylori infection on serum homocysteine level in patients with carotid atherosclerosis

LEI Ming, BAI Ju, WU Jianhua, XIA Danni, ZHENG Ruidong

2015, 30(9):  894-897.  DOI: 10.3969/j.issn.1673-8640.2015.09.007

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To investigate the influence of Helicobacter pylori (Hp) infection on carotid intima-media thickness(CIMT), plaque stability and the metabolism of homocysteine(Hcy) in carotid atherosclerosis patients.

According to carotid artery ultrasonography, 129 patients with carotid atherosclerosis were enrolled. ¹⁴C urea breath test was used to determine Hp infection. Meanwhile, enzymatic cycling method was used to determine serum Hcy levels .

Serum Hcy levels and CIMT of patients with carotid atherosclerosis in Hp infection group were higher than those in Hp non-infection group (P<0.01), and they increased with the severity of Hp infection with statistical significance (P<0.01). The plaque detection rate in Hp infection group was higher than that of Hp non-infection group(P<0.05), and the incidence of vulnerable plaques in Hp infection group was obviously higher than that in Hp non-infection group(P<0.05). Logistic regression analysis showed that the possibility of vulnerable plaques in Hp infection group was 2.35 times higher than that in Hp non-infection group. With serum Hcy increasing by 1 μmol/L, the possibility of vulnerable plaque increased by 9%.

Hp infection is likely to promote the development and progression of carotid atherosclerosis through influencing Hcy metabolism and increasing CIMT and the instability of carotid atherosclerotic plaque.

The expression and its difference of cornulin in cervical lesions

DIAO Wenjing, SUN Hong, GUO Qisang, WANG Li, TAO Xiang, SUI Long

2015, 30(9):  898-902.  DOI: 10.3969/j.issn.1673-8640.2015.09.008

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To study the cornulin expressions in cervicitis tissue, cervical intraepithelial neoplasia tissue and cervical cancer tissue, and to evaluate preliminarily the correlation of cornulin with cervical intraepithelial neoplasia tissue.

A total of 131 paraffin-embedded tissue samples were collected from cervical intraepithelial neoplasia and suspicious cervical cancer patients undergoing loop electrosurgical excision procedure (LEEP) for diagnostics or treatment. They included 39 patients with low-grade squamous intraepithelial lesion (LSIL), 49 patients with high-grade squamous intraepithelial lesion(HSIL), 43 patients with invasive cervical cancer(ICC), and 38 patients with cervicitis were enrolled as control group. Immunohistochemistry [streptavidin-peroxidase (SP)] using a rabbit polyclonal antibody against human cornulin was applied in all samples. Polymerase chain reaction (PCR) was used for the determination of cornulin mRNA expression in all 131 patients' cervical thinprep cytologic test samples prior to cervical LEEP procedure and control group's cervical thinprep cytologic test samples after histopathology diagnosis by colposcopy.

Immunohistochemistry showed that the expression levels of cornulin were significantly different with the other groups (P=0.000)except that between LSIL and control group(P=0.148). With the severity degree of cervical disease increasing, the expression of cornulin decreased, and it decreased significantly in ICC. The expression of cornulin mRNA was consistent with the results of cornulin expression level. If taking cornulin weakly positive(+)or negative (-)as the cut-off for screening cervical HSIL or higher, the sensitivity was 70.7%, and the specificity was 85.7%. And for screening cervial LSIL or higher, the sensitivity was 56.5%, and the specificity was 94.7%.

Cornulin might be one of the useful markers for cervical carcinogenesis.

Correlation study of serum ferritin and high-sensitivity C reactive protein in gestational diabetes mellitus

ZHANG Xianghui, ZHAO Yuanming, SUN Zhuoxiang, CHENG Panpan, JIN Chao

2015, 30(9):  903-905.  DOI: 10.3969/j.issn.1673-8640.2015.09.009

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To investigate the relationship between serum ferritin (SF) and high-sensitivity C reactive protein (hs-CRP) in patients with gestational diabetes mellitus (GDM) and their clinical significance.

