Objective To investigate the effect of anti-angiogenic drug Sitravatinib combined with poly(adenosine diphosphate[ADP]-ribose)polymerase inhibitor(PARPi)Niraparib on mucosal melanoma cell lines and its possible mechanism.Methods The CCK8 assay was used to detect the maximal half inhibitory concentration(IC50)of Sitravatinib and Niraparib targeting at mucosal melanoma(MM)cell lines.CompuSyn was used to detect the Combination Index(CI)in different concentrations of the two drugs.Flow cytometry was used to detect the effect of drugs on cell apoptosis.Colony formation assay was used to detect the effect of drugs on cell proliferation.Western blot was used to detect the protein expressions and RT-qPCR was used to detect mRNA expression.Results CI values was respectively 0.19 and 0.15 for Sitravatinib(2 μmol/L)in combination with Niraparib(20 μmol/L)in a human vaginal maligant melanoma cell line(HMVII)and a metastasis inguinal lymph node of vulvar malignant melanoma cell line(GAK).Compared with the control group and single-drug groups,the cell proliferation of the combination group was significantly reduced(P＜0.05 or P＜0.01 or P＜0.001).The cell apoptosis rate was signifi-cantly increased(P＜0.01 or P＜0.001).The protein and mRNA expression of apoptosis-related biomarkers signifi-cantly increased(P＜0.001);In addition,the protein and mRNA expression of cell autophagy biomarkers signifi-cantly increased(P＜0.01 or P＜0.001).The protein expression of DNA damage marker significantly increased.Moreover,compared with the control group,The expression of radiation sensitive protein 51(RAD51)recombinase in the Sitravatinib single-drug group and combination group significantly reduced.As the dose of Sitravatinib gradu-ally increased up to 2 μmol/L,the protein and mRNA expression of RAD51 both significantly reduced(P＜0.05 or P＜0.01),the mRNA expression of BRCA1 and BRCA2 also significantly reduced(P＜0.05 or P＜0.01 or P＜0.001).Conclusions Sitravatinib combined with Niraparib inhibits the proliferation of mucosal melanoma cells,induces cell apoptosis and promotes autophagy.The mechanism is potentially related to the inhibition of ho-mology-dependent recombination repairs(HRR).

Objective A competing endogenous RNA (ceRNA) regulatory network associated with long non-coding RNA (lncRNA) specific for lung adenocarcinoma (LUAD) was constructed based on bioinformatics methods, and the functional mechanism of actinfilament-associated protein 1-antisense RNA1 (AFAP1-AS1) in LUAD was analyzed, in order to provide a new direction for the study of LUAD therapeutic targets. Methods The gene chip of LUAD was downloaded from the Gene Expression Omnibus (GEO), and lncRNA and mRNA with differential expression between LUAD and normal tissues were screened using GEO2R online software, and their target genes were predicted by online databases to construct ceRNA networks and perform enrichment analysis. In cell experiments, AFAP1-AS1 was genetically knocked down and siRNA was constructed and transfected into LUAD cells A549 by cell transfection. CCK8, transwell, scratch assay and flow cytometry were used to detect the ability of cells to proliferate, invade, migrate and apoptosis. Results A total of 6 differentially expressed lncRNA and 494 differentially expressed mRNA were identified in the microarray of LUAD. The ceRNA network involved a total of 6 lncRNA, 22 miRNA, and 55 mRNA. Enrichment analysis revealed that mRNA was associated with cancer-related pathways. In cell assays, knockdown of AFAP1-AS1 inhibited cell proliferation, invasion, and migration, and AFAP1-AS1 promoted apoptosis. Conclusion In this study, we construct a lncRNA-mediated ceRNA network, which may help to further investigate the mechanism of action of LUAD. In addition, through cellular experiments, AFAP1-AS1 is found to have potential as a therapeutic target for LUAD.

