An iodine analysis instrument with sequential injection of urine samples was developed. A method for measurement of urine iodine was also developed by combining sequential injection and catalysis kinetics,and making use of catalysis facilitation of the iodine in the redox reaction of As~(3+) and Ce~(4+) . The sequential injection and stopped-flow stabilization determination were made possible by the program-controlled injector with controlled flow rate and the 16-hole program-controlled selection valves. The arsenic-cerium reaction with iodine-catalyzed at constant temperature state might proceed with the constant temperature flow cell. Using the syringes with program-controlled velocity,pushing and suction,Using the digital connected circuit and micro-iodine determination software,the reaction temperature of (32.0±0.1)℃,injection time of 45 s,stabilization time of 60 s,detection time of 20 s,injection volume of 400μL,linear range of 15 -600μg/L,detecting limitation of 5.01 μg/L(n=11) and recovery rate of 94. 1 % - 105. 1 % were obtained. With this method,the detecting result of the National Standard Reference (GBW09109 and GBW09110) materials was within a given standard range. Through this method,the detecting results had no significant differences comparing with those by standard method of National Health Service(P >0.05).

The purpose of this study was to optimize a fixation procedure for detection of cytoplasmic antigens by flow cytometry(FCM) and to evaluate the effect of intracellular CD3, CD22, CD79a and myeloperoxidase(MPO) in lineage assignment. Four kinds of fixation procedure and three or four color direct immunofluorescence staining were used to permeate cell membrane and label cell surface and intracellular antigens by means of FCM. Results showed that percentage of cytoplasmic antigens positive cells was the highest and cell scatter and fluorescence intensity of CD45 were not changed after using of FACS permeabilization solution. MPO protein was positive in 16/18 acute myeloid leukemia(AML) patients. 4 cases of T cell-acute lymphoblastic leukemia (T-ALL) cases were positive for cytoplasmic CD3(c CD3) but surface CD3 was negative. c CD22 was only detected in 9/13 of B-ALL and cCD79a was positive in 5/5 B-ALL. 18/38 cases of acute leukemia were expressed in more than one lineage marker, 8/21 cases of acute non-lymphocytic leukemia(ANLL) were CD7 positive. 7/17 cases of acute lymphocytic leukemia (ALL) expressed CD13. After further cytoplasmic antigen detection, one was considered to be a T/myeloid biphenotypic leukemia, another one was diagnosed as biclonal or mixed leukemia. The results suggest that intracellular CD3,CD22,CD79a and MPO are lineage specific markers, they are very important for biphenotypic and biclonal/mixed acute leukemia identification
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antigens, CD ; analysis ; biosynthesis ; Antigens, Differentiation, B-Lymphocyte ; biosynthesis ; CD3 Complex ; biosynthesis ; CD79 Antigens ; Cell Adhesion Molecules ; Child ; Child, Preschool ; Diagnostic Techniques and Procedures ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; methods ; Lectins ; biosynthesis ; Leukemia ; classification ; pathology ; Male ; Middle Aged ; Peroxidase ; biosynthesis ; Receptors, Antigen, B-Cell ; biosynthesis ; Sialic Acid Binding Ig-like Lectin 2

To prove the significance of 4-color fluorescence labeling flow cytometric analysis for leukemia immunophenotyping, bone marrow and peripheral blood samples from 88 cases with acute leukemia were analyzed by means of flow cytometry with 4-color fluorescence labeling monoclonal antibodies (McAbs). More than 90% of cases could be classified into acute myeloid, T- or B-lymphocytic leukemia by staining with 13 different kinds of monoclonal antibodies within 4 tubes of combination of antibodies. AML-M(2) - M(7) could also be identified simultaneously. Another one tube of combination of 4 McAbs could be used for further subtyping of those who were undetermined by the above 4 tubes. Patients with AML-M(0)/M(1) which may be negative of myeloperoxidase (MPO) in cytochemistry can be identified by labeling with both membrane and cytoplasmic antibodies. In conclusion, the accuracy of leukemia immunophenotyping has been improved by 4-color flow cytometry analysis.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Fluorescent Antibody Technique ; Humans ; Immunophenotyping ; Infant ; Leukemia ; immunology ; Leukemia, Myeloid, Acute ; immunology ; Leukocyte Common Antigens ; analysis ; Lipopolysaccharide Receptors ; analysis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; Proto-Oncogene Proteins c-kit ; analysis

BACKGROUND: The effect of pituitary adenylate cyclase activating polypeptide (PACAP) during traumatic brain injury (TBI) and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI. METHODS: Male Sprague Dawley rats were randomly divided into 3 treatment groups (n=6, each): sham-operated, vehicle (normal saline)+TBI, and PACAP+TBI. Normal saline or PACAP (1g/5L) was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4+ and CD8+ T cells in blood and the spleen were detected via flow cytometry. RESULTS: In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4+ CD8– lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI+vehicle, and CD4– CD8+ were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4+, and decreased CD8+, T lymphocytes in blood and the spleen. CONCLUSION: Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

