Objective To generate minichromosome maintenance protein 2(MCM2)gene knockout cervi-cal cancer HeLa cell lines using inducible CRISPR/Cas9 technology,and to explore the effect of MCM2 on DNA replication and replication stress.Methods The inducible CRISPR/Cas9 system,TLCV2,was used to construct MCM2 knockout HeLa cell lines.And the cell lines were divided into control group(Control),knockout group 1(KO1),and knockout group 2(KO2).Western blot,Edu incorporation experiment,real-time quantitative PCR(qPCR),immunofluorescence and MTT assay were used to analyze the effects of MCM2 knockout on DNA replica-tion and replication stress induced by hydroxyurea.Results The CRISPR/Cas9 system successfully knocked out the MCM2 gene after induction,and MCM2 knockout affected the stability of MCM2-7 complex.Compared with the control cells,MCM2 knockout cells had a dramatic decrease in the capacity of DNA replication,and the mRNA levels of Cyclin A1,Cyclin E1 and CDK4.Under DNA replication stress,MCM2 knockout cells decreased cell viability,DNA damage repair capacity,and increased genomic instability compared with control cells.Conclusion Knockout of MCM2 gene reduces the DNA replication capacity of HeLa cells under normal conditions and cell viability under replication stress.This study successfully generates MCM2 gene inducible knockout HeLa cell lines,laying the foundation for further research on the role and biological function of MCM2 gene in the occurrence and progression of cervical cancer.

BACKGROUND:At present,measurement osteotomy technique and gap balance technique are the two main surgical methods in total knee arthroplasty.Both methods have their advantages and disadvantages.By adjusting the osteotomy angle,the gap balance technique can reduce the release of soft tissue and obtain a more balanced flexion and extension space.The clinical efficacy of gap balance technique is superior,but this surgical method lacks tools and is easily influenced by the surgeon's surgical experience and subjective judgment,with more errors.Measurement osteotomy technique has a short learning curve,but it relies on anatomic markers and is prone to many complications due to inaccurate positioning.In recent years,many instrument companies have successively developed and launched tools that can improve the quality of surgery.However,there are still few reports about the tool. OBJECTIVE:To compare the clinical efficacy of gap balance technique and measurement osteotomy technique under Offset Repo-Tensor in total knee arthroplasty,and explore the value of Offset Repo-Tensor in total knee arthroplasty. METHODS:The medical records of 85 patients with total knee arthroplasty were collected and divided into two groups according to the operation method.Group A consisted of 44 patients who underwent total knee arthroplasty using a gap balance technique combined with Offset Repo-Tensor.Group B consisted of 41 patients who underwent total knee arthroplasty using measurement osteotomy technique.Surgical time,American Knee Society Score,knee range of motion,changes in lower limb alignment,and postoperative complications were compared between the two groups to evaluate the clinical efficacy of gap balance technique combined with Offset Repo-Tensors in total knee arthroplasty. RESULTS AND CONCLUSION:(1)All 85 patients were followed up.(2)Postoperative knee range of motion in both groups was higher than that before surgery(P<0.05),and the improvement in group A was more significant than that in group B(P<0.05).(3)American Knee Society Score in both groups after surgery was higher than that before surgery(P<0.05),and the American Knee Society Score in group A was higher than that in group B at 2 weeks,1,3 months,and the last follow-up(P<0.05).(4)The operation time of group A was slightly shorter than that of group B,but the difference was not significant(P>0.05).(5)The lower limb alignment in both groups was improved after operation,but there was no significant difference between the two groups(P>0.05).(6)The complication rate of group A(2%)was lower than that of group B(7%),but there was no significant difference between the two groups(P>0.05).(7)It is indicated that compared with the measurement osteotomy technique,the combination of Offset Repo-Tensor and gap balance technique can elevate the clinical effect and improve the function and motion range of the knee joint.

