【Objective】 To study and compare the effects of different storage temperature and time on coagulation factor after cryoprecipitated antihemophilic factor(CAF) melting, and to provide reference for the establishment of industry standards. 【Methods】 From June 2021 to May 2023, a total of 96 bags of CAF were sampled in 4 bags per month, and timely detected in the same month. After the CAF was melted in a 37℃ water bath, the mild to moderate lipemic blood was labeled. Each bag of CAF and two 50 mL transfer bags were divided into two bags and two groups of 20 mL each using a sterile adapter. One group was placed in a 4℃ refrigerator and the other in a 22℃ water bath for 0 h, 4 h, 8 h, 12 h, 24 h and 48 h. Then 2 mL of aseptic sample was taken separately and put into the test tube, and 1mL of sample and 3 mL of buffer were added into the other test tube with the sampling gun and mixed on the machine for testing. The experimental data of 60 bags without mild to moderate lipemic blood cryoprecipitation and coagulation factor were randomly selected and statistically analyzed by SPSS21.0. 【Results】 After melting, CAF was stored for 0 h, 4 h, 8 h, 12 h, 24 h and 48 h to detect the average content and growth rate of coagulation factor in the two groups: 1) Storage at 4℃, factor Ⅷ content was 118.62, 111.57(-5.95%), 105.51(-11.05%), 103.30(-12.92%), 94.35(-20.46%) and 83.25(-29.82%) IU/ bag, respectively; Storage at 22℃, the factor Ⅷ content was 118.62, 112.69(-5.00%), 111.41(-6.08%), 109.01(-8.10%), 101.55(-14.39%) and 92.75(-21.81%) IU/ bag, and the storage results of the two groups were compared. At 24 h at 4℃ and 48 h at 22℃, the content of factor Ⅷ had significant statistical significance(P<0.01), and when stored at 22℃, the decay rate of factor Ⅷ was slower; 2) When stored at 4℃, the content of factor V was 41.19, 41.31(0.29%), 40.52(-1.64%), 40.27(-2.23%), 39.05(-5.19%) and 36.99(-10.21%) IU/ bag, respectively; Stored at 22℃, the factor V content was 41.19, 41.71(1.25%), 42.54(3.28%), 41.94(1.80%), 39.21(-4.80%) and 35.64(-13.48%) IU/ bag, respectively. Comparison of storage results between the two groups showed that the content of factor V was statistically significant(P<0.05) and significantly significant(P<0.01) at 4℃48 h and 22℃48 h, respectively, and the decay rate of factor V was faster when stored at 22℃; 3) When stored at 4℃, the Fbg content was 268.86, 268.17(-0.26%), 262.46(-2.38%), 270.50(0.61%), 267.52(-0.50%) and 261.92(-2.58%) mg/ bag, respectively; Stored at 22℃, the Fbg content was 268.86, 265.86(-1.12%), 264.12(-1.77%), 265.89(-1.11%), 266.04(-1.05%) and 261.04(-2.91%) mg/ bag, respectively. There was no statistical significance between the 2 groups and the original 0 h content in each time period(P>0.05). 【Conclusion】 After CAF melting, coagulation factor decreased with the extension of storage time, especially the decrease of factor Ⅷ, followed by factor V, while Fbg basically unchanged. Comparison between the two groups showed that, factor Ⅷ decay rate is slower, factor V decay rate is faster of storage at 22℃. CAF should be transfused as soon as possible after melting. If the delay is unavoidable, for the delay time less than 12 h, storage at 4℃ is recommended, fot the delay time more than 12 h and less than 24 h, storage at 22℃ is recommended.

