In recent years, cytokinesis-block method was used to analyze cytomics indicators including micronucleus, nuclear bridge, nuclear bud, nuclear division index, cell apoptosis and cell necrosis. In public health, it has become the common method to explore the impacts of different population structure, environment and occupational exposure for genomic instability, chromosome breakage, chromosome loss and cell proliferation. This article reviews and discusses the application of using cytokinesis block method to analyze cytomics indicators in public health field.

Objective To evaluate the femoral neck anteversion (FNA) of patients with unilateral developmental dysplasia of the hip (DDH) and reveal the developmental regularity of the proximal femur.Methods 366 three-dimensional CTs of unilateral DDH patients were categorized into three age groups:＜18 months, 18 months-3 years,and＞3 years.The femoral neck anteversion of both sides were measured and a statistical comparison was performed between them.The line chart showing the relationship of femoral neck anteversion and age was drawn to reveal the developmental regularity of the proximal femur.Results In total,the affected side of unilateral DDH had an femoral neck anteversion 1°～5° significantly larger than the unaffected side (P＜0.05).There was no statistical difference between affected and unaffeted sides in age group＜18 months and 18 months-3 years,while it was significant in age group＞3 years.Conclusion Pathological changes of the proximal femur were observed not only the affected side in unilateral DDH but also the unaffected side.The FNA symmetrically developed with age older.

Objective To define the normal reference values of acetabular index and Sharp angle through 2333 standard anterior-posterior pelvic radiographs. Methods In our study, 2333 normal anteriorposterior pelvic radiograph images with standard exposure were selected. All the images were diagnozed normal by two radiologists and two pediatricians according to the criteria of T(o)nnis. All subjects were without any neuromuscular diseases and congenital defoemity. The acetabular index was measure in the subjects between age 0 to 10 years, and the groups were divided monthly within 1 year and yearly between 1 to 10years. The Sharp angle was measure in the subjects after 10 years, and the groups were divided yearly in adolescence and decadely in adults. Normal values of each age groups and the correlation charts were completed according to statistical analysis. Results The mean acetabular index was 28.39° in neonates followed by a steep decrease in the first three months after born. It decreased to 22.17°in the 1st year, 12.80°in the 10th years and then kept constant. Acetabular index of the female was generally 1°-2°larger than that of the male with statistical significance. The mean Sharp angle was 46.72°in the 10th years, which decreased to 39.10°in the 18th years and 35.69°in elderly people. Another declination was observed after age 40.Generally no gender difference was observed. Conclusion Acetabular index and Sharp angle vary with age.They reflect a dramatic morphological change in the acetabulum before adulthood and stay constant afterwards. Gender difference is obvious in many age groups but not all. Normal reference ranges of both gender at all age groups should be considered in clinical evaluation.

Objective:To explore the feasibility of the optimized cytokinesis-block (CB) assay on radiation-induced nucleoplasmic bridge (NPB), and to provide a scientific basis for the application of NPB in biological dose estimation.Methods:Human peripheral blood in vitro was irradiated with 2 Gy 60Co γ-rays at a dose rate of 1 Gy/min (0 Gy control group). According to the culture time after irradiation, blood samples were divided into group 48, 56, 68 and 72 h. Cytochalasin-B (Cyt-B) with a concentration of 6 μg/ml was added into the samples at 28 h and harvested at 48, 56, 68 and 72 h after irradiation, respectively. On the other hand, the blood samples were treated with different concentration of Cyt-B i. e., 0.6, 1, 2, 6 and 10 μg/ml at the beginning of culture (0 h) and harvested at 68 h after irradiation. The proportion of mononucleated, binucleated and multinucleated cells, radiation-induced NPB and micronucleus (MN) frequencies were analyzed. Results:The nuclear division index (NDI) and proportion of binucleated cells at 2 Gy and 0 Gy had tendency of increasing with cell culture time. NPB frequencies (0.023 0-0.033 0/cell) and MN frequencies had no significantly difference ( P> 0.05). With the increase of Cyt-B concentration, NDI and the proportion of binucleated cells in group 2 Gy and 0 Gy also increased, but NPB frequencies (0.023 0-0.047 0/cell) had no significant difference ( P> 0.05). MN frequencies of group 10 μg/ml were significantly lower than that of group 6 μg/ml ( U=2.74, P< 0.01). Conclusions:Cell culture time and Cyt-B concentration had no significant influence on radiation-induced NPB frequencies, suggesting that NPB could be obtained by appropriately reducing cell culture time and Cyt-B could be added into blood samples at the beginning of culture. But this protocol reduced the number of cells for further analysis, and thus its feasibility for dose estimation still need to be studied.

