Objective To investigate the application value of single nucleotide polymorphism(SNP)linkage analysis based on next-generation sequencing(NGS)technology in preimplantation genetic testing(PGT)of families with autosomal recessive polycystic kidney disease(ARPKD).Methods A family with ARPKD was selected,where the female member had a pregnancy ultrasound revealing polycystic kidney in the fetus.Genetic testing showed compound heterozygous mutations of the polycystic kidney/polycystic liver disease 1 gene(PKHD1),c.10444C>T(paternal)and c.4303del(maternal),with the c.4303del mutation being reported for the first time.Targeting the coding region of the PKHD1 gene,335 high-density tightly linked SNP sites were selected in the upstream and downstream 2M regions using multiplex polymerase chain reaction(PCR)and NGS.The couple′s SNP risk haplotypes carrying gene mutations were constructed.After in vitro fertilization,blastocyst culture was performed.Trophoblastic cells obtained from the biopsy were subjected to whole-genome amplification,and NGS was used for linkage analysis and low-depth chromosomal aneuploidy screening of the embryos.Sanger sequencing was used to verify the results of embryo linkage analysis.Results Among the 6 biopsied embryos,4 were mutation-free and euploid,1 exhibited heterozygous for the mutation and mosaic while another unstable sequencing data,making it impossible to judge.One of the mutation-free and developmentally healthy euploid embryos was implanted into the maternal uterus,resulting in the full-term delivery of a healthy baby.Conclusion Application of NGS-based SNP linkage analysis in PGT can effectively blocking the vertical transmission of ARPKD within families,while avoiding abortion issues caused by aneuploid embryos.This study is also the first PGT report target-ing the PKHD1 gene c.4303del mutation.

Objective:To investigate the clinicopathological features and differential diagnosis of eosinophilic vacuolated tumor (EVT).Methods:Seven cases of EVT with characteristic morphology and unequivocal diagnosis from the Affiliated Hospital of Qingdao University (6 cases), Qingdao, China and the 971 Hospital of PLA Navy (1 case), Qingdao, China between January 2010 and December 2021 were subject to morphological and immunohistochemical analyses. Additionally, whole exome sequencing (WES) was performed in two cases. Twenty-two cases of renal oncocytoma (RO) and 17 cases of eosinophilic chromophobe renal cell carcinoma (eChRCC) diagnosed at the same time were used as controls.Results:Four males and three females with a mean age of 42 years (range: 29-61 years) were included in the study. The tumors were nodular and well-circumscribed, with sizes ranging from 1.5 to 4.5 cm. On cross-section, they appeared gray-red or gray-white, solid, and soft. Tumor cells were arranged in nests, solid sheets, and acinar or small vesicular structures. These cells exhibited eosinophilic cytoplasm with large, prominent clear vacuoles and round nuclei with prominent nucleoli. Perinuclear halos were focally present in four cases, while small tumor cells with sparse cytoplasm and hyperchromatic nuclei were seen in one case. No necrosis or mitosis was noted. Edematous stroma was detected in three cases. All tumors were positive for CD117 and Cathepsin K, but negative for vimentin and CK7. CK20 was positive in scattered individual cells, and Ki-67 positivity ranged from 1% to 4%. Point mutations in MTOR were identified in both patients who were subject to the molecular analysis. Statistical differences in the expression of Cathepsin K, CD10, S-100A1, and Cyclin D1 between EVT and RO ( P<0.05) were significant, so were the differences in the expression of Cathepsin K, CD10, CK7 and claudin 7 between EVT and eChRCC ( P<0.001). Seven patients were followed up for 4 to 96 months (mean, 50 months), with no recurrences or metastases. Conclusions:EVT is a rare renal tumor that shares morphological and immunophenotypic features with RO and eChRCC, and it is closely linked to the TSC/MTOR pathway. The presence of large prominent transparent vacuoles in eosinophilic cytoplasm along with conspicuous nucleoli is its key morphological characteristics. The use of combined immunohistochemical stains greatly aids in its diagnosis. Typically, the tumor exhibits indolent biological behaviors with a favorable prognosis.

