Objective To construct expression vectors of human ribosomal protein S 3(RPS3) and RPS3Ser209 mutant in orcler to investigate the effect of RPS3Ser209 mutant on NF-κB signaling pathway and DNA binding capacity .Methods The vector RPS3-myc was amplified by polymerase chain reaction ( PCR) from the human liver cDNA and subcloned into pcDNA-3.1myc-HisB.RPS3S209A represented mutant RPS3 expression vectors, in which the designated amino acid was mutated to an alanine residue .Dual luciferase reporter gene assay was used to detect the NF-κB transcription activity in HEK293 cells,immunofluorescence to detect RPS3 location, and EMSA to examine NF-κB DNA-binding activity.Results The expression vectors of RPS3-myc and RPS3S209A-myc were constructed.Compared with wild-type RPS3,the nucleus translocation, transactivation activity of NF-κB and DNA binding ability of RPS3S290A were reduced significantly .Conclu-sion The impact of RPS3 on NF-κB signaling pathway depend on its serine 209.

Objective Acute liver failure is one of the significant causes of death clinically.It is important to explore new serum markers of liver injury for early diagnosis and prognosis prediction of severe liver disease.Hepassocin ( HPS) is a liver-specific mitogenic growth factor.Our study is intended to investigate the correlation between HPS serum levels and the degree of liver injury.Methods Firstly, a mouse model of acute liver injury was constructed via intraperitoneally injection with different doses of CCl4 .Then the survival rate, alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST) and the pathological changes in the liver were detected.Meanwhile, ELISA assay was performed to detect the serum level of HPS.In addition, the mRNA level and protein level of HPS were detected by real-time PCR and Western blotting, respectively.Results The mortality rate was increased and the liver damage was aggravated with the increase of the CCl4 dose.Besides, the ALT and AST levels were also increased in a dose-dependent manner.Additionally, the mRNA and protein levels of HPS were significantly up-regulated and closely related to the degree of the liver injury in the model. Conclusion HPS can be used as a new marker of liver injury in mice.

Objective To investigate the regulation of T cells in the process of liver regeneration using a model of mice after 70% liver resection.Methods We performed 70% hepatectomy in T-cell-deficient mice and control mice.The liver mass and body mass ratio, BrdU infiltration level, proliferating cell nuclear antigen (PCNA),expression of M phase marker protein p-HDAC3, and serum transaminase levels were measured.Results The recovery of liver mass and body mass ratio of thymus-deficient mice occurred significantly later than that of control mice.The peak time of BrdU infiltration levels and the expression of PCNA and p-HDAC3 in T-cell-deficient mice were later than in control mice, but the degree of liver injury was lower.Conclusion T cells are involved in the regulation of liver regeneration, and the absence of T cells delays the process of liver regeneration.

Objective To measure the placing position error of different body position by analog positioning machine in patients with different body mass index (BMI) during radiotherapy for rectal cancer, then calculate the clinical target volume (CTV) to plan target volume (PTV) margins (Mptv). Methods Thirty-six patients with rectal cancer were selected, and the patients were divided into vacuum bag group (18 cases) and routine belly board group (18 cases) according to method of placing position. The BMI was calculated. Each patient was treated with positive and lateral position X-ray film before treatment and once a week during the radiotherapy. The data were contrasted with digitally rendered radiographs (DRR) of three-dimensional conformal radiotherapy plan system, then the placing position error on X, Y and Z axis was calculated, and the Mptv on X, Y and Z axis of patients with different BMI was calculated. Results The placing position error on X, Y and Z axis gradually increased with the increase of BMI in vacuum bag group and routine belly board group, and there were statistical differences (P<0.01 or <0.05). The Mptv on X, Y and Z axis in patients with normal BMI were 4.60, 5.51 and 5.29 mm respectively. The Mptv on X, Y and Z axis in patients with overweight were 5.48, 6.81 and 6.16 mm respectively. The Mptv on X, Y and Z axis in patients with obesity were 8.92, 8.59 and 7.02 mm respectively. Conclusions The rectal cancer patients with overweight and obesity have more placing position error. The influence of BMI should be considered when the patient's Mptv is determined.

OBJECTIVETo study the effect of E3 ubiquitin ligase RNF31 knockdown on nuclear factor-κB (NF-κB) pathway activation and cell apoptosis.

