Objective To establish a simple and sensitive method for the determination of serum copper by spectrophotometry.Methods Nitro-PAPS was used as a coloring agent for serum copper in the presence of surfactants Tween-80 and Triton X-100 and the formed complex was measured by spectrophotometry.Results The maximum absorption wavelength of the complex was 570 nm and the molar absorption coefficient was 7.95?10~4 L/(mol?cm).The lineafity of the method was up to 63.0 ?mol/L and the recoveries ranged from 98.6% to 103.1%.The within-run and between-run CVs were 2.1%-3.3% and 2.7%-3.8%.The method(Y)was compared with an AAS method(X)and a correlation of Y=1.01X -0.27(r=0.998 2)was obtained.A reference interval(x~-?2s)determined with this method on 68 individuals was 9.7-24.1 ?moL/L.Conclusions A simple and sensitive method for serum copper has been established.It may used for the analysis of serum copper in clinical laboratories.

BACKGROUND: Vitamin A deficiency disorders (VADD) is a healthy problem of children in the world, especially in the west of China and remote areas, and the nutritional intervention is needed.OBJECTIVE: To compare the improved effects of biscuits fortified with three different doses of vitamin A on the vitamin A status in children aged 3-6 years and explore ap ideal dose of vitamin A supplement for preventing VADD.DESIGN: Randomized controlled observation.SETTING: Health Surveillance Institute, Chongqing Municipal Health Bureau; Staff Room of Nutrition and Food Hygiene, College of Public Health, Chongqing University of Medical Sciences; Center for Children Nutrition, Children's Hospital, Chongqing University of Medical Sciences.PARTICIPANTS: The investigation was done between March and December 2002. 753 children aged 3-6 years from 8 kindergartens in Banan district of Chongqing city were enrolled with the agreement of their guardians. They were divided randomly into 4 groups: 30% recommended intake group, 100% recommended intake group, 20 000 international unit (IU) and 200 000 IU groups.METHODS: ① The biscuits fortified with 30% recommended intake of vitamin A (500 IU) were once given to people of the 30% recommended intake group (177 cases) every day. ②The biscuits fortified with 100% recommended intake of vitamin A (1666 IU) were once given to people of the 100% recommended intake group (173 cases) every day. ③The biscuits fortified with 20 000 IU of vitamin A were once given to people of the 20 000 IU group (209 cases) every week. ④The soft gelatin capsule with 200 000 IU of vitamin A were once given to people of the 200 000 IU group (194 cases).Height, body mass, serum retinal, prealbumin, haematoglobin and retinal binding protein of all children were measured before intervention and after intervention for 3 months. Above indexes were rechecked after supplement for 9 months in 87 children of 30% recommended intake groupMAIN OUTCOME MEASURES: ①Prevalence rate of VADD before in-tervention and after intervention for 3 months in children of every group.② Serum retinal, serum prealbumin, serum retinal binding protein,haematoglobin, height and body mass of children before intervention and after interventional for 3 months in every group.RESULTS: Because of lose of samples and detective technology, only 580 children' examination results were got by rechecking. ①Comparison of the prevalence rate of VADD of children in every group: Three months supplementation later, the prevalence of VADD in every group all decreased sig nificantly [1.48%,1.42%,1.21%, 2.16% ;6.78%,6.54%,8.61%,8.25%(χ2=3.86-8.57, P ＜ 0.05-0.01 )]. ②Comparison of serum retinal, serum prealbumin, serum retinal binding protein, haematoglobin, height and body mass of children of every group: After supplement for 3 months, except prealbumin and haematoglobin in the 30% recommended intake group ,other indexes in each group all increased significantly (t=2.52-37.44, P＜ 0.05-0.01 ). The increase of serum vitamin A in the 20 000 IU group was larger than that in the other groups (F=4.62,P＜ 0.01 ). The increases of haematoglobin, prealbumin and height in the 30% recommended intake group were less than those in the other groups (F=5.04-7.78 ,P ＜ 0.01 ).After supplement for 9 months, the increases of haematoglobin and prealbumin in the 30% recommended intake group were larger than those in the other groups (F= 11.62,10.21 ,P ＜ 0.01). The increase of serum retinal was still lower than that in the 20 000 IU group (F=4.21 ,P ＜ 0.01 ).CONCLUSION: Supplement with biscuits fortified with 3 different doses of vitamin A and capsule with 200 000 IU of vitamin A can improve obviously the nourished status of vitamin A and the level of ferrohemoglobin, in which 30% recommended intake and 100% recommended intake have small risk, and everyday supplement can maintain stably the level of vitamin A. That may be suitable for the long-term supplement for children, and the effect of 30% recommended intake was better than that of 100% recommended intake.