Fluid resuscitation after burns is very crucial to the survival and prognosis of children. Although the principles of fluid resuscitation in children are similar to those in adults, the unique physiological needs of children must be fully considered to prevent insufficient or excessive fluid resuscitation. In clinical practice, due to the popularization of various monitoring techniques, insufficient fluid resuscitation can be observed in time, but excessive fluid resuscitation is often overlooked. Excessive fluid resuscitation can lead to fluid extravasation, increasing the probability of pediatric burn complications and poor prognosis. This review focuses on the unique pathophysiological characteristics of children with burns and how to avoid excessive fluid resuscitation in order to provide reference for rational fluid rehydration.

Objective:To investigate the prevalence and related influencing factors of thyroid nodules in different occupational groups in Shijiazhuang City.Methods:Different occupational groups who underwent physical examination at the Second Hospital of Shijiazhuang City from May to December 2015 and had lived there for more than 10 years were selected as the investigation subjects, including workers, farmers, science and technology education personnel, medical and health personnel, civil servants and individual businessmen. Questionnaire survey and thyroid examination were conducted, blood lipid, blood glucose and thyroid hormone levels were tested, and related influencing factors of thyroid nodules were analyzed.Results:A total of 1 440 different occupational subjects were surveyed, and the prevalence of thyroid nodules was 20.35% (293/1 440). The prevalence of thyroid nodules in females (22.59%, 159/704) was significantly higher than that in males (18.21%, 134/736), and the difference was statistically significant (χ 2 = 4.26, P = 0.039). There was a statistically significant difference in the prevalence of thyroid nodules in different age groups (χ 2 = 73.87, P < 0.001), with the highest prevalence in the 70 - 80 age group (43.75%, 21/48). There was a statistically significant difference in the prevalence of thyroid nodules among different occupational groups (χ 2 = 36.56, P < 0.001), with the highest prevalence in individual businessmen (31.84%, 78/245). By univariate analysis, the prevalence of thyroid nodules in people with good economic status [27.45% (42/153) vs 19.48% (248/1 273)], radiation exposure history [26.39% (109/413) vs 17.92% (184/1 027)], and diabetes history [33.71% (30/89) vs 19.47% (263/1 351)] was higher (χ 2 = 5.35, 13.06, 10.45, P < 0.05). Conclusions:The prevalence of thyroid nodules in occupational group in Shijiazhuang City is relatively high, and the prevalence of thyroid nodules in females is significantly higher than that in males. Economic status, radiation exposure history and diabetes history are the main influencing factors.

Premature ovarian insufficiency(POI),also known as"ovarian insufficiency",has an incidence of 1％～5％.The incidence has been on the rise in recent years,seriously affecting women's physical and mental health and quality of life.At present,the cause and mechanisms of POI are still unclear,and the method and applications of model construction are also confusing.Most models have some shortcomings in pertinence and stability.The limitations greatly limit research into the clinical diagnosis and treatment of POI.This paper summarizes and discusses the etiology and pathogenesis of POI and the construction of POI animal models to provide a comprehensive reference for those studying POI.

Objective:To investigate the current situation of moral resilience of elderly care workers in elderly care institutions and analyze its influencing factors.Methods:A total of 225 elderly care workers from 7 private elderly care institutions in Zhangzhou City were selected from Sep 2023 to Nov 2023 by convenience sampling.The general information questionnaire,Rushton moral resilience scale(RMRS),nurses'moral dilemma scale,nurses'moral courage scale,social support rating scale(SSRS),and hospital ethical atmosphere scale were used to investigate.Spearman correlation analysis was used to analyze the correlation of moral resilience with moral dilemma,moral courage,social support and ethical climate.Generalized linear regression was used to analyze the main factors affecting the moral resilience of elderly care workers.Results:The total score of moral resilience of elderly care workers in elderly care institutions was 41.00(35.00,47.00).Spearman correlation analysis showed that moral resilience was positively correlated with moral dilemma score and moral courage score(r=0.364 and 0.279,respectively,P＜0.01),and weakly correlated with social support score and ethical atmosphere score(P＞0.05).Generalized linear regression showed that moral courage and moral dilemma were the influencing factors of moral resilience of elderly care workers in elderly care institutions(β=0.041 and 0.216,respectively,P＜0.01).Conclusions:The moral resilience of elderly care workers in elderly care institutions is at a medium level.Managers of elderly care institutions should formulate corresponding measures to improve the ability of elderly care workers to deal with moral dilemmas,enhance the inherent potential of moral courage,and then improve the moral resilience of elderly care workers.

