Objective: To investigate the effects of different storage temperatures and durations on the activities of coagulation factor Ⅷ (Factor Ⅷ, FⅧ) and coagulation factor Ⅸ (Factor Ⅸ, FⅨ) after whole blood collection, so as to provide data support for the optimal storage conditions. Methods: A total of 16 mL of whole blood was collected from each of the 20 healthy volunteers at our blood center and aliquoted into 8 sodium citrate anticoagulant tubes. Two tubes were immediately centrifuged for the measurement of FⅧ and FⅨ activity levels. The remaining 6 tubes of whole blood were respectively stored under room temperature and low-temperature conditions. At 2, 4, and 6 h, the whole blood samples were centrifuged and analyzed for FⅧ and FⅨ activity levels. The mean values of the two immediately tested tubes were used as the control group, while other tubes were designated as the experimental groups for comparison. Statistical analysis was performed using SPSS 26.0. Results: The activity of FⅧ in whole blood remained stable after 4 hours of storage at both room temperature and low temperature (116.53±25.95 vs 125.22±27.33, 109.77±23.23 vs 125.22±27.33) (P>0.05 for both). However, by 6 hours, FⅧ activity showed a statistically significant decline compared to the control group (108.65±22.92 vs 125.22±27.33, 100.46±20.19 vs 125.22±27.33) (P<0.05 for both), though the room temperature group results were closer to the control values. The activity of FⅨ in whole blood remained stable after 6 hours of storage under both conditions (97.14±19.48 vs 96.76±19.67, 97.10±17.45 vs 96.76±19.6) (P>0.05 for all comparisons). Conclusion: For whole blood samples after collection, storage at either room temperature or low temperature for up to 4 hours does not compromise the accuracy of test results. When stored for 6 hours, FⅨ activity remains stable, whereas FⅧ activity decreases significantly. Notably, FⅧ activity demonstrates better stability at room temperature than under low-temperature conditions within the 6-hour storage.

Objective: To study the correlation between activated partial thromboplastin time (APTT) mixing test results and the inhibitor titers in hemophilia A inhibitor-positive patients. Methods: In this cross-sectional study, 41 patients with severe hemophilia A and inhibitors (and negative for lupus anticoagulant) were included from the hemophilia clinic of Shandong Blood Center from February 2022 to February 2024. All patients underwent APTT mixing test. The Rosner's index (RI, including the immediate RI and the RI after 2-hour water bath incubation [water bath 2h RI]), the time-dependent difference (Δ value), and the corrected percentage were calculated based on results of APTT mixing test. The median (interquartile range) of the corresponding indexes were calculated, and the ROC curves for identification of high inhibitor titers using the four indexes (the immediate RI, the water bath 2h RI, the Δ value, and the corrected percentage) were plotted, The correlations between APTT mixing test and inhibitor titers for coagulation factor Ⅷ (Factor Ⅷ, FⅧ) were investigated. Results: The median (lower quartile, upper quartile) of immediate RI, water bath 2h RI, Δ-value and corrected percentage for FⅧ inhibitor positive patients were 11.0 (5.4, 29.3)%, 45.0 (25.7, 75.0)%, 26.2 (7.6, 41.8) s, and 82.2 (58.5, 91.6)%, respectively. The median (lower quartile, upper quartile) of the immediate RI, water bath 2h RI, Δ-value and corrected percentage were 25.2 (13.0, 37.5)%, 64.1 (44.6, 72.6)%, 38.0 (14.3, 38.3) s, and 66.5 (50.1, 82.1)% for the high-titer inhibitor group, and 5.2 (4.2, 9.4)%, 17.9 (8.8, 28.0)%, 13.0 (7.6, 25.4) s, and 92.3 (88.0, 94.3)% for the low-titer inhibitor group. The AUCs of the ROC curves for discrimination between high and low titer inhibitor were: 0.9105 for immediate RI, 0.9118 for water bath 2h RI, 0.8873 for correcter percentage, and 0.6532 for Δ-value. Conclusion: High-titer inhibitors can be highly suspected in hemophiliac patients with an immediate RI >10% and a water bath 2h RI >45%, and the presence of low-titer inhibitors is suspected in patients with a 4-second < immediate RI <10% and a 13% < water bath 2h RI <45%.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide. Fat accumulation "sensitizes" the liver to insult and leads to nonalcoholic steatohepatitis (NASH). G protein-coupled receptor 35 (GPR35) is involved in metabolic stresses, but its role in NAFLD is unknown. We report that hepatocyte GPR35 mitigates NASH by regulating hepatic cholesterol homeostasis. Specifically, we found that GPR35 overexpression in hepatocytes protected against high-fat/cholesterol/fructose (HFCF) diet-induced steatohepatitis, whereas loss of GPR35 had the opposite effect. Administration of the GPR35 agonist kynurenic acid (Kyna) suppressed HFCF diet-induced steatohepatitis in mice. Kyna/GPR35 induced expression of StAR-related lipid transfer protein 4 (STARD4) through the ERK1/2 signaling pathway, ultimately resulting in hepatic cholesterol esterification and bile acid synthesis (BAS). The overexpression of STARD4 increased the expression of the BAS rate-limiting enzymes cytochrome P450 family 7 subfamily A member 1 (CYP7A1) and CYP8B1, promoting the conversion of cholesterol to bile acid. The protective effect induced by GPR35 overexpression in hepatocytes disappeared in hepatocyte STARD4-knockdown mice. STARD4 overexpression in hepatocytes reversed the aggravation of HFCF diet-induced steatohepatitis caused by the loss of GPR35 expression in hepatocytes in mice. Our findings indicate that the GPR35-STARD4 axis is a promising therapeutic target for NAFLD.

