Objective:To study the clinical characteristics and risk factors of necrotizing enterocolitis (NEC) in one of the premature twins.Methods:A retrospective study was conducted on twin premature infants who were admitted to the Department of Neonatology at the Third Affiliated Hospital of Zhengzhou University from January 2017 to December 2022 and only one got NEC. The twins were divided into NEC group and control group, the clinical data were collected and analyzed by SPSS 26.0 statistical software.Results:This study enrolled 109 pairs of premature twins, 109 cases in the NEC group, and 109 cases in the control group. Univariate analysis showed that birth weight, pre NEC white blood cell count were lower in NEC group than those in the control group, while the proportion of smaller than gestational age (SGA), donor of twin-to-twin transfusion syndrome, feeding intolerance, incomplete enteral feeding, start feeding time >48 h, red blood cell transfusion 72 h before NEC onset and the neutrophils ratio were higher in the NEC group than that of the control group, the difference was statistically significant ( P<0.05). Multivariate logistic analysis showed that low birth weight ( OR=1.558, 95% CI1.197-2.142), SGA ( OR=1.721, 95% CI 1.217-2.536), feeding intolerance ( OR=3.798, 95% CI 1.347-10.706), and incomplete enteral feeding ( OR=4.319, 95% CI 1.673-11.149) were independent risk factors for NEC ( P<0.05). Conclusions:Low birth weight, small for gestational age, feeding intolerance, and incomplete enteral feeding are independent risk factors for NEC in one of the premature twins.

Objective:To describe the current status and trends in the outcomes and care practices of extremely preterm infants at 22-25 weeks′ gestation age from the Chinese Neonatal Network (CHNN) from 2019 to 2021.Methods:This cross-sectional study used data from the CHNN cohort of very preterm infants. All 963 extremely preterm infants with gestational age between 22-25 weeks who were admitted to neonatal intensive care units (NICU) of the CHNN from 2019 to 2021 were included. Infants admitted after 24 hours of life or transferred to non-CHNN hospitals were excluded. Perinatal care practices, survival rates, incidences of major morbidities, and NICU treatments were described according to different gestational age groups and admission years. Comparison among gestational age groups was conducted using χ2 and Kruskal-Wallis tests. Trends by year were evaluated by Cochran-Armitage and Jonckheere-Terpstra tests for trend. Results:Of the 963 extremely preterm infants enrolled, 588 extremely preterm infants (61.1%) were male. The gestational age was 25.0 (24.4, 25.6) weeks, with 29 extremely preterm infants (3.0%), 88 extremely preterm infants (9.1%), 264 extremely preterm infants (27.4%), and 582 extremely preterm infants (60.4%) at 22, 23, 24, and 25 weeks of gestation age, respectively. The birth weight was 770 (680, 840) g. From 2019 to 2021, the number of extremely preterm infants increased each year (285, 312, and 366 extremely preterm infants, respectively). Antenatal steroids and magnesium sulfate were administered to 67.7% (615/908) and 51.1% (453/886) mothers of extremely preterm infants. In the delivery room, 20.8% (200/963) and 69.5% (669/963) extremely preterm infants received noninvasive positive end-expiratory pressure support and endotracheal intubation. Delayed cord clamping and cord milking were performed in 19.0% (149/784) and 30.4% (241/794) extremely preterm infants. From 2019 to 2021, there were significant increases in the usage of antenatal steroids, antenatal magnesium sulfate, and delivery room noninvasive positive-end expiratory pressure support (all P<0.05). Overall, 349 extremely preterm infants (36.2%) did not receive complete care, 392 extremely preterm infants (40.7%) received complete care and survived to discharge, and 222 extremely preterm infants (23.1%) received complete care but died in hospital. The survival rates for extremely preterm infants at 22, 23, 24 and 25 weeks of gestation age were 10.3% (3/29), 23.9% (21/88), 33.0% (87/264) and 48.3% (281/582), respectively. From 2019 to 2021, there were no statistically significant trends in complete care, survival, and mortality rates (all P>0.05). Only 11.5% (45/392) extremely preterm infants survived without major morbidities. Moderate to severe bronchopulmonary dysplasia (67.3% (264/392)) and severe retinopathy of prematurity (61.5% (241/392)) were the most common morbidities among survivors. The incidences of severe intraventricular hemorrhage or periventricular leukomalacia, necrotizing enterocolitis, and sepsis were 15.3% (60/392), 5.9% (23/392) and 19.1% (75/392), respectively. Overall, 83.7% (328/392) survivors received invasive ventilation during hospitalization, with a duration of 22 (10, 42) days. The hospital stay for survivors was 97 (86, 116) days. Conclusions:With the increasing number of extremely preterm infants at 22-25 weeks′ gestation admitted to CHNN NICU, the survival rate remained low, especially the rate of survival without major morbidities. Further quality improvement initiatives are needed to facilitate the implementation of evidence-based care practices.