In order to find a method suitable for purifying large amount of CD34(+) cells, from 5 cases who accepted autologous peripheral blood stem cell transplantation, CD34(+) cells were collected and enriched by using Isolex 300i (Nexell). Phenotypes were detected by flow cytometry and the biological viability were assayed by the colony-forming experiments and cell expansion experiment in vitro. The results showed that the number of mononuclear cells first collected was about (3.5 - 6.0) x 10(10) and (0.55 - 1.2)% of cells were CD34 positive. The number of positive production was about (2.0 - 3.0) x 10(8); the CD34(+) cells purity was (75 - 85)% and the yield was (40 - 65)%. The CD34(+) cells of positive production could expand up to 2 - 3 times when cultured with SCF + IL3 + FL + TPO + EPO in vitro. The results of colony-forming experiments demonstrated that the CD34(+) cells collected has enough colony-forming ability. All results showed the enriched CD34(+) cells with biological viability. In conclusion, the CD34(+) immunomagnetic isolation apparatus Isolex300i is suitable to clinical application for a large amount of CD34(+) cell enrichment.
Antigens, CD34 ; immunology ; Colony-Forming Units Assay ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Immunomagnetic Separation ; instrumentation ; methods ; standards ; Neoplasms ; blood

Objective To explore the influences of the final concentration and adding time of Cytochalasin-B (Cyt-B) on radiation-induced nucleoplasmic bridges (NPB) in cytokinesis-block assay.Methods Hunan peripheral blood samples were divided into 5 final concentration groups (group 2,4,6,8,10 μg/ml) according to different final concentrations of Cyt-B.Moreover,blood samples were divided into 4 adding time groups (group 0,28,40,44 h) according to different adding times of Cyt-B.Blood samples were irradiated with 0 (sham irradiation) and 2 Gy 60Co-rays in vitro,at a dose rate of 1 Gy/min.A cytokinesis-block assay was carried out to prepare NPB samples.The percentages of mononucleated,binucleated and multinucleated cells,as well as the frequencies of NPB and micronucleus (MN) in binucleated cells were analyzed using an optical microscope.Results Nuclear division index (NDI) and the percentages of binucleated cells increased with increased concentration of Cyt-B,and decreased with delayed adding time of Cyt-B (except group 0 h) in both final concentration groups and adding time groups.After exposed to 2 Gy,NPB frequencies were no significant difference (except group 0 h).MN frequencies had the trend of decreased with the increased concentration of Cyt-B,but no significant difference with adding time of Cyt-B.Conclusions In cytokinesis-block assay,different final concentration and adding time of Cyt-B may induce to the variation of NPB frequencies,but there was no significant difference.Appropriate increased final concentration or ahead adding time of Cyt-B can increase the percentage of binucleated cells that help to improve the efficiency of analysis.

Objective:To screen radiosensitive lipid metabolites in rat small intestine and analyze their metabolic pathways, in order to provide scientific basis for radiation enteropathy biomarkers.Methods:The total body irradiation of 60Co γ rays was performed to rats with different doses of 0, 1, 2, 3, 5 and 8 Gy. The changes of lipids in small intestine were studied by targeted lipidomics method based on liquid chromatography coupled mass spectrometry (LC-MS). Results:Fifteen lipids in small intestine were screened as radiosensitive metabolites at 3 d after irradiation, including 4 up-regulated lipids and 11 down-regulated lipids( t=-6.395, 5.998, 5.836, -5.503, -5.449, -5.422, 4.841, 4.802, 4.621, 4.457, 4.426, 4.373, 4.110, 3.945, 3.902, P< 0.05 and FDR < 0.05). The metabolic pathways of sphingolipid, glycerophosphoplipid were significantly enriched. Four phosphatidyl serines (PS)increased while 1 phosphatidic acid(PA), 2 sphingomyelins(SM) and 4 fatty acids(FA)decreased in a good dose-response manner( R2> 0.80, P< 0.05), which were more potential radiation enteropathy biomarkers. Conclusions:Lipid metabolites in rat small intestine were significantly changed after the rat was total body irradiated with 60Co γ-rays.Eleven lipids with good dose-response relationship were more potential to be radiation enteropathy biomarkers.

