Establishment and validation of OX40/FcγR-humanized mice for the study of agonistic anti-OX40 antibody / 上海交通大学学报（医学版）

Objective • To establish a rapid method to evaluate the activity of agonistic antibody using OX40 (tumor necrosis factor receptor superfamily member 4)/FcγR (Fcγ receptor)-humanized mice. Methods • Bone marrow cells from OX40-humanized mice and FcγR-humanized mice were collected and mixed with equal ratio. Then the mixed bone marrow cells were administrated into irradiated wild-type mice through the tail veins. The reconstruction efficiency of the immune system was confirmed by detecting the expression of hOX40 and hFcγR in the immune cells of chimera mice. After the chimera mice were generated successfully, they were used to evaluate the immunostimulatory activity of anti-hOX40 antibodies to CD4+ or CD8+ T cells. The results of flow cytometry were statistically analyzed. The unpaired t-test was used to compare the means between the two groups, and oneway ANOVA was used to compare the means between multiple groups. Results • Flow cytometry analysis showed that wild-type recipient mice were efficiently reconstituted with hFcγR expressing cells and hOX40 expressing cells to generate OX40/FcγR-humanized bone marrow chimera mice. In these mice, B cells and myeloid cells expressed hFcγRs (P<0.05), and T cells expressed hOX40 upon in vitro stimulation (P<0.05). When these mice were used to evaluate the immunostimulatory activity of anti-hOX40 antibody, significant expressions of IFN-γ and hOX40 were observed (P<0.05). Conclusion • OX40/FcγR-humanized bone marrow chimera mice are generated based on hFcγR expressing cells and hOX40 expressing cells, suggesting a rapid method to build a mouse model with both hFcγR and hOX40 expression. These mice are suitable for evaluating the immunostimulatory activity of agonistic human anti-hOX40 antibodies.

Clinical Significance of FGFR1 Gene Abnormalities in Blood Tumors / 中国实验血液学杂志

OBJECTIVE@#To study the potential significance and clinical application of FGFR1 gene abnormality in the diagnosis, clinical features, pathological mechanism and treatment in hematological tumors.@*METHODS@#Clinical data of total of 29 patient with chromosome of 8 short arm (8P) abnormality who had more comprehensive medical history from 2013 to 2018 were collected. The karyotype analysis of bone marrow chromosomes in patients was carried out by using chromosome R band banding technique. FGFR1 gene was detected by using fluorescence in situ hybridization (FISH).@*RESULTS@#Seven cases of FGFR1 gene abnormalities were decteted, including 3 cases of FGFR1 gene amplification, 2 cases of translocation, and 2 cases of deletion. Five patients with FGFR1 gene amplification or deletion not accompaned with eosinophilia, moreover the chromosome was a complex karyotype with poor prognosis; Two cases of FGFR1 gene translocation were non-complex chromosomal translocation and one of which survived for 6 years after bone marrow transplantation, the other chromosome karyotype showed no rearrangement of 8 short arm. However, FGFR1 gene rearrangement was confirmed by FISH analysis, which was a rare insertional translocation.@*CONCLUSION@#FGFR1 gene amplification or deletion often occur in cases with complex karyotype, which not accompany eosinophilia, moreover have poor prognosis. The patients with FGFR1 gene translocation accompany eosinophilia which is consistent with the clinical characteristics of myeloid / lymphoid neoplasms with FGFR1 abnormality. Karyotype analysis combined with FISH method can improve the detection of abnormal clones.

Effect of Tanshinone II A on Cytokines of Rats with Severe Acute Pancreatitis Lung Injury / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of Tanshinone II A on severe acute pancreatitis (SAP) lung injury (ALI) rats and its possible mechanism.</p><p><b>METHODS</b>SD rats were injected with sodium taurocholate to induce SAP group, and then intervened with sodium tanshinone II A sulfonate ( STS group). Simultaneously a sham-operation group (SO group) was set up. There were 24 rats in each group. The survival state and wet-to-dry weight ratio of lung tissues were observed. Activities of myeloperoxidase (MPO) in lung were determined by MPO reagent kit. Pathologic changes of lung tissues were determined by Hofbuaer method. Expression levels of three cytokines, TNF-alpha, IL-1beta, and intercellular cell adhesion molecule-1 (ICAM-1) were detected by ELISA.</p><p><b>RESULTS</b>The survival state of rats in the SAP group was deteriorated. The wet-to-dry weight ratio, MPO activities, pathologic changes in lung tissues, and expression levels of TNF-alpha, IL-1beta, and ICAM-1 increased significantly more in the SAP group than in the SO group (P < 0.05). Compared with those in the SAP group, the survival state of rats in the STS group was improved; the wet-to-dry weight ratio, MPO activities, pathologic changes in lung tissues, and expression levels of TNF-alpha, IL-1beta, and ICAM-1 obviously decreased in the STS group (P < 0.05).</p><p><b>CONCLUSION</b>Tanshinone II A had remarkable effect on SPA LI rats, which might be associated with changing cytokines levels and attenuating infiltration of lung inflammatory cells.</p>