Objective This article aimed to explore the inhibitory effect of β-ionone(BI)on the proliferation of breast canc-er cells through the nuclear factor kappa-B(NF-κB)pathway and its possible mechanism.Methods The methylene blue assay and MTT assay were used to determine the viability of breast cancer cells.The malachite green phosphate assay was used to detect the ac-tivity of protein phosphatase 2A(PP2A).Western blot was used to detect the levels of phosphorylated P65(s534 and s311)(p-P65),PP2A(A,B and C),and phosphorylated ataxia telangiectasia mutant(p-ATM)(s1981)protein.Results BI could significant-ly inhibit the proliferation of human breast cancer BT549 cells and MCF-7 cells in a time-and dose-dependent manner,and the difference was statistically significant(P<0.01).After treated with BI,NF-κB activity was significantly inhibited in MCF-7 cells,as shown by a significant decrease in the level of phosphorylated P65(s311 and s534)protein and an increase in the level of PP2A pro-tein,and the difference was statistically significant(P<0.05).In addition,BI also significantly reduced the phosphorylation of P65 protein and ATM protein in MCF-7 cells by the PP2A inhibitor-okada acid(OA).Conclusion This study shows that BI inhibits the proliferation of breast cancer cells by inhibiting NF-κB activity,and its mechanism may be achieved by increasing PP2A activity to regulate the NF-κB pathway.

Objective:To analyze the risk factors of bronchopulmonary dysplasia(BPD)in very preterm infants(VPI), and to provide scientific basis for the prevention and treatment of BPD in VPI.Methods:A prospective multicenter study was designed to collect the clinical data of VPI in department of neonatology of 28 hospitals in 7 regions from September 2019 to December 2020.According to the continuous oxygen dependence at 28 days after birth, VPI were divided into non BPD group and BPD group, and the risk factors of BPD in VPI were analyzed.Results:A total of 2 514 cases of VPI including 1 364 cases without BPD and 1 150 cases with BPD were enrolled.The incidence of BPD was 45.7%.The smaller the gestational age and weight, the higher the incidence of BPD( P<0.001). Compared with non BPD group, the average birth age, weight and cesarean section rate in BPD group were lower, and the incidence of male infants, small for gestational age and 5-minute apgar score≤7 were higher( P<0.01). In BPD group, the incidences of neonatal respiratory distress syndrome(NRDS), hemodynamically significant patent ductus arteriosus, retinopathy of prematurity, feeding intolerance, extrauterine growth restriction, grade Ⅲ~Ⅳ intracranial hemorrhage, anemia, early-onset and late-onset sepsis, nosocomial infection, parenteral nutrition-associated cholestasis were higher( P<0.05), the use of pulmonary surfactant(PS), postnatal hormone exposure, anemia and blood transfusion were also higher, and the time of invasive and non-invasive mechanical ventilation, oxygen use and total hospital stay were longer( P<0.001). The time of starting enteral nutrition, cumulative fasting days, days of reaching total enteral nutrition, days of continuous parenteral nutrition, days of reaching 110 kcal/(kg·d) total calorie, days of reaching 110 kcal/(kg·d) oral calorie were longer and the breastfeeding rate was lower in BPD group than those in non BPD group( P<0.001). The cumulative doses of amino acid and fat emulsion during the first week of hospitalization were higher in BPD group( P<0.001). Multivariate Logistic regression analysis showed that NRDS, invasive mechanical ventilation, age of reaching total enteral nutrition, anemia and blood transfusion were the independent risk factors for BPD in VPI, and older gestational age was the protective factor for BPD. Conclusion:Strengthening perinatal management, avoiding premature delivery and severe NRDS, shortening the time of invasive mechanical ventilation, paying attention to enteral nutrition management, reaching whole intestinal feeding as soon as possible, and strictly mastering the indications of blood transfusion are very important to reduce the incidence of BPD in VPI.