Objective:To gain a deeper understanding of the fear of recurrence in patients with atrial fibrillation during the "blanking period" after catheter ablation.Methods:The interview outline was developed based on common-sense model of self-regulated. From July to September 2023, purposive sampling was used to select 15 patients with atrial fibrillation in the "blanking period" after catheter ablation at the Cardiovascular Outpatient of the Affiliated Hospital of Qingdao University as research subjects, and semi-structured interviews were conducted. The targeted content analysis method was used to analyze data.Results:Four themes were extracted, namely triggering factors of fear of recurrence, perception of atrial fibrillation recurrence, negative emotional distress, and insufficient ability to cope with atrial fibrillation recurrence.Conclusions:For patients with atrial fibrillation during the "blanking period" after catheter ablation, medical and nursing staff should closely monitor the patient's cognitive level and psychological state, and carry out targeted health education to meet the patient's needs for postoperative disease management, life adaptation, and other aspects, so as to reduce the fear of recurrence in atrial fibrillation patients during the "blanking period" after surgery.

Objective To investigate the changes and clinical significance of γδT cells and their major functional subsets γδT1 and γδT17 cells in the peripheral blood of patients with systemic lupus erythematosus (SLE). Methods Ratios and absolute numbers of γδT cells, γδT1 cells and γδT17 cells in peripheral blood were detected by flow cytometry in 21 SLE patients (SLE) group and 16 healthy controls (HC group). Correlations of γδT cells, γδT1 cells and γδT17 cells in the peripheral blood with clinical indicators were analyzed by Pearson correlation analysis. Results In SLE patients, the percentage of γδT cells to lymphocytes in the peripheral blood was (2.30±1.10)%, which was significantly lower than (3.30±1.51)% in the HC group (P < 0.05); the percentage of γδT1 cells to lymphocytes in peripheral blood was (0.93±0.67)%, which was significantly lower than (2.09±1.21)% in the HC group (P < 0.001); the ratio of γδT17 cells to lymphocytes in the peripheral blood was (1.53±2.82)‰, which showed an increasing trend when compared to (0.18±0.12)‰ in the HC group (P>0.05). The absolute number of γδT cells in the peripheral blood in SLE patients was (2.88±2.18)×10⁴/mL, which was significantly lower than (7.96±3.96)×10⁴/mL in the HC group (P < 0.000 1); the absolute number of γδT1 cells in the peripheral blood in SLE patients was (1.20±1.17)×10⁴/mL, which was significantly lower than (5.05±3.04)×10⁴/mL in the HC group (P < 0.000 1); the absolute number of γδT17 cells in the peripheral blood in SLE patients was (1.03±1.08)×10³/mL, which was significantly higher than (0.40±0.24)×10³/mL in the HC group (P < 0.05). The proportion of γδT17 cells in the peripheral blood of SLE patients was positively correlated with the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) (r=0.59, P < 0.01) and erythrocyte sedimentation rate (r=0.65, P < 0.01), while the absolute number of γδT17 cells was positively correlated with the SLEDAI score (r=0.59, P < 0.01) and negatively correlated with the complement C3 level (r=-0.53, P < 0.05). Conclusion The significant decrease in the number of γδT cells in the peripheral blood of SLE patients is mainly due to the reduction of its γδT1 cell subset; γδT17 cells play an important role in the pathogenesis and activity of SLE.

