Objective To investigate the clinical and laboratory characteristics of patients with chronic lymphocytic leukemia (CLL). Methods 503 patients with CLL admitted from October 1998 to February 2015 were retrospectively analyzed. Baseline characteristics were compared using Chi-square test and Kaplan-Meier methodology was undertaken for survival analyses. Results The median age was 58 years (26-86 years):335 cases were male and 168 cases were female. 204 cases (40.5%) were at the clinical stage of Binet A, followed by Binet B (148 cases, 30.1%) and Binet C (151 cases, 29.3%). 108 cases (21.1%) had anemia at diagnosis, while 113 cases (26.5 %) had an elevated level of lactate dehydrogenase and the expression of CD38 was detected among 100 cases (29.1 %). Clonal abnormalities were observed using fluorescence in situ hybridization (FISH) analysis. Those involving 13q deletion were the most frequent (156 cases, 47.3 %), followed by IgH translocation (22.4 %), trisomy 12 (21.2 %) and 17p deletion (14.5 %). The mutational status of immunoglobulin heavy chain variable region was determined among 230 cases, 165 cases (71.7%) of which were found to be with mutated status. The most frequently encountered gene was V4-34 (28 cases, 12.4 %). The median progression-free survival (PFS) was 89.0 months (95 %CI 75.0-103.0 months), while the median overall survival was 129.0 months (95 %CI 106.9-151.1 months). Conclusion Compared with patients in the western world, CLL patients in this study are younger at diagnosis and have longer overall survival, which, to some extent, could reflects the characteristics of CLL patients in China.

Objective To use diagnosis related group (DRGs ) for the first time in overall evaluation of inpatient service performance evaluation of major diagnostic category (MDC)for all the Beijing municipal hospitals,and recommend how to strengthen Beijing municipal hospitals system in diagnosis and treatment ability of main diseases and improve inpateint service performance.Methods BJ-DRGs burster software was used to analyze the first page information of the medical records of cases discharged from all the Beijing municipal hospitals between 2012 and 2014 to determine the weight of each DRG,and based on such weight the related indicators of such hospitals and central hospitals in 2012, 2013,2014 were compared and analyzed.Results Improvements were found in such indices as diagnosis and treatment difficulty of 50% MDC,time efficiency of 81.8% MDC,cost efficiency of 77.3% MDC, and general capacity of 54.5% MDC for all Beijing municipal hospitals.In addition,the municipal hospitals were found superior to the central hospitals in such indices as cost efficiency of 68.2% MDC, and time efficiency of 59.1% MDC.On the other hand however,they were found inferior to the central hospitals in such indices as diagnosis and treatment difficulty of 72.7% MDC,and the comprehensive ability index of the two systems were found equivalent.Another finding was that there was no obvious improvement of the coverage of disease types at major tertiary hospitals in Beijing for the past three years.Municipal hospitals of greater contribution of MDC weight were highly consistent with the hospitals assigned with national key projects of disciplinary developments. Conclusion The comprehensive evaluation results of inpatient service performance of main diseases at Beijing’s municipal hospitals based on DRGs system,showed that the Beijing’s hospital authority had played an important role in improving inpatient service performance especially in reducing the burden of patients,improving the service efficiency through increasing government investment,optimizing service organization and implementation of performance management.But it also suggested that measures such as collectivize construction and management should be taken to improve municipal hospitals’linical specialty ability, improve the MDC diagnosis and treatment difficulty,and resume their functions of tertiary hospitals.

