This study was performed to screen the sequence of specific nanobody targeting SARS-CoV-2 S protein,and to express and purify recombinant anti-SARS-CoV-2 nanobodies,and evaluate their characteristics and application potential.Based on the previously constructed natural bacteriophage nanobody display library,the anti-SARS-CoV-2 S protein nanobody sequences were screened,with biotinylated SARS-CoV-2 S protein receptor-binding domain antigen as target,by biotin-streptavidin liquid phase screening method,phage-ELISA and sequencing.Recombinant anti-SARS-CoV-2 S protein nanobody expression vectors were constructed,the protein was induced by IPTG and then purified,and their characteristics and application potential were analyzed by SDS-PAGE,Westem blot and ELISA methods.Two anti-SARS-CoV-2 S protein nanobody sequences were successfully obtained,and the corresponding prokaryotic expression bacteria were constructed.The recombinant anti-SARS-CoV-2 S protein nanobody was expressed and purified successfully,which can specifically bind to SARS-CoV-2 S protein.In conclusion,the recombinant anti-SARS-CoV-2 S protein nanobodies have screened and expressed successfully,which provides a basis for its application in SARS-CoV-2 detection and treatment,and facilitates related researches.

Objective:To explore the clinical and genetic features of two Chinese pedigrees affected with Autosomal dominant intellectual developmental disorder 49 (MRD49).Methods:Two MRD49 pedigrees which were admitted to the Children′s Hospital Affiliated to Shandong University respectively on January 28, 2021 and November 10, 2022 were selected as the study subjects. Clinical data of the two pedigrees were collected and analyzed. Genomic DNA was extracted from peripheral blood samples of the probands and their family members. The probands were subjected to mutational analysis by high-throughput sequencing. Candidate variants were validated using real-time fluorescence quantitative PCR (q-PCR) or Sanger sequencing and bioinformatic analysis. This study was approved by the Medical Ethics Committee of the Children′s Hospital Affiliated to Shandong University (No. SDFE-IRB/T-2022002).Results:Proband 1 had presented with language delay, motor retardation and intellectual disability, and his maternal grandmother, mother, aunt and cousin all had various degrees of intellectual disability. Sequencing results showed that proband 1 had deletion of exons 3 ~ 7 of the TRIP12 gene. q-PCR verification showed that his mother, aunt, maternal grandmother and cousin had all harbored the same deletion. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was classified as pathogenic (PVS1+ PM2_Supporting+ PP1). Proband 2, who had mainly presented with language delay, motor retardation and intellectual disability, and was found to harbor a heterozygous c.3010C>T (p.Arg1004*) variant of the TRIP12 gene, which was verified to be de novo in origin. Based on the guidelines from the ACMG, the variant was classified as pathogenic (PVS1+ PS2+ PM2_Supporting). Conclusion:This study had diagnosed two MRD49 families through high-throughput sequencing. Above findings have enriched the phenotypic and mutational spectrum of MRD49 in China, which has also facilitated genetic counseling for the two pedigrees.

Background Environmental pollutants can affect N⁶-methyladenosine (m⁶A) level in the body, but the change of m⁶A level in kidney after being exposed to cadmium (Cd) and the molecular mechanism of renal injury need to be further studied. Objective To analyze the associations of m⁶A modification and methyltransferases/demethylases with microRNA-21 (miR-21) and transforming growth factor- β1 (TGF - β1) in kidney of rats exposed to Cd. Methods Twenty-four SPF male SD rats were divided into 4 groups, with 6 rats in each group, and were exposed to Cd by subcutaneous injection of 2.0, 1.0, and 0.5 mg·kg⁻¹ cadmium chloride (CdCl₂) and equal volume of normal saline for 2 weeks, 7 d a week, respectively. The levels of N-acetyl-β-D-glucosidase (UNAG) and albumin (UALB) in urine, and the levels of m⁶A methylation and TGF-β1 in kidney were detected by enzyme-linked immunosorbent assay (ELISA). The level of blood urea nitrogen (BUN) was measured by urease method. The levels of renal oxidative stress indicators such as malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) were detected by total bile acid method, water-soluble tetrazolium asssay, and colorimetric method respectively. The relative levels of TGF-β1, methyltransferases, and demethylases in kidney were measured by reverse transcription-polymerase chain reaction. The expression of miR-21 in kidney was detected by fluorescent quantitative polymerase chain reaction. Results After 2 weeks of exposure to Cd, the body weights of rats in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups decreased, and the ratio of kidney/body weight and the levels of BUN, UNAG, and TGF-β1 mRNA and protein increased in the 2.0 mg·kg⁻¹ cadmium chloride group (P<0.05). The expression levels of m⁶A modification, methyltransferases METTL3, METTL14, Wilms’ tumor 1-associated protein (WTAP), and miR-21 were increased both in the 2.0 and 1.0 mg·kg⁻¹ cadmium chloride groups, with significant differences compared with the control group (P<0.05). The results of correlation analysis showed that the m⁶A modification level was negatively correlated with SOD (r=−0.4489, P<0.05) and GSH-Px (r=−0.4874, P<0.05), METTL3 was negatively correlated with MDA (r=−0.5158, P<0.05), while there was a positive correlation between FTO and GSH-Px (r=0.4802, P<0.05). In addition, miR-21 was positively correlated with METTL3 (r=0.7491), METTL14 (r=0.6157), and WTAP (r=0.6660) (P<0.05), TGF-β1 was positively correlated with METTL3 (r=0.5025, P<0.05) but negatively correlated with FTO (r=−0.5634, P<0.05) . Conclusion Cd can induce m⁶A methylation and up-regulation of METTL3, METTL14, WTAP, and miR-21 expression levels in rat kidney tissues, indicating that m⁶A and miR-21 may be associated with Cd-induced renal fibrosis.

