OBJECTIVE: To establish a method for simultaneous determination of 13 constituents including isovalerylshikonin, acetylshikonin, β, β-dimethylacrylshikonin, isobutyrylshikonin, β-acetoxy-isovaleryshikonin, β-hydroxyisovalerylshikonin, propio-nylshikonin, 1-methoxyacetylshikonin, shikonin, shikonofuran A, shikonofuran B, shikonofuran D, and shikonofuran E in Arnebia plants by HPLC-MS. METHODS: The determination was performed on a Waters Sun Fire C18 column (2.1 mm×150 mm, 3.5 μm) with mobile phase A consisting of water containing 0.1% formic acid and 10 mmol·L-1 ammonium formate and mobile phase B of acetonitrile under negative ionization MRM mode. The qualities of 22 batches of Arnebia plants samples obtained from medicinal materi-alsmarkets were investigated and evaluated after methodology validations. RESULTS: There were good linear relationships between the peak areas and predetermined content ranges of these compounds. The average recoveries varied from 87.3% to 107.6%. The contents of shikonins and shikonofurans of the 22 batches of Arnebia plants samples showed significant difference. CONCLUSION: The established method is feasible and simple for the quality control of Medicinal Arnebia plants and their related plants and preparations.

Objective To evaluate the values of CD64 expression in diagnosis of infected patients referred to intensive care unit.Method Sixty febrile children referred to the hospital intensive care unit from 2009.11 to 2010.03 were enrolled for a retrospective study.Fever was defined as a body temperature reaching 38℃ or higher with specifically bacterial infection or highly suspected with bacterial infection or viral infection.There were 28 patients with bacterial infection and 32 with viral infection.The non-infectious diseases such as juvenile rheumatoid arthritis and Kawasaki disease were excluded.The controls were 50 healthy children asking for physical examination.On admission,CD64 were measured by using flow cytometry,and blood routine examination,ESR,PCT,blood cultures and sputum cultures were simultaneously detected in all febrile patients.Data were statistically analyzed by using SAS 16.0 software.Data are given as means±SE.Categorical variables were analyzed using X2 test and continuous variables were compared by applying paired 1-tailed t test,Significance level was set at less than 0.05.Results of them,57.1%bacterial infection patients and 71.9%viral infection patients contracted pneumonia.CD64 in bacterial infection patients、viral infection patients and the subjects of control group were(12.6±9.7),(5.4±2.42)and (2.9±0.77),respectively.The CD64 in the bacterial infection patients were significantly higher than those in the virus infection patients(F=11.002,P=0.004).Conclusions CD64 in infected children referred to a hospital intensive care unit can be clearly distinguished between bacterial infections and viral infections, providing an important guidance and a flexible strategy for clinical treatment and determine the timing of withdrawal.

Objective To study the genes which simultaneously participate in different carcinogenesis progression in lung adenocarcinoma for biomarkers identification. Methods 10 lung adenocarcinoma samples including pathologic stage Ⅰ,Ⅱ,Ⅲ were chosen for experiment and their matched normal tissues for control. After hybridization on 20 slides of microarray with 13824 genes, we analyze the expression profiles combined with pathologic stage and clinical prognosis by data mining. The genes differentially coexpressed in different stage and different prognosis samples were the target. Results 119 genes were identified. Among these targets, 26 genes have known to be related to lung cancer, 46 genes were unreported and 47 gene were new. Conclusions The 119 genes were very important during cancer occurrence and development and were the candidate biomarkers in lung adenocarcinoma.

Objective To investigate CD38 expression characteristic and the relation of clinic prognosis in children with acute lymphoblastic leukemia,in order to improve individual treatment.Methods Seventy-nine patients with childhood acute lymphoblastic leukemia(B-lineage) were enrolled into this study.Four-color fluorochrome labeled monoclonal antibodies were applyed to analyze the cell immunophenotypes and minimal residual disease screening.When CD38 low-expression was considered to be the effective screening marker and be used to continue monitoring.All patients were divided into CD38 low-expression groups and CD38 high-expression groups,to compared the immunophenotyping characteristic,risk stratification and survive rate of the two groups.All datas were assessed by means of SPSS16.0 and a P value of 0.05or less was considered to indicate statistical significance.Results All of 79 newly diagnosed ALL-B,The group of CD38 low-expression were 50/79 (63.3％) while the other group were 29/79(36.7％).of all patients,11 chilldren showed only a screening indicator-CD38/CD10/CD34/CD19,while 46 belonged to more than one markers (Such as TdT/CD10/CD34/CD19,CD66c/CD10/CD34/CD19 and CD45/CD10/CD34/CD19) and 18 no markers.The stratification of CD38 low-expression and CD38 high-expression groups as follows:21/5 patients with low-risk,14/15 with medium risk and 15/9 with high-risk.In the CD38 low-expression group,Early Pre-B 33,Pre-B 12,Mature-B 5,while in the CD38 high-expression group,Early Pre-B 21,Pre-B 5,Mature-B 3.This study showed that the high-risk stratification in the CD38 high-expression group was obviously higher than the CD38 low-expression group(F=6.24,P=0.044),but the survival time was signicantly shorter than CD38 low-expression group (x2 ＝ 5.22,P =0.022) and the difference was statistically significant.Conclusion CD38 as a MRD monitoring indicator of most acute lymphoblastic leukemia when it low-expression,CD38 high-expression in newly diagnosis childhood acute lymphoblastic leukemia(B-lineage) may be an independent risk factor for predicting poor prognosis.

