1.The repair function of united endothelial progenitor cells transplantation on injured liver endothelium by bone marrow transplantation conditioning
Kunming QI ; Shijuan XU ; Lu JIA ; Guoliang SONG ; Zhiling YAN ; Chong CHEN ; Kailin XU ; Lingyu ZENG
Chinese Journal of Organ Transplantation 2011;32(2):82-86
Objective To study the repair function of united endothelial progenitor cells (EPC)transplantation on injured liver endothelium by bone marrow transplantation (BMT) conditioning.Methods C57BL/6 mice were divided into four groups randomly: normal control group, without any treatment; irradiation alone group, administered a total body irradiation(TBI) pretreatment, without BMT; (3) BMT alone group: C57BL/6 mice were infused with bone marrow mononuclearcells (MNC) 5 × 106/only through caudal vein not more than 4 h after the same TBI pretreatment as the irradiation alone group; united transplantation group: receiving the same way as the BMT alone group, but C57BL/6 mice were infused with EPC 5 × 105/only at the same time. Two, 4, 7, 14, and 21 days after the TBI, the changes of the liver weight were observed regularly. The histopathological examination of liver was done at the 4th, 7th, 14th, and 21st day after the TBI. Results In irradiation alone group, BMT alone group and united transplantation group the liver weight began to increase significantly on the day 2 and peaked at 14th day after the TBI, and the peaks were respectively (1.65±0. 15) times (P<0. 05), (1.61 ±0.06) times (P<0.05), and (1.11 ±0.40)times (P<0. 05) of those in normal control group. At the day 14, the liver weight in irradiation alone group, BMT alone group and united transplantation group began to decrease, and on the day 21 the liver weight in united transplantation group had been completely restored to normal level, however the liver weight in irradiation alone group and BMT alone group were still significantly heavier than that in normal control group (P<0. 05). Liver histopathological examination revealed that there were obvious sinusoidal endothelial cells (SEC) injury, hepatocyte edema and severe inflammatory cell infiltration in irradiation alone group, and on the day 7 the hepatocyte edema and necrosis were significantly worse than before, and almost no alive SEC were found. On the day 14 the injury of SEC in BMT alone group was lighter than before, but on the day 21 the injury had not returned to normal. On the day 7 the injury of SEC, hepatocyte edema and necrosis were alleviated in united transplantation group as compared with irradiation alone group and BMT alone group, and on the day 14 the injury had returned to normal basically. Conclusion The transplantation conditioning could damage recipient liver endothelium and the injury would persist, and united EPC infusion could repair the injured SEC following BMT.
2.Relationship between graft-versus-host disease and endothelium injury following hematopoietic stem cells transplantation in mice
Zhiling YAN ; Lu JIA ; Shijuan XU ; Kailin XU ; Bin PAN ; Guoliang SONG ; Chong CHEN ; Lingyu ZENG
Chinese Journal of Organ Transplantation 2010;31(12):720-723
Objective To study the relationship between graft-versus-host disease (GVHD) and endothelium injury following hematopoietic stem cells transplantation in mice. Methods C57BL/6 mice as donors and Balb/c mice as recipients were randomly divided into 4 groups: control group, bone marrow transplantation group, GVHD group, GVHD mitigation group. The clinical manifestations,circulating endothelial cells and tissue pathological changes were observed at different time points after transplantation. Results No manifestations of GVHD were found in each group at the day 5, while those were found in GVHD group at the day 9 and all died within 15 days. The counts of endothelial cells in peripheral blood showed no significant difference at the day 5 between GVHD group (7. 34 ±1.26 cells/μl) and bone marrow transplantation group (11.51 ± 7. 40 cells/μl) or GVHD mitigation group (7. 36 ± 0. 16 cells/μl), while among three groups there was statistically significant difference at the day 9 (GVHD group: 153. 64 ± 35. 35 cells/μl vs bone marrow transplantation group: 10. 49 ±5. 61 cells/μl and GVHD mitigation group: 47. 82 ± 4. 69 cells/μl). The scores of pathological aGVHD had no significant difference at the day 5 between GVHD group (4. 33± 1. 53) and bone marrow transplantation group (3. 33 ± 0. 58) or GVHD mitigation group (4. 00 ± 1.73), while among three groups there was statistically significant difference at the day 9 (GVHD group: 10. 0 vs bone marrow transplantation group: 3. 33 ± 1.15 or GVHD mitigation group: 4. 33 ± 0. 58) and at the day 14 (GVHD group: 10. 33 ± 2. 58 vs bone marrow transplantation group: 2. 33 ± 1.25 or GVHD mitigation group 3. 33 ± 1.15). Conclusion Occurrence of GVHD causes endothelial damage again and injured endothelium worsens the GVHD.
