Objective To investigate the effects of anti-HLA donor-specific antibodies(DSA)and desensitization for DSA+patients on engraftment of haploidentical hematopoietic stem cell transplantation(haplo-HSCT).Methods The patients who underwent haplo-HSCT and examinations for HLA antibodies and DSA in our department from March 2017 to July 2023 were recruited in this study.The effects of desensitization measure on engraftment in the DSA+patients after haplo-HSCT were analyzed.Results Among the 70 patients who underwent haplo-HSCT and test for HLA antibodies,15(21.4％)patients were DSA positive,including 7(46.7％)cases of strong positive,3(20.0％)cases of moderate positive,and 5(33.3％)cases of weak positive.The median duration for neutrophil implantation was significantly extended in the DSA+patients than the negative patients(P=0.027).For the 6 patients developed graft failure(GF),4 were DSA+which was statistically higher than the DSA-patients(P=0.025).Multivariate regression analysis showed that DSA was an independent factor affecting GF(HR=9.273,95％CI:1.505～57.124,P=0.016).Among the 10 patients(7 strong positive and 3 moderate positive DSA)received desensitization therapy,4 patients received combination desensitization,with a 100％rate of successful transplantation,and 6 received single desensitization,with 4(66.7％)experiencing GF,so the GF rate was obviously lower in the combination than the single desensitization(P=0.008).Conclusion In haplo-HSCT patients,DSA is an important factor leading to implantation delay and GF.While,single desensitization treatment has limited efficacy.In combined DSA desensitization therapy,the decrease of antibody titer should be dynamically monitored to ensure the successful implantation of stem cells and reduce GF rate.

Objective To explore the source of endothelial cells in tumor vessels by A 549 tumor model with GFP nude mouse.Methods To establish the A 549 lung cancer models with GFP nude mice, expression of CD 31 was determined by immunofluorescence to label tumor vessels; to observe and take a picture of the tumor frozen section by confocal microscopy and invert microscope;expression of GFP in tumor vessels was determined by immunohistochemistry. Result The results of immunofluorescence showed:Tumor interstitial vascular endothelial cells or endothelial cells clusters and micro-vascular lumen size and shape are clearly visible by immunofluorescence, and part of vessels with no obvious lumen or irregular lumen.We can see green fluorescent in tumor cells of tumor tissue and endothelial cells which form of tumor vessels.The results of immunohistochemistry showed: expression of GFP was determined in cytoplasm of tumor stromal cells and endothelial cells in tumor vessels.Conclusion The endothelial cells which formed tumor neovessels that derived from GFP nude mice partly and the other part derived from tumor cells.

Objective There are a few reports on rats with spontaneous congenital cataract in China .The purpose of this study was to investigate the morphological changes of lens in rats with spontaneous congenital cataract . Methods 24 d, 1-year rats with cataract and microphthalmos cataract and normal rats (n=5) were selected as research objects .Their lens were observed by a slit lamp microscope and taken photos in front of them , followed by examination through light micrograph and transmission electron micros-copy. Results Rats with microphthalmos cataract showed narrowed palpebral fissure and broaden nucleus while rats with cataract showed normal palpebral fissure and narrowed nucleus .As for 24 d,1-year rats with microphthalmos cataract , the fibers of their lens showed derangement and vacuole-like degeneration by light microscope , in addition, the abnormal connection between fiber cells were observed by electron microscopy .As for 1-year normal rats , the fibers were in consistent structure and regular arrangement without cell ingredient . Conclusion The appearance and morphological changes of the lens in rats with spontaneous congenital cataracts are in consistence with the pathological changes of cataracts , which is appli-cable in further research on the pathogenesis of cataract .