Objective:To investigate the clinical features, treatment, and prognosis of transient hyperammonemia of the newborn (THAN).Methods:Data of two infants with severe THAN admitted to the Department of Neonatology of Shanghai Children's Hospital in September 2021 and August 2022 were retrospectively investigated. Clinical data of confirmed THAN cases (blood ammonia>400 μmol/L) were collected from relevant literature retrieved from the Wanfang Database, China National Knowledge Infrastructure, Chinese Medical Journal Database, and PubMed up to July 2022. A descriptive method was used for statistical analysis.Results:A total of 24 cases were involved (two in the present study, and 22 in 12 retrieved articles), including 19 (79.2%) premature newborns and five term infants. The average birth weight was (2 237±608) g and the average onset time was 27 h (4-55 h) after birth. The early clinical symptoms included respiratory distress and hyporesponsiveness (drowsiness, lethargy, coma or hypotonia) in 18 cases (75.0%), metabolic acidosis in 11 cases (45.8%), hypocalcemia in seven cases (29.2%), pupil fixation/dilation in six cases (25.0%), convulsion in five cases (20.8%), apnea in three cases (12.5%) and sinus bradycardia in one case (4.2%). The serum ammonia levels were 1 422.8 μmol/L (547.2-4 494.1 μmol/L). Treatments included peritoneal dialysis plus exchange transfusion in eight cases (33.3%), exchange transfusion in seven cases (29.2%), continuous renal replacement therapy (CRRT) in four cases (16.7%), arginine in two cases (8.3%), peritoneal dialysis in two cases (8.3%), and CRRT+peritoneal dialysis in one case (4.2%). During follow-ups of four months (one month to six years), 13 cases (54.2%) showed no abnormalities in development; two (8.3%) had a neurodevelopmental delay, and six (25.0%) died. The follow-up of the other three cases (12.5%) were not reported in the literature.Conclusions:The early clinical manifestation of severe THAN is atypical. A good prognosis can be expected through early exclusion of possible hyperammonemia-related genetic metabolic diseases and lowering the serum ammonia level. Long-term follow-up is needed for neurological evaluation.

【Objective】 To perform serological screening of K antigen in Qingdao area and analyze the results. 【Methods】 From March to June 2023, 16 201 samples of patients treated in our hospital and applied for blood grouping, and 4 153 samples of blood donors sent to the clinic by Blood Transfusion Department during the same period were detected for Rh and K antigen by Ortho BioVue System Rh/K Cassette. 【Results】 A total of 18 out of 16 201 patient samples and 8 out of 4 153 blood donor samples were positive for K antigen, with the positive rate at 0.127 7%. 【Conclusion】 There was K antigen in Qingdao population, and the positive frequency of K antigen was different from previous reports. It is of great significance to construct rare blood group bank in this area and formulate reasonable blood donation and transfusion strategy for population with K antigen, so as to avoid unexpected antibody production of Kell blood group system, prevent hemolytic disease of newborn and ensure the safety of blood use.

Whether direct manipulation of Parkinson's disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6-10 months), and thus provides a practical transgenic monkey model for future PD studies.
Animals ; Brain ; CRISPR-Cas Systems/genetics* ; Dependovirus/genetics* ; Haplorhini ; Phenotype ; Protein Kinases/genetics*

The TEA domain (TEAD) family proteins (TEAD1‒4) are essential transcription factors that control cell differentiation and organ size in the Hippo pathway. Although the sequences and structures of TEAD family proteins are highly conserved, each TEAD isoform has unique physiological and pathological functions. Therefore, the development and discovery of subtype selective inhibitors for TEAD protein will provide important chemical probes for the TEAD-related function studies in development and diseases. Here, we identified a novel TEAD1/3 covalent inhibitor (DC-TEADin1072) with biochemical IC

