Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

Objective To observe the value of shear wave elastography(SWE)for evaluating hepatic iron overload in children with β-thalassemia major(β-TM),as well as the correlations of relative parameters with MR T2*value and serum ferritin.Methods Totally 96 children with β-TM and 100 healthy children(control group)were retrospectively enrolled.Children with β-TM were divided into hematopoietic stem cell transplantation(HSCT)group(n=41)or non-HSCT group(n=55)according to underwent HSCT or not.SWE parameters were compared among groups.Spearman correlation was performed to observe the correlations of liver shear wave velocity with MR T2*value and serum ferritin,as well as Young's modulus with MR T2*value and serum ferritin in children with β-TM.Results Liver shear wave velocity(LSWV)and Young's modulus in HSCT group and non-HSCT group were all higher than those in control group(all P＜0.001).No significant difference of LSWV nor Young's modulus was found between HSCT group and non-HSCT group(both P＞0.05).SWE parameters of children with β-TM were moderately and negatively correlated with MR T2*value(r=-0.501,P＜0.05;r=-0.514,P＜0.05),while weakly and positively correlated with serum ferritin(r=0.488,P＜0.05;r=0.470,P＜0.05).Conclusion SWE was helpful for evaluating hepatic iron overload in children with β-TM,with parameters being negatively correlated with MR T2*value and positively correlated with serum ferritin.

Objective:To explore the expressions of programmed death-1 (PD-1) and programmed death-ligand 1 (PD-L1) and clinicopathological characteristics in post-transplant lymphoproliferative disorder (PTLD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children, with the aim of clarifying whether checkpoint inhibition of PD-1/PD-L1 inhibitors may serve as a therapy option.Methods:The clinical data of 13 cases of PTLD after allo-HSCT pathologically confirmed in Shenzhen Children′s Hospital from January 1, 2012 to December 30, 2019 were retrospectively analyzed.The detection was performed by immunohistochemical staining by MaxVision? method, Epstein-Barr virus(EBV) in situ hybridization and lymphoma gene rearrangement.The relationship between the expression of PD-1 and PD-L1 and the clinicopathological characteristics of PTLD were analyzed.Results:The expression of PD-1 was not correlated with gender, age, primary diseases, histopathological types, transplantation mode and the expression of EBV in situ hybridization (all P>0.05). The expression of PD-L1 was correlated with histopathological types ( P<0.05). Furthermore, the expression rate of PD-L1 on severe β-thalassemia was significantly higher than that of severe aplastic anemia [90.0%(9/10 cases) vs. 66.7%(2/3 cases)] and monomorphic PTLD was higher than that of polymorphic PTLD [100.0%(2/2 cases) vs. 83.3%(5/6 cases)]. Moreover, the positive PTLD in EBV was higher than the negative PTLD in EBV [90.9%(10/11 cases) vs. 50.0%(1/2 cases)]. The positive rates of PD-1 and PD-L1 in 13 cases with PTLD were 46.2%(6/13 cases) and 61.5%(8/13 cases) in tumor cells, 92.3% (12/13 cases) and 76.9% (10/13 cases) in microenvironmental cells, and 84.6%(11/13 cases) in EBV, respectively. Conclusions:PD-L1 has a higher positive rate in tumor cells with monomorphic PTLD; and routine staining for PD-1 and PD-L1 can be performed in all types of PTLD when standard immunotherapy and chemotherapy are ineffective.