[Abstract ] Objective Evidence from previous studies indicated that over-activation of C3a/C3aR axis existed in the renal tubular epithelial cells of patients with renal diseases including diabetic nephropathy .However , the pathological significance of over-ac-tivating C3a/C3aR axis still remains to be elucidated .In this study, we constructed a renal tubular epithelial cell line over-expressing C3a in a secretory manner in order to provide a cell model to investigate the pathological significance of over -activating C3a/C3aR axis under various pathological scenes . Methods We designed a synthesized C3a secretory expression unit and cloned it into the multi-clonal site of lentivirus expression vector pLenti 6.3-MCS-IRES2-EGFP.After identification by sequencing , recombinant lentivirus was packaged by using pLenti 6.3-C3a-IRES2-EGFP and packaging plas-mid in 293T cells.Then, the recombinant lentivirus was used to in-fect HK2, a cell line of human renal tubular epithelial cells .After screening in medium with blasticidin , blasticidin resistant cell clones were obtained .Real-time PCR and ELISA method were applied to analyze the expression and secretion of stable transfected cells cloned C3a and identify renal tubular epithelial cell lines with stable over-activating C3a. Results ①C3a secretory expression unit was suc-cessfully synthesized and correctly cloned into the multi-clonal site of pLenti6.3-IRES2-EGFP; ②C3a secrectory expression recombi-nant lentivirus LV-C3a was successfully packaged with a high titer of 5 ×108/mL;③HK2 Cell clones resistant for blasticidin were ob-tained;according to the analysis of Real-time PCR and ELISA, the C3a mRNA level in HK2-C3a cell lines was significantly higher than that of HK2 cells(1.0 ±0.5 vs 1321.0 ±18.0, P<0.01) and the secreted C3a level increased significantly ([0.3 ±0.2]ng/mL vs [249.0 ±37.0] ng/mL, P<0.01). Conclusion The present study successfully constructed C 3a secretory expression vector pLenti6.3-C3a-IRES2-EGFP and C3a over-expression renal tubular epithelial cell line HK 2-C3a, which is very useful in further study of the function and significance of C 3a/C3aR axis not only in renal tubular epithelial cells but also in other cell types .

Objective To analyze the clinical characteristics ,diagnosis and treatment of acute leukemia complicating deep ve‐nous thrombosis(DVT) to deepen the cognition on this complication .Methods The clinical data of consecutive patients with acute leukemia were performed the retrospective analysis .The occurrence situation of deep venous thrombosis was investigated and the a‐broad related literatures were reviewed .Results A total of 116 cases of acute leukemia in our department from July 2011 to March 2014 were treated ,in which 85 cases were acute myeloid leukemia and 31 cases were acute lymphoblastic leukemia;5 cases devel‐oped DVT with the proportion of 4 .31% (5/116) .Of these cases ,3 cases were acute promyelocytic leukemia and 2 cases were acute lymphoblastic leukemia .Conclusion The occurrence rate of DVT in the patients with acute promyelocytic leukemia is relatively higher ,if the patients have the corresponding symptom ,timely diagnosis and treatment should be conducted .

Objective To measure the range of routine blood counts and blood biochemical parameters ,to analyze hemorheology of the rats with congenital cataract .Methods Blood samples were taken from 90 rats with congenital cataract weight about 185 ~211 g.Routine blood analysis was performed and blood biochemical and hemorheology paramenters were determind using an automatic blood biochemical and hemorheology analyzer .Results There were no significant difference ( P >0.05 ) between the cataract rats and the normal rats in Blood test results; but there were significant difference between the microphthalmos cataract rats and the normal same -sex rats ( P <0.01 or P <0.05 ) . The biochemical results is the cataract rats and the normal rats were different significantly in ALB group ( P <0.01 or P<0.05), and the female microphthalmos cataract rats compared with the control rats had significant difference in Ure (P<0.01) , the female cataract rats ompared with the normal rats were very significant difference in Cr group ( P <0.05 or P <0.01).The erythrocyte counts of the male cataract rats and male microphthalmos cataract rats were significantly lower than that in the female ones, respectively(P <0.05, P <0.01).The platelet counts of the male cataract rats and the male microphthalmos cataract rats were significantly higher than that in the female ones , respectively(P <0.01), and the creatinine of the male cataract rats and the male microphthalmos cataract rats were significantly lower then that in the female ones, respectively(P <0.01).There were no significant difference in every group on hemorheology .Conclusions There were significant differences in some blood indexes between the congenital cataract rats and the normal rats .These data may become useful reference for biomedical researcher in this field .

