Abstract OBJECTIVE To investigate the specific binding of nucleic acid aptamers(k-α2ct) with plasminogen Kringle 5(K5) on the function of K5 in inhibiting proliferation and migration of vascular endothelial cells and promoting their apoptosis. METHODS The cloning and expression of recombinant K5 protein were performed by using a prokaryotic system, and the isolation and purification of the expressed K5 protein were performed by affinity chromatography. The affinity and specificity of K-a2ct and K5 were verified using isothermal titration calorimetry(ITC) and enzyme-linked oligonucleotide adsorption assay(ELONA). The effect of K-a2ct on the function of K5 in inhibiting the proliferation and migration of human umbilical vein endothelial cells(HUVEC) was investigated by CCK-8 and cell scratch assay. The apoptotic morphology of HUVEC cells stained with Hoechst 33342 was observed by laser confocal microscopy, and the effect of K-a2ct on the apoptosis- promoting function of K5 in HUVEC cells was also examined by Annexin V/PI double-stained flow cytometry. RESULTS Recombinant protein K5 was efficiently expressed in Escherichia coli and purified by affinity chromatography, identified as having a relative molecular weight of 12 kDa and a concentration of 0.32 mg·mL-1. ITC and ELONA results demonstrated that K-a2ct had a strong affinity and good selectivity for K5, showing the affinity specificity of a typical nucleic acid aptamer. CCK-8 and cell scratching assays showed that K-a2ct could inhibit the anti-proliferation and anti-migration effects of K5 on HUVEC cells in a dose-dependent manner. The laser confocal and flow cytometry results showed that K-a2ct inhibited the apoptosis-promoting function of K5 on HUVEC cells, mainly affecting late apoptosis of HUVEC cells effected by K5 but having little effect on early apoptosis. CONCLUSION The nucleic acid aptamers K-a2ct binds to K5 with high affinity and specificity, and inhibits its anti angiogenic function in a dose-dependent manner. It has great potential in targeting the regulation of K5 concentration and function in vivo and promoting angiogenesis.

OBJECTIVE: To develop a cell-based system for the diagnosis of vitamin K-dependent coagulation factor deficiency 1 (VKCFD1). METHODS: In HEK293 cells stably expressing the reporter gene FIX-Gla-PC, the gamma-glutamyl carboxylase (GGCX) gene was knocked out by using CRISPR/Cas9 technology. Enzyme-linked immunosorbent assay (ELISA), DNA sequencing and Western blotting were used to identify the GGCX gene knockout cells. A quickchange point variant method was used to construct the GGCX variant. ELISA was used to assess the influence of GGCX variant on the activity of reporter gene. RESULTS: Two monoclonal cell lines with no reporter activity by ELISA was identified. Edition and knockout of the GGCX gene was confirmed by DNA sequencing and Western blotting. The activity of the reporter gene was recovered by transfection of the wild-type GGCX gene. Thereby two monoclonal cells with GGCX knockout were obtained. By comparing the wild-type and pathogenic GGCX variants, the reporter activity was decreased in the pathogenic variants significantly. CONCLUSION A cell-based system for the detection of GGCX activity was successfully developed, which can be used for the diagnosis of VKCFD1 caused by GGCX variants.

Cleft lip and palate is a common congenital malformation, and its sequential treatment time is long and complicated. Many factors affect the treatment result . Operation is the main treatment for cleft palate. Its main goal is to close the abnormal defect to prevent disorders of speech development and eating, while ensuring the minimum impact on the growth of the maxilla. Cleft lip repair has been carried out for many years at home and abroad by using vomer bone flap to repair the hard palate at the same time, and a large number of research reports have been reported. This article reviews the effect of the treatment mode on the related anatomical structures of cleft lip repair with vomer bone flap repairing the hard palate, in order to explore the ideal treatment plan for cleft lip and palate.

Cleft lip and palate is a common congenital malformation, and its sequential treatment time is long and complicated. Many factors affect the treatment result . Operation is the main treatment for cleft palate. Its main goal is to close the abnormal defect to prevent disorders of speech development and eating, while ensuring the minimum impact on the growth of the maxilla. Cleft lip repair has been carried out for many years at home and abroad by using vomer bone flap to repair the hard palate at the same time, and a large number of research reports have been reported. This article reviews the effect of the treatment mode on the related anatomical structures of cleft lip repair with vomer bone flap repairing the hard palate, in order to explore the ideal treatment plan for cleft lip and palate.

Cleft lip and palate is a common congenital malformation, and its sequential treatment time is long and complicated. Many factors affect the treatment result . Operation is the main treatment for cleft palate. Its main goal is to close the abnormal defect to prevent disorders of speech development and eating, while ensuring the minimum impact on the growth of the maxilla. Cleft lip repair has been carried out for many years at home and abroad by using vomer bone flap to repair the hard palate at the same time, and a large number of research reports have been reported. This article reviews the effect of the treatment mode on the related anatomical structures of cleft lip repair with vomer bone flap repairing the hard palate, in order to explore the ideal treatment plan for cleft lip and palate.