A total of 86 patients with GDM and 120 pregnant women with normal glucose tolerance (NGT) were enrolled as GDM and NGT groups. SF and hs-CRP were determined. The relationship between SF and hs-CRP in the 2 groups was evaluated,

The SF and hs-CRP in GDM group were higher than those in NGT group (P=0.031 and 0.007). Meanwhile, serum SF was positively correlated with hs-CRP (GDM group: r=0.352, P=0.003; NGT group: r=0.285, P=0.001).

The levels of SF and hs-CRP may play important roles in the progress of GDM. SF and hs-CRP can be used as predictive factors for the prevention and treatment of GDM.

Significance of homocysteine and blood lipid detection in the diagnosis of sudden sensorineural hearing loss

CHEN Hong, YANG Hong, QI Guorong

2015, 30(9):  906-910.  DOI: 10.3969/j.issn.1673-8640.2015.09.010

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To study the significance of homocysteine(Hcy) and blood lipid detection [total cholesterol(TC), triglyceride(TG) and low-density lipoprotein cholesterol(LDL-C)] in the diagnosis of sudden sensorineural hearing loss(SSHL).

A total of 80 patients with SSHL were enrolled, and 78 subjects without SSHL matched with SSHL group for sex, age and race were enrolled as control group. The serum levels of Hcy, TC, TG and LDL-C were detected. The relationship with SSHL was analyzed.

The serum Hcy, TC, TG and LDL-C levels in SSHL group [(16.16±3.94) μmol/L, (5.38±0.99)mmol/L, 1.66(1.23-5.70)mmol/L and (3.49±0.78)mmol/L] were significantly higher than those in control group [(12.59±2.24)μmol/L, (4.51±0.91)mmol/L, 1.48(0.69-3.01)mmol/L and (2.59±0.80)mmol/L] (P<0.01). The correlation analysis showed that serum Hcy was positively correlated with TC, TG and LDL-C(r=0.669, 0.514 and 0.704, P<0.01). Logistic regression analysis showed that Hcy and LDL-C were important risk factors for SSHL[OR(95% confidence interval) were 1.85(1.256-2.725) and 4.412(1.134-17.170)]. The positive rates of Hcy and LDL-C were 47.5% and 51.3% in SSHL group and 9.0% and 14.1% in control group (P<0.01).The positive rate of the combined detection of Hcy and LDL-C was 66.3%, which was higher than any single-positive rate.

The combined detection of Hcy and blood lipid for SSHL has significance in preventing,diagnosing and risking assessment .

Influence of different processing techniques on peripheral blood monocyte-platelet aggregation by flow cytometry

LI Linyun, MEI Bing, WANG Changfu

2015, 30(9):  916-920.  DOI: 10.3969/j.issn.1673-8640.2015.09.012

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To investigate the influence of different processing techniques on monocyte-platelet aggregation(MPA) by flow cytometry in order to provide the reference for the determination of MPA.

Sodium citrate anticoagulation whole blood samples were collected randomly. CD14-phycoerythrin(PE) and CD61-fluorescein isothiocyanate (FITC) were used to label monocytes and platelets. CD62P-PE was used to label activated platelets. The percentages of CD14-PE/CD61-FITC-double-positive MPA in monocytes and CD62P-positive platelets were determined by flow cytometry. Different processing techniques were performed as follows. Whole blood samples were prepared immediately after collection or delayed for different times. After preparation, samples were analyzed immediately or delayed for different times. Antibody immunolabeling was performed before fixation of whole blood samples, immediately after fixation or at different delayed times after fixation. Before immunolabeling, whole blood samples were centrifugated or not. Different antibodies, anti-CD14-PE with anti-CD61-FITC, anti-CD14-PE with anti-CD41-ECD and anti-CD45-ECD with anti-CD61-FITC, were used to label MPA. The results of MPA from different processing techniques were compared.

