In order to obtain polyclonal antibodies against the fibrillar(Fiber)protein of fowl ade-novirus serotype 11(FAdV-11)and investigate its cross-reactivity to different serotypes of FAdV Fiber,the gene encoding the FAdV-11 Fiber protein was cloned into a prokaryotic expression vec-tor pET-32a by homologous recombination technology,then the plasmid was transformed into BL21(DE3)receptor cells,and the purified recombinant protein was used as an immunogen to im-munize rabbits to prepare polyclonal antibody after induced expression,and the cross-reactivity of the polyclonal antibody against different serotypes of FAdV Fiber proteins was identified by West-ern blot and indirect immunofluorescence(IFA).The results showed that the His-FAdV-11-Fiber recombinant protein was mainly expressed as inclusion bodies and was well expressed.Western blot and IFA showed that the prepared polyclonal antibody reacted with the Fiber proteins of FAdV-8a,FAdV-8b,and FAdV-11,but did not with the 2 Fiber of FAdV-4(Fiber 1 and Fiber 2)proteins.In conclusion,in this study,we successfully prepared rabbit polyclonal antibodies against FAdV-11 Fiber and showed that it specifically recognized the Fiber proteins of FAdV-8a,FAdV-8b and FAdV-11,which lays the foundation for further establishment of serological differential diag-nosis of FAdV-11.

Objective:To evaluate the role of minimal residual disease (MRD) monitoring during early induction therapy for the treatment of childhood acute lymphoblastic leukemia (ALL).Methods:This was a multicenter retrospective cohort study. Clinical data of 1 164 ALL patients first diagnosed between October 2016 and June 2019 was collected from 16 hospitals in South China Children′s Leukemia Group. According to MRD assay on day 15 of early induction therapy, they were divided into MRD<0.10% group, MRD 0.10%-<10.00% group and MRD≥10.00% group. According to MRD assay on day 33, they were divided into MRD<0.01% group, MRD 0.01%-<1.00% group and MRD≥1.00% group. Age, onset white blood cell count, central nervous system leukemia (CNSL), molecular genetic characteristics and other data were compared between groups. Kaplan-Meier method was used for survival analysis. Cox regression model was used to analyze prognostic factors.Results:Of the 1 164 enrolled patients, there were 692 males and 472 females. The age of diagnosis was 4.7 (0.5, 17.4) years. The white blood cell count at initial diagnosis was 10.7 (0.4, 1 409.0) ×10 9/L. Among all patients, 53 cases (4.6%) had CNSL. The follow-up time was 47.6 (0.5, 68.8) months. The 5-year overall survival (OS) and 5-year relapse-free survival (RFS) rates were (93.1±0.8) % and (90.3±1.1) %. On day 15 of early induction therapy, there were 466 cases in the MRD<0.10% group, 523 cases in the MRD 0.10%-<10.00% group and 175 cases in the MRD≥10.00% group. The 5-year OS rates of the MRD<0.10% group, MRD 0.10%-<10.00% group and MRD≥10.00% group were (95.4±1.0) %, (93.3±1.1) %, (85.4±2.9) %, respectively, while the RFS rates were (93.2±1.6) %, (90.8±1.4) %, (78.9±4.3) %, respectively ( χ2=16.47, 21.06, both P<0.05). On day 33 of early induction therapy, there were 925 cases in the MRD <0.01% group, 164 cases in the MRD 0.01%-<1.00% group and 59 cases in the MRD≥1.00% group. The 5-year RFS rates in the MRD 0.01%-<1.00% group was lowest among three groups ((91.4±1.2) % vs. (84.5±3.2) % vs. (87.9±5.1) %). The difference between three groups is statistically significant ( χ2=9.11, P=0.010). Among ALL patients with MRD≥10.00% on day 15 of induction therapy, there were 80 cases in the MRD <0.01% group on day 33, 45 cases in the MRD 0.01%-<1.00% group on day 33 and 45 cases in the MRD≥1.00% group on day 33. The 5-year RFS rates of three groups were (83.9±6.0)%, (67.1±8.2)%, (83.3±6.9)% respectively ( χ2=6.90, P=0.032). Univariate analysis was performed in the MRD≥10.00% group on day 15 and the MRD 0.01%-<1.00% group on day 33.The 5-year RFS rate of children with CNSL was significantly lower than that without CNSL in the MRD≥10.00% group on day 15 ((50.0±20.4)% vs. (80.3±4.4)%, χ2=4.13, P=0.042). Patients with CNSL or MLL gene rearrangement in the MRD 0.01%-<1.00% group on day 33 had significant lower 5-year RFS rate compared to those without CNSL or MLL gene rearrangement ((50.0±25.0)% vs. (85.5±3.1)%, χ2=4.06, P=0.044;(58.3±18.6)% vs. (85.7±3.2)%, χ2=9.44, P=0.002). Multivariate analysis showed that age ( OR=0.58, 95% CI 0.35-0.97) and white blood cell count at first diagnosis ( OR=0.43, 95% CI 0.27-0.70) were independent risk factors for OS. The MRD level on day 15 ( OR=0.55,95% CI 0.31-0.97), ETV6-RUNX1 fusion gene ( OR=0.13,95% CI 0.03-0.54), MLL gene rearrangement ( OR=2.55,95% CI 1.18-5.53) and white blood cell count at initial diagnosis ( OR=0.52,95% CI 0.33-0.81) were independent prognostic factors for RFS. Conclusions:The higher the level of MRD in early induction therapy, the worse the OS. The MRD levels on day 15 is an independent prognostic factor for RFS.The MRD in early induction therapy guided accurate risk stratification and individualized treatment can improve the survival rate of pediatric ALL.

