AIM:To investigate effects of thrombopoietin(TPO) and TPOⅡ on human platelet activation in vitro. METHODS:Human platelets were incubated in the phosphate-buffered saline containing rhTPO or TPOⅡ at the concentration of 100 μg/L for five minutes. In order to determine the rate of platelet activation. The CD62P and CD41 expressions on platelets were analysed by flow cytometry using fluorescence labelled monoclonal antibody to CD62P and CD41. RESULTS:The results demonstrated that expression of CD62P on platelets which were incubated with rhTPO or TPOⅡ didn't increase compared with that of contrast group. CONCLUSION:Both rhTPO and TPOⅡdidn't cause the disorder of platelet activation.

AIM: To investigate the biological activity of thrombopoietin Ⅱ(TPOⅡ) in vivo , which consists of two new kinds of ligand binding with thrombopoietin receptor. METHODS: Purified ligandⅠof TPOⅡ, artificial compound ligandⅡ of TPOⅡand rhTPO were injected into purebred Babl/c mice respectively in 7 days by intraperitoneal injection once for a day. Then the biological activity of TPOⅡ was analyzed by measuring peripheral platelet counts by the end of the seventh day. RESULTS: On the seventh day, the platelet counts of mice treated by ligandⅠof TPOⅡ were higher than that in the negative control group( P 0.05). On the fourteenth day, the platelet counts increased in two all experimental groups of TPOⅡcompared with negative control group( P 0.05). Moreover the platelet counts of mice in two experimental groups of TPOⅡ and the positive group showed increase with experimental days. CONCLUSION: The purified ligandⅠof TPOⅡ had obvious activity in increasing platelet production, which is not different from the effect of rhTPO.

AIM: To investigate effects of thrombopoietin(TPO) and TPOⅡ on human platelet activation in vitro. METHODS:Human platelets were incubated in the phosphate-buffered saline containing rhTPO or TPOⅡ at the concentration of 100 ?g/L for five minutes. In order to determine the rate of platelet activation. The CD62P and CD41 expressions on platelets were analysed by flow cytometry using fluorescence labelled monoclonal antibody to CD62P and CD41. RESULTS: The results demonstrated that expression of CD62P on platelets which were incubated with rhTPO or TPOⅡ didn't increase compared with that of contrast group. CONCLUSION: Both rhTPO and TPOⅡdidn't cause the disorder of platelet activation.

OBJECTIVE： Te analyze the problems of clinical use in pediatric drug in China， and to put forward related countermeasures and suggestions. METHODS： Using “children” and “pediatrics” as retrieval words， registered drug information were retrieved from the website of China Food and Drug Administration； the data of pediatric drug use prescription was retrieved from hospital prescription system of 9 hospitals； all drug information were retrieved from national drug data management system； off-label drug use investigation and the literatures of pediatric drug use in medical institutions （5 representative third grade general medical institutions） were retrieved from CNKI and Wanfang database. General information and problems of pediatric drug use in China were investigated. RESULTS： A total of 170 009 items of registered drug information were retrieved， including 2 784 drug information items labeled with ”children” or ”pediatrics”， accounting for 1.64％；320 000 drug prescriptions from hospital prescription system of 9 hospitals covered 22 treatment areas involving 1 186 drugs and 51 dosage forms. Only 10％ suitable for children. The retrieval results of database showed that the incidence of children off-label drug use in outpatients prescriptions of 5 hospitals was in high level， mainly manifesting as without the information of pediatric drug use， hyper-indication drug use， hyper-dosage drug use. There were many problems in clinical pediatric drug use in China， such as less variety for children， single dosage form and specification， widespread off-label drug use， lack of scientific reference for drug use， difficulty in developing pediatric drug clinical trials， etc. CONCLUSIONS： Although the policies to protect children’s clinical drug use have been introduced in China， the problems facing children’s clinical drug use in China are still very serious. While further implementing relevant policies， it is necessary to establish a linkage management system led by government departments， with the full participation and mutual cooperation of society， enterprises， medical institutions and patients so as to guaratee the safety of pediatric drug use in clinic.

Objectives:To explore the GAA varient spectrum and the genotype-phenotype correlations in patients with glycogen storage disease type Ⅱ (Pompe disease, PD), as well as to estimate the disease incidence based on carrier rate of GAA varients in Guangzhou population.Methods:A total of 57 PD cases were retrospectively enrolled at Guangzhou Women and Children′s Medical Center from January 1, 2010 to May 31, 2020. All patients presented symptoms before the age of 18 years. Each diagnosis was further confirmed by GAA enzyme activity and GAA variants. The carrier rate of GAA varients was calculated based on variants detected by whole exon sequencing among 2 395 healthy children in Guangzhou.Results:Among the 57 PD patients (including male 26, female 31),twenty-eight patients with infantile onset PD (IOPD) presented with progressive general muscle weakness and cardiomyopathy. The mean ages of symptom onset and diagnosis were (2.5±1.4) and (5.0±3.0) months, respectively. Twenty-six cases died in the first year after birth.Twenty-three patients with late onset PD (LOPD) presented with progressive muscle weakness. Seven of them had respiratory failure at diagnosis. The mean ages of symptom onset and diagnosis were (12.0±5.0) and (17.0±7.5) years, respectively. Six children with atypical IOPD showed motor delay, muscle weakness and cardiomyopathy. Their diagnosis was confirmed at 2.5-7.0 years of age. Among the 57 patients, 47 different variants were identified in the GAA gene. Three variants: c.797C>T, c.1109G>A and c.1757C>T were novel. c.1935C>A (25/114, 21.9%) and c.2238G>C (15/114, 13.2%) were the most common variants, detected in 57.1% of IOPD and 65.2% (15/23) of LOPD patients, respectively. Among the 28 IOPD patients, 26 cases (92.9%) carried at least one missense variant which indicated positive cross-reactive immunologic material (CRIM). The carrier rate of pathogenic variants in GAA gene among healthy children was 24/2 395. The estimated incidence of PD in this population is about 1/40 000. The frequencies of pseudodeficiency variants c.1726G>A and c.2065G>A homozygotes were 26.3% (15/57) and 35.1% (20/57) in PD patients, which were significantly higher than those (1.7% (40/2 395) and 3.9% (94/2 395)) in healthy children (χ2=151.2, 121.9; both P<0.01). Conclusions:PD presents as a spectrum, some as atypical IOPD. The c.1935C>A and c.2238G>C are common variants, correlated with IOPD and LOPD respectively. The c.796C>T and c.1082C>T are usually found in atypical IOPD. The majority of IOPD patients is predicted to be CRIM positive. The estimated incidence of PD is about 1/40 000.