Influence of short-time pasteurization on essential bioactive components and immune cells in human breast milk / 中华围产医学杂志

Objective:To explore the influence of short-time pasteurization (62.5±0.5℃ for 5 s) on the main bioactive components and immune cells in human breast milk.Methods:Fresh breast milk was collected from 53 women whose premature infants were admitted to the neonatal intensive care unit of the Children's Hospital of Fudan University from May 2020 to October 2020. Each sample (20 ml) was divided into unsterilized, Holder pasteurized (62.5 ℃ for 30 min), or short-time pasteurized groups. The concentration of secretory immunoglobulin A (sIgA), lactoferrin (LTF), lysozyme (LZM), and insulin-like growth factor binding protein-3 (IGFBP-3) in breastmilk whey were detected by enzyme-linked immunosorbent assay and the number of viable immune cells (leukocytes, monocytes, T cells, and B cells) in breastmilk by flow cytometry.Results:(1) A total of 87 breast milk samples were collected. The levels of sIgA, LTF, and LZM were the highest in the unsterilized group, followed by the short-time and Holder pasteurized group [0.42 mg/ml (0.33-0.65 mg/ml) vs 0.40 mg/ml (0.28-0.62 mg/ml) and 0.25 mg/ml (0.17-0.37 mg/ml); (3.57±1.06) vs (3.53±1.11) and (0.85±0.58) mg/ml; 128.60 μg/ml (77.18-203.00 μg/ml) vs 121.70 μg/ml (68.66-188.20 μg/ml) and 83.40 μg/ml (47.40-151.40 μg/ml); all P<0.05]. There was no significant difference in the level of IGFBP-3 among the groups. The median retention rates of sIgA, LTF, and LZM in the Holder pasteurized group were all lower than those in the short-time pasteurized group [55.87% (46.01%-71.41%) vs 96.93% (83.03%-115.90%); 21.72% (12.54%-29.42%) vs 97.88% (88.98%-104.30%); 69.26% (49.42%-89.08%) vs 93.80% (74.85%-110.20%); all P<0.05]. No significant difference in the level of preserved IGFBP-3 was observed between the three groups ( P>0.05). (2) The number of viable leukocytes, monocytes, T cells, and B cells in the Holder pasteurized group were lower than those in the unsterilized group [leukocytes: 185.50 (87.00-356.50) vs 1 271.00 (540.50-2 283.00); monocytes: 12.00 (6.00-16.75) vs 266.00 (137.30-518.80); T cells: 1.00 (0.00-2.00) vs 47.50 (28.50-116.00); B cells: 1.00 (0.00-1.75) vs 21.00(10.00-41.50); all P<0.05]. The percentage of viable leukocyte to the total leukocyte and the viable monocytes, T cells, and B cells to the viable leukocytes were lower in the Holder pasteurized group than those in the unsterilized group [24.80%(16.00%-36.80%) vs 74.20%(63.55%-86.45%); 5.91%(4.09%-8.77%) vs 21.90%(17.40%-29.30%); 0.31%(0.00%-1.31%) vs 4.00%(2.69%-6.43%); 0.30%(0.00%-0.86%) vs 1.27%(0.57%-2.85%); all P<0.05]. A similar trend was observed between short-time pasteurization and unsterilized groups (all P<0.05). (3) The percentages of viable monocytes, T cells, and B cells in their subsets were lower in both Holder and short-time pasteurized groups than those in the unsterilized group [2.94%(1.33%-7.14%) vs 9.72%(5.77%-16.00%) and 52.60%(31.35%-68.75%); 0.00%(0.00%-1.61%) vs 0.49%(0.00%-2.53%) and 28.10%(10.55%-57.00%); 0.00%(0.00%-0.83%) vs 0.24%(0.00%-2.47%) and 13.80%(3.27%-41.00%); all P<0.05].The number and percentage of viable leukocytes in total leukocytes and viable monocytes in total monocytes [leukocytes: 279.50(116.80-548.50), 32.20%(20.70%-45.75%); monocytes: 32.00(21.00- 83.75),15.60%(11.10%-19.15%)] were higher than those in the pasteurized group (all P>0.05). The short-time pasteurized group was noted only for a higher percentage of the viable monocytes to viable leukocytes than the Holder pasteurized group (all P<0.05). Conclusions:Compared with the Holder pasteurization, sIgA, LTF, LZM level, and monocyte activity in breast milk can be better preserved by short-time pasteurization.

Evaluation system for promoting learning based on the phased examination of medical practitioners and its effect evaluation / 中华医学教育探索杂志

We strive to explore a student-centered evaluation system focused on improving students' learning efficiency and learning effect. Through classroom questioning, discussion of difficult cases, new progress report, question bank test, writing of clerkship notes, simulated inquiry between students with bedside inquiry, physical examination between students with bedside physical examination, operation training of basic clinical skills, direct observation and evaluation of clinical skills, mini-clinical exercise evaluation, graduation examination and other measures, an evaluation system for promoting learning by class evaluation, clerkship evaluation and practice evaluation has been established to cultivate the connotation construction of medical moral quality and clinical practice ability of medical students, and further promote the mastery of basic theoretical knowledge and clinical operation skills. By comparing the results of the phased examination for medical practitioners, the passing rate of 2018 is significantly higher than that of 2017, with 23.31% increased passing rate of theoretical results, and 0.79% increased passing rate of skills operation results. It can be seen that the evaluation system for promoting learning has played a guiding role in the teaching practice of clinical courses in our hospital.