Objective:To evaluate the efficacy of allogeneic hematopoietic stem cell transplantation for the treatment of combined immunodeficiency (CID) and explore prognostic risk factors.Methods:In this retrospective cohort study, clinical characteristics, laboratory tests and prognosis of 73 CID children who underwent allogeneic hematopoietic stem cell transplantation from February 2014 to April 2022 in the Children′s Hospital of Fudan University were analyzed. Based on the subtypes of diseases, all patients were divided into severe combined immunodeficiency disease (SCID) group and other CID group. Based on the types of donors, all patients were divided into matched sibling donor group, matched unrelated donor group, unrelated cord blood group, and haploidentical donor group. Kaplan-Meier method and Log-Rank test were used to analyze the survival data. Cox regression was used to analyze prognostic factors.Results:Among the 73 patients, there were 61 (84%) males and 12 (16%) females. Fifty-five (75%) patients were SCID, and 18 (25%) patients were other CID. Donor source included 2 (3%) matched sibling donors (MSD), 3 (4%) matched unrelated donors (MUD), 64 (88%) unrelated cord blood (UCB), and 4 (5%) haploidentical donors. The age at transplant was 10.7 (5.9, 27.5) months, and the follow-up time was 36.2 (2.5, 62.9) months. The 3-year overall survival rate of 73 patients with CID was (67±6) %. No significant difference was found in the 3-year overall survival rates between patients with SCID (55 cases) and other CID (18 cases) ((64±7) % vs. (78±10) %, χ2=1.31, P=0.252). And no significant difference was found in the 3-year overall survival rates among patients who received MSD or MUD (5 cases), UCB (64 cases), and haploidentical donor (4 cases) transplant (100% vs. (66±6)% vs. (50±25) %, χ2=2.30, P=0.317). Cox regression analysis showed that the medical history of sepsis ( HR=2.55, 95% CI 1.05-6.20, P=0.039) and hypoalbuminemia at transplant ( HR=2.96, 95% CI 1.14-7.68, P=0.026) were independent risk factors for the prognosis of allogeneic hematopoietic stem cell transplantation in pediatric patients with CID. Conclusions:Allogeneic hematopoietic stem cell transplantation is an effective treatment for CID. The medical history of sepsis and hypoalbuminemia at transplant were risk factors for prognosis. Enhancing infection prevention and nutritional intervention before transplant can improve patient prognosis.

Objective:To investigate the efficacy and safety of protein A immunoadsorption (PAIA) combined with rituximab (RTX) in highly sensitized patients who underwent haplo-hematopoietic stem cell transplantation (haplo-HSCT) .Methods:The clinical data of 56 highly sensitized patients treated with PAIA and RTX before haplo-HSCT at the First Affiliated Hospital of Soochow University and Soochow Hopes Hematonosis Hospital between March 2021 and June 2023 were retrospectively analyzed. The number of human leukocyte antigen (HLA) antibody types and the mean fluorescence intensity (MFI), humoral immunity, adverse reactions during adsorption, and survival within 100 days before and after adsorption were measured.Results:After receiving the PAIA treatment, the median MFI of patients containing only HLA Ⅰ antibodies decreased from 7 859 (3 209-12 444) to 3 719 (0-8 275) ( P<0.001), and the median MFI of HLA Ⅰ+Ⅱ antibodies decreased from 5 476 (1 977-12 382) to 3 714 (0-11 074) ( P=0.035). The median MFI of patients with positive anti-donor-specific antibodies decreased from 8 779 (2 697-18 659) to 4 524 (0–15 989) ( P<0.001). The number of HLA-A, B, C, DR, and DQ antibodies in all patients decreased after the PAIA treatment, and the differences were statistically significant (A, B, C, DR: P<0.001, DQ: P<0.01). The humoral immune monitoring before and after the PAIA treatment showed a significant decrease in the number of IgG and complement C3 ( P<0.001 and P=0.002, respectively). Forty-four patients underwent HLA antibody monitoring after transplantation, and the overall MFI and number of antibody types decreased. However, five patients developed new antibodies with low MFI, and nine patients continued to have high MFI. The overall survival, disease-free survival, non-recurrent mortality, and cumulative recurrence rates at 100 days post-transplantation were 83.8%, 80.2%, 16.1%, and 4.5%, respectively. Conclusions:The combination of PAIA and RTX has a certain therapeutic effect and good safety in the desensitization treatment of highly sensitive patients before haplo-HSCT.

