OBJECTIVE To optimize hematopoietic stem cell transplantation therapy and provide support for drug research by investigating the dynamic process of hematopoietic and immune system reconstitution after bone marrow transplantation(BMT)in mice.METHODS CD45.2+C57BL/6 mice were used as recipient mice and randomly divided into the normal control group and transplantation group,with 30 mice in each.The transplantation group was irradiated by a lethal dose of cobalt-60 rays.Bone marrow cells were prepared from CD45.1+C57BL/6 mice and transfused into recipient mice through the tail vein.Peripheral blood,spleens,lymph nodes,thymuses and bone marrow were collected at 1,2,4,8 and 16 weeks after transplantation.Blood routine examination was performed with peripheral blood and total cell numbers in suspensions of other organs were counted by an automated cell counter.Cell classification analysis of white blood cells in peripheral blood,cell suspensions of other organs was performed by flow cytometry.RESULTS Four weeks after BMT,the numbers of white blood cells and red blood cells in peripheral blood of recipient mice returned to the same level of or higher level than normal control(P<0.05).Although the number of platelets recovered significantly,it was still mark-edly lower than that of normal control until 16 weeks post BMT(P<0.05).In addition,the percentages of myeloid leukocytes and B cells in peripheral blood,spleens,lymph nodes,and bone marrow,as well as megakaryocytes and erythrocyte progenitor cells in bone marrow also returned to normal,and the majority of myeloid leukocytes and B cells were CD45.1+cells from the donors.Eight weeks after BMT,T cells in peripheral blood,spleens,lymph nodes,thymuses,and bone marrow of recipient mice returned to normal,and CD45.1+T cells were dominating.CONCLUSION The hematopoietic and immune reconstitution of recipient mice is nearly completed eight weeks after BMT.However,the reconstruction speed of different kinds of cells and the reconstruction status of same kind of cell in different organs vary widely.

Objective:To investigate the effects of methyltransferases like 3(METTL3)mediated m6A modification of double homology cassette A pseudogene8(DUXAP8)on the proliferation,migration and invasion of salivary adenoid cystic carcinoma SACC-LM cells and its potential molecular mechanisms.Methods:Whole-transcriptome sequencing showed that DUXAP8 was highly ex-pressed in SACC than in para-cancerous tissues(P＜0.05).The m6A modification sites on DUXAP8 were predicted using the SRAMP website,and the mRNA and protein expression of m6A-modified genes and the genes associated with the epithelial-mesen-chymal transition(EMT)was measured by qRT-PCR and Western blot,respectively.METTL3 and DUXAP8 was knocked down or overexpressed in SACC-LM cells,and the proliferation,migration,and invasion of the cells were assessed by CCK-8,scratch and Transwell assays.The correlation between METTL3 and DUXAP8 was evaluated using MeRIP-qPCR.Results:The expression of DUXAP8 in SACC tumor was higher than that in para-cancerous tissues(P＜0.05).Knockdown of DUXAP8 reduced proliferation,migration and invasion of SACC-LM cells,as well as the expression of EMT-related genes(P＜0.05).Multiple m6A modification sites of high confidence were found on DUXAP8.METTL3 was highly expressed in tumor tissues,more than other related genes(P＜0.05)and enzyme-encoding genes in SACC-LM cells(P＜0.05).METTL3 was found to function as a methyltransferase to regulate the expression of DUX-AP8,and downregulation of METTL3 inhibited prolifera-tion,migration and invasion of SACC-LM cells and partially reversed the promotion of these activities induced by DUX-AP8 overexpression(P＜0.05).Conclusion:METTL3-me-diated m6A modification upregulated DUXAP8 expression,which promotes the proliferation,migration and invasion of SACC cells.

Objective:To investigate the current situation of expected sadness of family caregivers of first stroke patients and analyze its influencing factors, so as to provide reference for the formulation of psychological decompression intervention program for family caregivers of stroke patients.Methods:Convenience sampling method was used to conduct cross-sectional survey of 212 family caregivers of first stroke patients who were hospitalized in the First Affiliated Hospital of Shandong First Medical University from March 2020 to May 2021, using general information questionnaires and Anticipatory Grief Scale and Connor-Davidson Resilience Scale to investigate and anlalyzed its influencing factors.Results:The scare of anticipatory grief of the caregivers of first stroke patients was 87.93 ± 17.36. The results of multiple stepwise regression analysis showed that the age of family caregivers, psychological elasticity, the degree of relationship with patients, patients′ cognitive ability and self-care ability were the influencing factors of anticipatory grief of family caregivers of first stroke patients ( t values were -6.73 -11.77, P <0.05). Conclusions:The caregivers of first stroke patients have serious anticipatory grief. Staff should pay attention to the psychological conditions of the family caregivers and take effective measures to promote their physical and mental health.