【Objective】 To investigate the possible molecular pathogenesis of a child with hemophilia A accompanied by coagulation factor Ⅺ reduction by testing coagulation-related indicators and genotyping in the child and his family. 【Methods】 Peripheral blood from the patient and his parents for detection of coagulation factors Ⅷ, Ⅸ, Ⅺ, Ⅻ, VWF∶Ag, lupus anticoagulants and F VIII, F XI inhibitors were collected. All exons and flanking sequences of the genes encoding FⅧ and FⅪ were sequenced and bioinformatically analyzed. 【Results】 The child had low FⅧ and FⅪ activity and no parental abnormalities were observed. The sequencing results showed that there was a c. 1834(exon12) C>T heterozygous mutation in the FⅧ gene and a c. 1817 (exon15) G>A heterozygous mutation in the FⅪ gene, which was de novo. Bioinformatics analysis shows that the FⅪ mutation changes the original protein structure and increases the number of carboxyl groups. 【Conclusion】 For patients with prolonged APTT, in addition to excluding factors that interfere with APTT testing, all coagulation factors related to APTT should be tested to clarify the diagnosis.

Objective:To analyze the clinical and pathological features of adolescent- onset primary nephrotic syndrome (PNS) in children (10 years≤age≤18 years), so as to explore the renal biopsy indications in adolescent-onset PNS.Methods:It was a single-center retrospective observational study. The clinical and pathological data of adolescent-onset PNS (age≥10 years) who underwent renal biopsy in Children's Hospital Affiliated to Nanjing Medical University from December 2004 to June 2022 were analyzed retrospectively.Results:A total of 110 children were included in the study, including 76 males (69.1%) and 34 females (30.9%), with the onset age ranging from 10 years to 14 years and 9 months. Forty-nine cases (44.5%) were accompanied by hematuria, including 14 cases (12.7%) of gross hematuria and 35 cases (31.8%) of microscopic hematuria. Twenty-five cases (22.7%) had hypertension, 19 cases (17.3%) had renal insufficiency, and 4 cases (3.6%) had low complement C3 at the onset. Fifty-two cases (47.3%) were steroid sensitive nephrotic syndrome and 58 cases (52.7%) were steroid resistant nephrotic syndrome. Biopsy results showed that minimal change disease(MCD) was the most common histopathological subtype (47.3%, 52 case), followed by focal segmental glomerulosclerosis (FSGS) in 22 cases (20.0%), IgA nephropathy (IgAN) in 17 cases (15.5%), membranous nephropathy (MN) in 7 cases (6.4%), mesangial proliferative glomerulonephritis in 5 cases (4.5%), IgM nephropathy in 4 cases (3.6%), membranous proliferative glomerulonephritis in 2 cases (1.8%), and C1q nephropathy in 1 case (0.9%). Among 44 children with simple type nephrotic syndrome, the pathological type was mainly MCD (77.3%), and 66 children with nephritic type nephrotic syndrome were mostly non-MCD (72.7%), such as IgAN, FSGS, MN, etc. If there are two or more clinical manifestations of persistent hematuria, hypertension, renal insufficiency or low C3 levels, the proportion of non-MCD would further increase to 92.0%(23/25). The pathological type of patient with gross hematuria or low C3-emia was non-MCD. The frequency of hematuria (69.0% vs. 17.3%, χ2=29.619, P<0.001), hypertension (31.0% vs. 13.5%, χ2=4.821, P=0.028) and renal insufficiency (24.1% vs. 9.6%, χ2=4.047, P=0.044) in non-MCD group was significantly higher than those in MCD group. Conclusions:If the clinical manifestation of PNS in adolescent over 10 years old is simple type nephrotic syndrome, the histopathological lesion is mostly MCD, and most of them are steroid sensitive. It is recommended to give hormone treatment first, and then perform renal biopsy if steroid resistance occurs; If the clinical manifestation is nephritic type nephrotic syndrome, the histopathological lesion is mostly non-MCD, especially those with gross hematuria or low C3-emia, or those have two or more clinical manifestations of persistent hematuria, hypertension, renal insufficiency and hypocomplement C3-emia, a kidney biopsy should be performed at onset.