Objective:To explore the research status, research focus and development trend of nursing human resource allocation in nursing homes in China.Methods:The article on nursing human resource allocation in nursing homes in China was systematically retrieved from China National Knowledge Infrastructure (CNKI), VIP and WanFang Data, and the retrieval time limit was from the establishment of the database to June 2022. CiteSpace was used to analyze the amount of articles, authors, institutions, funds and keywords, and draw a knowledge map.Results:A total of 890 articles were included. The number of articles and the support of national level funds were growing. The cooperation network among authors was extensive but not deep enough, and a core group of authors was not formed. A core journal group was formed. The research institute with the largest number of articles was the School of Nursing of Peking University. The article was mainly distributed in Beijing, Shanghai, Guangzhou and other first tier cities. The high-frequency keywords were "combination of medical care and nursing care" "supply" "demand" "talent training" and " care service for the aged", which could be clustered into 11 tags. The keyword cluster atlas showed that the combination of medical care and nursing care had always been the mainstream research direction in the field of nursing human resource allocation in nursing homes in China. "Qualitative research" "rural care for the aged " "care model for the aged " and "care industry for the aged " would become the emerging development trend in this field.Conclusions:The research on the nursing human resource allocation in nursing homes in China is growing. The research focuses on the combination of medical care and nursing care, talent training, care industry for the aged, and the supply and demand of care service for the aged. In the future, we should take the combination of medical care and nursing care as the leading mode, constantly improve our elderly care service system, build a nursing human resource allocation mode with Chinese characteristics in nursing homes, and promote the healthy aging development of society.

Objective:To understand the incidence of post-traumatic stress disorder(PTSD) in fathers with premature infants and analyze the risk factors.Methods:Conveniently, the 203 fathers of premature infants in NICU of Affiliated Hospital of Jining Medical University of Shandong Province from May to August 2021 were selected as the subjects to fill in the general data questionnaire, Perinatal Post-traumatic stress disorder Questionnaire-Chinese edition (PPQ-C), Parents, Perception of Uncertainty Scale, Social Support Rating Scale, Simplified Coping Style Questionnaire. Logistic regression was used to analyze the risk factors of PTSD and establish a nomogram model. ROC curve was used to verify the discrimination ofthe model. Hosmer-Lemeshow goodness-of-fit test and Calibration Plot were used to verify the calibration.Results:PPQ-C total score of 203 fathers was 17.17 ± 8.77, 81 fathers, symptoms were positive and the incidence of PTSD was 39.90%(81/203). Logistic regression analysis showed college degree ( OR = 0.297, 95% CI 0.116 - 0.763, P<0.05), very low birth weight ( OR = 2.491, 95% CI 1.027 - 6.044, P<0.05), sense of disease uncertainty ( OR = 1.038, 95% CI 1.012 - 1.066, P<0.05), negative coping style ( OR = 1.871, 95% CI 1.127 - 3.108, P<0.05) were risk factors of PTSD in fathers with premature infants. The nomogram model was established basing on the results of the Logistic regression analysis, and the ROC curve proved (AUC = 0.751) the model having a good discrimination.The Hosmer-Lemeshow goodness-of-fit test ( P = 0.974) and the calibration plot demonstrated that the prediction values tends to coincide with the actual monitoring values. Conclusions:A higher incidence of PTSD was observed in fathers with hospitalized premature infant. It was related to education level, the infants′s birth weight, disease uncertainty and coping style. Therefore, the nurses should give information support and psychological guidance according to the individual situation of the infants′ father to reduce the incidence of PTSD.