Objective:To explore the risk factors affecting the incidence of acute kidney injury(AKI)after liver transplantation(LT).Methods:From November 2019 to November 2022, clinical data were retrospectively reviewed for 105 recipients of classic orthotopic LT.There are 89 males and 16 females with an age range of(50.52±10.35)years.They are assigned into two groups of AKI(66 cases)and non-AKI(39 cases)according to the AKI diagnostic and staging criteria of Global Kidney Disease Prognosis Organization in 2012.General profiles and clinical data(e.g.previous medical history, MELD score, total bilirubin, albumin, serum creatinine level, coagulation function, anhepatic phase and time to surgery)of two groups of recipients are compared.The factors with statistically significant differences are included into multivariate Logistic regression analysis for obtaining independent risk factors for early AKI post-LT.Results:Among them, 66 patients developed AKI within 7 days post-operation with an incidence rate of 62.86%(66/105).The clinical stages of AKI are Ⅰ(46 cases, 69.70%), Ⅱ(10 cases, 15.15%)and Ⅲ(10 cases, 15.15%).Statistically significant inter-group differences exists in age, abdominal surgery history, preoperative serum level of creatinine, operative duration, anhepatic phase and intraoperative plasma transfusion(all P<0.05).Multivariate Logistic regression analysis indicated that abdominal surgery history( OR=5.803, 95% CI: 1.008~33.401, P=0.049), anhepatic phase( OR=1.054, 95% CI: 1.008~1.101, P=0.020)and preoperative serum level of creatinine( OR=0.968, 95% CI: 0.943~0.994, P=0.016)are independent risk factors for early AKI after classical orthotopic LT recipients. Conclusions:Abdominal surgery history, anhepatic phase, and preoperative serum level of creatinine are independent risk factors for early AKI in classic orthotopic LT recipients.

Objective:To discuss the value of real-time three-dimensional speckle tracking imaging(RT3D-STI) in the evaluation of the left atrial (LA) volume and function in end-stage renal disease (ESRD) patients with different dialysis methods.Methods:Eighty-three ESRD patients who were treated in the Affiliated Hospital of Qingdao University from February to August 2019 were enrolled and divided into non-dialysis group ( n=25), hemodialysis group (HD group, n=31) and peritoneal dialysis group (PD group, n=27). And 30 healthy volunteers were enrolled as control group. Subsequently, the LA volume indexs including LA maximal volume index (LAVImax), LA minimal volume index (LAVImin) and LA presystolic volume index (LAVIpre), the LA function parameters including LA ejection fraction (LAEF), LA passive ejection fraction(LApEF) and LA active ejection fraction (LAaEF) and the LA global strain parameters including global longitudinal strain(GLS), global radial strain(GRS) and global circumferential strain(GCS) were obtained by RT3D-STI. The differences of the parameters among these groups were compared and the correlations between the myocardial strain parameters and dialysis time were analyzed. Results:①The LAVImax, LAVImin and LAVIpre of the non-dialysis group, PD group and HD group were higher than those in the control group, while the LAEF, LApEF, GLS, GRS and GCS were lower than those in the control group(all P<0.05); the LAVImin of the PD group as well as the LAVImin, LAVIpre and LAVImax of HD group were higher than those of non-dialysis group, while the LAEF, LApEF, GLS, GCS of the PD group and the LAEF, LApEF, GLS, GRS, GCS of HD group were lower than those of non-dialysis group (all P<0.05). The LAVImax, LAVImin, LAVIpre of HD group were higher than those of PD group, while the LAEF, LApEF, GLS, GRS and GCS of HD group were lower than those of PD group (all P<0.05). ②The GLS and GCS in PD group were negatively correlated with the dialysis time( r=-0.670, -0.596, all P<0.05), and the GLS, GCS and GRS in HD group were negatively correlated with the dialysis time( r=-0.624, -0.631, -0.551, all P<0.05). Conclusions:RT3D-STI can assess the LA volume and function of ESRD patients with different dialysis methods effectively and can further evaluate the prognosis of patients and guide timely clinical intervention.