An on-line two dimensional liquid chromatographic (2D-LC) method was established by using an ultimate dual gradient liquid chromatography, three chromatographic columns and valve-switching technology to detect 2460A in trichoderma hazianum fermentation liquor. MF C8 column (10 mm×4. 6 mm, 5. 0 μm) was used as purification column and MG C18 column (20 mm×4. 6 mm, 5. 0 μm) was used as enriching column. Methanol and water were used as mobile phase with a gradient elution at a flow rate of 2. 0 mL/min. The sample was separated on the Thermo Hypersil GOLD C18 column (250 mm×4. 6 mm, 5. 0μm) maintained at 40 ℃ using methanol and water. The flow rate was 1. 0 mL/min and 1. 0 mL sample was injected into the 2D-LC system. The detection wavelength was 424 nm. The whole analytical time was less than 60 min. The standard curve was linear over the 2460A concentration range of 0. 0025-10. 0 mg/L(r=0. 9981, n=8). The limit of detection was calculated to be 1 . 2μg/L ( S/N=3 ) and the limit of quantification was calculated to be 2 . 5 μg/L ( S/N=10 ) . The average recoveries varied from 88 . 0% to 104 . 4%.

This study investigated the effects of hyperoxia on dynamic changes of thioredoxin-1 (Trx1) and thioredoxin reductase-1 (TrxR1) in alveolar type II epithelial cells (AECII) of premature rats. Pregnant Sprague-Dawley rats were sacrificed on day 19 of gestation. AECII were isolated and purified from the lungs of premature rats. When cultured to 80% confluence, in vitro cells were randomly divided into air group and hyperoxia group. Cells in the hyperoxia group were continuously exposed to 95% O(2)/5% CO(2) and those in the air group to 95% air/5% CO(2). After 12, 24 and 48 h, cells in the two groups were harvested to detect their reactive oxygen species (ROS), apoptosis, TrxR1 activity and the expressions of Trx1 and TrxR1 by corresponding protocols, respectively. The results showed that AEC II exposed to hyperoxia generated excessive ROS and the apoptosis percentage in the hyperoxia group was increased significantly at each time points as compared with that in the air group (P<0.001). Moreover, TrxR1 activity was found to be markedly depressed in the hyperoxia group in comparison to that in the air group (P<0.001). RT-PCR showed the expressions of both Trx1 and TrxR1 mRNA were significantly increased in AECII exposed to hyperoxia for 12 and 24 h (P<0.01), respectively. At 48 h, the level of Trx1 mRNA as well as that of TrxR1 mRNA in the hyperoxia group was reduced and showed no significant difference from that in the air group (P>0.05). Western blotting showed the changes of Trx1 protein expressions in the hyperoxia group paralleled those of Trx1 mRNA expressions revealed by RT-PCR. It was concluded that hyperoxia can up-regulate the protective Trx1/TrxR1 expressed by AECII in a certain period, however, also cause dysfunction of the cytoplasmic thioredoxin system by decreasing TrxR1 activity, which may contribute to the progression of oxidative stress and cell apoptosis and finally result in lung injury.

Objective:To explore the intervention effect of motivational interviews based on timing theory on self-efficacy, negative affect and coping styles of parents with infantile spasms children.Methods:Cluster sampling was used to select 82 parents of infantile spasms hospitalized in the Department of Neurology of a children’s hospital, a three-A hospital from January 2019 to October 2019. They were divided into control group and observation group with 41 cases each according to random number table. The control group received routine health education, and the observation group received five motivational interviews based on timing theory interventions on the basis of routine care. The effect of the intervention was evaluated by General Self-Efficacy Scale (GSES), Hospital Anxiety and Depression Scale (HADS), and the Chinese version of Coping Health Inventory for Parents (CHIP) before intervention, on the day of discharge, and 3 months after discharge.Results:Before the intervention, there was no significant difference in the scores of GSES, HADS and CHIP scales between the parents of the two groups ( P>0.05). After intervention, The GSES scores of the observation group on the day of discharge and 3 months after discharge were (19.63±0.87) and (22.58±1.28) points, which were significantly higher than (18.92±0.74) and (19.46±1.25) points of the control group. The difference between both groups was statistically significant ( t values were -3.865, -10.926, P<0.01). HADS-A/HADS-D scores of the observation group on the day of discharge and 3 months after discharge were (12.50±0.82), (10.50±0.87) and (9.78±0.80), (8.63±0.87) points, respectively. The HADS-A/HADS-D scores of the control group on the day of discharge and 3 months after discharge were (12.92±0.74), (11.72±0.99) and (10.23±0.78), (9.38±1.04) points, respectively. The difference was statistically significant ( t values were 2.412-5.764, P<0.05 or 0.01). The observation group scores on CHIP subscales on the day of discharge and 3 months after discharge are higher than the control group, the difference was statistically significant (t values were -7.93--2.490, P<0.05 or 0.01). Conclusions:Motivational interviews based on timing theory can enhance parents’ self-efficacy, improve their negative emotions and family coping styles, and thereby promote the recovery of children.