Objective:To compare the clinical outcomes between OTA/AO-C open and closed fractures of the distal humerus treated by open reduction and internal fixation.Methods:The clinical data were retrospectively analyzed of the 70 patients who had been treated at Department of Traumatology and Orthopedics, Beijing Jishuitan Hospital for OTA/AO-C fractures of the distal humerus from January 2014 to June 2017. Of them, 22 suffered from open fractures (Gustilo types Ⅰ/Ⅱ) and 48 closed fractures. There were 18 males and 4 females with an age of (42.6±13.0) years in the open group and 21 males and 27 females with an age of (42.2±17.1) years in the closed group. Analyzed were interval from injury to surgery, hospitalization time, injury energy and functional outcomes which included range of motion (ROM) in elbow flexion and extension, ROM in elbow rotation, Mayo elbow performance score (MEPS), Disabilities of the Arm, Shoulder and Hand (DASH), complications and rate of secondary surgery.Results:There was no significant difference between the 2 groups in age, injury energy or interval from injury to surgery ( P>0.05), but there were significantly more males in the open group than in the closed group ( P=0.011). The follow-up time for all the patients averaged 34.0 months (from 25 to 54 months). There were no statistically significant differences between the 2 groups in hospitalization time [9.5(6.0, 13.0) d versus 8.5 (6.0, 11.0) d], ROM in flexion and extension [120.0° (100.0°, 137.8°) versus 128.5° (110.0°, 140.0°)], ROM in rotation [155.0° (151.3°, 155.0°) versus 155.0° (155.0°, 155.0°)], MEPS [95.0 (80.0, 100.0) versus 95.0 (80.0, 100.0)] or DASH [2.6 (0.63, 9.2) versus 1.7 (0.0, 8.5)] ( P>0.05). There were no statistically significant differences between the 2 groups either in rate of secondary surgery [36.4% (8/22) versus 33.3% (16/48)], ulnar nerve symptoms [54.5% (12/22) versus 60.4% (29/48)], local irritability in the region of internal fixation [9.1% (2/22) versus 6.3% (3/48)] or elbow stiffness [13.6% (3/22) versus 10.4% (5/48)] ( P>0.05). Conclusion:Open reduction and internal fixation can lead to similar clinical outcomes in the treatment of both open (Gustilo types Ⅰ/Ⅱ) and closed distal humeral fractures of OTA/AO-C, with no significant differences in postoperative ROM, functional scores or complications.

Objective:To analyze the risk factors of medical adhesive-related skin injury (MARSI) in preterm infants to establish a nomogram model to predict the risk of MARSI.Methods:From July to September 2018, 268 premature infants who were hospitalized in NICU were enrolled in this study. Their clinical data were analyzed and univariate analysis was used to detemine the risk factors related to MARSI, The significant variables were included in the multivariate logistic regression analysis to analyze the independent risk factors for MARSI. Then the R software was used to establish a predictive nomogram model, Bootstrap method was used to validate the nomogram model and the consistency test of the correction curve was used to explore the predictive efficacy of the model in predicting the MARSI.Results:A total of 64 cases out of 268 premature infants had MARSI. The prevalence rate was 23.9 % (64/268). Birth age (28 to 32 weeks) ( P value was 0.021, OR value was 2.736, 95 % CI 1.163-6.435), edema ( P<0.01, OR value was 33.782, 95 % CI 10.510-108.583), maternal diabetes ( P value was 0.039, OR value was 16.011, 95 % CI 1.146-223.692), easy to tear tape ( P value was 0.027, OR value was 13.567, 95 % CI 1.340-137.311) were all independent risk factors for MARSI. The nomogram model showed a conformance-index of 87.29 %, while the consistency test of the correction curve showed that the prediction probability is consistent with the actual occurrence probability. Conclusions:The nomogram built based on the indexes of fetal age, skin state, maternal disease, mucous product has good discrimination and accuracy which could be helpful for screening the patients with high risk, with potentially high clinical application value.

Objective:To evaluate the effect of oropharyngeal colostrum administration on building adequate enteral feeding process of very low birth weight neonates and extremely low birth weight neonates.Methods:A total of 62 very/extremely low birth weight neonates in NICU were selected from a tertiary hospital in shandong province. They were randomly assigned to the observation group ( n=32) and the control group ( n=30) according to the random number table. The observation group was given oropharyngeal colostrum administration. The control group was given oral care with 0.9% sodium chloride solution, and the wiping method and frequency was the same as the observation group. Gastric retention, enteral feeding time, adequate enteral feeding time, the weight at the time of discharge, etc. were compared between two groups. Results:Gastric retention times on the 10th day in the observation group (1.90±1.30) was significantly lower than that in the control group (2.77±1.50), and the two groups were statistically significant ( t value was -2.449, P<0.05). The duration of 100 ml·kg -1·d -1 enteral feeding and adequate enteral feeding (150 ml·kg -1·d -1) in the observation group [(24.63±9.42) days, (29.75±10.15) days] were significantly shorter than those in the control group [(33.90±2.73) days, (35.13±9.29)days], and the two groups were statistically significant ( t value was-4.621, P<0.01; t value was -2.362, P<0.05). The weight of the observation group was higher than that in the control group at the time of discharge ( t value was 4.677, P<0.05). Conclusion:Oropharyngeal colostrum administration can reduce gastric retention times, shorten the adequate enteral feeding time, promote growth and development in very low birth weight neonate and extremely low birth weight neonate.But it had not been proved to reduce the incidence of related infection indicators.