Objective To analyzed the expression and clinical characteristics of CD 20 marker in children with B-lineage acute lymphoblastic leukemia ( B-ALL) and evaluated its medical significance in assessing the prognosis of disease.Methods From November 2008 to July 2012,125 cases of children with B-lineage acute lymphoblastic leukemia were collected from Shanghai Children ′s Hospital,including 79 males and 46 females, aged between 2 months to 14 years old.Flow cytometry based immunophenotyping and Minimal Residual Disease ( MRD) screening were applied to these children when newly diagnosed ,and MRD monitoring was again carried out after 35 days of induction remission therapy to those bears the MRD markers.These 125 patients were divided into CD20-positive group and CD20-negative group, and the corresponding clinical characteristics ,stage of immunophenotype ,MRD,risk stratification,and overall survival rates were recorded and compared.Data were statistically analyzed by using SPSS 16.0 software including χ2 test,t-test,standard deviation test and survival test.Results A total of 125 children with ALL-B,the group of CD20-positive were 48 while CD20-negative groups were 77,with a median age of 6 years old,and the median follow-up time of 30 months.Multivariate Cox regression Analysis showed that there was no clear correlation between CD20 expression level with age ,sex,white blood cell count at diagnosis ,fusion-gene,the stage of immunophenotype as well as risk stratification.The MRD-positive incidence at 35 days in the CD20 positive group was 35.4%,much higher than that of the CD20-negative group (16.9%),which is statistical significance (χ2 =5.236,P<0.05),while the overall survival rate (OS) for the CD20 positive group is 75.0%,much lower than that of the CD20 negative group (84.4%,χ2 =4.160,P<0.05).Conclusions CD20 positive expression level in children with B-lineage acute lymphoblastic leukemia at diagnosis demonstrates negative correlation with the overall survival rate of the patient ,indicating its usefulness as an additional joint marker for the current regimens to incorporate CD 20-targeted monoclonal therapy.

BACKGROUNDRRM1 may be a prognostic factor in non-small cell lung cancer (NSCLC). The aim of this study is to evaluate RRM1 expression and prognosis in NSCLC by the means of tissue microarray.

METHODSA total of 417 paraffin-embedded specimens of NSCLC from Lung Cancer Study Center in Guangdong Provincial People's Hospital were collected and tissue microarray was constructed. RRM1 expression was detected by SP method and its correlation with prognosis was evaluated.

RESULTSNo statistic difference was found in RRM1 expression in different gender, age, tumor site, histology, differentiation, T stage, N stage, M stage and pTNM stage groups (P > 0.05). Univariate analysis showed that RRM1 was not an independent prognostic factor (P > 0.05). At the multivariate analysis, differentiation and N stage were considered independent prognostic factors.

CONCLUSIONSRRM1 expression detected by immunohistology is not an independent prognostic factor in NSCLC. TNM stage is still the best prognostic factor up to now.

Objective To observe the glymphatic system functional changes of bilateral cerebral hemispheres in early-stage Parkinson disease(PD)patients complicated with unilateral limb motor symptoms.Methods A total of 52 early-stage PD patients complicated with unilateral limb motor symptoms were prospectively enrolled,including 21 cases of left-onset PD(LPD group),31 cases of right-onset PD(RPD group).Meanwhile,28 healthy individuals were recruited as healthy controls(HC group).Clinical data and indices of diffusion tensor imaging analysis along the perivascular space(DTI-ALPS)were compared among 3 groups and between each 2 groups.The correlations of DTI-ALPS indices and clinical scales were analyzed.Results Significant difference of Hamilton depression scale(HAMD)score was found among 3 groups(P＜0.05).HAMD score of LPD group and RPD group were higher than that of HC group(both P＜0.017).There were significant differences of left and right cerebral hemispheres DTI-ALPS(DTI-ALPS_L,DTI-ALPS_R)indices among 3 groups(both P＜0.05).DTI_ALPS_R in LPD group was lower than that in HC group(P＜0.017),and DTI_ALPS_L in RPD group was lower than that in HC group(P＜0.017).No obvious correlation of bilateral cerebral hemisphere DTI-ALPS indices with clinical scales was detected in PD patients(all P＞0.05).Conclusion Like that of motor symptoms,glymphatic system dysfunction of PD patients had characteristics of laterality,manifested as DTI_ALPS_R decreased in LPD patients while DTI_ALPS_L decreased in RPD patients.

Objective:To investigate the clinical and genetic features of patients with ACTL6B gene variations, and to report novel pathogenic variations of the ACTL6B gene, summarize the clinical phenotypes and genotypes of the gene. Methods:The clinical phenotypes and genotypes of a infant with developmental epileptic encephalopathy carrying the ACTL6B gene variations, who visited the Department of Neurology, Capital Institute of Pediatrics-Peking University Teaching Hospital on March 12, 2021 were analyzed. The phenotypes and genotypes of patients carrying the ACTL6B gene variations reported in the literature were also summarized and analyzed. Results:The proband was a 2-month-old male presented with convulsive seizures, development delay, dystonia, and cherry erythema in the fundus. The whole exome sequencing of his family showed that he carried compound heterozygous variation c.937-2A>G(p.?), c.11delG(p.G4Afs *86) which derived from his parents respectively. These 2 genotypes had not been reported. A total of 42 cases with ACTL6B gene variation were reported in the literature and in this study. There were 11 de novo heterozygous variations and 31 bi-allelic variations inherited from the parents (24 homozygous and 7 compound heterozygous). Individuals with variations tended to have epilepsy, development delay, ambulation disability, speech disability and dystonia. Minor facial dysmorphisms and autism spectrum disorder also can be seen. Conclusion:This paper summarizes the clinical and genetic features of patients with ACTL6B gene variations, reports 2 novel variations and a novel combination of this gene with cherry erythema in the fundus.