Objective To analyze the clinical characteristics of Klebsiella pneumoniae infection in preterm infants. Methods Clinical data of 75 preterm infants infected with Klebsiella pneumoniae treated in BaYi Children's Hospital from February 6,2008 to February 10,2010 were retrospectively analyzed.The difference of auxiliary examination between early-onset and late-onset infection group were compared by two independent samples t test.Spearman correlation analysis and non-conditional Logistic regression analysis were used to analyze the high risk factors and the prognostic factors of Klebsiella pneumoniae infection in preterm infants. Results The incidence of Klebsiella pneumoniae infection was 2.8％ (75/2721) in preterm infants,and the mortality rate was 9.3％ (7/75). There were 71 cases of Klebsiella pneumoniae sepsis and 4 cases of Klebsiella pneumoniae pneumonia.Among 75 cases,63 cases were early-onset infection (onset age≤72 h) and 12 were late-onset infection (onset age＞72 h).All patients presented with poor response,heart rate during quiet sleep ＞ 160/min and low oxygen saturation.The mean corpuscular volume and mean corpuscular hemoglobin concentration in early-onset Klebsiella pneunoniae infection cases were higher than those in late-onset neonates [(128.87±24.60) fl vs (113.72±13.54) fl,t=-2.07,P＜0.05and (38.11±2.15) pg vs (36.98±1.05) pg,t=-2.76,P＜0.05].Low birth weight and caesarean section were associated with early-onset Klebsiella pneumoniae sepsis (r=0.250 and -0.240,P＜0.05). The prognosis of Klebsiella pneumoniae infection was associated with hospital stay and duration of premature rupture of membranes (r=0.368 and 0.318,P＜0.05). Conclusions There were no specific clinical manifestations for Klebsiella pneumoniae infection in preterm infants.Preterm infants with low birth weight,long duration of premature rupture of membranes,delivered by caesarean section and received invasive operation are likely to develop Klebsiella pneumoniae infection.

Objective To investigate the incidence and high risk factors of brain injury in very low birth weight infants(VLBWI),to reduce the morbidity of brain injury,and improve the developmental outcome of VLBWI. Methods Data of 181 VLBWI admitted in the neonatal intensive care unit(NICU)between October 2008 and September 2009 were retrospectively analyzed. The difference in basic information,maternity diseases,treatment and complication were analyzed between two groups(brain injury group and normal newborn group),and Logistic regression analysis was adopted to analyze the risk factors for brain injury. Results Seventy-eight of the 181 neonates(43.09%)were found to have brain injury,including 67 neonates(37.01% )with periventricular/intraventricular hemorrhage(43 with intraventricular hemorrhage(IVH)gradeⅠ,12 with IVH grade Ⅱ,10 with IVH grade Ⅲ,and 2 with IVH grade Ⅳ)and 12 neonates(6.63%,one complicated with IVH grade Ⅲ)with periventricular leukomalacia. The younger the gestational age,the higher the brain injury rate was observed. Concerning the brain injury rate,there were no differences in gender,single birth/plural births,birth weight,the mode of delivery,fetal distress,premature rupture of membrane,hypertension during pregnancy,placenta abruption,and intrauterine growth restriction(IUGR)between these two groups(P ＞ 0.05). The difference in therapeutic measures such as pulmonary surfactant therapy,nasal continuous positive airway pressure(nCPAP),conventional mechanical ventilation,and high-frequency oscillatory ventilation was significant(P ＜ 0.05),except aminophylline therapy(P ＞ 0.05). As to the complication,there were significant differences in the incidences of asphyxia,neonatal respiratory distress syndrome(NRDS),hypercapnia,metabolic acidosis,hyperglycemia,anemia,and personal digital assistant(PDA)(P < 0.05). However,there was no difference in the incidences of hypoglycemia,sepsis,thrombocytopenia,apnea,pulmonary hemorrhage,and hyperbilirubinemia between these two groups(P ＞ 0.05). Further Logistic regression analysis showed that NRDS,high-frequency oscillatory ventilation,and PDA were the main risk factors for brain injury in VLBWI. Conclusions VLBWI is the high-risk population of brain injury. Pulmonary surfactant therapy,nCPAP,conventional mechanical ventilation,high-frequency oscillatory ventilation,asphyxia,NRDS,hypercapnia,metabolic acidosis,hyperglycemia,anemia,and PDA were confirmed to be the high-risk factors for brain injury in VLBWI. And,NRDS,high-frequency oscillatory ventilation and PDA were main risk factors.