OBJECTIVE： To establish a method for the determination of levofloxacin concentration in human pleural effusion， to study its pharmaceutical characteristics. METHODS： Totally 6 patients with infectious pleural effusion received levofloxacin 0.4 g qd intravenous drip. Pleural effusion was collected at 0.5， 1， 2， 4， 8， 12 and 24 hours after administration. After treated with methanol precipitation protein， HPLC was used to determine the concentration of levofloxacin. The determination was performed on Agilent ZORBAX SB-C18 column with mobile phase consisted of methanol-0.02 mmol/L KH2PO4 buffer （containing 0.3％ triethylamine， 70 ∶ 30， V/V at the flow rate of 1.0 mL/min. The detection wavelength was set at 294 nm， and the column temperature was 35 ℃. The sample size was 20 μL. The pharmacokinetic parameters were calculated with WinNonlin 5.2 software. RESULTS： Under this chromatogram condition， retention time of levofloxacin was about 4.9 min， the peak shape was good， the baseline was stable， and the determination of endogenous substances in pleural effusion had no interference. The linear range of levofloxacin were 0.625-20 μg/mL（R2＝0.998 9）. The relative recovery rates were （83.75±1.66）％-（87.73±2.43）％ for low， medium and high concentration samples （n＝3）； RSDs of intra-day were 2.23％-4.96％ （n＝5）； RSDs of inter-day were 4.10％-4.78％（n＝5）； the accuracy ranged （97.76±4.85）％-（100.87±2.25）％（n＝5）； RSD of concentration was no more than 5％ in stability test （n＝3） for low， medium and high quality control sample. The pharmacokinetic parameters of levofloxacin included cmax were （2.21±0.87） μg/mL； AUC0-24 h were （37.31±11.94） μg·h/mL； t1/2 were （4.50±0.21） h. CONCLUSIONS： Established method is simple， reliable and sensitive， and can be used for the determination of levofloxacin concentration in human pleural effusion and its pharmacokinetic study.

