Correlation between umbilical artery erythropoietin level and perinatal factors in premature infants and its clinical significance / 第二军医大学学报

Objective: To explore the correlation between umbilical artery blood erythropoietin (EPO) level and perinatal factors in premature infants and its clinical significance. Methods: Umbilical artery blood samples from 107 premature infants born in the Eastern Branch of Shanghai First Maternity and Infant Hospital of Tongji University between Jan. 2019 and Jun. 2019 were collected. The levels of EPO and ferritin were measured by ELISA and chemiluminescence assay, respectively. The 107 infants were divided into three groups according to the quartile EPO level: low level group, medium level group and high level group. The relationship between umbilical artery blood EPO level and gestational age, birth body mass and other perinatal factors, the incidence of anemia of prematurity (AOP), necrotizing enterocolitis (NEC), patent ductus arteriosus (PDA) and atrial septal defect (ASD) in premature infants, and the clinical characteristics of pregnant mothers was analyzed. Results: The EPO level of umbilical artery blood in 107 newborn premature infants was 5.94-137.18 mU/mL, and the median level was 23.51 (14.60, 51.28) mU/mL. There were 26 cases in the low level group (the EPO level 14.60 mU/mL), 54 in the medium level group (14.60-51.27 mU/mL), and 27 cases in the high level group (≥51.28 mU/mL). Univariate analysis showed that the gestational age of the infants in the low level group was significantly lower than those in the medium level group and the high level group (both P < 0.05), the age of the pregnant mothers was significantly higher than those in the medium level group and the high level group (both P < 0.05), the natural pregnancy rate was significantly lower than that in the high level group (P < 0.05), and the continuous positive airway pressure (CPAP) usage rate of the infants was significantly higher than that in the medium level group (P < 0.05). The ferritin level of umbilical artery blood was significantly higher in the midium level group than that in the high level group (P < 0.05). The incidence of AOP in the high level group was significantly higher than that in the midium level group (P < 0.05). Multiple linear regression analysis showed that the EPO level of umbilical artery blood was positively correlated with the gestational age of newborn premature infants and the natural pregnancy rate of pregnant mothers (both P < 0.01). Multivariate logistic regression analysis showed that the higher the natural pregnancy rate, the higher the level of EPO in umbilical artery blood, and the higher the natural delivery rate, the lower the level of EPO in umbilical artery blood. The risks of PDA and NEC decreased and the risk of ASD increased with the increase of EPO level in umbilical artery blood (all P < 0.05). Conclusion: Conception method and delivery mode are the influencing factors of EPO level in umbilical artery blood. Monitoring the EPO level of umbilical artery blood is helpful to diagnose the common complications such as AOP, PDA, ASD and NEC in premature infants.

Role of miR-21 in rat hepatic oval cell proliferation and activation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the possible mechanisms of miR-21-mediated regulation of proliferation and activation of hepatic oval cells.</p><p><b>METHODS</b>The 2-acetamidofluorene/partial hepatectomy (2-AAF/PH) method was applied to generate hepatic oval cell activation model in male Sprague-Dawley rats; after the 7 days of 2-AAF/PH or PH alone (control), the rats were sacrificed at 0 h, 6 h, 12 h, 24 h, 72 h and 168 h. Expression of miR-21 was detected by real-time PCR and differences between groups were evaluated using the two-sample t-test. Differential transcription of miR-21 target genes was assessed bioinformatically, and with western blotting to detect changes in protein expression of the target gene.</p><p><b>RESULTS</b>The rat hepatic oval cell activation model was successfully established.The 2-AAF/PH rats showed miR-21 expression beginning to increase at 12 h, peaking at 24 h, and decreasing thereafter until an increase at 168 h.For the control group, the miR-21 expression began to increase at 6 h, until 24 h when expression began steadily declining to reach the original level.Compared to the control group, the experimental group showed expression of miR-21 that was significantly less at 6 h (P=0.039, t =3.029) and significantly more at 24 h and 168 h (P=0.026, t =-3.433 and P=0.007, t =-5.105). Among the predicted target genes of miR-21 were WW domain containing E3 ubiquitin protein ligase 1 (WWD), Smad family member 7 (Smad7), and polybromo-1 (Pbrm1).Smad7 protein expression began to decrease at 6 h in the control group, until reaching its minimum at 24 h when it increased; in the experimental group, SMAD7 expression increased at 6 h, then began to decrease with the minimum detected at 168 hour.In the control group, the Smad7 mRNA expression decreased slightly at 6 h, then began to increase, reaching its peak at 24 h when the expression fell to the original level. In the experimental group, the Smad7 mRNA expression began to increase at 6 h and reached its peak at 24 h when it decreased; the expression was little more than its original level at 168 h.Smad7 protein expression was negatively correlated with miR-21, and Smad7 mRNA expression was positively correlated with miR-21 but negatively correlated with Smad7 protein expression.</p><p><b>CONCLUSION</b>miR-21 may play a vital role in the activation and proliferation of hepatic oval cells.As a target gene of miR-21, Smad7 might be involved in the process.</p>

Establishment of a real-time PCR assay for simultaneously detecting human BKV and CMV DNA and its application in renal transplantation recipients / 病毒学报

To establish a fluorescent quantitative PCR method (FQ-PCR) with TaqMan probe for simultaneous detection of polyomavirus (BKV) and cytomegalovirus (CMV) and to evaluate its clinical application in the renal transplantation recipients. The conservative sequences of BKV and CMV were targeted and amplified by nested PCR technique. The PCR products were cloned into the plasmids pcDNA3. 1(+). The recombinant plasmid containing target sequences of BKV and CMV were constructed as external standards. The TaqMan-based assay was optimized. For evaluating the assay, the sensitivity was determinated by diluted standard (5 X 103-10icopies/mL), and the specificity was verified by negative control and positive control, and the precision was assessed by intra-assay coefficient of variation (ICV) through detecting standard repeatedly (20 times). A total of 480 blood samples of renal transplantation recipients were used to detect BKV and CMV DNA simultaneously with FQ-PCR, and the concentrations of FK506 were measured by ELISA. The association of DNA copy and concentrations of FK506 was analyzed. The cloned target BKV and CMV DNA was confirmed by sequencing and analysis. The sensitivity of the FQ-PCR assay reached 5 X 103 copies/ml in detecting BKV or CMV DNA. Control DNA verified the assay specifically detecting target DNA. The precision of the assay to quantif target DNA copies was acceptable (Intra-assay CV was 3.44% for BKV and 2.23% for CMV; Inter-assay CV was 4. 98% for BKV and 3.76% for CMV;). Of 480 samples, 130 samples (27. 08%) were CMV DNA positive, significantly higher than the BKV DNA positive (13.33%, 64/480, P<0.05). The positive BKV or CMV DNA was found to be associated with high concentrations of FK506 (P<0. 05). In conclusion, the developed real-time PCR assay for detecting both CMV and BKV DNA simultaneously was s high sensitive, precise and time-effectiveand could be applied in the monitoring of the CMV and BKV infection in the renal transplantation recipients.