Antiviral Agents ; therapeutic use ; Hepatitis B, Chronic ; drug therapy ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferons ; therapeutic use ; Treatment Outcome

Objective To study the interobserver variabilities and the differential diagnosis value of Breast Imaging Reporting and Data System-Ultrasound (BI-RADS-US) lexicon for small ( ≤ 2 cm) breast nodules. Methods Between January 2009 and December 2011, 289 patients with small (≤2 cm) breast nodules (n=317) were included. According to sizes, the lesions were divided into two groups, i.e., 0-1 cm (n=160) group and 1-2 cm (n=157)group. Each lesion was described independently by 3 radiologists using BI-RADS-US lexicon. Interobserver variabilities were assessed by Kappa test. Chi-square test was used to compare the frequency difference of the descriptors between malignant and benign lesions. Sensitivity, speciifcity, accuracy, positive predictive value and negtive predictive value were calculated. Results (1)Moderate agreements were obtained for lesion shape, orientation, margin, echo pattern, surrounding tissue and calciifcations (κ=0.44, 0.57, 0.48, 0.43, 0.51 and 0.57) in 0-1 cm group. Substantial agreements were obtained for lesion shape, orientation, margin and echo pattern (κ=0.65, 0.61, 0.64 and 0.63) in 1-2 cm group. (2)Irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo and microcalciifcations were more frequently found in malignant nodules than in benign nodules in 0-1 cm group [52.3% (34/65) vs 20.0% (19/95), 38.5%(25/65) vs 13.7%(13/95), 75.4%(49/65) vs 32.6%(31/95), 18.6%(12/65) vs 0 (0/95) and 10.8%(7/65) vs 2.1%(2/95);χ2=18.19, 13.08, 28.22, 16.39 and 3.95;P=0.000, 0.000, 0.000, 0.000 and 0.047]. Similarly, irregular shape, non-parallel orientation, non-circumscribed margin, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were signiifcantly more frequent found in malignant nodules than in benign nodules in 1-2 cm group [74.2%(49/66) vs 12.1%(11/91), 36.3%(24/66) vs 5.5%(5/91), 93.9%(62/66) vs 22.0%(20/91), 37.9%(25/66) vs 3.3%(3/91), 30.3%(20/66) vs 7.7%(7/91), 15.2%(10/66) vs 0 (0/91) and 16.7%(11/66) vs 4.4%(4/91);χ2=62.59, 24.21, 79.40, 31.22, 13.73, 12.30 and 6.67;P=0.000, 0.000, 0.000, 0.000, 0.000, 0.000 and 0.010]. (3)In both groups, a good sensitivity was demonstrated (75.4%&93.9%) when using the non-circumscribed margin as a criterion for malignancy, and high speciifcity was achieved in two groups (80.0%-100%and 87.9%-100%) when other descriptors including irregular shape, non-parallel orientation, echogenic halo, shadowing, changes of Cooper′s ligament and microcalciifcations were used as differentiation criteria. Conclusions Good interobserver agreement can be achieved using the BI-RADS-US lexicon in the diagnosis of small breast nodules. Non-circumscribed margin are proved as the most valuable sign for screening malignant breast lesions ≤ 2 cm. High speciifcity was found for irregular shape, nonparallel orientation, echogenic halo, shadowing, Cooper′s ligament changes and microcalciifcations, which can help biopsy and preoperative diagnosis.