Objective:To explore the influencing factors of severe pneumonia in children with respiratory syncytial virus (RSV) infection.Methods:A retrospective case-control study was used to collect 210 children with RSV infected pneumonia admitted to Hebei Children′s Hospital from October 2017 to October 2020. Among them, 70 children with severe pneumonia were included in the severe pneumonia group, and 140 children with common pneumonia were included in the common pneumonia group; the baseline data and relevant laboratory indicators of the two groups were compared; Logistic regression was used to analyze the influencing factors of severe pneumonia in children infected with RSV.Results:The proportions of wheezing, congenital heart disease, respiratory failure, heart failure and pleural effusion of children in severe pneumonia group were higher than those in common pneumonia group, and the forced vital capacity (FVC) and forced expiratory volume in the first second (FEV 1) were lower than those in common pneumonia group (all P<0.05); the levels of C-reactive protein (CRP), CD8 + cells, RSV load and Beclin-1 in severe pneumonia group were higher than those in common pneumonia group, and the levels of CD4 + cells and 1, 25-dihydroxyvitamin D [1, 25-(OH) 2D] were lower than those in common pneumonia group (all P<0.05). After treatment, the levels of CRP, CD8 + cells and Beclin-1 in children with severe pneumonia were lower than those before treatment, and the levels of CD4 + cells and 1, 25-(OH) 2D were higher than those before treatment (all P<0.05). Multiple regression model analysis was established. The results showed that congenital heart disease, high CRP level, high CD8 + cells, high RSV load and high Beclin-1 level were risk factors for severe pneumonia in children with RSV infected pneumonia (all OR>1, P<0.05), and high CD4 + cells and 1, 25-(OH) 2D level were protective factors (all OR<1, P<0.05). Conclusions:Severe pneumonia in children with RSV infected pneumonia may be affected by congenital heart disease, CRP, CD4 + cells, CD8 + cells, 1, 25-(OH) 2D, RSV load and Beclin-1.

Objective:To explore the application of collaborative nursing combined with medical integration model in minimally invasive surgery for pediatric urinary calculi, to provide clinical guidance for minimally invasive surgical nursing of urinary calculi in children.Methods:From December 2020 to July 2022, 90 children who underwent minimally invasive surgery for urinary calculi in Children′s Hospital Affiliated to Zhejiang University Medicine College were selected as subjects by quasi experiment. According to the time sequence of enrollment, they were divided into observation group ( n = 45) and control group ( n = 45). The control group was given routine nursing mode nursing, and the observation group was given collaborative nursing combined with medical integration mode nursing. The clinical related indicators, pain, anxiety and complications were compared between the two groups. Results:In the observation group, the operation time, the amount of bleeding during operation, the first time to get out of bed after operation, the rate of stone removal immediately after operation and the rate of stone removal 1 month after operation were (75.48 ± 5.92) min, (168.75 ± 17.28) ml, (6.12 ± 1.05) h, 84.44%(38/45) and 97.77%(44/45), respectively. The control group was (78.39 ± 6.75) min, (178.30 ± 19.25) ml, (6.84 ± 1.23) h, 64.44 % (29/45) and 82.22 %(37/45), respectively, the differences were statistically significant ( t/ χ2 = 2.17, 2.48, 2.99, 4.73, 6.05, all P<0.05). The Face, legges, Activity, Cry, Consolability Behavioral Tool scores of the observation group at 1 d, 3 d and 7 d after operation were 6.28 ± 1.21, 5.35 ± 1.37 and 2.87 ± 0.65, respectively, and those of the control group were 7.05 ± 1.55, 5.95 ± 1.25 and 3.21 ± 0.73, respectively, the differences were statistically significant ( t = 2.63, 2.17, 2.33, all P<0.05). The Modified Yale Preoperative Anxiety Scale and Frankl Compliance Scale Scores in the observation group at the day before discharge hospital were 55.28 ± 5.52, 3.60 ± 0.40, while in the control group were 60.02 ± 8.56, 2.82 ± 0.61, the differeces were significat ( t = 3.12, 3.02, both P<0.05). The incidence of complications was 2.22 %(1/45) in the observation group and 15.56 %(7/45) in the control group, the difference was significant ( χ2 = 4.94, P<0.05). Conclusions:The application of collaborative nursing combined with medical care integration model in minimally invasive surgery of urinary calculi in children has a significant effect.