Neutrophil phenotypes in peripheral blood of neonates with late-onset sepsis / 中华围产医学杂志

Objective To investigate the changes in neutrophil immunophenotypes in neonates with late-onset sepsis and their clinical significance.Methods A total of 42 neonates with late-onset sepsis were enrolled prospectively as sepsis group from Children's Hospital of Fudan University from January 2015 to October 2016,which included 26 preterm infants and 16 term infants.Another 33 neonates without infectious diseases consisting of 20 preterm infants and 13 term infants were selected as control group.According to the severity of sepsis,neonates in the sepsis group were further divided into severe (n=11)and mild sepsis (n=31) subgroups.Expression of CD16 and CD62L on neutrophils was measured by flow cytometry to determine the distribution of neutrophil subsets in neonatal peripheral blood.Differences in the distribution of neutrophil subsets between the two groups and two subgroups were compared using Wilcoxon rank-sum test.Multivariate logistic regression analysis was used to investigate the relationship between neutrophil subsets and the severity of neonatal sepsis.Receiver operating characteristic (ROC) curves were used to assess the diagnostic value of neutrophil subsets for the severity of sepsis.Results Four neutrophil subsets were identified in neonatal peripheral blood,including early-stage neutrophils (CD16-/CD62L-),immature neutrophils (CD16-/CD62L+),mature neutrophils (CD16+/CD62L+) and activated neutrophils (CD16+/CD62L-).Activated [preterm:61.1％ (20.2％-79.4％),term:47.6％ (15.2％-70.1％)] and mature neutrophils [preterm:35.7％ (19.9％-75.8％),term:52.0％ (25.6％-82.8％)] were the dominant subsets in the control group.In preterm infants,the proportion of early-stage [3.5％ (1.7％-9.4％) vs 1.9％ (0.6％-4.0％),Z=-2.501,P=0.012] and immature neutrophils [6.3％ (0.7％-45.5％) vs 0.4％ (0.3％-0.7％),Z=-3.878,P＜0.001] were higher,but that of activated neutrophils [8.3％ (2.3％-49.2％) vs 61.1％ (20.2％-79.4％),Z=2.991,P=0.002] were lower in the sepsis group than those in the control group;same differences were found in the absolute counts of each neutrophil subsets.Among term infants,more immature neutrophils were found in the sepsis group than those in the control group [49 (18-200) vs 13 (5-36)/μl,Z=-2.193,P=0.028].The proportion and the absolute counts of early-stage and immature neutrophils in the severe sepsis subgroup were all higher than those in the mild cases [early-stage neutrophils:5.8％ (3.4％-17.8％) vs 3.0％ (1.4％-7.3％),304 (137-1478) vs 158 (53-321)/μl;immature neutrophils:23.0％ (6.3％-47.0％) vs 0.9％ (0.5％-6.8％),1003 (487-2818) vs 85 (18-275)/μl;all P＜0.05].Multivariate logistic regression analysis showed that the proportion of early-stage neutrophils was associated with the severity of sepsis (OR=1.2,95％CI:1.0-1.4,P=0.012).In addition,the diagnostic value of the proportion of early-stage neutrophils for severe sepsis was the highest when the cut-off value was 3.3％,with the area under the ROC curve was 0.7 (95％CI:0.6-0.9),sensitivity of 81.8％ (95％CI:48.2％-97.7％) and specificity of 62.3％ (95％CI:42.2％-78.2％).Conclusions There are four neutrophil subsets in the peripheral blood of neonates and autoactivation of neutrophils may exist.With the onset of sepsis,neutrophil subsets react differently between preterm and term infants.The proportion of early-stage neutrophils may be correlated with the severity of neonatal sepsis,which may have a predictive value for severe sepsis.

Retrospective analysis of 76 neonates with invasive fungal infection in 2004-2014 / 中华围产医学杂志