Objective : To analyze the data related to the clinical outcome of preimplantation genetic testing (PGT) for double frozen , double biopsied blastocysts and double frozen , once biopsied blastocysts , in order to expand the existing data and provide some guidance for the clinical value and safety of PGT for frozen⁃thawed embryos . Methods : Retrospective analysis was made on the 38 PGT cycles of frozen⁃thawed blastocysts . According to the frequency of biopsy , cases in the study were divided into two groups : double frozen , double biopsy ( DFDB) group and double frozen , single biopsy ( DFSB) group . The freezing method was vitrification . Results : There were 24 patients in DFDB group , 34 blastocysts were not diagnosed in the last PGT cycle , 32 blastocysts survived after thawing , and the survival rate of thawed blastocysts was 94. 12% . After the second biopsy of these 32 blastocysts , genetic testing was performed , and all of them were definitely diagnosed , including 15 normal blastocysts (46. 88% ) and 17 abnormal blastocysts (53 . 13% ) . There were 14 patients in DFSB . The remaining 50 blastocysts in the last ICSI cycle were thawed and all blastocysts survived after thawing . Biopsy of these 50 blastocysts and genetic analysis showed that 47 blastocysts were diagnosed , including 9 normal blastocysts (18 . 00% ) , 28 abnormal blastocysts (56. 00% ) , 10 mosaic blastocysts (20. 00% ) , and 3 undiagnosed blastocysts (6. 00% ) . In DFDB group and DFSB group , 8 patients and 5 patients transferred the normal blastocystswhich all survivedafter thawing . There were 5 clinical pregnancies and 3 clinical pregnancies , respectively . One healthy live birth was obtained respectively in each group . Conclusion Acceptable pregnancy rate can be obtained whatever DFSB or DFDB blastocyst , which is of clinical value . However , due to the small sample size , we need to expand the sample size to further explore its safety .

Abstract Allergic diseases can occur in all systems of the body, covering the whole life cycle, from children to adults and to old age, can be lifelong onset and even fatal in severe cases. Children account for the largest proportion of the victims of allergic disease, Children s allergies start from scratch, ranging from mild to severe, from less to more, from single to multiple systems and systemic performance, so the prevention and treatment of allergic diseases in children is of great importance, which can not only prevent high risk allergic conditions from developing into allergic diseases, but also further block the process of allergy. At present, there is no consensus on the management system of allergic children in kindergartens and primary schools. The "Consensus on Allergy Management and Prevention in Kindergartens and Primary Schools", which includes the organizational structure, system construction and management of allergic children, provides evidence informed recommendations for the long term comprehensive management of allergic children in kindergartens and primary schools, and provides a basis for the establishment of the prevention system for allergic children.

Objective:To investigate the clinical diagnostic value of MRI retention enema cannula enhanced scanning in the high complex anal fistula.Methods:The clinical data of 60 anal fistula patients underwent surgery treatment from May 2020 to May 2022 in Affiliated Hospital of Shanxi University of Traditional Chinese Medicine were retrospectively analyzed. All patients underwent MRI plain scanning and enhanced scanning before operation. Compared with the surgical results, the difference between MRI plain scanning and enhanced scanning in the diagnosis of high complex anal fistula was compared.Results:All of the 60 patients successfully completed surgical treatment, and 58 cases internal orifices, 55 cases complex anal fistulas and 53 cases high anal fistulas were found intraoperatively. MRI plain scanning results showed 32 cases internal orifices, 46 cases complex anal fistulas and 42 cases high anal fistulas were found. MRI enhanced scanning results showed 54 cases internal orifices, 53 cases complex anal fistulas and 50 cases high anal fistulas were found. Based on surgical results, the coincidence rates of internal orifice, complex anal fistula and high anal fistula in MRI enhanced scanning were significantly higher than those in MRI plain scanning: 93.10% (54/58) vs. 55.17% (32/58), 96.36% (53/55) vs. 83.64% (46/55) and 94.34% (50/53) vs. 79.25% (42/53), and there were statistical differences ( χ2 = 21.76, 4.95 and 5.27; P<0.01 or <0.05). Conclusions:The MRI retention enema cannula enhanced scanning has obvious advantages in the diagnosis of high complex anal fistula, which provides scientific reference value for the diagnosis and operation of high complex anal fistula in clinic.