Objective:To comparae the accuracy of imaging examination in preoperative parathyroid localization, and the safety and effect of total parathyroidectomy on secondary hyperparathyroidism complicated by chronic renal failure.Method:A total of 257 patients with secondary hyperparathyroidism who underwent total parathyroidectomy at the Department of General Surgery , Zhongshan Hospital, Dalian University from Mar 2012 to Mar 2022 were analyzed retrospectively.Result:Six hundred fourty parathyroid glands were found by color Doppler ultrasound and 954 parathyroid glands were by enhanced CT before the operation. Among them, the number of patients with accurate location of all 4 parathyroid glands by color Doppler ultrasound was 54, while that by enhanced CT was 216. The parathyroid detection rate by enhanced CT was significantly higher than that of color Doppler ultrasound ( χ2=325.480, P<0.001), and the accuracy rate was significantly higher tnan that of color ultrasound ( χ2=215.146, P<0.001). The average values of iPTH before operation, on the day after operation, on the 1st day and 7th day after operation were (1 880±890), (137±82), (66±46) and (34±23) pg/ml, respectively. The clinical symptoms of all patients were significantly relieved. Conclusions:Enhanced CT is superior to color Doppler ultrasound in the overall detection rate and individual localization accuracy of preoperative parathyroid localization. Total parathyroidectomy is safe and reliable in the treatment of secondary hyperparathyroidism complicated by chronic renal failure.

Objective:To summarize the clinical phenotype and genotypic characteristics of children with truncation variation in SMC1A gene. Methods:The clinical data of a child with late-onset cluster seizures caused by truncation variation in SMC1A gene diagnosed in February 2021 in Children′s Hospital Affiliated to Zhengzhou University were collected. The relevant literature was reviewed to summarize the clinical characteristics. Results:The proband was a 5-year-old girl, presenting with first seizure at the age of 5 and cluster seizures. She had poor response to multiple antiepileptic drugs, and had normal neurodevelopment before seizures. Whole exome sequencing results revealed a spontaneous heterozygous nonsense variation c.55C>T in SMC1A gene, causing a nonsense variant in the amino acid sequence p.Gln19Ter(p.Gln19 *), which has not been reported. There were a total of 14 relevant literatures, and there were in total 32 cases with truncation variation in SMC1A gene including this case. All children were female and 30 children had early-onset intractable epilepsy, and first seizure median age was 5 months (range: 4 weeks to 40 months); 78.1% (25/32) of them had cluster seizures; 93.8% (30/32) had mental retardation; Cornelia de Lange syndrome clinical score in 68.8% (22/32) of them was≥4. The truncation variations in SMC1A gene of 31 children were de novo, and there were 16 children with frameshift variation (16/32), 12 children with nonsense variation [12/32; 3 children (9.4%, 3/32) with c.2923C>T], 4 children with splice variation (4/32). Conclusions:This study further expands the clinical phenotype and genotype of cases with truncation variation in SMC1A gene. Case presenting with female late-onset cluster seizures has not been reported in China, and genetic testing can be beneficial for early diagnosis of hereditary epilepsy and precision treatment.