Establishment of Internal Quality Control Method for Mixed and Split Samples by NAT in Blood Donor Screening / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To establish and evaluate the methods of internal quality control in blood donor screening by nucleic acid test (NAT).</p><p><b>METHODS</b>After HBV-DNA standard quality control (QC) sample (60 IU/ml) was diluted by pooling 6 samples, the concentration was 10.0 IU/ml, which was approach twice of the low limit. When the pooling result turned out reactive, the pooling samples need to be split into single sample to process. Meanwhile, the standard QC samples were tested as well. The same batch QC samples were tested 20 times respectively, calculate the mean (x̄), standard deviation (SD) and CV. Make Levey-Jennings QC curves by setting x̄±2SD as warning, x̄±3SD as rejected. The Levey-Jennings quality controls chart was mapped by using Microsoft Excel.</p><p><b>RESULTS</b>After 20 times test of mixed/split samples, the x̄±2SD were 33.03±1.47 and 30.08±0.98, the x̄±3SD were 33.03±2.20 and 30.08±1.47, the CV were 2.22% and 1.63%, respectively. The P value of t test was 0.08 and 0.17 respectively, there was no statistically significant difference between the 2 group.</p><p><b>CONCLUSION</b>When establish an internal QC system in the screening laboratory by nucleic acid testing, the concentration of the QC samples should be equal to normal specimens. This type of QC system may validate the extraction and amplification of the nucleic acid, and improve the stability of the test results.</p>

Infection status of HBV, HCV and HIV in voluntary blood donors of Chinese Nanjing area during 2010-2013 / 中国实验血液学杂志

This study was purposed to understand the infection of HBV, HCV, HIV among the voluntary blood donors and the epidemic trend in infectious population in Chinese Nanjing area, and to guide the mobilization and recruitment of blood donors. A total of 199777 whole blood samples of voluntary blood donors were tested by ELISA, the nucleic acid technology (NAT) combined detection (HBV-DNA, HCV-RNA, HIV-RNA) was added for detection of the samples with HBsAg,anti-HCV, anti-HIV at least unilateral negative donors from June 10, 2010 to June 9, 2013 years, and these statistic data were analyzed. Every HIV reactive sample(HIV-antibody and/or HIV-RNA) was sent to be confirmed in the Centers for Disease Control and Prevention in Nanjing. The results showed that the voluntary donors' infection rate of HBsAg, anti-HCV, anti-HIV were 0.45%, 0.28%, 0.11% respectively; NAT positive rate was 0.07%, 32 cases were confirmed with anti-HIV positive, in which 30 cases were male (6 cases were repeated blood donors) and 2 cases were female, 3 cases were unconfirmed, in which 2 cases were males and 1 case was female. The statistical analysis demonstrated that the difference of unqualitative rate of HBsAg, anti-HCV, anti-HIV was statistically significant between the first-time and repeated blood donors.It is concluded that the positive rate of anti-HCV and anti-HIV displayed a declining trend year by year in Nanjing voluntary blood donation population from June 10,2010 to June 9, 2013 years. The unqualitative rate of HBsAg and NAT increased with the age increasing, while that of anti-HCV, anti-HIV decreased with age increasing. The unqualitative rate of the repeated blood donors is far lower than that of the first-time blood donors. The ELISA positive rate of anti-HIV testing in females is higher than that in males, but the confirmed positive rate of male is significantly higher than that of female. Therefore the consulting skills before donating should be improved, concerning the link of recruiting donors, focusing on strengthening the first-time donors' consultation, evaluating and developing the fixed voluntary blood donors, and vigorously popularizing NAT technology in blood screening to improve the blood safety effectively.

Hematopoietic growth factors expressed in human aorta-gonad-mesonephros (AGM)-derived stromal cells in vitro / 中国实验血液学杂志

This study was aimed to investigate the hematopoietic growth factors expressed in human aorta-gonad-mesonephros (AGM)-derived stromal cells in vitro in order to provide the basic data for elucidating the role of AGM -derived-stromal cells in embryo-hematopoiesis and its hematopoietic suppoitive effect. RT-PCR was used to analyze the expression of IL-6, SCF, Flt3-L, oncostatin M (OSM), IL-3, TPO, M-CSF and LIF in human aorta-gonad-mesonephros-derived stromal cells (hAGMS1-S5) at mRNA level. IL-6, SCF and Flt3-L levels were detected in the supernatant of hAGMS1-S5 stromal cells by ELISA assay. Umbilical cord blood CD34(+) cells were cocultured with hAGMS1-S5 feeder cells, and hematopoietic cells were collected at day 14 for colony analysis in methylcellulose semisolid medium. The results showed that human aorta-gonad-mesonephros-derived stromal cells S1-S5 expressed IL-6, SCF, Flt3-L and OSM mRNA, but did not express IL-3, TPO, M-CSF and LIF mRNA. In the supernatant of hAGMS1-S5 cells, IL-6, SCF and Flt3-L could be detected by ELISA assay at different levels, while there was no significant difference between groups of hAGMS1-S5 (P > 0.05). When cocultured with umbilical cord blood CD34(+) cells, hAGMS1-S5 could support the expansion of CFU-GM, BFU-E, and CFU-Mix. The supportive effects of hAGM S1-S5 were significantly different (P < 0.05), hAGM S3 and S4 were better than hAGM S1, S2, and S5. It is concluded that detection of hematopoietic growth factors expressed in human aorta-gonad-mesonephros-derived stromal cells provided a solid foundation to elucidate the mechanism of hematopoiesis and the hematopoietic supportive effect of these stromal cells.