Cleft lip and palate is a common congenital malformation, and its sequential treatment time is long and complicated. Many factors affect the treatment result . Operation is the main treatment for cleft palate. Its main goal is to close the abnormal defect to prevent disorders of speech development and eating, while ensuring the minimum impact on the growth of the maxilla. Cleft lip repair has been carried out for many years at home and abroad by using vomer bone flap to repair the hard palate at the same time, and a large number of research reports have been reported. This article reviews the effect of the treatment mode on the related anatomical structures of cleft lip repair with vomer bone flap repairing the hard palate, in order to explore the ideal treatment plan for cleft lip and palate.

Objective:To construct and validate a prediction model for the risk of intermediate cesarean delivery for primiparous women with failed vaginal trial of labor.Methods:Clinical data of 6 128 pregnant women who gave birth in the Affiliated Hospital of Jining Medical College from January 2019 to December 2020 were collected. The puerpera was randomly divided into train set ( n=4 290) and validation set ( n=1 838). The factors influencing the conversion to cesarean section in primiparous women with failed vaginal trial of labor were analyzed with univariate and binary multivariate logistic regression, and a risk prediction model was established based on the influencing factors. The predictive power of the model was assessed by receiver operating characteristic (ROC) curve and the Hosmer-Lemeshow goodness-of-fit test in train set and validation set. Results:Among 6 128 pregnant women 1 042 cases failed in vaginal trial of labor and were transferred to cesarean section. Univariate analysis showed age, occupation, gestational weight gain, days of gestation, body temperature before delivery, fetal heart condition at delivery, fetal abdominal circumference, Bishop score, premature rupture of membranes, gestational illness, mode of induction of labor, labor analgesia, and fetal orientation were significantly associated with converting to cesarean delivery (all P<0.05). The multivariate binary logistic regression analysis showed that the age, gestational weight gain, body temperature, gestational co-morbidities, days of gestation, premature rupture of membranes, amniotic fluid contamination, induction of labor, and abnormal occipital position were independent risk factors for intermediate cesarean delivery ( OR=1.03-8.06, all P<0.05); while height, occupation, Bishop score, and labor analgesia were protective factors for intermediate cesarean delivery ( OR=0.17-0.96, all P<0.05). A risk prediction model was constructed based on the risk factors and protective factors. In train set, the area under the ROC curve(AUC) of the model was 0.902 (95% CI: 0.89-0.92, P<0.001), with the best cutoff value of 0.138, the sensitivity and specificity were 0.837 and 0.825, respectively; and the Hosmer-Lemeshow goodness-of-fit test showed P=0.192. In validation set the AUC of the model was 0.917 (95% CI: 0.90-0.93, P<0.001), and the sensitivity and specificity were 0.826 and 0.851, respectively; the total correct rate of the model was 87.21% (1 603/1 838). Conclusion:The risk prediction model of failed vaginal trial of labor in primiparous women for intermediate cesarean delivery constructed in this study has good clinical prediction efficacy and high correctness rate.

Objective:To analyze the application of indocyanine green (ICG) fluorescence imaging in laparoscopic resection of pancreatic cancer.Methods:Data of 15 patients undergoing laparoscopic surgery for pancreatic cancer in the Department of Hepatobiliary Surgery of the First Affiliated Hospital of Wannan Medical College from June 2022 to March 2023 were retrospectively analyzed, including 13 males and 2 females, aged (67.0±8.6) years. ICG were intraoperatively injected to visualize the lesion and guide surgical resection. The surgical methods, postoperative pathology, ICG fluorescence imaging and tumor margins were reviewd.Results:Among the patients, seven underwent laparoscopic pancreaticoduodenectomy, seven underwent laparoscopic radical antegrade modular pancreaticosplenectomy, and one conversed to open pancreaticoduodenectomy due to combined superior mesenteric vein reconstruction. Postoperative pathology confirmed pancreatic moderately differentiated adenocarcinoma in nine cases, pancreatic moderately-low differentiated adenocarcinoma in four cases, pancreatic follicular cell carcinoma in one case, and inflammatory lesion in one case. Negative surgical margins were confirmed in all cases. Pancreatic lesion were visualized in 14 cases (fluorescent delineation of the tumor capsule) but not well visualized in one case (with moderately differentiated adenocarcinoma). In the case of inflammatory disease, the lesion parenchyma were visualized.Conclusion:ICG injection in laparoscopic surgery enables visualization of pancreatic tumor, which facilitates tumor localization and margin determination.