The patient, an 84-year-old man, was admitted to the hospital with "low back pain with limitation of movement for more than half a year". Admission examination: mild kyphotic deformity of the spine, significant tenderness and percussion pain in the lower back, bilateral lower limb muscle strength graded 5, normal skin sensation. Lumbar MRI and CT revealed a compressive fracture of the L 4 vertebra. Dual-energy X-ray absorptiometry (DEXA) indicated a bone mineral density T-score of -2.6, suggesting osteoporosis. Admission diagnosis: osteoporotic compressive fracture of the L 4 vertebra. The patient underwent thorough examinations to exclude surgical contraindications. On the fourth day of admission, the patient underwent percutaneous vertebroplasty of the L 4 vertebra. At the end of the operation, the patient became unresponsive, with a blood pressure drop to 94/63 mmHg and oxygen saturation falling to 80%. Cranial CT showed multiple punctate gas density shadows within the brain. Lumbar CT revealed gas accumulation in the soft tissue adjacent to the lumbar spinous processes, localized intraductal gas, and punctate gas density shadows within the vessels in both groin areas. The diagnosis was intracranial arterial gas embolism. The patient's condition deteriorated further, with loss of consciousness, neck stiffness, increased muscle tone of both lower limbs, and positive Babinski's sign on both sides. Symptomatic treatments included brain protection, maintaining cerebral perfusion, and improving collateral cerebral circulation, but the patient did not regain consciousness. The patient developed a pulmonary infection one month postoperatively and died three months postoperatively due to respiratory failure. This case highlights the potential risk of gas embolism during vertebroplasty. Measures to reduce such complications should be implemented, such as minimizing the duration of venous blood-air contact, pre-filling the cannula with saline to reduce the venous blood-air interface, and appropriately increasing venous pressure to reduce the risk of gas entry. It is recommended to use smaller diameter catheters. For patients with pre-existing cardiac conditions or elderly patients, preoperative cardiac Doppler ultrasound should be performed to exclude anatomical abnormalities such as patent foramen ovale.

Objective:To study the effects of plan-do-check-action (PDCA) cycle in quality improvement of neonatal resuscitation.Methods:From 2016 to 2020, the clinical data of neonates born in our hospital were analyzed. Neonates born during 2016 to 2017 were pre-PDCA group and neonates born during 2018 to 2020 were post-PDCA group. PDCA quality improvement included step-by-step, high-frequency and low-dose training, strengthening teamwork and adding equipment.Results:A total of 7 728 live-birth neonates were delivered before PDCA with 319 cases (4.1%) of asphyxia. 10 174 live-birth neonates were delivered after PDCA with 422 cases (4.1%) of asphyxia. The asphyxia rates showed no significant difference between the two groups ( P>0.05). The incidences of severe asphyxia before and after PDCA were both 0.8% without significant difference ( P>0.05). The success rates of resuscitation for severe asphyxia before and after PDCA was 27.9% and 44.9%, respectively, and the differences were statistically significant ( P<0.05). The mortality rates within 7 d before and after PDCA were 0.5‰ and 0.1‰ respectively, without significant differences ( P>0.05). Conclusions:The implementation of PDCA cycle and step-by-step, high-frequency, low-dose neonatal resuscitation training can effectively improve the success rate of resuscitation in newborns with severe asphyxia.

Objective:To explore the construction and implementation effect of clinical-thinking patterned curriculum for parallel graduate students with "running through clinical diagnosis and treatment process."Methods:In this study, 94 Batch 2016-2017 graduate students with clinical medical professional degree in Shanxi Provincial People's Hospital were selected as the research subjects. Among them, 48 Batch 2017 parallel graduate students were selected as the experimental group. The curriculum of clinical-thinking patterned training of "running through the clinical diagnosis and treatment process" was used. In addition to participating in the degree courses and residency courses, a series of training to improve the clinical-thinking ability were introduced. And 46 Batch 2016 parallel graduate students were divided in the control group, using a traditional curriculum and only participating in degree courses and residency courses. The differences among mini-clinical evaluation exercise (Mini-CEX) scores, direct observation of procedural skills (DOPS) scores, objective structured clinical examination (OSCE) scores and students' satisfaction were compared by t test, chi-square test and repeated measures analysis of variance. Results:The Mini-CEX scores showed the average scores of other aspects except humanistic care were higher than those of the control group, and the score of communication skills was significantly higher than that of the control group [(6.55±0.98) vs. (5.77 ±1.12)], with significant differences ( t=3.62, P<0.001). In the DOPS scores, except for the skills of communication with patients, ability to consider patient's feelings and practice of occupational literacy, the average scores of other aspects of the experimental group were higher than those of the control group, and the real operation-ability score of clinical skills was significantly higher than that of the control group [(6.38 ± 1.38) vs. (5.53±1.23)], with a significant difference ( t=3.12, P=0.002). In terms of the outpatient receiving station, the emergency treatment station, the clinical thinking station①, the clinical thinking station②, the specialist skill station, the auxiliary examination station and the case writing station, their OSCE scores at different stages in the experimental group were higher than those in the control group, and in terms of clinical thinking station①, the score of the experimental group was significantly higher than that of the control group, with a significant difference ( F=6.51, P=0.012). The satisfaction rate of the experimental group in terms of curriculum was higher than that in the control group except for the future career development, and in improving logical-thinking ability, the score of the experimental group was significantly higher than that of the control group, with a significant difference ( χ2=19.18, P<0.001). Conclusion:The curriculum of clinical-thinking ability with "running through the clinical diagnosis and treatment process" can enhance the clinical practical ability of parallel graduate students, making them meet the residential academic standards as soon as possible and effectively promoting the training quality of the students.