Objective:To investigate the clinical manifestations, pathological characteristics, treatment and prognosis of anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV) in 13 children.Methods:The clinical and pathological data of 13 cases of AAV in children′s Hospital of Nanjing Medical University from June 2000 to December 2021 were retrospectively analyzed.Results:Among the 13 cases, 12 cases were diagnosed with microscopic polyangiitis (MPA) and 1 case was granulomatosis with polyangiitis (GPA), including 10 females and 3 males. The onset age ranged from 3 years and 11 months to 13 years and 10 months. The most frequently involved organ was the kidney (12 cases, 92.3%), followed by respiratory system (7 cases, 53.8%), skin (5 cases, 38.5%), digestive system (4 cases, 30.8%), nervous system (4 cases, 30.8%) and cardiovascular system (3 cases, 23.1%). There were 10 cases with orthotic anemia, 7 cases with positive antinuclear antibody, and 3 cases with mildly decreased complement C3. Among the 12 children with renal impairment, 9 cases were accompanied by abnormal renal function at the beginning of the disease. Renal biopsy was classified according to the Berden as follows: sclerotic in 5 cases, crescentic 3 cases, focal in 2 cases and mixed in 2 cases. All children were treated with glucocorticoid combined with immunosuppressant. During the follow-up time from 8 months to 128 months, 4 cases acquired complete remission, 8 cases achieved partial remission and 1 case recurred after complete remission, and 7 cases progressed to chronic kidney disease stage 5. Three children with complete remission underwent repeated renal biopsy, including 2 cases of mixed type and 1 case of crescent type initially, and all changed to focal type.Conclusions:AAV in children occurs mainly in school-age female, and most of AAV in children is MPA. The clinical manifestations are various. Most of them have renal damage and anemia, and lung damage is also common. Patients with skin purpura onset may be misdiagnosed as Henoch-Schonlein purpura, and AAV with ANA positive or complement reduction should exclude systemic lupus erythematosus. Once the renal function is abnormal in AAV, especially estimated glomerular filtration rate<60 ml·min -1·(1.73 m 2) -1 and the pathological classification is sclerotic type or crescent type, it is difficult to reverse even after active treatment. Early diagnosis and treatment are very important for AAV.

Objective:To investigate the microstructural abnormalities of the white matter in first-episode medication-free patients with schizophrenia (FESZ) based on inter-voxel diffusivity metric local diffusion homogeneity (LDH) and traditional intra-voxel diffusivity metric.Methods:A total of 56 FESZ patients admitted to our hospital from January 2020 to December 2021 were selected as research subjects, and 64 healthy volunteers recruited during the same period were selected as healthy controls. All subjects accepted magnetic resonance diffusion tensor imaging, and the LDH, anisotropy fraction (FA), mean diffusivity (MD), axial diffusivity (AD) and radial diffusivity (RD) were obtained by FSL and PANDA software. Tract-based spatial statistics (TBSS) method was employed to analyze and compare the differences in the diffusivity metrics of white matter fibers between the two groups. For white matter fiber tracts with significant inter-group differences, receiver operating characteristic(ROC) curve was used to evaluate the diagnostic value of each diffusivity metric in FESZ, and Spearman partial correlation was used to analyze the relation between each diffusivity metric and positive and negative symptom scale (PANSS) scores.Results:As compared with that in the heathy control group, LDH in partial voxel clusters in bilateral anterior thalamic radiation of the FESZ group was statistically increased ( P<0.05, FWE correction). As compared with that in the heathy control group, the mean LDH in the tracts of interest (TOI) of the right inferior longitudinal fasciculus and the right uncinate fasciculus in the FESZ group was significantly increased ( P<0.05, FDR correction). In the above 4 fasciculi, ROC analysis showed that LDH had diagnostic efficacy ( P<0.05), and the diagnostic efficacy of LDH was the highest when the fasciculi structures were combined (AUC=0.733, P<0.001). LDH in white matter fiber tracts with significant inter-group differences had no statistical correlation with PANSS scores ( P>0.05). No voxel clusters or TOIs enjoyed statistical differences between groups were noted in the FA, MD, AD and RD parameter maps ( P>0.05). Conclusion:The inter-voxel diffusivity metric LDH can detect the compensatory changes of white matter microstructure in FESZ patients that could not be detected by traditional intra-voxel diffusivity metric.