Objective Increased renal mast cells have been found in significant correlation with clinicopathological features of diabetic nephropathy ( DN) .However, the exact pathological significance of mast cells still remained to be elucidated.As one of the most abundant serine proteases, tryptase is specifically expressed by mast cells.The study was to observe the expression of tryptase in the urine of patients with diabetic nephropathy and realize the pathological significance of urinary tryptase and renal mast cells in the development of diabetic nephropathy. Methods Urine samples from 103 patients with diabetic nephropathy who were hospitalized at the clinical unit of the nephrology centre of Jingling Hospital from January 2012 to June 2013 were collected.According to concentra-tion of urinary albumin excretion rate, the patients were conducted to early-stage DN group (microalbuminuria,n=10), middle-stage DN group (proteinuria stage, n=31) and end-stage DN group (renal insufficiency stage,n=62).Meanwhile, urine samples from 20 normal persons were taken as the normal control group.Tryptase level in each urine sample was measured by using an ELISA kit.The average tryptase levels were compared among different groups and the correlation between urinary tryptase level and clinical indices was analyzed. Results ①In most cases, tryptase was not detected in the urine samples from normal persons(0.7 ±1.4 ng/mg creati-nine).However, the urinary trypatse level increased significantly with the development of diabetic nephropathy (11.6 ±10.5 ng/mg creatinine for early stage, 29.7 ±19.2 ng/mg creatinine for middle stage, and 44.6 ±43.4 ng/mg creatinine for late stage group).②The urinary tryptase level was found in significant correlation with ser-um creatinine (r=0.325, P<0.01), serum cystatin C (r=0.391, P<0.01), serum urea nitrogen (r=0.27, P<0.01), 24 hour uri-nary protein (r=0.389, P<0.01), urine C3 (r=0.276, P<0.05), urine retinol-binding protein (r=0.365, P<0.01), urine lysozyme (r=0.461, P<0.01), urine N-acetyl-β-glucosaminidase level (r=0.568, P<0.01), urinary kidney injury molecule-1 (r=0.434, P<0.05) and interleukin-18 level (r=0.375, P<0.05).No significant correlation was found between urinary tryptase level and age, gender, fasting blood sugar, postprandial blood sugar, HbA1c, triglyceride, high density lipoprotein or low density lipo-protein. Conclusion Mast cells play an important role in the development of diabetic nephropathy and might be a novel and effective target for the treatment of DN.

Objective To study the effects of toys for laboratory animals on reproductive performance and growing development of experimental mice.Methods ICR mice were fed with toys, the informations of reproductive performance and growing development were recorded.Results All the data of reproductive performance of the test group were higher than the control group except the number of newborn mice, and showed significant difference ( P <0.05 ) .The data of growing development of the test group were higher than the control group too, but showed no significant difference ( P >0.05).Conclusion Toys for laboratory animals have good effects on reproductive performance and growing development of mice, and suggested to be used into the process of experimental mice raising.

Objective To study whether the green fluorescent protein ( GFP) gene can be successfully expressed in green fluorescent nude mice and the tissue distribution characteristics.Methods Small animal imaging system and RT-PCR assay were used to detect the GFP tissue distribution and fluorescence expression level.Results The GFP can be expressed in multiple tissues in green fluorescent nude mice.A higher expression was observed in the pancreas, heart, brain, and skin.Conclusion Exogenous GFP can be stably expressed and inherited in green fluorescent nude mice, with the highest expression in the pancreas.

Objective:To determine the immunological effects of a new type adjuvant CTA 1-DD/IgG.Methods:Intranasal im-munization of BALB/c mice with OVA in combination with adjuvant CTA 1-DD or CTA1-DD/IgG.Blood samples were subjected to OVA-specific IgG Abs detection and IgG subclass profiles assessment using ELISA.Splenic plasma cells secreting anti-OVA Abs were detected by ELISPOT.Results:CTA1-DD/IgG showed stronger activity in inducing OVA-specific IgG Abs and splenic Abs-secreting cells compared with CTA1-DD.The enhancement of IgG1,IgG2a and IgG2b Abs were observed,and the ratio IgG2a/IgG1 were >1 in both groups.Conclusion: CTA1-DD/IgG can exert enhanced adjuvant effects compared with CTA 1-DD.Mixed IgG subclasses are induced,indicating a more balanced Th1/Th2 response.