Compared with immediate preparation after blood collection, the percentages of MPA and CD62P-positive platelets increased with the delayed time at room temperature(18-25℃) and low temperature (2-8℃) (P<0.05), and there was a positive correlation(room temperature: r=0.82, P<0.05; low temperature: r=0.83, P<0.05). Compared with immediate determination after preparation, the percentages of MPA of samples stored at low temperature for 24h after preparation did not alter significantly(P>0.05). Compared with immunolabeling samples before fixation with 1% paraformaldehyde, no significant change was observed in MPA percentages from immediate immunolabeling after fixation or immunolabeling samples stored at low temperature for 24 h after fixation(P>0.05). Centrifugation of whole blood samples before immunolabeling resulted in significant increase of the percentages of MPA and CD62P-positive platelets(P<0.05).MPA percentages resulted from different antibodies had no significant variance (P>0.05).

Whole blood samples should be handled as soon as possible. Fixation with 1% paraformaldehyde at low temperature could store samples for 24h before handling. Centrifugation should be avoided before immunolabeling. After preparation and fixation, samples could be stored at low temperature for 24h before analysis. Different antibody combinations, including anti-CD14-PE/anti-CD61-FITC, anti-CD14-PE/anti-CD41-ECD and anti-CD45-ECD/anti-CD61-FITC, are all suitable for immunolabeling MPA.

Performance evaluation of SUCCEEDER SF-8000 coagulation testing system

WU Qijiao, TONG Yi, CAO Liyan, YANG Deqin, XU Junquan, CHANG Jun, FAN Linxia

2015, 30(9):  921-925.  DOI: 10.3969/j.issn.1673-8640.2015.09.013

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To evaluate the performance of domestic SUCCEEDER SF-8000 coagulation testing system(SF-8000).

The precision, accuracy, linearity, comparability and so on of SF-8000 were evaluated.The 200 g/L hemoglobin solution was added to 12 cases of fresh plasma in the proportions of 1:200, 2:200, 3:200, 4:200 and 5:200, and the experimental plasma was prepared. The differences of the results in the experimental plasma, the original plasma and the brine-diluted control plasma were analyzed. The differences of 15 cases of bubble specimens and control specimens' results were analyzed by paired t-test. The 12 fresh specimens were measured with fresh dissolved-reagents and 24h-refrigerated reagents separately, and the differences of the 2 sets of results were analyzed. A total of 13 cases of 4h-refrigerated specimens, 24h-refrigerated specimens and fresh specimens were measured, and the differences were analyzed.

The basic properties of the instrument were all in accordance with the requirements of manufacturers. Hemolysis and bubbles affected the results of SF-8000 in different levels. When hemoglobin concentration reached 1-1.5 g/L, the results of activated partial thromboplastin time(APTT)and prothrombin time(PT) had significantly statistical differences from those of the original plasma group(P<0.01), when it reached 4 g/L, they had statistically significant differences from the control plasma group(P<0.01). In addition to the reagent of APTT(P<0.01), the results of 24 h-refrigerated reagents and fresh refrigerated reagents had no statistical differences with good stability. The results of 4h-refrigerated specimens and fresh specimens had no statistical difference, but the results of 24 h-refrigerated specimens and fresh specimens in APTT, PT and fibrinogen(FIB) had statistical significance(P<0.01).

The performance of SF-8000 meets the national and industrial standards. The manual operation should be strictly carried out to avoid appearing confounding factors, in order to ensure the results with clinical applicability.

Performance evaluation of PL-11 platelet analyzer in the detection of platelet count and platelet aggregation

CAI Yuchan, ZHAO Xuhong, HAN Ping, CHEN Changming, LI Zhi

2015, 30(9):  926-930.  DOI: 10.3969/j.issn.1673-8640.2015.09.014

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To investigate the performance of PL-11 platelet analyzer in the detection of platelet count and platelet aggregation.