Objective To study the relationships of TGFβ1 (-509C/ T, +869T/ C) and TGFβR2 (-875 G/ A) single nucleotide polymorphisms (SNPs) with colorectal cancer (CRC) in Chinese Han popu-lation in Shandong. Methods TGFβ1 -509C/ T and +869T/ C SNPs in a total of 490 patients with CRC were detected using gene chip. TGFβR2 -875 SNPs was analyzed using PCR-RFLP. TGFβ1 concentrations in serum samples were measured by ELISA. Immunohistochemistry was used to detect the expression of TGFβR2. The relationships of TGFβ1 (-509C/ T, +869T/ C) and TGFβR2 (-875 G/ A) SNPs with CRC were analyzed through a case-control study. Chi-square test or t test was used for statistical analysis. Rela-tive risk was estimated by odds ratio (OR) and 95％ confidence interval (95％ CI). Results No signifi-cant difference in genotype or allele frequency at TGFβ1 -509 / +869 was found between patients with CRC and healthy subjects (P>0. 05). The frequencies of TGFβR2 -875GG genotype and -875G allele in pa-tients with CRC were significantly higher than those in healthy subjects (-875GG: χ2 = 4. 65, P = 0. 031, OR=1. 32, 95％ CI=1. 03-1. 71; -875G: χ2 =4. 95, P=0. 026, OR=1. 29, 95％ CI=1. 03-1. 61). Com-pare with the healthy control group, higher frequencies of TGFβR2 -875GG genotype and -875G allele were also detected in rectal cancer ( -875GG: P = 0. 04, OR = 1. 39, 95％ CI = 1. 02-1. 95 and -875G: P =0. 045, OR=1. 32, 95％ CI = 1. 01-1. 73), tubular adenocarcinoma ( -875GG: P = 0. 004, OR = 1. 51, 95％ CI=1. 14-2. 00 and -875G: P=0. 003, OR=1. 45, 95％ CI=1. 14-1. 85) and highly differentiated tu-bular adenocarcinoma (-875GG: P=0. 003, OR=1. 68, 95％ CI=1. 19-2. 38 and -875G: P=0. 002, OR=1. 62, 95％ CI=1. 18-2. 21) groups. The serum TGFβ1 levels in TGFβR2 -875G carriers with CRC were significantly higher than those in TGFβR2 -875AA carriers in both CRC (t= -3. 42, P<0. 05) and healthy control (t= -5. 09, P<0. 001) groups. TGFβR2 expression in -875G carriers with rectal cancer was signifi-cantly lower than that in -875AA carriers with rectal cancer (P=0. 047) and healthy subjects (P=0. 027).Conclusion TGFβR2 -875GG might be a potential risk factor for CRC in Chinese Han population in Shandong and TGFβR2 - 875G might be a risk factor for rectal cancer and highly differentiated tubular adenocarcinoma.

Objective To investigate the protective effect of caffeine citrate on white matter damage in 2-day-old neonatal rats induced by postnatal infection.Methods Forty-eight 2-day-old SD rats were randomly divided into the control group (group A,n =16),lipopolysaccharide(LPS) infection group (group B,n =16),and caffeine citrate intervention group (group C,n =16) according to the random table method.The newborn rats of group B and C were continuously injected LPS 0.6 mg/kg intraperitoneally for 5 days from 2 days old,and the newborn rats of group A were continuously injected by an equal volume of 9 g/L saline intraperitoneally.Group C was continuously injected by caffeine citrate 10 mg/kg intraperitoneally for 7 days from 4 days old;equal volume of 9 g/L saline was injected into group A and B for 7 days continuously.At 12 days old,8 rats of each group were sacrificed randomly to evaluate the expression of myelin basic protein(MBP) subcortical white matter by immunohistochemical method.Both sides of hippocampus of the rest 8 rats of each group were taken out in ice surface rapidly.The left hippocampus was used to detect the expression of MBP and Caspase-3 by Western blot method,and the right hippocampus was used to evaluate the MBP and Caspase-3 protein level by quantitative real-time PCR(qPCR) method.Results The mean integral optical density (IOD) of subcortical MBP positive expression in group A,group B and group C were 132.64 ± 1.94,102.43 ± 2.12,114.25 ± 2.04,and the difference was statistically significant among 3 groups (F =22.912,P ＜ 0.05).The relative expression levels of MBP mRNA of 3 groups in hippocampus were 0.79 ± 0.01,0.39 ± 0.03,0.55 ± 0.02,and the difference was statistically significant among 3 groups (F =18.584,P ＜ 0.05).The relative expression levels of MBP protein of 3 groups in hippocampus were 0.64 ± 0.03,0.31 ± 0.03,0.51 ± 0.05,and the difference was statistically significant among 3 groups (F =25.780,P ＜ 0.05).The relative expression levels of Caspase-3 mRNA of 3 groups in hippocampus were 0.34 ± 0.02,0.74 ± 0.03,0.57 ± 0.04,and the difference was statistically significant among 3 groups (F =6.105,P ＜ 0.05).The relative expression of Caspase-3 protein of 3 groups in hippocampus were 0.11 ± 0.03,0.36 ± 0.02,0.23 ± 0.03,and the difference was statistically significant among 3 groups (F =40.541,P ＜ 0.05).Conclusions Caffeine citrate has showed protective effect on white matter damage in neonatal rats of 2 days old induced by postnatal infection.The mechanism may be related to inhibition of inflammatory response and reducing apoptosis.