Objective:To explore the application effects of dual probiotic preparation and colostrum oral immunotherapy on improving feeding intolerance in very low birth weight infants.Methods:A systematic sampling method was used to select 173 very low birth weight infants who were admitted to the Neonatal Intensive Care Unit (NICU) from December 2017 to December 2018. According to the random number table method, they were divided into in group A ( n=56) , group B ( n=59) and group C ( n=58) . Group A was given routine nursing, parenteral nutrition and enteral nutrition and minimal breast feeding. Group B was given dual probiotic preparation on the basis of routine nursing. Group C was given colostrum oral immunotherapy on the basis of routine nursing. After 7 days of intervention, the improvement of feeding intolerance was observed, and the feeding intolerance rates were compared among the three groups after 3 d, 5 d and 7 d of intervention. Intestinal flora, incidence of complications, length of NICU stay and speed of birth weight recovery were compared. Results:The intake of milk in group B and group C were higher than those in group A, and the retention milk volume, retention times and duration of abdominal distension were all lower than those in group A, while retention times and duration of abdominal distension in group C were shorter than those of group B, and the differences were statistically significant ( P<0.05) . On the 3rd, 5th and 7th day of intervention, the feeding intolerance rates of the three groups continued to decrease. The feeding intolerance rates of group B and C were lower than those of group A, and the feeding intolerance rate of group C was lower than that of group B on the 3rd day of intervention , and the differences were statistically significant ( P<0.05) . The number of Enterococcus faecium, the number of Bacillus subtilis and the ratio of the logarithmic number of bifidobacteria to the logarithmic number of Escherichia coli (B/E value) in groups B and C were all higher than those in group A, and the incidences of sepsis and necrotizing enterocolitis in group B and group C were lower than those in group A, and the differences were statistically significant ( P<0.05) . The length of stay in NICU was ranked from long to short as Group A, Group B, and group C, and the speed of birth weight recovery was ranked from slow to fast as group A, Group B, and group C, and the differences were statistically significant ( P<0.05) . Conclusions:Colostrum oral immunotherapy can improve the feeding intolerance of very low birth weight infants, reduce complications related to premature delivery, promote the growth and development of neonates and shorten the length of stay in NICU. The clinical effect is slightly better than that of probiotics.

Objective: To explore the risk factors for elbow stiffness after open reduction and internal fixation (ORIF) for intercondylar fractures of the distal humerus. Methods: From January 2013 to May 2017, 159 patients underwent ORIF for intercondylar fractures of the distal humerus with dual plating. They were 83 males and 76 females with a mean age of 42.6 years (from 14 to 79 years). They were divided into 2 groups according to their range of motion at the latest follow-up and the secondary elbow arthrolysis they had undergone or not. The stiffness group had extension-flexion and/or pronation-supination <100° and secondary elbow arthrolysis while the non-stiffness group had extension-flexion and pronation-supination ≥100° and no secondary elbow arthrolysis. Age, gender, fracture side, mechanism of injury, AO fracture classification, open/close fracture, additional fracture, preoperative nerve injury, time from injury to surgery, surgical approach, configuration of plating, medication for anti-heterotopic ossification and implant removal were analyzed as risk factors for elbow stiffness using Logistic regression analysis. Results: The mean follow-up period for this cohort was 32.0 months (from 10 to 63 months). The latest follow-up showed fracture union in all the patients. The stiffness group had 38 patients and the non-stiffness group 121. Multivariate regression analysis showed that high energy trauma (OR=3.141, 95%CI 1.396~7.070, P=0.006) and time from injury to surgery >one week (OR=2.596, 95%CI 1.123~6.000, P=0.026) were independent risk factors for elbow stiffness after ORIF for intercondylar fractures of the distal humerus. Conclusion The patients with high energy trauma and time from injury to surgery >one week should be treated with caution and special care in clinical practice because the 2 factors are closely related to posttraumatic elbow stiffness after ORIF for intercondylar fracture of the distal humerus.