3.Construction of periodic Brugia malayi cysteine protease inhibitor and glyceraldehydes-3-phosphate dehydrogenase eukaryotic recombinant plasmid and its immunogenicity
Shijuan LU ; Zheng FANG ; Sainan ZHANG ; Hui WANG ; Hao FANG ; Haiyan TONG ; Bangsheng XU
Chinese Journal of Infectious Diseases 2012;30(5):259-263
ObjectiveTo construct the eukaryotic expression plasmids containing cysteine protease inhibitor (CPI) and glyceraldehydes-3-phosphate dehydrogenase (GAPDH) gene from periodic Brugia malayi (Bm),and to observe its cellular immune response in mouse.Methods pcDNA3.1 (+)-BmCPI/BmGAPDH was constructed.The recombinant plasmids were screened and identified by digestion with restriction enzyme.BALB/c mice were injected intramuscularly with a dosage of 100 μg purified recombinant plasmid DNA with GpG oligodeoxynucleotide (CpG ODN) and two same doses were administrated at 2-week intervals.pcDNA3.1 (+) and phosphate buffered solution (PBS) were used as controls.The tissue of muscles at 4 weeks after the third injection was collected and the target gene was detected by reverse transcription-polymerase chain reaction (RTPCR).Two weeks after the third immunization,the stimulation index (SI) of spleen lymphocytes of immunized mice was measured by methylthiazolyldiphenyl-tetrazolium bromide (MTT) method and the serum levels of interleukin (IL)-4 and interferon (IFN)-γ were detected by enzyme-linked immunosorbent assay (ELISA).The data were analyzed by t test.ResultsBmCPI/BmGAPDH gene in the injected muscle of the immunized mice was detected by RT-PCR. At 6 weeks after immunization,the SIot spleen T lymphocytes in pcDNA3.1 (+)-BmCPI/CpG group and pcDNA3.1 (+)-BmCPI/BmGAPDH/CpG group were 1.466 ± 0.635 and 1.610 ± 0.112,respectively,which were both higher than PBS group and pcDNA3.1( +)-CpG group (1.004 ± 0.019 and 1.078 ± 0.129,respectively) (t=64.438,45.318,42.749 and 34.314,respectively; all P<0.05).At 4 weeks after immunization,the serum levels of IL-4 and IFN-γ of mice in pcDNA3.1 ( + )-BmCPI/BmGAPDH/ CpG group were significantly higher than those in pcDNA3.1 (+)-CpG group (t=288.053 and 76.453,respectively; both P<0.05),while the serum level of IFN-γ was also higher than that in pcDNA3.1 (+)-BmCPI/CpG group (t=129.642,P<0.05). ConclusionThe recombinant eukaryotic plasmid pcDNA3.1 (+)-BmCPI/BmGAPDH could be expressed in mice,and could elicit specific cellular immune responses in immunized mice.
4.Clinical analysis of 182 cases of traumatic and hemorrhagic shock
Yunxia WANG ; Shijuan HE ; Xia KANG ; Hui ZHONG ; Hong HE ; Hui XU
Chongqing Medicine 2013;(22):2624-2625,2628
Objective To discuss the importance of saving the life of patients and the severity assessment in early emergency for the trauma and hemorrhagic shock patients in the first-aid of pre-hospital and emergency department.Methods Retrospective anal-ysis of the 182 patients data.Results With early and aggressive treatment,177 cases of survival,5 cases of death,and the survival rate 97.3%.Conclusion The disease of traumatic shock patients is complex,high mortality.the early detection,taking timely and effective rescue measures are the key to increase the survival rate.