This study was aimed to investigate the expression of FLT3 internal tandem duplication (FLT3-ITD) in pediatric patients with acute myeloid leukemia (AML) and to analyse the clinical features of patients with mutations and the relation of FLT3-ITD with multidrug resistance gene 1 (mdr1). RT-PCR was used to determine the expressions of FIT3-ITD and mdr1 gene in bone marrow samples from 81 new diagnosed pediatric patients with AML, the cytogenetics and immunophenotypes of bone marrow cells were routinely examined. The results indicated that the FLT3-ITDs were detected in 8 out of 81 pediatric patients (9.88%) and all mutations detected were hybrid, while less frequently this mutation was detected in adult patients. Although they were irrelevant with sex and immunophenotypes, the mutations seemed predominant in older pediatric patients. The leukocyte counts and bone marrow blast cell counts in pediatric patients with FLT3-ITD at diagnosis were higher than those in pediatric patients without FLT3-ITD (p = 0.001 and p = 0.041 respectively), but the normal chromosomes were found in most pediatric patients with FLT-ITD. The patients with FLT3-ITD had lower induction remission rate (only 25%), but the patients without FLT3-ITD had higher remission rate (76.1%). According results detected by RT-PCR, the mdr1 gene was found in 27 pediatric patients, but only 3 out of 8 pediatric patients with FLT3-ITD were detected to express both FLT3-ITD and mdr1, which suggests unrelation between FLT3-ITD occurrence and mdr1 expression. It is concluded that the FLT3-ITD is frequent mutation in pediatric patients with AML, the prognosis is worse and the induction remission rate is lower in these patients, but the FLT3-ITD not relates with the mdr1, which suggests that the common MDR modulators may be un effective for therapy of the patients with FLT3-ITD.
ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Adolescent ; Child ; Child, Preschool ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Female ; Gene Duplication ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Male ; Mutation ; Prognosis ; Tandem Repeat Sequences ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVEThe hypoxic-ischemic encephalopathy caused by asphyxia in peripartum is a serious disease in newborn infants, with a high disability and mortality rate. Lack of regenerative ability in central nervous system after injury is considered as the fundamental cause. However, in recent years many studies have revealed that there are myelin-associated neurite growth inhibitory factors that exert inhibiting effect through the Nogo receptor (NgR). This study aimed to investigate the expression level of NgR and the possible neuroprotective effect of NEP1-40 in newborn rats with hypoxic ischemic brain damage (HIBD).

METHODEighty healthy Wistar rats aged 7 days were randomly divided into 4 groups; 8 in control group, 24 in HIBD model group, 24 in GM-1 group and 24 in NEP1-40 group. The rats of the control group and HIBD group were injected with normal saline (0.25 ml/kg) intraperitoneally, while those in NEP1-40 group and GM-1 group with NEP1-40 12.5 microg/d, GM-1 10 mg/(kg.d) for continuous 3 days of 72-hour group or 7 days of 168-hour group, respectively. In situ hybridization was adopted for detecting the expression of NgR in the brain of the rats at the time point of 24 hours, 72 hours and 7 days. Meanwhile histopathological changes of neurons and axon were detected by transmission electron microscopy (TEM). The SPSS statistical software package for Windows, version 10.0, was used to run Chi-square tests and least significance difference (LSD-t) on the data presented, and P value of less than 0.05 was regarded as statistically significant.

RESULTThe expression level of Nogo-A receptor in the control group was higher than that of the other groups at different time point (t value was 5.48, 6.11, 6.96, 8.24, 5.99 and 5.34, respectively, and all P values were less than 0.05). There were no significant differences in Nogo-A receptor level among the HIBD group, the GM-1 group and the NEP1-40 at 24 hours (t was 1.48, 2.76 and 1.29, respectively, and all P > 0.05), while the expression of Nogo-A receptor of NEP1-40 at 72 hours and 7 days was lower than that of the HIBD group and the GM-1 group at the same time point, respectively (all P < 0.05). Repair of neurons in damaged brain to some extent was found after GM-1 treatment and satisfactory repair of neurons and axon regeneration was obtained with NEP1-40 administration as shown by TEM.

CONCLUSIONHypoxic ischemic brain damage can down-regulate the expression of Nogo-A receptor in the central nervous system. NEP1-40 contributes to the regeneration of axon and repair of brain damage, thus exerts neuroprotective effect.