Objective To study the correlations between genetic polymorphisms of TNF-α as well as IL-6 and susceptibility to colorectal cancer among Chinese Han people in Shandong province.Methods Single nucleotide polymorphisms (SNPs) of TNF-α-238G/A,-308G/A and IL-6-174G/C,-572G/C,-597G/A in 490 patients with colorectal cancer were analyzed by using gene chip.Concentrations of TNF-α and IL-6 in serum samples were measured by ELISA.A case-control study was conducted to analyze the correlations between SNPs of TNF-α-238G/A,-308G/A as well as IL-6-174G/C,-572G/C,-597G/A and susceptibility to colorectal cancer.Chi-square test or t test was used for statistical analysis.Relative risks were estimated based on the values of odds ratio (OR) and 95% confidence interval (95%CI).Results The frequency of TNF-α-308AA in patients with colorectal cancer was significantly higher than that in healthy subjects (x2 =6.15, P<0.05, OR=2.08, 95%CI=1.17-3.71), while the frequency of IL-6-572CC in patients with colorectal cancer was significantly lower than that in healthy subjects (x2 =4.97, P<0.05, OR=0.73, 95%CI=0.55-0.96).The frequency of TNF-α-308AA in patients with colon cancer (OR=2.31, 95%CI=1.17-4.55), tubular adenocarcinoma (OR=2.32, 95%CI=1.28-4.21), high (OR=2.05, 95%CI=1.01-4.15) or moderately differentiated adenocarcinoma (OR=5.88, 95%CI=1.79-19.30) was significantly higher than that in healthy subjects.The levels of serum TNF-α in TNF-α-308AA carriers with colorectal cancer were significantly higher than those in TNF-α-308G carriers with colorectal cancer (t=2.13, P<0.05) as well as those in healthy TNF-α-308AA carriers (t=2.13, P<0.05).The levels of serum IL-6 in colorectal cancer group were significantly higher than those in control group (t=6.74, P<0.001).Conclusion The SNPs of TNF-α-308 and IL-6-572 are associated with the occurrence and development of colorectal cancer in Chinese Han people in Shandong province.

Objective To compare the effect of treating cricopharyngeal achalasia in stroke survivors using transnasal or transoral balloon dilatation.Methods Thirty stroke survivors with cricopharyngeal achalasia were randomly divided into a transnasal and a transoral balloon dilatation group (group N and group O),each of 15.Both groups were given routine swallowing rehabilitation training as well as the transnasal or transoral balloon dilatation.Their heart rate was monitored during the dilatation.Nasal bleeding,mucous membrane swelling and pain were also observed.Their swallowing function was evaluated using the Fujishima Ichiro swallowing efficacy score (FISE) and videofluoroscopy (VFSS) before and after the intervention.Results After the treatment,the average FISE and VFSS scores of both groups had improved significantly comnpared to before the treatment but there were no significant differences between the groups.During the treatment,the average heart rate of group O increased significantly less than that of group N.The treatment acceptance of group O was 98.2％,significantly higher than that of group N (80.1％).One case of mucosal bleeding was observed in group O,and laryngeal edema occurred significantly less often than in group N (9 cases vs.7).The average pain score was also significantly lower in group O.Conclusions Balloon dilatation facilitates swallowing among stroke survivors with cricopharyngeal achalasia.The transoral approach can help to reduce the occurrence of complications such as mucosal bleeding,laryngeal edema and pain,and has better patient acceptance.

Eight newborn screening quality indicators were developed though expert discussing organized by National Center for Clinical Laboratories according to NBS quality management in USA .These indicators can not only be used to evaluate the performance throughout critical aspects of pre -examination, examination and post-examination processes for routine tests , but also be applied to monitor a series of processes from specimen collecting to results interpretation in newborn screening .Meanwhile, NCCL can use these quality indicators to carry out External Quality Assessment programs in order that the total service quality of newborn screening can be improved.