According to the instrument performance verification standards in the Clinical and Laboratory Standards Institute (CLSI) and the Health Industry Standard of the People's Republic of China, WS/T 406-2012: the Quality Standard of Routine Tests in Clinical Hematology, the performance of PL-11 platelet analyzer in the detection of platelet count and platelet aggregation were evaluated. Platelet aggregation rate in healthy subjects was compared for correlation, which was detected by PL-11 platelet analyzer, LBY-NJ4 platelet tester and TEG-5000 thromboelastogram.

The within-run and inter-day precisions of PL-11 platelet analyzer were<1/3 of total error (7%), and the carry-over rate was 0.32%. In (4.12-1 380.4)×10⁹/L linear range, the regression equation slope was 1.03 (R²=0.993). The coincidence rate for platelet count by PL-11 platelet analyzer with reference method was 84%. There was no statistical significance of platelet aggregation rate among PL-11 platelet analyzer, LBY-NJ4 platelet tester and TEG-5000 thromboelastogram in 79 patients with type 2 diabetes mellitus (P>0.05). However, there was statistical significance in platelet aggregation rates before and after taking clopidogrel hydrogen(P<0.01).

PL-11 platelet analyzer can provide accurate and reliable results for platelet count and aggregation.

Preparation of polyclonal antibody and development of a competitive ELISA for fungal (1,3)-beta-D-glucan

YAN Jun, YANG Wei, ZHAI Shuanzhu, XUE Zhixin, YU Peng, ZHOU Zeqi

2015, 30(9):  931-933.  DOI: 10.3969/j.issn.1673-8640.2015.09.015

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To develop a competitive enzyme-linked immunosorbent assay (ELISA) with high specificity and sensitivity for fungal (1,3)-beta-D-glucan.

Rabbit polyclonal antibody was produced by immunization with protein-conjugated (1,3)-beta-D-glucan and fungal extract. The highest titer and little cross-reactivity polyclonal antibody was labeled with horseradish peroxidase (HRP) and purified. Finally, a competitive ELISA was established for fungal (1,3)-beta-D-glucan.

The linear range was 3.125-200 pg/mL. The recovery range for 100, 25 and 6.25 pg/mL serum antigen was 97.8%-113.6%, and the coefficient of variation for repeated test was <15%. The detection system could effectively detect low concentrations of Aspergillus fumigatus and Candida albicans and high concentration of Cryptococcus neoformans from serum samples. Meanwhile, the detection system demonstrated little interference against 5 kinds of pathogenic bacteria, including Mycobacterium tuberculosis, Escherichia coli, Salmonella, Klebsiella and Staphylococcus aureus.

A competitive ELISA is developed successfully for the detection of invasive fungal disease with (1,3)-beta-D-glucan used as a coated antigen and enzyme-labeled antibody HRP-Ab3B used as a detection antibody.

The establishment and clinical application of Ureaplasma parvum relative quantitative assay

ZHAO Zhen, LIU Lu, ZHAO Fang, CAO Guojun, HUANG Yanqun, JI Yuhua

2015, 30(9):  934-938.  DOI: 10.3969/j.issn.1673-8640.2015.09.016

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To establish Ureaplasma parvum (Up) polymerase chain reaction (PCR) relative quantitative assay, to avoid the effect of different vaginal sampling ways for determination results, and to investigate the role of Up in bacterial vaginosis (BV).

The primers and probes for Up were designed according to DNA topoisomerase Ⅳ gene sequences of 4 Up serovars to detect Up in vaginal secretion samples. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was analyzed by PCR quantitative assay to determine host cells. The Up relative quantification was calculated as Up copies in 10³ host cells. Up standard isolate and clinical isolates, Ureaplasma urealyticum standard isolate and clinical isolates and 11 common vaginal microorganisms were determined to identify the specificity of the assay. The vaginal secretion samples were collected from 532 BV patients and 276 healthy women for the analysis of Up relative quantification.