Animals ; Animals, Newborn ; Brain ; drug effects ; metabolism ; pathology ; GPI-Linked Proteins ; Hypoxia-Ischemia, Brain ; metabolism ; pathology ; Myelin Proteins ; pharmacology ; Nogo Receptor 1 ; Peptide Fragments ; pharmacology ; Rats ; Rats, Wistar ; Receptors, Cell Surface ; Receptors, Peptide ; metabolism

OBJECTIVETo observe and investigate the risk factors and pathogen diversification of nosocomial lower respiratory infections in patients with hematological malignancy after chemotherapy.

METHODSRespiratory tract microbial population of fifty patients with different kinds of hematological malignancy and para-prepared to chemotherapy was quantitatively analyzed before and after chemotherapy at an arranged time from April, 2004 to December, 2005. Susceptibility test was determined for bacterium of nosocomial infection, and the homology of the same species of the bacteria was analyzed by a pulsed field gel electrophoresis (PFGE).

RESULTSIncidence rate of lower respiratory infections in patients with the hematological malignant after chemotherapy was 16%. The major nosocomial infectious pathogens were Acinetobacter spp; Escherichia coil and Fungus. Among them, Acinetobacter spp, were highly resistant to cephalosporins, quinolones, aminoglycosides, carbapenems and antibiotic with enzyme inhibitor, respectively but susceptible to Cefoperazone/Sulbactam belonging to antibiotic with enzyme inhibitor. And it was shown that there were two clones by the pulsed field gel electrophoresis (PFGE).

CONCLUSIONFollowing-up of nosocomial lower respiratory infection in patients with hematological malignancy after chemotherapy might offer theoretical evidence for the rational use of antibiotics and the control of nosocomial infections.

Acinetobacter baumannii ; drug effects ; isolation & purification ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Cross Infection ; epidemiology ; Escherichia ; drug effects ; isolation & purification ; Female ; Follow-Up Studies ; Hematologic Neoplasms ; drug therapy ; immunology ; Humans ; Leukocyte Count ; Male ; Middle Aged ; Opportunistic Infections ; epidemiology ; Respiratory Tract Infections ; epidemiology

Objective To investigate the expression levels of CD40L in CD4 + T cells from systemic sclerosis (SSc) patients.Methods PBMC ( peripheral blood mononuclear cells) cells were isolated from the peripheral venous blood of SSc patients (16females,10males) and healthy donors (15females,10males) by density gradient centrifugation.CD4 + T cells were isolated using magnetic beads.mRNA levels of CD40L in CD4 + T cells were measured by real-time quantitative polymerase chain reaction (RTPCR).Flow cytometric analysis was used to detect the CD40L protein on the surface of CD4 + T cells.Results Both CD40L mRNA and protein expression in CD4 +T cells was significantly elevated in female SSc patients compared with female healthy controls [5.61 ± 1.86 vs 2.80 ±0.94,P ＜0.01 ; (6.70 ±3.55)％vs (2.37 ± 1.39)％,P ＜ 0.05,respectively].No significant increase in CD40L mRNA and protein expression was observed in male SSc patients compared with male controls [2.59 ± 0.89 vs 1.92 ± 0.56,P ＞0.05; (2.06±1.09)％ vs (2.13±0.87)％,P ＞0.05,respectively].Conclusions CD40L was overexpressed in CD4 + T cell of female patients but not males,which maybe one of the important reasons for female susceptibility to SSc.