OBJECTIVE:To establish a method for simultaneous determination of fatty acids in Isaria cicadae,Cordyceps militaris and C.sinensis,and to compare the difference of the contents of fatty acids among above medicinal herbs.METHODS:GC-MS method was adopted.Chromatographic condition:the determination was performed on TG-5MS gas phase capillary column with carrier gas of nitrogen at the flow rate of 1.2 mL/min.The inlet temperature was 290 ℃ by splitlesssampling;valve opening time was 1 min,and volume of sample was 1 μL.Mass spectrometry condition:the ion source is an electrospray ionization source.The temperatures of ion source and transmission line were 280 ℃,the initial temperature of the chromatographic column was 80 ℃ (gradient elution),ionization voltage was 70 eV.Solvent delay time was 5 min,and scan mass range was m/z 30-400.RESULTS:The linear ranges of tetradecanoic acid,pentadecanoic acid,palmitic acid,palmitoleic acid,heptadecanoic acid,heptadecenoic acid,docosahexaenoic acid,methyl oleate,linoleic acid,arachidonic acid,eicosenoic acid,diolefinic acid,eicosatrienoic acid,heneicosanoic acid,behenic acid,tricosanoic acid,lignoceric acid were 1.400-44.520 μg/mL(r=0.999 8),2.091-93.721 μg/mL(r=0.999 7),3.146-85.856 μg/mL(r=0.998 2),1.664-61.444 g/rnL(r=0.998 7),1.773-64.983 g/mL(r=0.999 5),1.781-68.421 μg/ mL (r=0.999 7),1.706-55.606 μg/mL (r=0.999 8),1.439-47.989 μg/mL (r=0.999 6),1.738-66.908 μg/mL (r=0.999 6),2.086-94.206 μg/mL(r=0.999 5),1.356-44.966 μg/mL(r=0.999 4),1.444-56.814 μg/mL(r=0.999 7),1.375-52.335 μg/mL(r =0.999 8),1.512-60.312 μg/mL(r=0.999 5),1.450-59.760 μg/mL(r=0.999 7),1.427-58.757 μg/mL(r=0.999 1),1.269-58.109 μg/ mL(r=0.999 3),respectively.The limit of quantitation was no more than 1 764.71 μg/mL,and the limit of detection was no more than 529.42 μg/mL.RSDs of precision,stability and reproducibility tests were all lower than 2.0％.The recoveries were 84.87％-108.93％ (RSD ranged 0.19％-2.23％,n=6).There were 17 fatty acids in C.militaris,16 fatty acids in Ⅰ.cicadae and 16 fatty acids in C.sinensis.The contents of unsaturated fatty acids in above medicinal herbs were higher than that of saturated fatty acids.The content of fatty acids in artificial cultivated Ⅰ.cicadae was mostly higher than other medicinal herbs.COCLUSIONS:The method is simple,accurate,stable and reproducible.It can be used for simultaneous determination of fatty acids in I.cicadae,C.militaris and C.sinensis.Above 3 kinds of medicinal materials.From the perspective of fatty acid content,the quality of artificial cultivated I.cicadae is best.

Objective:To explore the curative effect of anterior cervical surgery for lower cervical dislocation combined with spinal cord injury.Methods:A retrospective case series study was carried out to analyze the clinical data of 72 patients with lower cervical dislocation and spinal cord injury admitted in Beijing Boai Hospital, China Rehabilitation Research Center from January 2011 to August 2018, including 55 males and 17 females, with an average age of 35.8 years (range, 18-63 years). The segments of injury were at C 3 in 2 patients, at C 4 in 16, at C 6 in 24 and at C 7 in 30. Bases on American Spinal Injury Association (ASIA) grade, there included grade A in 43 patients, grade B in 22, grade C in 6 and grade D in 1. All patients were treated with anterior decompression and fusion. Operation time, intraoperative hemorrhage, decompression and fusion, complications and ASIA grade were observed. Results:All patients were followed up for 3-72 months (average 24.2 months). Operation time was 90-180 months [(118±58)minutes] and intraoperative bleeding volume was 50-180 ml [(104±30)ml]. Spinal cord was decompressed in all patients. The reduction was excellent in 58 patients (81%) and fair in 14 patients (19%). Fusion was confirmed in 48 patients (67%). The complications were improved after timely treatment, including cervical edema in 1 patient, laryngeal edema in 1, cerebrospinal fluid leakage in 4 and axial pain in 2. ASIA grade change of spinal cord injury: grade A were recovered to grade B in 2 patients, grade A to grade C in 1, grade B to grade C in 1, grade C to grade D in 2 ( P>0.05). Conclusion:For dislocation of lower cervical spine combined with spinal cord injury, anterior surgery can obtain good decompression, reduction and fusion effect, but it has limited help for improvement of neurological function.