Objective: To study the characteristics of clinical diagnosis and treatment for 3 children with phytosterolemia. Methods: The different clinical manifestations of 3 children with phytosterolemia were retrospectively reviewed. The case 1 and case 2, who were 7 years and 2 months old twin sisters, hospitalized for frequent epistaxis and abdominal pain. The case 3, who was 5 years and 7 months old male, came to the hospital for cutaneous xanthoma. The phytosterol levels in serum of the children were analyzed by gas chromatography-mass spectrometry, and the second generation sequencing method was used to analyze the disease-causing gene. Sanger sequencing method was used to verify the ABCG5 gene mutation and parental source. Results: (1) The case 1 and case 2 showed moderate anemia, raised reticulocytes, total bilirubin and indirect bilirubin as well as splenomegaly. The blood smear showed that there were more irregular red blood cells, such as oral red blood cells, increased large/giant platelets, and ristomycin-induced platelet aggregation test was decreased. The urine routine examination indicated that there was bleeding in the urinary system. The results of blood lipid test were almost normal. The case 3 showed mild anemia with normal shape of erythrocyte and normal size of spleen. The large/giant platelets increased. The results of platelet aggregation test, bilirubin and urine routine examination were in normal range, but the levels of total cholesterol and low-density lipoprotein cholesterol increased significantly. (2) The levels of serum phytosterol were significantly increased in all the 3 children. (3) Two heterozygous mutations were detectable in ABCG5 gene of case 1 and 2 which were complex heterozygous mutation, i.e., c.9041G＞A and c.751C＞T. The variations were from their father and mother respectively. In case 3, only one homozygous mutation was detectable in ABCG5 gene which originated from their parents. Conclusion When the child showed increased large/giant platelets, hemolytic anemia, erythrocytosis or xanthoma of skin and rised total cholesterol and low-density lipoprotein cholesterol at first visit, the possibility of phytosterolemia should be considered. The blood phytosterol content and gene detection should be carried out as early as possible in order to treat early and improve prognosis.

OBJECTIVE:To establish a method for the concentration determination of voriconazole in human plasma.METHODS:Plasma samples were precipitated with acetonitrile.Using ketoconazole as internal standard,HPLC method was adopted.The determination was performed on Dionex U-3000 Dimonsil C18 column with mobile phase consisted of triethylamine-glacial acetic acid-water mixed solution (1 ∶ 1∶98,V/V/V,pH was about 4.0)-acetonitrile (40∶60,V/V) at the flow rate of 1.0 mL/min.The detection wavelength was set at 255 nm.The column temperature was 40 ℃,and sample size was 20 μL.RESULTS:The linear range of voriconazole was 0.2-20.0 μg/mL.The limits of quantification was 0.2 μg/mL,and the minimum detection limit was 0.03 μg/mL.RSDs of inter-day and intra-day were lower than 10％.The method recoveries were 92.06％-106.26％ (RSD＜5％,n=5),and extraction recoveries were 75.62 ％-90.59 ％ (RSD ＜ 5 ％,n=5).The plasma concentration of voriconazole in 10 children ranged 0.22-4.90 μg/mL (n =10).CONCLUSIONS:The method is simple,rapid,specific and can be used for drug monitoring of voriconazole.

Objective To study effect of a novel schiff base ruthenium coordination compound on cell proliferation and apoptosis of gastric cancer SGC-7901 cell.Method Gastric cancer SGC-7901 cell were divided into four groups according to different treatment of novel schiff base ruthenium coordination compound (concentration of 10, 30, 50μmol/L) and blank group with DMSO.Cell proliferation was detected by MTT assay, cell apoptosis and cell cycle were analysed by flow cytometry.ResuIts MTT results showed the inhibitory effect of novel schiff base ruthenium coordination compound on SGC-7901 cell enhanced with the increase of its concentration, and inhibitory rates were higher than that of blank group at 24, 48, 72 h.Flow cytometry results showed the apoptosis rate of novel ruthenium coordination compound groups of 10, 30, 50μmol/L were (17.64 ±1.21)%, (26.47 ± 0.61)%, (55.63 ±1.49)%, respectively, all higher than that of blank group (P<0.05).The cell proportion of G1 phase increased with the increasing of the novel schiff base ruthenium coordination.ConcIusion A novel schiff base ruthenium coordination compound could inhibit the growth of gastric cancer SGC-7901 cells, promote apoptosis and arrest gastric cancer SGC-7901 cells at G1.

OBJECTIVETo observe the status of iron deposition in patient with β thalassemia major, and to formulate appropriate treatment strategies.