Objective:To evaluate the efficacy of disposable disinfection cap in preventing catheter-related blood stream infections (CRBSIs).Methods:Literature on the prevention of CRBSIs by disposable disinfection caps were retrieved from CNKI, Wanfang Database, VIP database, Chinese Biomedical Literature Database, PubMed, Cochrane Library, Embase and Web of Science databases. The retrieval period was from the database construction to June 30, 2022. After literature screening, data extraction and quality evaluation were independently carried out by 2 researchers, RevMan5.4 software was used for analysis.Results:A total of 12 articles were included, including 9 832 patients. Meta-analysis results showed that compared with conventional manual disinfection, disposable disinfection cap could reduce the incidence of CRBSIs, and the difference was statistically significant ( RR = 0.58, 95% CI 0.43-0.79, P<0.01). Disposable disinfection cap could reduce the incidence of CRBSIs in adults, but there was no significant difference in the incidence of CRBSIs in children ( P>0.05). It could reduce the incidence of CRBSIs in patients with indwelling vascular catheters in ICU ( RR = 0.58, 95% CI 0.38-0.89, P<0.05), but there was no statistical difference in the incidence of CRBSIs in patients without indwelling vascular catheters in ICU ( P>0.05). In addition, the compliance of the use of disinfection caps by nursing staff was improved by 80% - 90% and the treatment cost was saved by about 282 - 464 dollars. Conclusions:Disposable disinfection caps can reduce the occurrence of CRBSIs, improve the disinfection compliance of nursing staff, and save the hospitalization cost of patients. It is suggested to be popularized in clinical practice.

Objective:To compare the genotypes and phenotypes between the monozygotic twins via whole genome sequencing to further clarify the autosomal dominant inherited neurofibromatosis type 1 (NF1) variants related to congenital pseudarthrosis (CP).Methods:According to the diagnostic criteria of congenital tibial pseudarthrosis and the clinical diagnostic criteria of NF1, two pairs of monozygotic twins with NF1 were included. Both were female and only one of each pair had congenital pseudarthrosis. The other did not have congenital pseudarthrosis. Whole genome sequencing was performed using the peripheral blood of the two pairs of monozygotic twins. Customized bioinformatics analysis was then performed to identify single nucleotide variants (SNVs), short insertion deletion variants (InDel), copy number variants (CNVs), and structural variants (SVs). Classified the variants according to the American College of Medical Genetics and Genomics (ACMG) and ClinGen criteria. The germline variants within the monozygotic twins were compared to identify the CP patients' unique variants. The shared pathogenic or likely pathogenic germline variants between the unique variants in the CP patients from the twins were also analyzed. Further, the identified disease-causing variants were validated by Sanger sequencing in the family of the twins and their parents. Finally, the genotypes and phenotypes regarding the pathogenic variants of the NF1 gene among the twins were characterized. Results:Both the two monozygotic twins were identified pathogenic variants in the NF1 gene. One with c.3047_3048del (p.Cys1016SerfsTer4), and the other with c.4267A>G (p.Lys1423Glu). By Sanger sequencing validation in family quads, the two CP patients and their siblings harbored de novo heterozygous variants of the NF1 gene. In addition to the NF1 gene, no other genes were identified pathogenic or likely pathogenic variants uniquely in the CP patients compared with their twin sisters, as well as SVs and CNVs. In addition, by analyzing the rare and damaging variants in the two CP patients from the two twins, they had no overlapping genes against the SNVs, InDels, SVs, or CNVs. Conclusion:Whole genome sequencing revealed that both the two monozygotic twins with NF1 were detected pathogenic variants of gene NF1. No other pathogenic variants specific to the CP patients among the twins were identified. The two CP patients shared no other common genes from the detected likely pathogenic variants.