ObjectiveTo investigate the application of next-generation sequencing (NGS) in determining the full-length sequence and baseline resistance-associated substitution (RAS) of hepatitis C virus (HCV) subtype 3b. MethodsNucleic acid was extracted from plasma of a HCV RNA-positive blood donor, and after sequence-independent amplification, a sequencing library was constructed and NGS was performed using Illumina Hiseq. Bioinformatics methods were used to analyze full-length HCV sequence, viral genotype, and baseline RAS to direct-acting antivirals (DAAs). ResultsA total of 8.4 Gb data with more than 56 million reads were obtained. The full-length HCV sequence was obtained by bioinformatics analysis, with an average sequencing depth of 488 007 and a genotype of 3b subtype. A total of 12 RASs were identified in HCV amino acid sequence, i.e., Y56H, Q80K, Q80R, and A156G located in NS3, M28G, Q/A30G, Q/A30K, L31F, L31M, and Y93H located in NS5A, and S282T and V321A located in NS5B, among which Q/A30K and L31M located in NS5A had high frequencies of 99.16% and 98.37%, respectively, while the other 10 RASs had low frequencies of ＜0.5%. ConclusionNGS can be used to determine the full-length sequence and genotype of HCV subtype 3b and identify baseline RASs, which has great significance in the epidemiological study of HCV subtype 3b and the development of DAA treatment regimens.

Objective:To explore the association between interleukin(IL)-28B single nucleotide polymorphisms and natural outcome of hepatitis C virus.Methods:The IL-28B rs12979860 locus was genotyped in 266 HCV infected volunteer blood donors(107 spontaneous cleared and 159 chronic infection) and 97 healthy controls using Sanger sequencing assay.The difference in rs12979860 genotypes and allele frequencies between the six groups(107 spontaneous cleared and 159 chronic infection,266 HCV infection and 97 healthy controls,159 chronic infection and 97 healthy controls) were analyzed by statistics.Results:159 HCV chronic infection,107 spontaneous cleared and 97 healthy controls,were shown more CC genotype,accounting for 83.6%,95.3%and 86.6%,respectively, while the CT genotype accounted for 16.4%,4.7%and 13.4%respectively.No TT genotype was found.The CC/CT genotype was not significant difference between HCV infection and healthy controls,chronic infection and healthy controls(χ2=0.204,P=0.652;χ2=0.406,P=0.524),but between chronic infections and spontaneous clearance had statistically significant(χ2=8.474,P=0.004),the frequence of C allele in spontaneous cleared was higher than HCV chronic infection(χ2=7.949,P=0.005).Conclusion: The gene polymorphism of IL-28B rs12979860 is not related to HCV susceptibility,but there are differences in chronic infection and spontaneous cleared,showing the C allelic in favor of HCV spontaneous cleaed.

OBJECTIVETo observe the effect of intrapartum and postpartum factors on abnormal neurological findings in the extremely preterm infants.

METHODClinical data of 62 premature infants (33 of male, 29 of female) were retrospectively analyzed. None of the premature infants had birth defect; their gestational ages were all less than 28 weeks (23(+ 6)-27(+ 6) weeks). They were hospitalized within 12 hours after birth in the neonatal intensive care unit (NICU) of BAYI Children's Hospital from November 2010 to June 2013. The blood gas, birth condition, complications, the mechanical ventilation and the ultrasonic encephalography were recorded. The 62 cases were divided into 2 groups, alive group and died group. Meanwhile, all cases of survial were divided into brain injuries group and normal brain group. Data were analyzed with t-test, Chi square test and Spearman correlation analysis.

RESULTFifty-six cases were alive, and 6 cases died (3 were during the treatment and 3 were after parents gave up). The average birth weight of brain injuries group was (954 ± 182) g; and that of the normal brain group was (1 071 ± 136) g. There were significant differences between the two groups in gender (χ(2) = 4.314, P = 0.038), gestational age (χ(2) = 11.622, P = 0.001), birth weight (t = 2.728, P = 0.009), which had significant correlation with neurological outcomes. The Spearman correlative coefficients were -0.278, 0.456 and 0.364 respectively. And P values were 0.038, 0.000 and 0.006. The rates of multiple pregnancy, lung hemorrhage and surgical operation in brain injuries group were 45%(9/20), 55%(11/20), 40%(8/20), which were significantly higher than those in normal brain group, 3%(1/36), 17%(6/36), 11%(4/36)(χ(2) = 12.800, 8.936, 4.773, P all < 0.05). These three factors were the high risk factors for adverse neurological outcomes, the odds ratios were 28.64, 6.11 and 5.33 respectively. There was no significant difference in delivery mode, amniotic fluid, maternal infection, asphyxia, necrotizing enterocolitis, patent ductus arteriosus, sepsis, mechanical ventilation, inhaled nitric oxide therapy, blood glucose, blood gas analysis, doses of dopamine between brain injuries group and normal brain group. The birth weight in alive group was (1 029 ± 163) g, which was significantly higher than those in died group (870 ± 144)g (r=0.29, P=0.022). There was no significant difference in other factors between alive group and died group(P all>0.05).