Objective: To observe the difference in clinical efficacy between acupuncture with point selection based on syndrome differentiation along the meridians and acupuncture at non-meridian and non-acupoint points for functional dyspepsia (FD). Methods: A total of 74 FD patients were randomized into an observation group and a control group, with 37 cases in each group. Both groups received acupuncture treatment. Zusanli (ST 36) and Neiguan (PC 6) were selected in the observation group, with Taichong (LR 3) and Neiting (ST 44) added for excess syndrome, and Gongsun (SP 4) and Yinlingquan (SP 9) added for deficiency syndrome. Four non-meridian and non-acupoint points were selected in the control group. The treatments in both groups were performed once a day with a 2-day break after 5 consecutive treatments, which constituted one treatment course. A total of 4 courses were performed. The scores of Nepean dyspepsia index (NDI) and Leeds dyspepsia questionnaire (LDQ) were recorded before and after treatment, and during follow-up (8, 12, 16, 20 and 24 weeks after recruitment) to assess the clinical efficacy. Results: The NDI scores in the two groups after treatment and at each time point during follow-up were higher than those before treatment (all P<0.05), and the LDQ scores were lower than those before treatment (all P<0.05). The NDI scores after treatment and at each time point during follow-up in the observation group were higher than those in the control group (all P<0.01); the total LDQ score and scores of upper abdominal pain, postprandial satiety and upper abdominal burning sensation after treatment and at each time point during follow-up in the observation group were significantly lower than those in the control group (P<0.01 or P<0.05).. Conclusion: Acupuncture with point selection based on syndrome differentiation along the meridians has a better curative effect than acupuncture at non meridian and non-acupoint points in the treatment of FD.

Objective To investigate the expression of SP 100 protein in ATRA-treated NB4 cells and its effect on pro-liferation in NB4 cells.Methods Q-PCR was employed to measure the expression of SP 100 mRNA;Western blot was used to detect the expression of SP 100 protein; Immunofluorescence was adopted to determine the location of SP100;Cell viability was analyzed by CCK 8;Flow cytometry was used for cell cycle analysis .Results ATRA may induce the expression of mRNA and protein of SP 100.ATRA changes the location of SP100 from a micro-punctate pattern into a punctate nuclear pattern in NB 4 cells.SP100-shRNA promotes the proliferation of NB 4 cells and in-creased the cells in G2/M phase.Conclusions The expression of SP100 was significantly increased in ATRA-treated NB4 cells, and SP100 may be involved in the regulation of proliferation activity of NB 4 cells.

To investigate whether ABL specific tyrosine kinase specific inhibitor STI571 can restore beta1 integrin mediated negative effect on Ph(+) chronic myeloid leukemia(CML), the inhibitory effect of beta 1 integrin activator (beta1 integrin activating antibody 8A2, cytokines such as GM-CSF, G-CSF and SCF) and/or FN on the granulocyte-macrophage colony forming unit (CFU-GM) from 16 patients with Ph(+)CML and 13 normal individuals were examined; the bone marrow mononuclear cells (BMMNC) before and after ABL kinase specific inhibitor STI571 pretreatment (0.1 micro mol/L for 30-60 minutes) were target cells in this study. The roles which VLA4 and VLA5 played in this process were evaluated through blocking assay. The results showed: (1) beta1 integrin activator(s) or FN alone have no effect on CFU-GM from CML or normal bone marrow mononuclear cells before or after STI571 pretreatment, nor STI571 pretreatment itself. (2) The inhibitory effect of beta1 integrin activator(s) plus FN on CML CFU-GM are significantly lower than that on normal CFU-GM. (3) The inhibitory effect of beta1 integrin activator(s) plus FN on CML CFU-GM after STI571 pretreatment is comparable to that on normal CFU-GM. (4) Monoclonal antibody to VLA4 and VLA5 or to total beta1 integrins almost completely abrogate the above effect of STI571. The results suggested enhancing beta1 integrin mediated negative effect on myeloid progenitors in Ph(+)CML is one of the therapeutic mechanisms of STI571 on Ph(+)CML.