A case-control study on small-for-gestational-age in relation to obstetrical risk factors / 中华流行病学杂志

<p><b>OBJECTIVE</b>Previous studies suggested that a number of obstetrical factors were associated with small-for-gestational-age (SGA) infant. However, it remained uncertain which obstetrical factors might increase the risk of SGA due to limitations of small sample size and poor study designs in the previous studies in China. We assessed the association of a few maternal factors, such as pregnancy-induced hypertension (PIH), oligohydramnios, infectious diseases of reproductive system, abnormal conditions of placenta or umbilical cord, previous adverse pregnancy outcomes, hysteromyoma and maternal education level, and the risk of SGA.</p><p><b>METHODS</b>We examined the association in a case-control study, in which the SGA group included all of 834 (443 males and 391 females) singleton alive SGA deliveries in our hospital during January 2000 to October 2004. 2502 (1329 males and 1173 females) appropriate-for-gestational-age (AGA) infants (1:3, SGA/AGA) matched by gender- and gestational-age-stratified frequencies of SGA were randomly selected for controls from infants with sex-specific birthweight for gestational age ranged between 10th-90th percentiles. Odds ratios (95% confidence interval, 95% CI) of these factors for SGA were calculated in univariate and multivariate analyses. The maternal risk factors were diagnosed by the relevant criteria used in China. SGA was defined as sex-specific birthweight for gestational age that was less than 10th percentile cut-off of fetal growth reference of singleton alive infants delivered in our hospital during that period.</p><p><b>RESULTS</b>Results from univariate analysis showed that PIH, oligohydramnios, history of adverse pregnancy outcome, poor maternal education and viral hepatitis were significantly associated with SGA. The odds ratios (95% CI) of these five factors for SGA were 3.95(2.79-5.60), 2.94(2.27-3.80), 7.01(3.87-12.70), 3.62(2.92-4.49) and 0.52(0.32-0.87), respectively. In the multivariate logistic analysis, a similar result was shown. The ratios of maternal PIH, oligohydramnios, history of adverse pregnancy outcome and viral hepatitis in the SGA group were 4.00(2.81-5.71), 2.95 (2.27-3.83), 5.95 (3.05-10.64), 0.50(0.30-0.84) folds of those in the AGA group; the ratio of maternal low formal education (< or = 9 years) in the SGA group was 3.46(2.75-4.24) times of the AGA group.</p><p><b>CONCLUSION</b>PIH, oligohydramnios, poor maternal education and a history of adverse pregnancy outcome were significantly associated with an increased risk of SGA while viral hepatitis might be associated with a decreased risk of SGA.</p>

Validation study on the criteria for clinical classification of small for gestational age infants / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the validity of criteria currently used in China for the classification of symmetric small for gestational age infants (SGA) as compared with its definition.</p><p><b>METHODS</b>This study included 417 inpatients diagnosed as SGA in authors' hospital from January 1998 to June 2002. Symmetric SGA was diagnosed by the following three criteria: (1) the Ponderal Index (PI), (2) the crown-heel length-to-head circumference ratio (BL/HC) issued in Chin J Pediatr (1988;26:164 - 165), as well as (3) the SGA definition. The definition criterion was considered as the "gold standard". The sensitivity, specificity, false positive and negative values, positive and negative predictive values, exact agreement ratio, diagnosis index, and Cohen's Kappa value were used to evaluate the validity and agreement of the methods of PI and BL/HC. Receiver Operating Characteristic (ROC) analysis was used to evaluate the validity of the diagnosis.</p><p><b>RESULTS</b>Of 417 SGA infants, 376 (90.17%), 376 (90.17%) and 187 (44.84%) subjects were diagnosed as symmetric type with PI, BL/HC and the definition criteria, respectively. (2) The agreement rate and Kappa value between PI and BL/HC was 80.82% and -0.093 (SEM 0.026), respectively. And the agreement rates between PI or BL/HC and the definition criterion were 49.88% and 50.84%, respectively. As compared with the definition criterion, the PI and BL/HC methods had sensitivities of 91.8% - 96.4%, specificities of 9.3% - 25.9%, positive predictive values of 45.8% - 51.1%, negative predictive values of 72.7% - 82.8%, diagnosis indices of 4.9% - 17.7% and Kappa values of 0.070 - 0.167. (3) The areas under the ROC curves in full-term and preterm infants by PI method were 0.635 (95% CI, 0.573 - 0.697) and 0.698 (95% CI, 0.622 - 0.725), respectively. PI cutoffs at 2.47 in full-term SGA, at 2.43 in preterm SGA, and BL/HC cutoff at 1.43 produced the maximum diagnosis indices that were 24.7%, 39.6% and 33.7%, respectively. When the PI at 2.50 (full-term), PI at 2.31 (preterm) and BL/HC values at 1.46, the sensitivity closed mostly to the specificity. The sensitivities and specificities in full-term and preterm infants were 59.4% and 59.3%, 65.3% and 65.5%, and 66.3% and 65.5%, respectively.</p><p><b>CONCLUSION</b>In the classification of SGA, the results showed a poor agreement between PI or BL/HC and the definition criterion. The results suggested that the current cutoffs of PI and BL/HC might not be appropriate for the diagnosis of symmetric SGA. Low AUC suggested that PI and BL/HC could not give a valid diagnosis at any cutoffs.</p>