Objective:To study the correlations of neonatal hemodynamic parameters with gestational age (GA) and birth weight (BW) using non-invasive ultrasound cardiac output monitor (USCOM).Method:From March to September 2019, neonates with stable hemodynamics admitted to the Department of Neonatology of our hospital were enrolled in this prospective study. According to their GA, they were assigned into <29 w group, 29~33 w group, 34~36 w group and ≥37 w group. According to their BW, they were assigned into <1 000 g group, 1 000~1 499 g group, 1 500~2 499 g group and ≥2 500 g group. Cardiac output (CO), cardiac index (CI), stroke volume (SV), myocardial contractility (inotropy, INO), flow time corrected (FTC), systemic vascular resistance index (SVRI) and heart rate (HR) were measured using USCOM. The univariate linear regression method was used to analyze the correlation of hemodynamic parameters with different GA and BW.Result:A total of 120 neonates with stable hemodynamics were enrolled, including 69 males and 51 females. The average GA was (34.2±3.8)w and the average BW was (2 221±860) g. SV ( r=0.489, P<0.001), CO ( r=0.681, P<0.001), CI ( r=0.348, P<0.001), FTC ( r=0.266, P=0.003), INO ( r=0.446, P<0.001)and HR ( r=-0.322, P<0.001) showed significant linear correlations with GA. No linear correlation existed between SVRI ( r=-0.052, P=0.574) and GA. SV ( r=0.603, P<0.001), CO ( r=0.852, P<0.001), CI ( r=-0.390, P<0.001), INO ( r=0.576, P<0.001) and HR ( r=-0.440, P<0.001) showed significant linear correlations with BW. No significant linear correlations existed between SVRI ( r=-0.076, P=0.409) or FTC ( r=0.090, P=0.329) and BW. Conclusion:USCOM can monitor neonatal hemodynamic parameters in real-time.Hemodynamic parameters including SV, CO, CI and INO are significantly different among newborns with different GA and BW and these parameters are linearly correlated with GA and BW.

Whether direct manipulation of Parkinson’s disease (PD) risk genes in the adult monkey brain can elicit a Parkinsonian phenotype remains an unsolved issue. Here, we used an adeno-associated virus serotype 9 (AAV9)-delivered CRISPR/Cas9 system to directly co-edit PINK1 and DJ-1 genes in the substantia nigras (SNs) of two monkey groups: an old group and a middle-aged group. After the operation, the old group exhibited all the classic PD symptoms, including bradykinesia, tremor, and postural instability, accompanied by key pathological hallmarks of PD, such as severe nigral dopaminergic neuron loss (>64%) and evident α-synuclein pathology in the gene-edited SN. In contrast, the phenotype of their middle-aged counterparts, which also showed clear PD symptoms and pathological hallmarks, were less severe. In addition to the higher final total PD scores and more severe pathological changes, the old group were also more susceptible to gene editing by showing a faster process of PD progression. These results suggested that both genetic and aging factors played important roles in the development of PD in the monkeys. Taken together, this system can effectively develop a large number of genetically-edited PD monkeys in a short time (6–10 months), and thus provides a practical transgenic monkey model for future PD studies.

Angiogenesis is an essential process in tumor growth, invasion and metastasis. VEGF receptor 2 (VEGFR2) inhibitors targeting tumor angiogenic pathway have been widely used in the clinical cancer treatment. However, most of currently used VEGFR2 kinase inhibitors are multi-target inhibitors which might result in target-associated side effects and therefore limited clinical toleration. Highly selective VEGFR inhibitors are still highly demanded from both basic research and clinical application point of view. Here we report the discovery and characterization of a novel VEGFR2 inhibitor (CHMFL-VEGFR2-002), which exhibited high selectivity among structurally closed kinases including PDGFRs, FGFRs, CSF1R, etc. CHMFL-VEGFR2-002 displayed potent inhibitory activity against VEGFR2 kinase in the biochemical assay (IC = 66 nmol/L) and VEGFR2 autophosphorylation in cells (ECs ∼100 nmol/L) as well as potent anti-proliferation effect against VEGFR2 transformed BaF3 cells (GI = 150 nmol/L). In addition, CHMFL-VEGFR2-002 also displayed good anti-angiogenesis efficacy and exhibited good PK (pharmacokinetics) profile with bioavailability over 49% and anti-angiogenesis efficacy in both zebrafish and mouse models without apparent toxicity. These results suggest that CHMFL-VEGFR2-002 might be a useful research tool for dissecting new functions of VEGFR2 kinase as well as a potential anti-angiogenetic agent for the cancer therapy.