Up PCR quantitative assay had high sensitivity (100 copies/μL), without amplification to Ureaplasma urealyticum and 11 other vaginal microorganisms. The infection rates of Up in BV patients and healthy women were 49.8% (265/532) and 43.1% (119/267), respectively, without statistical significance (χ²=3.263, P=0.071). Up relative quantification in BV patients was 667 copies/10³ cells, which was significantly higher than that in healthy women (20 copies/10³ cells) (χ²=47.012,P=0.000 1). When using >50 copies/10³ cells as a reference, the positive rate of Up in BV patients was 39.3% (209/532). It was significantly higher than that in healthy women (17.0%, 47/276) (χ²=41.537,P=0.001).

The cell adhesion ability of Up is a key factor in the pathogenesis. Up PCR relative quantitative assay can overcome the influence of different vaginal sampling ways on the determination results and objectively reflect the pathogenicity of Up.

Multilocus sequence typing study of ESBLs-producing Escherichia coli in one hospital

PAN Jun, XU Qingxia, XIAO Weiqiang, CHANG Yanmin, SHEN Yong

2015, 30(9):  939-943.  DOI: 10.3969/j.issn.1673-8640.2015.09.017

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To investigate the multilocus sequence typing (MLST) and genetic evolution of extended-spectrum beta-lactamase (ESBLs)-producing Escherichia coli in the Affiliated Cancer Hospital of Zhengzhou University, and to provide the reference for the monitoring of ESBLs-producing isolates' epidemic and hospital infection control.

Gene amplification and sequence analysis were performed for 7 housekeeping genes of 26 isolates of ESBLs-producing Escherichia coli, and alleles were compared with those assigned at MLST databases.

A total of 16 sequence types (ST) were obtained in the 26 isolates of ESBLs-producing Escherichia coli, in which there were 3 ST69 and 3 ST131.

ESBLs-producing Escherichia coli has diversified genetic backgrounds, and ST131 and ST69 are the most prevalent types, while other types were scattered.

Observation of cucurbitacin E-induced egress of Toxoplasma gondii from host cells

QIU Jiayang

2015, 30(9):  944-947.  DOI: 10.3969/j.issn.1673-8640.2015.09.018

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To investigate the effect of cucurbitacin E on the egress of Toxoplasma gondii from host cells.

Toxoplasma gondii infected HFF cells were treated with cucurbitacin E (20,50 and 100 μmol/L) for 1 h, and the egress rate was evaluated statistically. Toxoplasma gondii infected HFF cells were pretreated with Cyto-D and 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis acetoxymethyl ester(BAPTA-AM), followed by 50 μmol/L cucurbitacin E treatment for 1 h and egress determination. The virulence change of egressing Toxoplasma gondii, such parasites were collected and used to re-infect HFF or infect Kunming mice, was analyzed.

Cucurbitacin E could induce the egress of Toxoplasma gondii from host cells, and the egress rate increased with the concentration of cucurbitaci E. Cucurbitacin E-induced ergess depended on the gliding ability of Toxoplasma gondii and intra-parasite calcium. There was no difference between cucurbitacin E-induced egressing parasite and naturally egressing parasite in virulence.

Cucurbitacin E could induce the egress of Toxoplasma gondii from host cells, which provides new direction to study the interactions between Toxoplasma gondii and its host cells.

Research progress of Shanghai Glycohemoglobin Harmonization Program

SHAO Wenqi, SUN Lin, WANG Beili, WU Jiong, SONG Binbin, ZHANG Chunyan, GUO Wei, PAN Baishen

2015, 30(9):  948-952.  DOI: 10.3969/j.issn.1673-8640.2015.09.019

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To improve the harmonization of hemoglobin A_1c (HbA_1c) results from laboratories participating in Shanghai Glycohemoglobin Harmonization Program(SHGHP)in order to improve the diagnosis of diabetes mellitus.

A total of 4 level Ⅰ laboratories certificated by the National Glycohemoglobin Standardization Program (NGSP) and a laboratory by the International Federation of Clinical Chemistry and Laboratory Medicine(IFCC)were comprised the proposed reference laboratory network. SHGHP website was established, and whole blood calibrators and comparative specimens were sent to all participating clinical laboratories. The harmonization of results before and after calibration was compared. The mean value, standard deviation and coefficient of variation(CV) of HbA_1c detected by the laboratories before and after calibration were calculated and analyzed for difference statistically.