Objective: To understand the level and risk factor of lead exposure among children in one city of Jiangsu. Methods: In northern Jiangsu Province, 373 children from 2 primary schools were enrolled and were tested for blood lead and heavy metal exposure. Lead exposure was tested in household dust of 46 children. A multivariate Logistic regression was used for lead exposure risk analysis. Spatial distribution of lead contamination in household dust was conducted and compared with the location of industrial enterprises. Results: The geometric mean of blood lead in 373 children was 25.80 mg/L,the blood lead of 3 children (0.8%) was more than 100 mg/L. Pencil biting ( OR=4.26, 95%CI=1.61-10.68, P <0.05) and lead contamination in surrounding environment ( OR=2.93, 95%CI=1.24-7.34, P =0.02) was positively related to high blood lead level in children. The geometric mean household dust lead concentrations in 46 children was 302.27 μg/mg, and household with high dust lead levels were mainly located around manufacturing enterprises. Conclusion Environmental factors correlate with blood lead level in children. Efficient strategies and public health policies are urgently needed to control and prevent environmental lead pollution. Families and schools should actively carry out health education to engourage children good hygiene habits, and effectively reduce lead exposure.

Cannula ; Catheters ; Drainage ; Duodenal Diseases ; Humans ; Intestinal Fistula/surgery*

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.
Apoptosis Regulatory Proteins ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; Transfection ; U937 Cells

OBJECTIVETo explore gene expression of NANOG in T-cell acute lymphoblastic leukemia (T-ALL) cell lines and the effects of NANOG gene down-regulation on apoptosis of leukemia cells.

METHODSReal-time PCR (RT-PCR) and Western blot were used to detect the expression level of NANOG gene and protein in MOLT-4, CCRF-HSB2 and Jurkat cells. To test the efficiency of RNA interference, MOLT-4 cells were firstly infected by lentiviral vectors, which were successfully constructed with NANOG specific shRNA. NANOG expression levels were subsequently re-evaluated by RT-PCR and Western blot. The percentages of early apoptotic cells (Annexin V⁺/7-AAD⁻) and late apoptotic cells (Annexin V⁺/7-AAD⁺) were analyzed by flow cytometry. The expression of apoptosis-related genes was also detected.

RESULTSBoth NANOG gene and protein expression was positive in MOLT-4 and CCRF-HSB2 cells. The lentiviral vectors pLB-shNANOG-1, pLB-shNANOG-2, and pLB-sh control were successfully constructed, as evidenced by the viral titers (1.83-3.12)× 10⁸ IU/ml. The experimental data on infection of MOLT-4 cells with such lentiviral vectors revealed that both shRNA interfering sequences (shNANOG-1 and shNANOG-2) could stably down-regulate NANOG gene and protein expressions. The percentages of early apoptotic cells in groups of shNANOG-1[(8.06 ± 1.61)%]and shNANOG-2[(5.67 ± 1.59)%]were significantly increased as compared to that of MOLT-4 group[(1.13 ± 0.40)%]or sh-control [(1.15±0.49)%](P<0.05). However, no statistical difference among them was observed for late apoptotic cells (P>0.05). The gene expression of TP53, PMAIP1, and CASP9 of either shNANOG-1 or shNANOG-2 group was augmented as compared to that of MOLT-4 group or sh-control (P<0.05). Reversely, a significant down-regulation of Bcl-2 gene expression was observed (P<0.05).