Objective:To investigate the changes and clinical significance of high mobility group protein B1 (HMGB1) in condyloma acuminatum (CA).Methods:Sixty four patients with initial CA(initial group) and 48 patients with recurrent CA(recurrent goup) treated in the Second Affiliated Hospital of PLA Air Force Military Medical University Hospital from January 2019 to November 2020 were included. In the same period, 31 patients who underwent circumcision and 19 female underwent sexual organ cosmetic plastic surgery were taken as the control group, and the normal foreskin and healthy vulva were collected. The expression of HMGB1 in wart was detected by real-time quantitative polymerase chain reaction(RT-PCR), and the expression of soluble apoptosis related factor ligand (sFasL), cell lymphoma-2 gene (Bcl-2), soluble apoptosis related factor (SFAS) and Survivin, caspase-3 were detected. At the same time, serum interleukin (IL) - 6, IL-17, IL-23 and tumor necrosis factor - α (TNF- α) were detected by enzyme-linked immunosorbent assay(ELISA).Results:The expression of HMGB1 mRNA in the warts of patients in the initial group, recurrence group and control group were 1.96 ± 0.20, 1.53 ± 0.14, 1.05 ± 0.11, there was statistical difference ( F = 15.20, P<0.05) ; the expression of HMGB1 mRNA in the warts of patients in the initial group was significantly higher than that in the recurrence group and the control group ( P<0.05), and the recurrence group was also significantly higher than that in the control group ( P<0.05). The mRNA expressions of sFas, Bcl-2, sFasL and caspase-3 in the warts of patients in the initial group were significantly lower than those in the recurrence group and the control group: 0.52 ± 0.08 vs. 0.82 ± 0.16, 1.10 ± 0.19; 0.50 ± 0.05 vs. 0.79 ± 0.13, 1.08 ± 0.21; 0.47 ± 0.06 vs. 0.81 ± 0.15, 1.01 ± 0.19; 0.35 ± 0.04 vs. 0.68 ± 0.09, 0.91 ± 0.16, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The expression of Survivin mRNA in the warts of patients in the initial group was significantly higher than those in the recurrence group and the control group: 2.14 ± 0.40 vs. 1.60 ± 0.27, 0.99 ± 0.18, P<0.05, and the recurrence group was also significantly higher than that in the control group ( P<0.05). The serum levels of TNF-α and IL-6 in the initial group were significantly lower than that in the recurrence group and the control group: (20.08 ± 1.95) μg/L vs. (26.93 ± 2.74), (37.65 ± 3.83) μg/L; (31.05 ± 3.24) μg/L vs. (38.13 ± 3.76), (45.98 ± 4.69) μg/L, P<0.05; and the recurrence group were also significantly lower than those in the control group ( P<0.05). The serum levels of IL-17 and IL-23 in the primary group were significantly higher than those in the recurrence group and the control group: (423.71 ± 28.68) ng/L vs. (384.26 ± 21.70) and (335.43 ± 19.65) ng/L; (289.50 ± 18.53) ng/L vs. (251.07 ± 15.96) and (214.67 ± 13.20) ng/L, P<0.05; and the recurrence group were also significantly higher than those in the control group ( P<0.05). The results of correlation analysis showed that the mRNA expression of HMGB1 in the warts of CA patients were negatively correlated with the mRNA expression of caspase-3, sFas, Bcl-2 and sFasL in the warts ( r = - 0.602, - 0.734, - 0.692, - 0.717, P<0.05), and was positive correlation with Survivin mRNA expression ( r = 0.645, P<0.05); and were positive correlation with the contents of IL-17 and IL-23 in serum ( r = 0.673, 0.685, P<0.05), and negatively correlated with the contents of TNF-α and IL-6 ( r = - 0.698, - 0.764, P<0.05). Conclusions:HMGB1 is obviously abnormal in the warts of patients with condyloma acuminatum, and is closely related to apoptosis, immune and inflammation-related factors, and may be jointly involved in the occurrence and recurrence of CA.