CONCLUSIONGender, gestational age and birth weight may have relation with the neurological outcomes of extremely preterm infants. Multiple pregnancy, pulmonary hemorrhage and surgical operation are the risk factors of brain injuries. Birth weight is related to the survival of extremely preterm infants.

Birth Weight ; Brain Injuries ; physiopathology ; Female ; Gestational Age ; Humans ; Infant, Extremely Premature ; Infant, Newborn ; Infant, Premature, Diseases ; Lung ; pathology ; Male ; Pregnancy ; Pregnancy, Multiple ; Retrospective Studies ; Risk Factors

Blood screening is essential for blood safety. Traditional methods are not very effective in unknown pathogen testing. The blood metagenomic analysis with high-throughput sequencing as the main method is not limited to a particular microbe but detects all microbes in the sample (total genome), which has advantages that traditional methods cannot compare with. With the increasingly maturation of technology and the gradual reduction of cost, metagenomic sequencing has been applied more and more widely. In this paper, the concept and research procedures of metagenomics, the current status and future prospect of metagenomics sequencing technology in the field of blood screening are reviewed.

Objective:To construct a scientific and practical ovarian tumor nursing quality sensitive indicator system so as to provide a reference for evaluating the quality of nursing care for patients with ovarian tumors.Methods:Based on Donabedian's three-dimensional theoretical model of structure-process-outcome quality management, we used literature review and Delphi expert correspondence consultation to construct a nursing quality sensitive indicator system for ovarian tumor patients. From February to April 2019, we selected 20 experts from 16 ClassⅢ Grade A general hospitals and 2 higher nursing institutions from 7 provinces/municipalities in Shandong Province, Jiangsu Province, Beijing, Jilin Province, Shanghai, Guangdong Province and Sichuan Province for consultation.Results:Among two rounds of consultation, valid recovery rates were 90.00% and 94.44% respectively; authority coefficients were all 0.92; familiarity coefficients were 0.89 and 0.91 respectively; judgment coefficients were 0.94 and 0.92 respectively; Kendall harmony coefficients were 0.204 and 0.426 respectively; the differences were all statistically significant ( P<0.05) . The final nursing quality sensitive indicator system for ovarian tumor patients included 3 first-level indicators, 12 second-level indicators and 23 third-level indicators. Conclusions:The nursing quality sensitive indicator system for ovarian tumor patients is highly scientific and practical which can be used to standardize clinical nursing care for patients with ovarian tumors by gynecological nurses and improve the nursing quality.

【Objective】 To investigate the CMV-IgG positive yeild among blood donors in Guangzhou and explore the differences in the efficacy of three test reagents, aimed at improving blood safety and service capacity of blood centers. 【Methods】 A total of 630 blood samples from eligible blood donors from July to October 2020 in our center were randomly selected and screened for CMV-IgG by one ELISA reagent.Among them, 180 samples were tested in parallel using three reagents (two ELISA reagents and one ECLIA reagent), and those tested negative were conducted quantitative CMV-DNA detection.The test results of different reagents were compared and analyzed. 【Results】 Out of the 630 samples, a total of 598 positive samples were screened out, including 180 samples yielded by three reagents, 171 and 175 by the two ELISA reagents, respectively, and 175 by ECLIA.The results given by three reagents were consistent (Kappa>0.4), and no significant difference in the positive yeild by three reagents was found.In the 180 samples, 11 were negative, among which 3, 2 and 6 samples were negative by all three reagents, two reagents and one reagent (ELISA), respectively.All the 11 samples were tested negative for CMV-DNA. 【Conclusion】 The yeild of positive CMV-IgG in blood donors was 94.9% (598/630), suggesting a high prevalence of CMV in Guangzhou. CMV serologically negative blood should be considered when providing blood products to immunocompromised patients to improve the safety of recipients.The detection results of ELISA reagents and ECLIA reagent for CMV- IgG are consistent, but ECLIA reagent has better detection efficacy.