CONCLUSIONNANOG can be expressed in various human T-ALL cell lines. Down-regulation of NANOG can trigger leukemia cellular apoptosis through mitochondria-dependent apoptosis pathway.

Apoptosis ; genetics ; Cell Line, Tumor ; Down-Regulation ; Gene Expression ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Nanog Homeobox Protein ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics

BACKGROUNDPatent ductus arteriosus (PDA) is one of the most common congenital heart diseases and began to get treated by transcatheter occlusion since 1997 in China. Since then, several devices have been invented for occluding PDA. This study aimed to evaluate the technical feasibility, safety, and efficacy of transcatheter occlusion of PDA with different devices.

METHODSOne thousand five hundred and twenty-six patients (537 boys, 989 girls) with PDA from January 1997 to September 2014 underwent descending aortogram and transcatheter occlusion procedure. We retrospectively analyzed data of these patients, including gender, age, weight, size and morphology of PDA, and devices used in transcatheter occlusion, outcomes, and postoperational complications.

RESULTSMedian age and median weight were 4.0 years (range: 0.3-52.0 years old) and 15.3 kg (range: 4.5-91.0 kg), respectively. Mean ductal diameter, aortic ductal diameter, ductal length, and pulmonary artery pressure were 3.50 ± 2.15 mm, 10.08 ± 2.46 mm, 7.49 ± 3.02 mm, and 30.21 ± 17.28 mmHg, respectively. Morphology of PDA assessed by descending aortogram was of type A in 1428 patients, type B in 6 patients, type C in 79 patients, type D in 4 patients, and type E in 9 patients according to the classification of Krichenko. Of all the 1526 patients, 1497 patients underwent transcatheter PDA closure, among which 1492 were successful. Devices used were Amplatzer duct occluder I (ADO I, 1280, 85.8%), Cook detachable coils (116, 7.8%), ADO II (ADO II, 68, 4.6%), muscular VSD occluder (12, 0.8%), and Amplatzer vascular plug (16, 1.0%).

CONCLUSIONSExcellent occlusion rates with low complication rates were achieved with all devices regardless of PDA types. With transcatheter occlusion technique and devices developing, more patients with PDA can be treated with transcatheter closure both safely and efficiently.

Adolescent ; Adult ; Cardiac Catheterization ; Child ; Child, Preschool ; Ductus Arteriosus, Patent ; surgery ; Female ; Heart Defects, Congenital ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Prosthesis Design ; Retrospective Studies ; Septal Occluder Device ; Treatment Outcome ; Young Adult

BACKGROUNDThis study was to examine the expression of total vascular endothelial growth factor (VEGF) and the anti-angiogenic VEGF 165 b isoform in the vitreous body of retinopathy of prematurity (ROP) patients, and to further study the role of the VEGF splicing in the development of ROP.

METHODSThis was a prospective clinical laboratory investigation study. All patients enrolled received standard ophthalmic examination with stage 4 ROP that required vitrectomy to collect the vitreous samples. The control samples were from congenital cataract patients. The expression of total VEGF and the anti-angiogenic VEGF 165 b were measured by enzyme-linked immunosorbent assay. Results were analyzed statistically using nonparametric tests.

RESULTSThe total VEGF level was markedly elevated in ROP samples while VEGF 165 b was markedly decreased compared to control group. The relative protein expression level of VEGF 165 b isoform was significantly decreased in ROP patients which were correlated with the ischemia-induced neovascularization.

CONCLUSIONSThere was a switch of VEGF splicing from anti-angiogenic to pro-angiogenic family in ROP patients. A specific inhibitor that more selectively targets VEGF 165 and controls the VEGF splicing between pro- and anti-angiogenic families might be a more effective therapy for ROP.

Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Infant, Newborn ; Infant, Premature ; Male ; Prospective Studies ; Protein Isoforms ; metabolism ; Retinopathy of Prematurity ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism ; Vitreous Body ; metabolism