Objective To investigate the clinical features of neonatal invasive fungal infection(IFI) so as to guide diagnosis,prevention and treatment of IFI.Methods Seventy-six neonates with IFI admitted to the Neonatal Intensive Care Unit (NICU) at Children's Hospital of Fudan University from 2004 to 2014 were included in the study.Pathogens,clinical manifestation,risk factor exposure,laboratory findings,complications,and clinical outcome of neonatal IFI were analyzed.Results Seventy-six cases were diagnosed as IFI between 2004 and 2014,with an yearly increasing trend.Sixty-eight patients were premature infants (89.5％).Of the 76 cases,except one with unknown birth weight,11(14.7％),34(45.3％),20(26.7％)and 10 (13.3％) cases had birth weight ＜ 1 000 g,(≥ 1 000-＜1 500) g,(≥ 1 500-＜2 500) g and ≥ 2 500 g,respectively.The pathogens were mainly Candida (74/76,97.4％),including 26 cases of Candida albicans (34.2％).However,the incidence of non-Candida albicans infection was increasing.Candida guilliermondii was the most common in nonCandida albicans,accounting for 29.2％ (14/48).All Candida albicans were sensitive to fluconazole.One strain of Candida glabrata was resistant to fluconazole.The most common risk factors included use of broad-spectrum antibiotics(93.3％,56/60),parenteral nutrition(70.0％,42/60),central vein catheterization(53.3％,32/60),invasive ventilation(40.0％,24/60) and history of abdominal surgery(21.7％,13/60).Clinical manifestations of IFI included temperature instability,frequent apnea,increased requirement of respiratory support and feeding intolerance.Among all cases,six were diagnosed as central nervous system infection.Of the patients who received cranial MRI,46.8％(22/47) showed multiple abnormal signals in cerebral parenchyma.Fiftytwo patients were cured and seven patients died before discharge,including one death due to fungal infection.Conclusions There is an increasing trend of IFI cases in NICU,especially in premature infants.Non-Candida albicans has become the main pathogenic fungus.There are no specific clinical manifestations in neonatal IFI.Use of broad-spectrum antibiotics,parenteral nutrition and central venous catheterization are common risk factors,and preventive measures should be taken in high-risk infants.In addition,IFI in neonates may affect important organs such as central nervous system,thus early treatment is necessary in suspected patients.

"Research on the training strategy of clinical practice ability of medical students based on ""three orientation""" / 中华医学教育探索杂志

Based on the particularity of the following medical disciplines, Three orientation is the higher medical personnel training mode which orients in the future development of medical students, and makes scientific allocation and combination to medicine curriculum system , personnel training system , management mode and evaluation system and so on. The clinical practice ability of medical students is the core of the professional quality training in the three orientation. In this article, the ability of professional quality, clinical practice ability and the ability to operate are clinical practice ability training goals and the training methods and evaluation system are divided into 2 stages, the fourth year and the fifth year, as well as three aspects such as professional quality ability training, clinical practice ability training and skills training.

Therapeutic efficacy of three bispecific antibodies on rheumatoid arthritis mice models / 药学学报

In order to obtain the lead compound for treatment of rheumatoid arthritis (RA), in this study, therapeutic efficacy of three bispecific antibodies (BsAB-1, BsAB-2 and BsAB-3) against both hIL-1beta and hIL-17 were compared on CIA model mice. First, by ELISA method we compared the binding capacity of the three bispecific antibodies to the two antigens. The results showed that all three antibodies could simultaneously bind both antigens, among these antibodies, BsAB-1 was superior over BsAB-2 and BsAB-3. CIA model was established with chicken type II collagen (CII) and developed RA-like symptoms such as ankle swelling, skin tight, hind foot skin hyperemia. The CIA mice were treated with three antibodies once every two days for total of 29 days. Compared with the CIA model mice, the RA-like symptoms of the antibody treated-mice significantly relieved, while the BsAB-1 treated-mice were almost recovered. CII antibody level in the serum and cytokines (IL-2, IL-1beta, IL-17A and TNF-alpha) expression in the spleen were examined. Compared with the CIA model mice, all three antibodies could significantly reduce CII antibody and cytokine expression levels. BsAB-1 antibody was more potent than BsAB-2 and BsAB-3. In summary, BsAB-1 is superior over BsAB-2 and BsAB-3 in amelioration of RA symptoms and regulation of CII antibody production and pro-inflammatory cytokine expression, therefore, BsAB-1 can be chosen as a lead compound for further development of drug candidate for treatment of RA.

Peripheral blood monocyte subpopulations in patients with syphilis / 中华皮肤科杂志

Objective To analyze the changes in peripheral blood monocyte subpopulations in patients with primary, secondary and latent syphilis. Methods Flow cytometry was used to detect CD14highCD16- and CD14+CD16+ monocyte subpopulations in peripheral blood from 58 patients with untreated syphilis, including 36 cases of latent syphilis,8 cases of primary syphilis and 14 cases of secondary syphilis, as well as from 65 normal human controls. Restflts Compared with the normal controls, the proportion of CD14+CD16+ monocytes among total monocytes was significantly elevated (12.0% ± 5.0% vs 6.0% ± 3.3%, t = 7.25, P < 0.01), while that of CD14highCD16- monocytes was down-regulated (88.0% ± 5.1% vs 94.0% ± 3.5%, t = -7.20, P < 0.01). No statistical difference was observed in the proportion of CD14+CD16+ or CD14hhighCD16- monocytes among the patients with primary syphilis, secondary syphilis and those with latent syphilis (all P > 0.05). Conclusions The changes in peripheral blood monocyte subpopulation in patients with untreated syphilis may be associated with the permanent infection of Treponema pallidum, but have no obvious correlation with clinical stage of syphilis.

Association betwen Mannose-binding lectin structural gene polymorphisms and susceptibility of SLE in Han Nationality population from Hubei province of China / 中国免疫学杂志

0.05).Conclusion:MBL B allele is not a risk component in the developing process of SLE Chinese patients.