Objective:To investigate the clinical effect of Ilizarov bone transport technique with assisted guiding pin in medullary cavity for treatment of posttraumatic bone defect of tibia.Methods:A retrospective case series study was conducted to analyze the clinical data of 17 patients with post-traumatic bone defect of tibia admitted to Third Hospital of Hebei Medical University form November 2014 to March 2018.There were 13 males and 4 females, aged 19-60 years [(37.2±13.4)years]. The bone defect length was 4.6-14.0 cm [(8.6±2.8)cm] after debridement. All patients underwent treatment with Ilizarov bone transport technique. The alignment of transport bone segment was controlled by a guiding pin in medullary cavity of tibia. Bone grafts were performed to accelerate fracture healing of docking point. The wound healing, bone healing, external fixation time, external fixation index (EFI), alignment recovery were recorded. Bone healing and functional results were evaluated according to the criteria given by Association for the Study and Application of the Method of Ilizarov (ASAMI). The complication was recorded according to Paley's criteria. The physical component summary score (PCS) and mental component summary score (MCS) ware recorded according to the MOS 36-item Short-form Health Survey (SF-36) questionnaire and compared with the national norm to evaluate the quality of life.Results:After removal of the apparatus, follow-up period was 12-37 months [(29.9±4.4)months]. Wound healing was achieved without flap transfer. At the latest follow-up, all patients achieved bone healing without recurrent infection. External fixation time was 242-801 days [(436.5±154.6)days] and external fixation index was 35.7-60.5 d/cm [(50.6±6.2)d/cm]. The affected extremity alignment was restored in all patients except for residual angular deformity in one patient. According to ASAMI, the excellent and good rate of bony results and functional results were 88% and 94%. According to Paley's criteria, complications included 12 problems, 7 obstacles and 1 sequelae. The PCS and MCS in SF-36 questionnaire were (85.8±11.6)points and (69.6±11.1)points. Compared with the national norm [PCS: (87.6±16.8)points, MCS: (78.8±15.4)points], PCS showed no statistical difference ( P>0.05), but MCS showed statistical difference ( P<0.05). Conclusions:Ilizarov bone transport combined with intramedullary guiding pin can accelerate wound and fracture healing, control infection and restore lower limb alignment. The physical function of the affected extremity can be restored in spite of some complications and psychological effects. The technique is an effective treatment for posttraumatic bone defect of tibia.

Objective:To analyze the expression level of malignant biological marker gene in locally advanced low rectal cancer tissues after neoadjuvant chemotherapy with XELOX, so as to provide guidance for clinical treatment.Methods:Based on the established inclusion and exclusion criteria, 104 patients with locally advanced low rectal cancer treated in the Surgery Department of the Affiliate Hospital of Qingdao University from January 2017 to January 2018 were included in the study. Based on the random number table method, they were divided into observation group and control group, with 52 patients in each group. Radical resection was performed in both groups, while radical resection was performed directly in the control group, and neoadjuvant chemotherapy with XELOX plus radical resection was performed in the observation group. The effect of tumor resection in the two groups was compared and analyzed, and the expression levels of tumor markers and malignant biological marker genes in the tumor tissues after surgery were determined. The 3-year postoperative survival rate of the two groups was followed up.Results:The R0 resection rate was 96.15% in the observation group and 80.77% in the control group, with statistically significant difference ( P<0.05). There were no significant differences in the serum levels of cancer antigen 19-9 (CA19-9), carcinoembryonic antigen (CEA) and carbohydrate antigen 50 (CA50) before the operation in the two groups ( P>0.05). One week after the operation, the serum levels of CA19-9, CEA and CA50 in the observation group were significantly lower than those in the control group ( P<0.05). Through detection, the relative mRNA expression levels of PPTG and Smad4 in the postoperative observation group were higher than those in the control group, while the relative mRNA expression levels of Runx3 and APC were lower than those in the control group, with statistically significant difference ( P<0.05). The 3-year follow-up survival rate in the observation group was 90.38%, significantly higher than 78.85% in the control group ( P<0.05). Conclusions:The application of XELOX neoadjuvant chemotherapy in radical resection of locally advanced low rectal cancer has important clinical value in reducing tumor malignancy, improving tumor resection effect and improving survival.