Hematopoietic stem cell transplantation related thrombotic microangiopathy (TA-TMA) is a clinical syndrome characterized by microvascular hemolytic anemia, thrombocytopenia and involvement of end organ.The pathogenesis of TA-TMA involves vascular endothelial cell injury and abnormal activation of complement system.The risk factors include conditioning regimens, graft-versus-host disease, immunosuppressants, infection and HLA compatibility.Timely diagnosis and early initiation of appropriate treatment are essential to prevent multiple organ dysfunction and eventual death.At present, the lack of clinical evidence of TA-TMA has led to the fact that its diagnostic criteria and treatment have not been unified.The diagnosis and treatment suggestions put forward by domestic and foreign experts are intended to help clinicians evaluate potential TA-TMA, establish diagnosis in time, and give reasonable treatment and management.

Objective:To explore the clinical characteristics of Diamond-Blackfan anemia (DBA) in children caused by RPL5 gene mutation, thus improving the understanding of the etiology of DBA.Methods:The clinical data and sequencing results of a child with DBA caused by RPL5 gene mutation treated in the Children′s Hospital of Fudan University were analyzed.In addition, through literature review of reported DBA cases at domestic and home, summarized the clinical features of DBA.Results:The patient was an 8-year-old male child.Bone marrow puncture examination of the child showed DBA, and a heterozygous mutation of RPL5 gene c. 657C>G, p.Y219X was identified for the first time in the DBA case.A total of 47 cases of DBA were retrieved from the online databases plus the one reported in this study (48 cases in total), and their clinical features were summarized as follows: the incidence of DBA was similar in men and women.The number of DBA patients in Asia was lower than that in Europe and the United States.DBA was mainly a sporadic disease.Among the exon mutations in European and American cases of DBA, 43.0% of them had mutations in Exon3.The malformation rate of DBA patients with RPL5 mutation was 81.3% (39/48 cases, excluding short stature cases), which was higher than that of patients with other mutation types.The response rate of glucocorticoid therapy for DBA was 46.0%, which was lower than that of the overall response rate.Conclusions:chr1: 93303142(c.657 C>G, p.Y219X) is a newly detected mutation of RPL5 gene in the DBA case, which expands the pathogenic gene spectrum of DBA.Patients with RPL5 mutation have higher rates of teratogenicity and multiple teratogenicity, and a lower response rate to hormone therapy.

Objective:To investigate the serum measles antibody in children with tumor and to provide the clinical evidence for measles vaccination strategy for this special population.Methods:From January 2016 to December 2018, the blood samples of children who were diagnosed with hematological malignancy or solid tumors and received chemotherapy in the Department of Hematology or Oncology Surgery of Children′s Hospital of Fudan University were collected. Enzyme-linked immunosorbent assay was used to quantitatively detect the level of measles IgG antibody, and dynamically monitor the changes of measles antibody level during chemotherapy. Kruskal-Wallis test and chi-square test were used for statistical analysis.Results:A total of 441 children with tumors were enrolled, with the positive rate of measles antibody of 79.1%(349/441), and only 43.3%(191/441) of children had the protective level of IgG antibody. There was a statistically significant difference of the antibody protection rate in children aged0.05). The protection rate of serum measles antibody in children with hematological malignancy and solid tumor were 45.6%(78/171) and 41.9%(113/270), respectively, and there was no statistically significant ( P>0.05). There were 16.3%(16/98) of children who were observed to lose the pre-existing protective antibody during chemotherapy. There was no statistically significant difference of the protection rate of serum among children who had finished chemotherapy 0.05). Conclusions:Serum measles antibody is below the protective level in more than 50% of children with malignancy after chemotherapy. Chemotherapy can compromise the protective antibody against measles. It is recommended for this special population to re-schedule measles vaccine after individualized evaluation to acquire the immuneprotection against measles.