METHODSHuman RNF31 siRNA sequences were cloned into the lentiviral vector pGreenPuro and transiently transfected in HEK293T cells to screen the most effective fragments, which were co-transfected along with the packaging plasmids PMD and SPA in 293T cells. The cell supernatant was collected at 24 h and 48 h after the transfection and the viral titers were determined with flow cytometry. Real-time PCR and Western blotting were used to evaluate the effect of RNF31 knockdown on the expression of NF-κB downstream target genes and IκBα activity; the changes of NF-κB pathway transcriptional activity were assessed with dual luciferase reporter gene. Hochest dying was used to examine the influence of RNF31 down-regulation on cell apoptosis.

RESULTSRNF31 knockdown mediated by the lentiviral vector pGreenPuro-RNF31 suppressed the transcriptional activity of NF-κB and the downstream target genes in HEK293 cells stimulated with TNF-α. RNF31 knockdown also resulted in suppression of NF-κB-stimulated expression of pIκBα and in increased apoptosis of cells stimulated with TNF-α for 24 h.

CONCLUSIONRNF31 down-regulation inhibits NF-κB pathway activation induced by TNF-α.

Apoptosis ; Down-Regulation ; Gene Expression Regulation ; Genetic Vectors ; HEK293 Cells ; Humans ; I-kappa B Proteins ; metabolism ; Lentivirus ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Transfection ; Tumor Necrosis Factor-alpha ; pharmacology ; Ubiquitin-Protein Ligases ; genetics ; metabolism

Objective:To establish a risk prediction model for neonatal asphyxia in cesarean section and test its application effect.Methods:This was a retrospective study. We retrospectively analyzed the clinical data of 2 244 infants (modeling group) who were delivered by cesarean section in Affiliated Hospital of Weifang Medical University from April 2021 to December 2021. Newborns were divided into asphyxia group ( n=176) and non-asphyxia group ( n=2 068) according to the occurrence of neonatal asphyxia. Logistic regression was used to screen the risk factors of neonatal asphyxia in cesarean section and a line chart model was established to predict the risk. Another 683 neonates were selected as validation group for external validation of the model from January to March in 2022. Results:Five factors including preterm birth, fetal distress, fetal growth restriction, abnormal S/D value of umbilical artery and umbilical cord around the neck were included in the prediction model. The area under ROC curve of the modeling group was 0.902, the Youden index was 0.687, the sensitivity was 0.837, and the specificity was 0.850. Hosmer-lemeshow test showed that χ2=1.79, and P=0.877. In the validation group, the area under ROC curve was 0.823, the Youden index was 0.555, the sensitivity was 0.835, and the specificity was 0.720. It showed that the model had a good fitting effect and identification validity. Conclusions:The risk prediction model has a good clinical application value in the prediction of neonatal asphyxia in cesarean section, and provides reference for obstetricians to take preventive management measures of neonatal asphyxia in time.

Objective:To systematically review qualitative research on maternal participation in skin-to-skin contact experience during cesarean delivery, the promotion of skin-to-skin contact in cesarean section.Methods:We searched databases including the PubMed, Embase, Web of Science, Cochrane Library, Ebsco, SinoMed, Wan Fang, CNKI, VIP. All of the qualitative research on the real experience of skin-to-skin contact from the participation of caesarean section women were collected. The search time limit was from the establishment of the databases to May 2022. The JBI Critical Appraisal Tool for qualitative studies from Australia was used to evaluate the methodology quality of the included research, and the meta-aggregation was used to conduct the synthesis.Results:Totally 11 qualified studies were included and integrated into 49 valuable findings. Similar results were summarized into 12 groups, and 4 integrated results were synthesized, including the needs of cesarean section mothers; positive experience after skin-to-skin contact in caesarean section; skin-to-skin contact practice strengthen the role of mothers and promotes breastfeeding; difficulties and challenges of skin-to-skin contact during cesarean section.Conclusions:Skin-to-skin contact positively affects mother and infant delivered by cesarean section. Medical staff need to pay attention to the feelings and needs of cesarean section women participating in skin-to-skin contact, optimize information, environment, and humanistic support, and improve skin-to-skin contact practices. Medical institutions should rationally allocate obstetric resources to alleviate the difficulties faced by skin-to-skin contact practices.