【Objective】 To explore the effectiveness of using donor plasma reinfusion to flush the leukoreduction system (LRS) chamber during the final reinfusion phase with the Trima Accel automated blood collection system in preventing the reduction of CD4+ and CD8+ T lymphocytes. 【Methods】 A longitudinal and cross-sectional study was designed. CD4+ count<200 cells/μL and CD8+ count<125 cells/μL were considered as the criteria for deficiency. Eighteen first-time platelet donors were followed up. The lymphocyte count was measured at 0, 3-6 and 7-14 times of blood donation in the last 300 days. 170 healthy blood donors who have not donated blood were selected as the control group. According to the cut-off point(October 2021), 88 blood donors who mainly used automatic blood collection system to donate platelet apheresis in the last 365 days(median blood donation times ≥17.5)were divided into three groups(A, B and C)and blood samples were obtained. The time for Groups A, B and C started donating platelet apheresis were as follows: Group A: before October 2019, Group B: from October 2019 to September 2021, Group C: after October 2021. Blood samples were analyzed to obtain blood counts including CD4 + and CD8 + T lymphocytes. Blood samples were analyzed to obtain blood cell counts including CD4+ and CD8+ T lymphocytes. Through a comparative analysis, this study aimed to determine if there are any statistical differences in the detection indices between the follow-up groups with varying frequencies of blood donation, the control group, and groups A, B, and C. This approach was employed to infer the efficacy of donor plasma reinfusion in flushing the leukoreduction system (LRS) chamber for preventing the decline of CD4+ and CD8+ T lymphocytes. 【Results】 Eighteen first-time blood donors who were converted to regular platelet apheresis donors did not show a decrease of CD4 + and CD8 + T lymphocytes in the 5 th and 11 th blood donation (median number of blood donation), and there was no significant difference between the above indexes and those in the 0 th blood donation. Among the previous frequent blood donors, the CD4+ and CD8+ T lymphocyte counts in Group B and Group C are both higher than the standard value, showing no statistical difference from the control group. Among regular blood donors, the CD4+ and CD8+ T lymphocyte counts in groups B and C were higher than the criteria values, and had no statistical difference compared to the control group.The CD4+ T lymphocyte count in Group A was normal, with only one donor in Group A having a CD8+ T lymphocyte count below 125 cells/μL. This donor has donated 281 times of platelet apheresis, and the group he belongs to has started blood donation 2-21 years(median of 5 years) before the adjustment of reinfusion mode. The CD4+ and CD8+ T lymphocyte counts in Group A showed significant differences compared to the control group, with median counts (Group A/Control Group) of 359/521 and 257/372, respectively, P<0.001. In Group A, 0%(0/35) had a CD4+ count below 200 cells/μL, and 2.85%(1/35) of donors had a CD8+ count below 125 cells/μL, which was far lower than the proportion of CD4+ and CD8+ T cell deficiency found in regular apheresis donors by John M. Gansner and Mahboubeh Rahmani. The study showed that the adjustment of the plasma reinfusion mode did not further reduce the T lymphocyte counts in blood donors, but instead further restored the T lymphocyte counts in regular blood donors. This indicated that after the adjustment of plasma reinfusion mode, blood donors might not have lost CD4+ and CD8+ T lymphocytes during blood donation, or only lost a small amount, and can recover even if they donate platelet apheresis frequently. 【Conclusion】 Trima Accel automated blood collection system has a good effect on preventing CD4 + and CD8 + T lymphocytes from being reduced by flushing the LRS chamber with donor plasma.

【Objective】 To study the CD4 T cell epitopes in Core and NS3 protein of genotype 1(GT1) and 6(GT6) of hepatitis C virus(HCV). 【Methods】 A total of 298 overlapping peptides(16-mer) spanning Core and NS3 protein of GT1 and GT6 HCV were synthesized. Peripheral blood mononuclear cells(PBMCs) from 17 HCV+ and 7 healthy blood donors were stimulated by peptide pools, followed by evaluating T cell response by IFN-γ ELISPOT, by which 21 peptides with positive results were found. These peptides were further applied to individually stimulate 20 HCV+ and 18 healthy PBMCs. The differences of responsive frequencies to the 21 positive peptides between the two study groups were compared. 【Results】 Pooled and individual peptide stimulation tests showed that HCV+ PBMCs were responsive to the stimulation of 5 peptides(GT1 NS31273-1288 and NS31315-1330; GT6 NS31033-1048, NS31087-1102 and NS31351-1366), with a responsive frequency ranging 18.9%-27.0%. In contrast, healthy PBMCs were not or low responsive(0%-4.0%) to these five peptides. The responsive frequencies were statistically different between the two groups(P<0.05). No reported epitopes in IEDB were found identical with these 5 peptides via sequence alignment. 【Conclusion】 Our study identified novel CD4 T cell epitopes in NS3 protein of GT1 and GT6 HCV, which has potential application value for the research and development of HCV vaccine.