Objective To explore the effects of psychological intervention in breast cancer patients. Methods From March 2016 to July 2017,this study selected 141 breast cancer patients with surgery at Jiangsu Taizhou People's Hospital as subjects by convenience sampling. All of patients were divided into control group(n=69)and observation group(n=72)based on admission order. Control group carried out routine nursing. On the basis of routine nursing,observation group implemented psychological nursing intervention. This study compared the treatment compliance,anxiety,depression and nursing satisfaction of patients between two groups. Results After intervention,the treatment compliance of patients in observation group was better than that in control group with a statistical difference(P<0.05);the scores of Self-rating Anxiety Scale(SAS) and Self-rating Depression Scale(SDS)of patients in observation group were lower than those in control group with statistical differences(P<0.05);patient satisfaction of observation group was higher than that of control group with a statistical difference(P< 0.05). Conclusions Positively and effectively psychological nursing intervention for breast cancer with surgery can improve their compliance,reduce negative emotions and promote harmonious nurse-patient relationship which is worthy of clinical generalization and application.

Objective To investigate the difference of the motif patterns in brain directed functional network between first-episode schizophrenia patients and healthy controls, and to analysis the alterations of the underlying information flow patterns in patient networks. Methods The resting-state functional MRI data were collected from 44 first-episode schizophrenia patients and 39 healthy controls. The convergent cross mapping approach was employed to measure the causality connections between brain regions, and the directed functional networks were constructed. The calculations of the frequency and probability spectrum of all motif classes were performed at both whole brain and modular connected level. The between-group difference was then calculated. Results Compared with healthy controls, the frequency spectrum values of all motif classes in schizophrenia were significantly reduced (P＜0.05, FDR corrected), the Z scores of frequency spectrum of were decreased in chain-like motifs and increased in loop-like motifs. In the two groups, the probability values were higher at modular level than at whole brain level in two loop-like motifs (P＜0.05). Conclusion The present study revealed a loss in brain directed functional connections and abnormal alterations in the basic information flow patterns in first-episode schizophrenia brain.

Objective To explore the relationships between the screen time with parent-child relations as well as social ability and behavioral problem in preschool children.Methods A total of 866 children aged 3-6 years old in Dazhou City conducted the sampling survey by adopting the children screen time questionnaire,parent-child relationship self-rating scale and young children's social ability and behavior assessment scale(SCBE-30).The single factor analysis and multinomial Logistic regression method were used to conduct the statistical analysis.Results The surveyed preschool children watching TV every day accounted for 99.31 ％,those playing mobile phones every day accounted for 81.87％ and those using computer accounted for 68.36％s.The average daily screen time at ordinary time was 1.75 h/d,those ≥2 h/d accounted for 16.05％;the average screen time at the weekend was 2.32 h/d,those ≥2 h/d accounted for 46.57％.The univariate analysis showed that the screen time in boy,left behind children,only child,mothers engaging in agriculture of peasant-worker or housewife was longer(P＜0.05);the screen time affected the social ability and behavior problems of children(P＜0.05).The longer the time children spent with their parents every day,the higher the parent-child relationship score(P＜0.05);the screen content affected the parent-child relationship and anxiety withdrawal behavior (P＜0.05).The polynormial regression analysis between the children's screen time with social ability,behavior problems,parent-child relation and contacting screen mode showed that the screen time had the negative effect relation with the social ability and parent-child relation(parents and children questionnaire) score(β=-1.115,-1.728,-1.909,P＜0.05),and had the positive effect relation with the scores of anger attack behavior and anxiety retreat behavior,and individual contacting screen (β=0.982,1.474,0.877,P＜0.05).Conclusion The screen time in preschool children is related with parent-child relation,social ability and behavioral problems.The parental accompany is beneficial to the development of parentchild relations in preschool children.