Objective:To investigate the clinical efficacy of vitamin D drops combined with insulin aspart in the treatment of gestational diabetes mellitus (GDM), and the effect of vitamin D drops on the serum levels of 1, 25 dihydroxyvitamin D 3 [1, 25(OH) 2D 3] and retinol binding protein 4 (RBP4). Methods:A total of 94 GDM patients admitted to the Baoding Second Central Hospital from March 2019 to March 2021 were selected and randomly divided into an observation group and a control group with 47 cases each using a random number table method. The control group received subcutaneous injection of insulin aspartate for treatment, while the observation group received oral vitamin D drops for treatment. After 4 weeks of continuous treatment, the blood glucose control effect and adverse reactions were observed in both groups. The glucose metabolism indicators of the two groups were compared before and after treatment, including fasting blood glucose (FPG), 2-hour postprandial blood glucose (2-hour PG), insulin resistance index (HOMA-IR), and pancreatic islets β Cell Function Index (HOMA-β) and serum levels of 1, 25(OH) 2D 3, RBP4, lipoprotein related phospholipase A2 (Lp-PLA2), and vascular cell adhesion molecule-1 (VCAM-1). All patients were followed up until the end of pregnancy, and Statistical analysis was conducted on the adverse outcomes of two groups of mothers and infants. Results:The time to reach the standard for FPG and 2-hour PG in the observation group, as well as the time for both to reach the standard were significantly shorter than those in the control group (all P<0.05). There was no statistically significant difference in the incidence of dawn phenomenon and hypoglycemia between the observation group and the control group (all P>0.05). After treatment, FPG and 2-hour PG in both groups were significantly reduced compared to those before treatment (all P<0.05); However, after treatment, there was no statistically significant difference between the groups (all P>0.05). Compared with before treatment, HOMA-IR in both groups significantly decreased (all P<0.05), All HOMA- β significantly increased (all P<0.05); And the improvement was more significant in the observation group (all P<0.05). After treatment, the serum levels of 1, 25(OH) 2D 3 in the observation group significantly increased compared to that before treatment ( P<0.05), but there was no significant change in the control group before and after treatment ( P>0.05). After treatment, the levels of serum RBP4, Lp-PLA2, and VCAM-1 in both groups significantly decreased compared to those before treatment (all P<0.05); After treatment, the serum levels of RBP4, Lp-PLA2, and VCAM-1 in the observation group were lower than those in the control group (all P<0.05). The incidence of adverse maternal and neonatal outcomes in the observation group was 14.9%(7/47) and 10.6%(5/47), respectively, which were lower than those in the control group [34.0%(16/47) and 27.7%(13/47)] (all P<0.05). There were 8 cases of hypoglycemia in 94 patients (3 in the observation group and 5 in the control group), and no other adverse events occurred. Conclusions:The combination of vitamin D drops and insulin aspartate in the treatment of GDM can safely, effectively, quickly, and steadily control patients′ blood sugar, improve IR and pancreatic islets β The effect of cell function on reducing the incidence of adverse maternal and fetal outcomes may be related to increasing serum 1, 25(OH) 2D 3 levels and down-regulating the expression levels of serum RBP4, Lp-PLA2, and VCAM-1.

Objective:To investigate the overall clinical performance of nursing students and analyze its influencing factors to provide evidence for the improvement.Methods:A questionnaire survey was carried out among 288 nursing students from Qilu Hospital of Shandong University in March to May 2020. The survey included a general data questionnaire, Holistic Clinical Assessment Tool, Clinical Learning Environment, Supervision and Nurse Teacher evaluation scale, Medical Student Safety Attitudes and Professionalism Survey of nursing students. The factors associated with overall clinical performance of nursing students were analyzed by t test, analysis of variance, Pearson correlation analysis and multiple linear regression. Results:The total score of Holistic Clinical Assessment Tool was 114.95 ± 22.91. The total score of Clinical Learning Environment, Supervision and Nurse Teacher evaluation scale was 156.70 ± 26.98. The total score of Medical Student Safety Attitudes and Professionalism Survey was 123.65 ± 31.00. The influencing factors included social practice experience and clinical learning environment, which accounted for 28.0% of the total variation of overall clinical performance of nursing students.Conclusions:Educators and managers of nursing students' schools and internship hospitals should actively explore the educational reform practice to promote the improvement of clinical ability, so as to promote the improvement of the overall clinical ability of nursing students.