Objective: To analyze the risk factors of adverse neurodevelopmental prognostic in very /extremely low birth weight infants (VLBWI/ELBWI). Methods: The 24 hours VLBWI/ELBWI admitted to the neonatal intensive care unit between January 2016 and October 2016 were enrolled.These infants were followed up and neurodevelopmental evaluation was performed at a corrected age of 12 months by using the Bayley Scales of Infant Development Ⅱ (BSID-Ⅱ). According to the neurodevelopmental outcomes, they were divided into normal and abnormal neurodeve-lopment groups.The data of prenatal, intrapartum and postpartum periods in the two groups were compared.Potential risk factors of neurodevelopmental impairment were analyzed with Logistic stepwise regression. Results: A total of 167 VLBWI/ELBWI were enrolled, among which 14 cases died during hospitalization or after giving up treatment.At the corrected age of 12 months, 140(91.5%) infants completed follow-up and 13(8.5%) were lost to follow-up.Among 140 infants who completed neurodevelopmental evaluation at the corrected age of 12 months, there were 86 males, 54 females, and in which 28 cases had extremely low birth weight, 112 cases with very low birth weight, with gestational age of (30.4±2.2) weeks (25.3-36.0 weeks). There were one hundred and twenty-five cases with normal neurodevelopmental outcomes and 15 cases with abnormal neurodevelopmental outcomes.The results of univariate analysis showed that birth weight<1 000 g, exposure to antenatal steroids, N -terminal brain natriuretic peptide (NT-proBNP) ≥35 000 ng/L and bronchopulmonary dysplasia (BPD) in the two groups were statistically significant(P<0.05). Logistic stepwise regression showed that NT-proBNP level≥35 000 ng/L may independently predict neurodevelopmental impairment among VLBWI/ELBWI (OR=22.774, 95%CI: 3.079-168.425, P=0.002). Exposure to antenatal steroids may be a protective factor for neurodevelopmental impairment (OR=0.125, 95%CI: 0.020-0.782, P=0.026). Conclusions Plasma NT-proBNP level ≥35 000 ng/L may independently predict neurodevelopmental impairment among VLBWI/ELBWI.Antenatal corticosteroids may be the protective factor of poor neurodevelopmental outcomes.

Objective To discuss the experience of surgical treatment for cerebral sparganosis and analyze the factors affecting the removal of live worms.Methods A retrospective analysis was performed on clinical data and treatment experience of 49 patients with cerebral sparganosis,admitted to and accepted surgery in our hospital from January 2001 to December 2017.These patients were divided into two groups according to whether the live worms were removed during the operation,namely live worms removal group and live worms non-removal group;factors affecting the removal of live worms were analyzed.Results A total of 36 live worms were removed from 49 patients,one of them took out two live worms in the same lesion.In 45 patients with seizures before surgery,39 of them had seizure-flee after surgery (Engel grading I).Re-examination MR imaging showed that lesions disappeared and no new lesions were found in the brains of the 39 patients.Another two with original lesion enhancement had shadow disappearance;however,new lesions appeared behind the lesion;one of them was re-operated in the contralateral temporal lobe of the original lesion,and the other one continued follow-up observation.No new hemorrhage lesion,infarction or meningitis after operation was noted in all patients.Statistical analysis showed that there were significant differences in times from lesion discovery to surgery and lesion locations between patients of the live worms removal group and the live worms non-removal group (P＜0.05).Conclusions Surgical treatment of cerebral sparganosis is safe and effective.Surgical treatment should be performed as soon as possible after the lesion is discovered.The times from lesion discovery to operation should be shortened to prevent the displacement of the worms.At the same time,it is necessary to select the specific surgical plan and timing of surgery in combination with lesion location to improve removal rate of live insects and reduce surgical complications.If necessary,surgery can be performed again.

Objective To investigate the normal development of anterior and posterior acetabulum in children through measuring anterior and posterior acetabular indexes with the baseline from the thinnest point of acetabulum to the center of femoral head.Methods MRI data in 165 normal children aged 0-12 years were collected.The baselines were drawn from the center of the femoral head to the thinnest point of acetabulum (method 1) and from the one midpoint of Y cartilage to the contralateral (method 2),then the anterior or posterior bony acetabular index (A/PBAI) and anterior or posterior cartilaginous acetabular indexes (A/PCAI) were measured.The consistency of above parameters measured using two methods and between two observers was observed,and the correlation with parameters-gestational ages was analyzed.Results The consistency of ABAI (ICC=0.832) measured with two methods was good,and the consistency of ACAI (ICC=0.535),PBAI (ICC=0.565) and PCAI (ICC=0.472) was fair.The consistency between two observers was good (all ICC＞0.75).ABAI,ACAI and PBAI were negatively correlated with age (r=-0.762,-0.475,-0.368,all P＜0.001),and PCAI had no correlation with age (r=-0.190,P＜0.005).Before 4 years old,ABAI gradually decreased with age and gradually stabilized after 4 years of age.ACAI and PBAI decreased slightly with aging.PCAI did not change obviously with aging.Conclusion The measuring method of anterior and posterior acetabular indexes with the baseline from the thinnest point of acetabulum to the center of femoral head can accurately evaluate the normal development of anterior and posterior acetabulum in children.