5.Prognostic value of APACHE Ⅱ score in patients with severe acute organophosphorus poisoning
Jiyuan ZHANG ; Jinhua GU ; Shijuan HE ; Guoping LIU ; Yunzhang WANG ; Hui XU
Chongqing Medicine 2014;(24):3174-3175
Objective To explore the prognostic value of APACHE Ⅱscore in patients with Severe acute organophosphorus poi-soning .Methods 42 patients with Severe acute organophosphorus poisoning ,in which 34 cases survived ,8 cases dead ,were select-ed .The APACHE Ⅱscores of patients in first 24 h of admission were collected ,and receiver operating characteristic curves (ROC curve) were drawn .Results APACHE Ⅱ score of the 42 patients with Severe acute organophosphorus poisoning was 18 ~30 (20 .11 ± 6 .32) ,in which the survival group was(16 .10 ± 3 .12) ,the dead group was(28 .01 ± 4 .46) (P<0 .01) .With the increase of APACHE Ⅱ score ,the fatality rate gradually increased .The total area under the ROC curves of APACHE Ⅱ score for death judgment was 0 .922 ,APACHE Ⅱ score of 21 .2 was the best diagnostic point ,the sensitivity was 95% ,and specificity was 89% . Conclusion The APACHE Ⅱscore could predict severity of patients with Severe acute organophosphorus poisoning ,and APACHEⅡscore ≥21 .2 could be used as the prognosis for death of the patients .
6.Effects of endothelial progenitor cell combined with allogeneic bone marrow transplantation on hematopoietic reconstitution in mice
Guoliang SONG ; Bin PAN ; Kunming QI ; Licai AN ; Lu JIA ; Shijuan XU ; Shiling YAN ; Kailin XU ; Lingyu ZENG
Chinese Journal of Organ Transplantation 2011;32(11):688-692
Objective To explore a proper dose of endothelial progenitor cells (EPCs)administration that can achieve optimal hematopoietic improving effectiveness in a murine allogeneic hernatopoietic stem cell transplantation (allo-HSCT) model.Methods Female Balb/c mice were lethally irradiated with 60Co source,and then were injected intravenously with 5 106 bone marrow cells from C57BL/6 mice (bone marrow transplantation group).In co-transfer experiments,5 × 104,1 ×105,5 × 105 or 1 × 106 donor EPCs (EPCs treated groups) were injected simultaneously with bone marrow cells.The recipients were monitored for survival,peripheral white blood cells,hematopoietic stem cells (HSCs) and bone marrow histology.Results Compared with bone marrow transplantation group,all EPCs treated groups had accelerated recovery of peripheral white blood cells (P<0.05),platelets (P<0.05) and HSCs (P<0.05).When infused with less than 5 × 105 EPCs,these effective hernatopoietic improving phenomena showed a positive correlation with the administrated doses of EPCs.However,when infused with 1 × 106 EPCs,the mice showed lower survival rate (P<0.05)and slower recovery of peripheral white blood cells (P<0.05),platelets (P<0.05) and HSCs (P<0.05) than 5 × 105 EPCs treated grpup.Bone marrow histopathology analysis confirmed the above findings.Conclusion Co-transfer with donor EPCs can improve survival rate and hematopoietic reconstitution of recipient mice in allo-HSCT,and 5 × 105 EPCs should be a proper dose to achieve the best effectiveness.
7.Sample pretreatment methods of pesticide residues in Panax notoginseng of Chinese traditional medicine.
Na WU ; Wei LIU ; Heping YAN ; Aiping FAN ; Dushu HUANG ; Bo ZHOU ; Xianlan CHEN ; Shijuan XU ; Ling SHI
Acta Pharmaceutica Sinica 2013;48(10):1585-9
Two sample pretreatment methods of pesticide residues in Panax notoginseng of Chinese traditional medicine were developed. For Method I, the residues were extracted from homogenized tissue with n-hexane-dichloromethane (6:4) by means of ultrasonication, the crude extract was purified by an Envi-carb/NH2 solid-phase extraction (SPE) column. For Method II, matrix solid-phase dispersion (MSPD) technique was used for extracting and cleaning up. The eluates were concentrated by rotary evaporation, and then were redissolved in dichloromethane prior to GC-MS determination. The determination was performed in selected ion monitoring (SIM) mode with the external calibration for quantitative analysis. Under the optimal conditions, the results indicated that the methods are easier and faster, the recoveries of method I for the spiked standards at concentration of 0.01, 0.5, and 2.0 mg x kg(-1) were 81.90%-102.10% with the relative standard deviations (RSDs) of 3.60%-7.10%. The recoveries of method II were 96.26%-104.20% with the RSDs of 3.52%-7.94%. The detection limits (S/N) for residues of pesticides were in the range of 0.48-1.34 ng x g(-1). The results indicated that these multiresidue analysis methods can meet the requirements for determination of residue pesticides and can be appropriate for trace analysis of residue pesticides in Panax notoginseng.