Objective:To analyze the impact of adolescent pregnancy on maternal and infant outcomes.Methods:A retrospective cohort study was conducted on 5 765 parturbirths in Jining Medical College Hospital from January 1, 2019 to December 31, 2020. The parturbirths were divided into adolescent group (maternal age<20 years, 280 cases), age group 1 (maternal age 20-24 years, 1 733 cases) and age groups 2 (maternal age 25-34 years, 3 752 cases). All information was collected through the hospital′s electronic case system. General data, pregnancy characteristics and outcomes were compared among the three groups by analysis of variance (ANOVA), χ 2 tests and binary logistics regression analysis was used to analyze the impact of adolescent pregnancy on maternal and infant outcomes. Results:In the adolescent group, the proportion of women with an education of junior high school or below, rural residence, no fixed income, unmarried, and no history of induced abortion were all significantly higher than those in age group 1 and age group 2 (82.50% vs 17.37%, 14.37%; 59.29% vs 42.70%, 43.36%; 80.71% vs 15.52%, 14.71%; 75.71% vs 12.23%, 9.97%; 82.50% vs 71.84%, 71.91%) (all P<0.05); there was no significant differences in age at menarche, body mass index before pregnancy, and weight gain during pregnancy among the three groups (all P>0.05). The proportion of preterm birth, low birth weight infants and transferring to neonatal intensive care unit (NICU) in the adolescent group were all significantly higher than those in age group 1 and age group 2 (5.36% vs 1.10%, 1.57%; 5.00% vs 0.23%, 0.05%; 21.79% vs 6.12%, 15.17%); the incidence of anemia in pregnancy in the adolescent group was significantly higher than that in age group 1 (15.36% vs 9.75%), and the incidence of postpartum hemorrhage was significantly higher than that in the age group 2 (10.71% vs 6.08%). The incidence of failed vaginal trials leading to cesarean section, amniotic fluid contamination, and episiotomy was significantly lower in the adolescent group than those in age group 2 (8.57% vs 15.22%, 10.71% vs 18.10%, 33.95% vs 40.01%) (all P<0.05). The incidence of failed vaginal trials leading to cesarean section was inversely associated with gestational age (adolescent group, OR=0.252, 95% CI: 0.123-0.515; age group 1, OR=0.673, 95% CI: 0.567-0.799) (both P<0.05); the risks of low birth weight infants (adolescent group, OR=7.440, 95% CI: 3.426-16.156; age group 1, OR=0.103, 95% CI: 0.032-0.330) and transferring to the NICU (adolescent group, OR=1.661, 95% CI: 1.120-2.463; age group 1, OR=0.360, 95% CI: 0.290-0.448) showed a U-shaped distribution in different pregnancy age groups, they were both higher in the adolescent group than those in the age group 2 (both P<0.05); the risk of episiotomy (adolescent group, OR=0.002, 95% CI: 0-0.016; age group 1, OR=1.308, 95% CI: 1.151-1.485) showed an inverted U-shape distribution across the different pregnancy age groups, it was lower in the adolescent group than that in age group 2 (both P<0.05). Conclusion:Adolescent pregnancy is associated with a lower risk of conversion to cesarean section and episiotomy due to failed vaginal delivery, but may increase the risk of low birth weight infants and transferring to NICU.

Objective:To investigate the current status of women′s level of birth experience and lactation initiation time and explore the effect of level of birth experience on lactation initiation time.Methods:This was a cross-sectional study. A convenience sampling method was used to select 622 maternal cases attending the Affiliated Hospital of Jining Medical College from November, 2020 to January, 2021, and the distribution of their labor experience level and lactation initiation time was investigated by questionnaire and follow-up assessment.Results:There were 622 women with transvaginal deliveries who had lactation initiation times of more than 72 h in 241 cases (38.75%). The scores for each dimension of the Childbirth Experience Questionnaire (CEQ) were (49.63 ± 8.58)points, and the scores for each dimension of CEQ were perceived safety, professional support, involvement and self-efficacy in descending order. The results of the correlation analysis showed that there was a negative correlation between the scores on each dimension of CEQ and the total score and lactation initiation time ( r values were -0.436 to -0.146, all P<0.01). Stratified regression analysis showed that after controlling for age, number of births, gestational weeks of labour, illness during pregnancy and labour analgesia as the underlying variables affecting lactation initiation time, the scores for self-efficacy, involvement, perceived safety and professional support in the CEQ all affected lactation initiation time after delivery ( t values were -6.76 to -2.02, all P<0.05). Conclusions:The birth experience and lactation of women who deliver via vaginal birth need to be taken into account. The more negative the birth experience, the longer the lactation initiation time. The women′s involvement in the birth process, their own competence, perceived safety and level of professional support are all valid influencing indicators of lactation initiation time.