Objective To explore the efficacy of continuous renal replacement therapy (CRRT) in the treatment of neonatal acute kidney injury (AKI).Methods Totally 17 critically ill neonates treated with CRRT were selected who were hospitalized at Department of Neonatology,Shanghai Children's Hospital,Children's Hospital Affiliated to Shanghai Jiaotong University,from June 2012 to June 2017,and among them there were 15 cases with AKI,and the clinical data of these 15 patients were retrospectively analyzed,while 15 AKI neonates were treated with CRRT combined with conventional treatment.The model for CRRT was continuous veno-venous hemofiltration dialysis (CVVH-DF) in 13 cases,plasma exchange (PE) in 2 cases.The changes of blood pressure(BP),renal function,electrolyte,acid-base balance index and hemodynamic indicators were analyzed respectively before CRRT treatment,12 h,24 h,48 h after treatment and by the end of CRRT treatment.The efficacy of CRRT treatment was evaluated in these 15 AKI neonates.Results Gestational age of 15 AKI newborns was 33 +4-40 +1 weeks,admission day age was 2-28 days,birth weight was 2.25-4.00 kg.Primary diseases were severe asphyxia in 6 cases,neonatal septicemia in 5 cases,congenital hereditary metabolic disease in 2 cases,traumatic asphyxia in 1 case,and liver failure in 1 case.CRRT treatment persisted for 49-190 hours.BP value [(50.8 ± 6.57) mmHg(1 mmHg =0.133 kPa)] could reach normal level after 12 h CRRT treatment,and blood pH value (7.31 ± 0.25) increased significantly after 12 h CRRT treatment,while blood K+[(5.51 ±1.86) mmoL/L],urea nitrogen (BUN) [(9.5 ±3.7) mmol/L],creatinine(Cr) [(93± 14)μmol/L] significantly decreased after 12 h CRRT treatment,and reached the normal range [K + (4.78 ± 2.95)mmol/L,BUN (7.5 ±2.1) mmol/L,Cr (54 ± 13) μmol/L] after 24 h treatment,but urine volume[(0.8 ±0.2)mL/(kg· h)] significantly increased after 24 h treatment.Partial pressure of oxygen/fraction of inspired oxygen reached 200 mmHg after 12 h treatment and more than 300 mmHg after 24 h treatment.CRRT treatment of 15 AKI neonates turned out to be effective.Conclusions CRRT can effectively improve the internal environment of AKI neonates and reduce the death rate of neonatal AKI,which can provide an effective adjuvant treatment measures for the treatment of AKI neonates.

Objective To study the effect of different breast milk enhancement strategies and the incidence of complications in premature infants.Method Premature infants whose gestational age less than 34 weeks and birth weight less than 2 000 g were prospectively enrolled from January 2017 to February 2018 at the Department of Neonatology of Huangshi Maternal and Child Health-Care Hospital.According to the odd even number at the end of the hospitalization admission number,participants were assigned into 50～＜70 ml/(kg· d) group and 70～＜90 ml/(kg· d) group,When the children reached the corresponding amount of breast-feeding to be given breast milk fortifier.The demographic information,incidence of complications,rate of weight gain,percentage of extrauterine growth retardation (EUGR) and decrease of Z score at discharge were compared between groups.Result A total of 140 cases were included,with gestational age (31.4±1.9) weeks and birth weight (1 402±213) grams.Among the participants,67 infants were assigned to 50～＜70 ml/(kg·d) group,and 73 infants were assigned to 70～＜90 ml/(kg·d) group.There was no statistical difference between two groups in gender,gestational age,birth weight,length,head circumference,rates of asphyxia,ratio of intrauterine growth retardation,Z score of weight at birth,age at which breast milk fortifiers were added,full enteral feeding time,duration of parenteral nutrition,average length of hospital stay and the time of restoration of birth weight (P＞0.05).The proportion of feeding intolerance in 50～ ＜70 ml/(kg· d) group was higher than that in 70～＜90 ml/(kg· d) group (11.9％ vs.4.1％),the difference was statistical significant (P=0.013).There was no statistical difference in other complications between the two groups (P＞ 0.05).The body weight increase rate of premature infants in 50～＜70 ml/ (kg· d) group was higher than that in 70～＜90 ml/(kg· d) group,and decrease of Z score at discharge in 50～＜70 ml/(kg· d) group was lower than that of 70～＜90 ml/(kg· d),the difference was significant (P＜0.05).Conclusion Adding breast milk fortifier earlier——when the breast feeding amount of 50～＜70 ml/(kg· d)——is more beneficial to the growth and development of premature infants,it also reduces the incidence of EUGR on discharge.However,during the feeding process,it was necessary to be aware of the complications.