METHODThe data of status of transfusion and chelation in 135 patients aged from 6 years and 4 months to 17 years and 11 months with β thalassemia major were collected and analyzed. Serum ferritin levels were determined and cardiac and hepatic iron deposition was determined using MRI T2(*) technology.

RESULTOf the 135 cases studied, 66 were male, and 69 were female, their average age was 12.1 years. Serum ferritin (SF) was determined for 111 cases, it varied from 1 086.8 µg/L to 15 011.5 µg/L. Among them, 16 cases had SF level <2 000 µg/L (14.5%) , in 41 cases SF were between 2 000 and 4 000 µg/L (36.0%) ;in 54 cases SF >4 000 µg/L (48.7%) . Liver MRI T2(*) results showed that in only 8 cases (5.9%) iron content in the liver was in normal range, 19 cases (14.9%) showed mild liver iron deposition;34 (25.2%) moderate and 74 (54.8%, the youngest one was only 6 years and 4 months of age) had severe iron deposition respectively. Cardiac MRI T2(*) showed that in 89 cases (65.9%) iron content in the heart was in normal range;19 cases (14.1%) had mild cardiac iron deposition and 27 (20.0%) presented severe iron deposition (the youngest one was only 9 years and 3 months of age) . SF level was obviously related to liver and cardiac iron deposition (MRI T2(*)) r and P value were -0.284, 0.003 and -0.374, 0.000 respectively. In 108 cases regular transfusion and chelation were delayed due to financial problem. The late and insufficient dosage administered and irregular chelation caused the higher SF level and the severe iron deposition.

CONCLUSIONThe survival status of β thalassemia major in China is worrisome. Majority of them had not received regular transfusion and chelation. Liver and cardiac iron deposition occur early and had a high incidence.

Adolescent ; Child ; Female ; Ferritins ; blood ; Humans ; Iron ; metabolism ; Iron Chelating Agents ; adverse effects ; therapeutic use ; Iron Overload ; epidemiology ; etiology ; Liver ; metabolism ; Magnetic Resonance Imaging ; Male ; Myocardium ; metabolism ; Radiography ; Retrospective Studies ; Transfusion Reaction ; beta-Thalassemia ; diagnostic imaging ; metabolism ; therapy

Objective To evaluate the therapeutic effect of spironolactone on schistosomal pulmonary arterial hypertension(SPAH). Methods A total of 62 patients suffered from hepatosplenic schistosomiasis with pulmonary arterial hypertension were divided into the spironolactone group(n=31) and control group (n=31). All the patients underwent serial echocardiography and the clinical effect before and after the treatment was evaluated by assessing the mean pulmonary arterial pressure (mPAP) and pulmonary arterial diameter (PAD). At the same time, the varieties of the clinical symptoms, signs and the distance of the 6-minute walking test (6-MWT) were investigated. Results In spironolactone group, mPAP(-x±s) decreased from (31.8±7.1) mmHg to (21.2±2.1) mmHg, PAD(-x±s) decreased from (28.0±5.0) mm to (20.0±3.5) mm before and after the treatment respectively(P<0.01). There were significant differences in mPAP, PAD, the distance of 6-MWT and the heart function before and after the treatment in the spironolactone group. However, the data did not show the significant difference in the control group. Conclusion The therapeutic effect of spironolactone in the treatment of SPAH is satisfactory.

Objective To analyze the associated factors on abnormal liver function(LF)in chronic schistosomiaisis japonica patients.MethodsThe clinical data of 120 inpationts with chronic schistosomiasis were collected and analyzed by logistic regression.The relevant treatment methods were summarized.ResultsThe associated factors on abnormal LF of the patients were hepatitis B,alcohol drinking,hyperlipidemia,nonalcoholic fatty liver disease(NAFLD),and obesity,respectively.The ordinary medication for improving liver function was effective to these causes.Praziquantal can be administrated successfully to these patients after LF normalized.ConclusionChronic schistosomiasis patients with abnormal LF but without severe vital organ diseases could be treated with praziquantal after their LF improving.