Objective: To investigate the dynamic changes of antibodies induced by leptospiral vaccines. Methods: Antigens for antibody detection were screened out. ELISA was used to analyze antibody responses induced at different time points after immunizing guinea pigs with different batches of leptospiral vaccines from different manufacturers. To investigate the relationship between antibody responses induced by leptospiral vaccines and their protective effects in animal model, guinea pigs were challenged with Leptospira after immunization. Results: There was no significant antigen-antibody reaction between the LigA protein or Patoc Ⅰ antigen and the serum samples of guinea pigs immunized with leptospiral vaccines. Notable IgG and IgM antibody reactions were observed in all vaccination groups when using bacterial proteins from seven Leptospira reference strains which were used for the preparation of leptospiral vaccines as envelope antigens. Antigen-specific IgG antibodies peaked at 35 d after the last immunization, and the highest peak of antigen-specific IgM antibodies was reached 11 d after the last immunization. Results of the challenge test showed that non-diluted leptospiral vaccines induced significant IgG and IgM antibody reactions in guinea pigs as compared with those diluted three or nine times, showing good protective effects. Conclusions Analysis of the dynamic changes of antibodies induced by leptospiral vaccines revealed that there was correlation between the induced serum antibody responses and the protective effects. This study provided reference for further study on alternative methods for evaluating leptospiral vaccine potency.

Collaboration of c-KIT mutations with AML1-ETO (AE) has been demonstrated to induce t(8; 21) acute myeloid leukemia (AML). Targeted therapies designed to eliminate AE and c-KIT oncoproteins may facilitate effective treatment of t(8; 21) AML. Homoharringtonine (HHT) features activity against tumor cells harboring c-KIT mutations, whereas oridonin can induce t(8; 21) AML cell apoptosis and AE cleavage. Therefore, studies should explore the efficacy of combination therapy with oridonin and HHT in t(8; 21) AML. In this study, we investigated the synergistic effects and mechanism of oridonin combined with HHT in t(8; 21) AML cell line and mouse model. The two drugs synergistically inhibited cell viability and induced significant mitochondrial membrane potential loss and apoptosis. Oridonin and HHT induced significant downregulation of c-KIT and its downstream signaling pathways and promoted AE cleavage. HHT increased intracellular oridonin concentration by modulating the expressions of MRP1 and MDR1, thus enhancing the effects of oridonin. The combination of oridonin and HHT prolonged t(8; 21) leukemia mouse survival. In conclusion, oridonin and HHTexert synergistic effects against t(8; 21) leukemia in vivo and in vitro, thereby indicating that their combination may be an effective therapy for t(8; 21) leukemia.

Objective To explore the clinical features and treatment of skin ulcers.Methods A total of 185 skin ulcer patients with clinical data was analyzed retrospectively in our hospital from October 2014-October 2016.Results Of the 185 patients with skin ulcers,84(45.4％) were 51-70 years old,diabetic ulcers were most common,and most occurred in ankle (44.3％) and limbs (35.7％).After further statistical analysis,the healing rate of patients treated by surgery was significantly higher than non-surgical patients.Surgical procedures of local flap transfer after ulcer resection were most commonly used and of the highest healing rate.The number of ulcers,the largest diameter of ulcers,scar ulcers and blood barrier ulcers were four factors that were independent of the outcome of skin ulcers.Conclusions Skin ulcers hospitalized patients are more common in patients over 50 years of age,occur most in the lower limbs.Surgical therapy is the preferred treatment for most ulcers,of which local flap transfer after ulcer resection has the best effect.Ulcer patients should actively treat the primary disease,strengthen the care,early diagnosis,and early treatment.