8.A meta-analysis of clinical efficacy on Wuling San treatment for chronic heart failure
Fei GAO ; Shijuan YI ; Xiaojie XU
International Journal of Traditional Chinese Medicine 2018;40(12):1160-1164
Objective To evaluate the clinical efficacy of Wuling San in the treatment of Chronic heart failure by meta analysis. Methods The randomized controlled trials were searched from CNKI, Wanfang Data, VIP Data, CBM Data, Cochrance Library, PubMed, EMbase. Review 5.2 Manager software, stata 14.0 software, GRADE profiler 3.6.1 software were used to bias risk assessment, meta analysis, and quality evaluation of the studies. Results Fifteen studies with 1160 patients were included. The overall quality of studies is low. The meta analysis results showed that the total efficiency of improving heart function in the treatment group were superior to the control group [OR=4.60, 95% CI (3.12, 6.77), Z=7.72, P<0.01]. Due to the heterogeneity, the studies of left ventricular ejection fraction (LVEF) were divided into two subgroups according to the course of treatment, one group with more 4 weeks course treatment and the other group with less 4 weeks. The LVEF in the treatment group was superior to the control group [WMD=9.53,95% CI (8.80, 10.28), Z=25.63, P<0.01]. The brain natriuretic peptide (BNP) iin the treatment group was superior to the control group [WMD=-65.52, 95% CI (-88.45, -48.59),Z=6.74,P<0.01]. Conclusions The present meta-analysis suggests that combination of conventional western medicine and Wuling San has better effect on heart failure. Due to the poor quality of included RCTs, the conclusions may exist bias. Thus, more RCTs with high quality are needed.
9.Study on UPLC fingerprint and content determination of mangiferin of Gentiana rhodantha
Chaokun YANG ; Shijuan XU ; Wenfen XU ; Qingwen SUN ; Bo WANG ; Jiangtao GUO ; Yongping ZHANG
China Pharmacy 2023;34(1):34-39
OBJECTIVE To provide reference for quality control of Gentiana rhodantha. METHODS Taking 52 batches of G. rhodantha as subject, ultra-high performance liquid chromatography (UPLC) fingerprint was adopted. The similarity of 52 batches of medicinal materials samples was evaluated by the Similarity Evaluation System for Chromatographic Fingerprints of Traditional Chinese Medicine (2004A edition); the content of mangiferin was determined; chemometric analyses [cluster analysis, principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA)] were performed. RESULTS UPLC fingerprints of 52 batches of G. rhodantha were established, 17 common peaks were identified, and 6 of them were identified, which were loganic acid (peak 1), neomangiferin (peak 3), swertiamarin (peak 5), dangyin (peak 6), mangiferin (peak 7) and isoorientin (peak 9). The similarities of 52 batches of medicinal materials samples were all greater than 0.9; cluster analysis showed that S1-S46, S48-S52 clustered into one class, and S47 alone; PCA results showed that the cumulative variance contribution rate of the first six principal components was 82.928%; OPLS-DA results showed that the corresponding components of swertiamarin, mangiferin and chemical composition represented by peak 4, 14, 15, 16 were the main iconic components affecting the quality differences of G. rhodantha medicinal materials. The contents of mangiferin in 52 batches of medicinal material samples ranged from 18.2 to 101.0 mg/g, mostly in accordance with 2020 edition of Chinese Pharmacopoeia. CONCLUSIONS The established UPLC fingerprint and chemometric analysis methods combined with content determination method of mangiferin can comprehensively evaluate the quality of G. rhodantha.