The CV of low-level and high-level HbA_1c by the 5 reference laboratories were 2.27% and 1.58%, respectively. The CV of HbA_1c detected by the third-grade hospitals after calibration decreased significantly than that before calibration (P<0.001). The CV after calibration ranged from 0.94% to 4.22%. The percentages of passed laboratories in SHGHP from 2012 to 2013 were above 90% in which the third-grade hospitals all passed the evaluation.

SHGHP calibrates the different methods by fresh whole blood with no significant matrix effects and improves the harmonization of HbA_1c significantly among the participating laboratories.

Application of allowable total error in sigma metrics for assessing the analytical quality of clinical chemistry determination

ZHANG Lu, WANG Wei, WANG Zhiguo

2015, 30(9):  953-957.  DOI: 10.3969/j.issn.1673-8640.2015.09.020

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To investigate the importance of allowable total error (TEa) source in sigma(σ) metrics for assessing the analytical quality of clinical chemistry determination.

In this study, the data were collected from the second internal quality control of routine chemistry and the first external quality assessment of routine chemistry in 2014 organized by the National Center for Clinical Laboratory. One of the laboratories was selected for its coefficient of variation (CV) and the bias of 19 clinical chemistry items from the data. σ of 2 runs were calculated by 5 different TEa. The σ metrics' performance for assessing the analytical quality of clinical chemistry determination was analyzed comparatively.

σ metrics varied with the changes of TEa and imprecision. Under the National Health Industry Standard, the major σ values(68.4%)for control 1 ranged from 2 to 4 and from 3 to 6 for control 2(58%). Under RiliBÄK, except triglyceride (negative) and alanine aminotransferase (ALT)(<3), others had a σ value from 3 to 6, even up to 14.78. Under the Clinical Laboratory Improvement Amendment of 1988 (CLIA'88), 89.47% of control 1 showed a σ value>3, up to 7.69, and 84.2% of control 2 showed a σ value > 3, up to 10.43. Under biological variability, the σ value of control 1 ranged from 1 to 5, and the most (63%) was < 3, and that of control 2 ranged from 1 to 6, but those of 9 from 19 were < 3. Under the TEa of Australian, the σ value of control 1 was <3, and that of 79% control 2 was <3 .The σ value of control 2 was generally higher than that of control 1.

The 6σ is an efficient way to control quality, but the lack of TEa for many analytes and inconsistent TEa from different sources are important variables for the interpretation of σ metrics in a routine clinical laboratory.

A case of bloodstream infection caused by Prevotella oralis through operation

GUO Jian, ZHI Yunqing, HE Lihua, LÜ Li, QIANG Sufeng, WU Wenjuan

2015, 30(9):  958-961.  DOI: 10.3969/j.issn.1673-8640.2015.09.021

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To report a bloodstream infection case of a female patient after genital tract abscess operation by Prevotella oralis.

Blood samples were collected by aseptic method, and were detected by VersaTREK240-10 automatic microbial detection system. The positive culture blood samples were smeared, inoculated of plates for pure isolates of pathogenic bacteria, identified by VITEK 2 Compact, and confirmed by 16S rRNA sequencing.

Two bottles of left side and right side of anaerobic blood culture were positive after 48 and 72 h, respectively. The results of VITEK 2 Compact and 16S rRNA sequencing results were both Prevotella oralis, and the identification rates were 96% and 99%, respectively.

A isolate of Prevotella oralis is identified from a female patient with bloodstream infection in East Hospital.

The historical evolution and development prospect of routine blood smear staining

SONG Xiaoying, YUAN Baojun, HAO Jihong

2015, 30(9):  962-967.  DOI: 10.3969/j.issn.1673-8640.2015.09.022

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The objection is to state of composition, function, history development and application prospect of routine blood smear staining, in order to stress that blood staining is the basis of morphological diagnosis.The staining effect determines the identification of blood cells and impacts on the level of blood system disease diagnosis and differential diagnosis.