[Abstract] Objective: To construct CD33-CAR modified NK92 cells based on CD33-scFv sequence, and to explore its killing effect on CD33+ AML (acute myeloid leukemia) cells. Methods: DNA fragment encoding CD33-CAR was synthesized by gene synthesis and molecular cloning technology and then cloned into lentiviral vector. Lentivirus were packaged and used to transfect NK92 cells. The transfection efficiency was detected by flow cytometry, and puromycin was used to screen NK92 cells stably expressing CD33-CAR (CD33-CAR-NK92). Killing effect of CD33-CAR-NK92 cells on AML cells in vitro was examined with calcein-AM release assays. IFN-γ secretions of NK92 cells and CD33-CAR-NK92 cells were measured by ELISA. Results: The pCDH-CD33-CAR lentiviral vector was successfully constructed. After lentiviral transfection, about 18.7% of NK92 cells express CD33-CAR (referred as CD33-CARNK92 cells). The percentage of CD33-CAR+ NK92 cells was about 86.3% after puromycin selection. In contrast to unmodified NK92 cells, significantly higher cytotoxic effect against CD33+ MOLM-13 cells was found in CD33-CAR-NK92 cells (P<0.01); however, there was no significant difference in cytotoxicity against CD33- JURKAT cells between NK92 cells and CD33-CAR-NK92 cells (P> 0.05). After co-culture at an effect-target ratio of 2∶1 for 6 hours, the level of IFN-γ secreted by the CD33-CAR modified NK92 cells was significantly higher than that of the unmodified ([190.97±11.52] vs [88.41±2.75]pg/ml, P<0.01). Conclusion: The CD33-CARNK92 cells could specifically recognize CD33 antigen and kill CD33+ AML cells in comparison with the unmodified NK92 cells, which provides experimental basis for clinical transformation of CD33-CAR-NK92 cells in treatingAML.

OBJECTIVETo measure levels of cytokines including IL-1β, IL-12, IL-18 and TNF-α in children with newly diagnosed type 1 diabetes and to analyze their correlation with clinical indices such as infection and onset time.

METHODSA total of 33 children with newly diagnosed type 1 diabetes were assigned to the case group, and 27 healthy children to the control group. The case group was further divided into increased white blood cell (WBC) and normal WBC subgroups according to peripheral WBC level. The serum levels of cytokines including IL-1β, IL-12, IL-18 and TNF-α were measured by enzyme-linked immunosorbent assay. Blood pH, blood sugar, blood lactate, fructosamine, peripheral leukocytes and neutrophils and some other clinical indices were also measured.

RESULTSThe level of IL-12 in the case group was higher than in the control group (P<0.001). In the case group, the level of IL-18 was negatively correlated with onset time (r=0.413, P=0.015), the neutrophil count was positively correlated with IL-1β level (r=0.413, P=0.023) and the WBC count was positively correlated with IL-18 level (r=0.352, P=0.038). IL-1β, IL-12 and IL-18 levels in the increased WBC subgroup were higher than in the normal WBC subgroup (P<0.05 for all comparisons).

CONCLUSIONSCytokine secretion disorders of Th1 cells exist in children with type 1 diabetes. Infections may induce cytokine secretion and might contribute to the early onset of diabetes.

Adolescent ; Child ; Child, Preschool ; Cytokines ; blood ; Diabetes Mellitus, Type 1 ; immunology ; Female ; Humans ; Infant ; Male ; Th1 Cells ; immunology

Objective:To evaluate the effect of blunt separation method on peripherally inserted central venous catheters.Methods:The randomized controlled trials regard to the effect of blunt separation method on peripheral central venous catheters were retrieved from CNKI, Wan fang Data Knowledge Service Platform, CQVIP, CBM, Cochrane, PubMed, EMBASE and Web of science from January 2010 to March 2021, two researchers performed quality assessments and extracted data according to Cochrane evaluation manual standards. RevMan 5. 4 software was used for statistical analysis.Results:A total of 11 randomized controlled trials were analyzed, including 1379 participants. The meta-analysis results showed that there was no statistical difference in the successful rate of one-time sheath delivery ( OR=1.62, 95% CI 0.92-2.86, P>0.05). However, patients in blunt separation group showed lower blood loss ( OR=0.24, 95% CI 0.11-0.50, P<0.05), lower the incidence of seepage at puncture site ( OR=0.18, 95% CI 0.09-0.37, P<0.05) than those in traditional skin expansion group at 24h after catheterization, and less the time of maintenance within 7 days after catheterization ( MD=-0.95, 95% CI-1.78--0.11, respectively, P<0.05). Conclusions:Blunt separation method can significantly reduce the incidence of complications and the time of catheter maintenance. Due to the limited quality of included studies, more high quality studies are needed to verify the conclusions.