Determination of cinobufagin and resibufogenin in liver tissue by HPLC-MS/MS / 法医学杂志

OBJECTIVE@#To develop a sensitive and accurate assay for detecting cinobufagin and resibufogenin in liver tissue using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).@*METHODS@#The homogenization of liver tissue with internal standard dexamethasone was extracted with dichloromethane. The extracts with methanol were purified through ProElut C18 solid phase extraction and tested in positive electrospray ionization with multiple reaction monitoring of HPLC-MS/MS.@*RESULTS@#The good linear relationship of cinobufagin and resibufogenin in liver tissue were 1-204 ng/g and 1-206 ng/g, respectively. The minimal detection threshold (S/N > or = 3) of this method was 0.3 ng/g for both cinobufagin and resibufogenin. The matrix effect was 96.5%-126.7%. The extraction recovery coefficient was 70.0%-82.3%. The precision of intra-day and inter-day was less than 10%.@*CONCLUSION@#This method is sensitive and reliable, and can be used in forensic toxicological analysis.

LC-MS/MS analysis of determination of strychnine and brucine in formaldehyde fixed tissue / 法医学杂志

OBJECTIVE@#To establish a method for determination of strychnine and brucine in formaldehyde fixed tissue by LC-MS/MS analysis.@*METHODS@#The samples were pretreated with solid phase extraction using SCX cartridges and separated on SB-C18 column with mobile phase 0.1% formic acid : 0.1% formic acid-acetonitrile (75:25). Electrospray ionization (ESI) source was utilized and operated in positive ion mode. Multiple reactions monitoring (MRM) mode was applied. External standard method was applied for quantitation.@*RESULTS@#The chromatographic separation of strychnine and brucine in formaldehyde fixed nephritic and hepatic tissues resulted successfully. The standard curve was linear in the range of 0.002-2.0 microg/g for strychnine and brucine in formaldehyde fixed tissues, and the correlation coefficient was more than 0.996. The limits of detection (LOD) of strychnine and brucine in nephritic tissues were 0.06ng/g and 0.03 ng/g, respectively. The LOD of both chemicals were 0.3 ng/g in hepatic tissues. The extraction recovery rate was more than 74.5%. The precision of intra-day and inter-day were both less than 8.2%.@*CONCLUSION@#Strychnine and brucine can be sensitive to be determined in formaldehyde fixed tissue by LC-MS/MS analysis. It can be applied in the forensic toxicological analysis.

Chemical composition of Galla chinensis extract and the effect of its main component(s) on the prevention of enamel demineralization in vitro / 国际口腔科学杂志·英文版

To determine the chemical composition of Galla chinensis extract (GCE) by several analysis techniques and to compare the efficacy of GCE and its main component(s) in inhibition of enamel demineralization, for the development of future anticaries agents, main organic composition of GCE was qualitatively determined by liquid chromatography-time of flight-mass spectrometry (LC-TOF-MS) and quantified by high-performance liquid chromatography-diode array detector (HPLC-DAD). Inorganic ions were tested by inductively coupled plasma-atomic emission spectroscopy and F was especially measured by ion chromatography. Then, bovine enamel blocks were randomly divided into four treatment groups and were subjected to a pH-cycling regime for 12 times. Each cycle included 5-min applications with one of four treatments: 4 g⋅L(-1) GCE solution, 4 g⋅L(-1) gallic acid (GA) solution, 1 g⋅L(-1) NaF solution (positive control), deionized water (DDW, negative control), and then 60-min application in pH 5.0 acidic buffer and 5-min application in neutral buffer. Acidic buffers were retained for calcium analysis. The main organic composition of GCE were GA and its isomer, and, to a lesser extent, small molecule gallotannins. The content of GA in GCE was 71.3%±0.2% (w/w). Inorganic ions were present in various amounts, of which Ca was (136±2.82) µg⋅g(-1), and Zn was (6.8±0.1) µg⋅g(-1). No F was detected in GCE. In pH cycling, GA showed an effect similar to GCE in inhibiting enamel demineralization (P>0.05). GA was found to be the main effective, demineralization inhibiting component of GCE and could be a promising agent for the development of anticaries agents.

New tactics of human red blood cells stored at 4 degrees C-protective effect of antioxidant solution on red blood cells damage / 中国实验血液学杂志

Aliquots of venous blood from healthy donor were collected in plastic blood storage bags with ACD, GMA or antioxidant solution (superoxide dismutase, SOD), respectively, and stored at 4 degrees C. After storage for varying periods, the parameters of the blood were detected in the blood samples. Results showed that the parameters of the blood stored at 4 degrees C for 75 days in SOD group were following: the recovery of RBC-Hb was 87.2%, plasma-Hb (mg/L) was 193.2, P50 (mmHg) was 34.0 (normal value was 33.1); deformability (DImax) was 0.2413 (74.3% of normal value). There was no evident hemolysis, color change, air bubble and clots. It was concluded that human RBC stored at 4 degrees C for 75 days with SOD solution, recovery of levels of RBC-Hb and plasma-Hb were accorded with the requirements of "Basic Demands of Blood Station" in China.