Objective To compare the effect of three-point positioning method and traditional puncture method in patients with arteriovenous fistula in hemodialysis.Methods A total of 800 hemodialysis patients with malignant tumor of urinary system in our hospital were divided into three-point positioning method group (observation group) and traditional puncture method group (control group), with 400 cases in each group.All the puncture was performed by the senior nurses with color Doppler, and vein intima thickness of artery fistula before puncture and 60 times after puncture were observed, puncture failure rate, incidence of venous fistula aneurysm, arteriovenous fistula stenosis and satisfaction were compared.Results There was no significant difference in vein intima thickness of artery fistula(P<0.05);The effect degree of vein intima thickness of artery fistula in the observation group after treatment was significantly lower than that of the control group, and the difference was statistically significant(P<0.05);The puncture failure rate, incidence of arteriovenous fistula aneurysm, and arteriovenous fistula stenosis rate in the observation group after 60 times puncture were significantly lower than that in the control group, and the differences were statistically significant(P<0.05);The satisfaction in the observation group was significantly higher than that in the control group, the difference was statistically significant (P<0.05).Conclusion Three-point positioning puncture combined with losartan has significant effect for arteriovenous fistula patients with the malignant tumor of urinary system, and it can effectively prolong longevity of vascular puncture, and reduce complications of vascular with fistula.

Objective To compare the effect of three-point positioning method and traditional puncture method in patients with arteriovenous fistula in hemodialysis.Methods A total of 800 hemodialysis patients with malignant tumor of urinary system in our hospital were divided into three-point positioning method group (observation group) and traditional puncture method group (control group), with 400 cases in each group.All the puncture was performed by the senior nurses with color Doppler, and vein intima thickness of artery fistula before puncture and 60 times after puncture were observed, puncture failure rate, incidence of venous fistula aneurysm, arteriovenous fistula stenosis and satisfaction were compared.Results There was no significant difference in vein intima thickness of artery fistula(P<0.05);The effect degree of vein intima thickness of artery fistula in the observation group after treatment was significantly lower than that of the control group, and the difference was statistically significant(P<0.05);The puncture failure rate, incidence of arteriovenous fistula aneurysm, and arteriovenous fistula stenosis rate in the observation group after 60 times puncture were significantly lower than that in the control group, and the differences were statistically significant(P<0.05);The satisfaction in the observation group was significantly higher than that in the control group, the difference was statistically significant (P<0.05).Conclusion Three-point positioning puncture combined with losartan has significant effect for arteriovenous fistula patients with the malignant tumor of urinary system, and it can effectively prolong longevity of vascular puncture, and reduce complications of vascular with fistula.

Objective To evaluate the effect of assertive case management on relapse and health economic evalua?tion in schizophrenics living in communities. Methods Two hundred outpatients were randomly divided into the study group (107 enrolled, 107 completed) which received assertive case management and the control group (93 enrolled, 91 completed) which received normal management treatment for 12 months. Clinical global impression scale (CGI) and the cost of treatment were assessed every three months. Medication adherence and family burden were evaluated before treat?ment and 3, 6 and 12 months after the treatment using Medication Adherence Rating Scale (MARS) and Family Burden Instructing, respectively. Results The study group was less likely to relapse compared with the control group over the 12-month follow-up and the relapse rates were 1.9%and 11.0%in study and control groups, respectively (P<0.01). The repeated-measures analysis of variance indicated that time main effect was significant in severity of illness factor score of CGI (P<0.01). The time main effect and group main effect in factor 1 and factor 3 scores of MARS were significant (all P<0.05) and there was an interaction effect in factor1 score of MARS (P<0.01). In the study group, time effect were signifi?cant in factor 1 score of MARS (P<0.01). The time main effects in indirect cost and total cost were significant and so were interaction effects in direct cost and total cost (P<0.05). In the study group, time effects were significant in direct cost and total cost (P<0.01). Comparison of FBI dimensions before and after the intervention showed that family relation?ship was much more decreased in the study group than in the control group (P<0.01). Conclusions Assertive Case Man?agement can reduce the recurrence of schizophrenia living in communities, improve compliance medication and family re?lationship as well as reduce the cost of treatment.