[Objective] To study the methylation status of CD40L gene regulatory regions in peripheral blood CD4+ T cells from patients with systemic sclerosis (SSc).[Methods] Peripheral blood mononuclear cells were isolated from the venous blood of 21 SSc patients and 20 healthy controls by density gradient centrifugation.CD4+ T cells were separated by using magnetic beads.Genomic DNA was isolated from the CD4+ T cells and treated with sodium bisulfite.Nested PCR was perfonned to amplify the desired regulatory sequences (including the promotor and enhancer) of CD40L,and the amplicons were transformed into the Escberichia coli DH5α.Subsequently,8 independent clones were selected and sequenced for each of the amplified fragments.[Results] In healthy female controls,half of the cloned fragments of CD40L regulatory sequences were unmethylated,and the other half were methylated.The mean methylation levels of CD40L promoter and enhancer from female SSc patients were significantly lower than those from healthy female controls (both P ＜ 0.01 ).Almost all of the cloned fragments of CD40L promoter and enhancer were unmethylated in healthy male controls and male SSc patients,with no significant difference in the methylation level between male SSc patients and healthy controls (both P ＞ 0.05,respectively).[Conclusion]s There is a low methylation level of CD40L regulatory elements on the inactive X chromosome in female SSc patients,which may contribute to the CD40L overexpression in CD4+ T cells.

OBJECTIVETo study the inhibitory effect of paeoniflorin (PAE) on TNF-α-induced TNF receptor type I (TNFR1)-mediated signaling pathway in mouse renal arterial endothelial cells (AECs) and to explore its underlying molecular mechanisms.

METHODSMouse AECs were cultured in vitro and then they were treated by different concentrations PAE or TNF-α for various time periods. Expression levels of intercellular cell adhesion molecule-1 (ICAM-1) were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 6-h TNF-α 30 ng/mL), the low dose PAE group (cultured by 2-h PAE 0.8 μmo/L plus 6-h TNF-α 30 ng/mL), the middle dose PAE group (cultured by 2-h PAE 8 μmol/L plus 6-h TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 6-h TNF-α 30 ng/mL) with Western blot analysis. Nuclear translocation of transcription factor NF-κB (NE-κB) was detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 45-mm TNF-α 30 ng/mL), and the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 45-min TNF-α 30 ng/mL) by immunofluorescent staining. Expression levels of the phosphorylation of extracellular signal-regulated (protein) kinase (ph-ERK) and p38 (ph- p38) were detected in the normal group (cultured by serum-free culture media) and the high dose PAE group (2-h PAE 80 μmol/L culture) by Western blot. NF-κB inhibitor-α (IκBα) protein expressions were detected in the normal group (cultured by serum-free culture media), the TNF-α group (cultured by 2-h serum-free culture media plus 30-min TNF-α 30 ng/mL), the high dose PAE group (cultured by 2-h PAE 80 μmol/L plus 30-min TNF-α 30 ng/mL), the p38 inhibitor group (SB group, pretreatment with SB238025 25 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min), the ERK inhibitor group (PD group, treated by PD98059 50 μmol/L for 30 min, then treated by PAE 80 μmol/L for 2 h, and finally treated by TNF-α 30 ng/mL for 30 min) by Western blot.

RESULTSCompared with the normal group, ICAM-1 protein expression levels obviously increased (P < 0.01). Compared with the TNFα group, ICAM-1 protein expression levels were obviously inhibited in the high dose PAE group (P < 0.05). Protein expression levels of ph-p38 and ph-ERK were obviously higher in the hIgh dose PAE group (P < 0.05). Compared with the normal group, IκBα protein expression levels obviously decreased in the TNF-α group (P < 0.01). Compared with the TNFα group, TNF-α-induced IκBα degradation could be significantly inhibited in the high dose PAE group (P < 0.01); the inhibition of PAE on IκBα degradation could be significantly inhibited in the SB group (P < 0.05). NF-κB/p65 signal was mainly located in cytoplasm in the normal group. NF-κB/p65 was translocated from cytoplasm to nucleus after stimulated by 45 min TNF-α in the TNF-α group, while it could be significantly inhibited in the high dose PAE group.

CONCLUSIONSPAE inhibited TNF-α-induced expression of lCAM-1. Its action might be associated with inhibiting TNFR1/NF-κB signaling pathway. p38 participated and mediated these actions.

Animals ; Cells, Cultured ; Endothelial Cells ; cytology ; drug effects ; Glucosides ; pharmacology ; Intercellular Adhesion Molecule-1 ; metabolism ; Mice ; Monoterpenes ; pharmacology ; NF-kappa B ; metabolism ; Receptors, Tumor Necrosis Factor ; metabolism ; Signal Transduction ; drug effects ; Tumor Necrosis Factor-alpha ; pharmacology