Preparation of biotin-grafted pullulan acetate nanoparticles conjugated with CD3 antibodies and their uptake of T cells / 医学研究生学报

Objective Cytotoxic T lymphocytes are the main effector cells of anti-tumor immunity. Active targeting of nanoparticles to T cells and activation of T cells can be achieved by conjugation with specific antibodies. We prepared the biotin-grafted pullulan acetate nanoparticles conjugated with CD3 (Bio-PA-CD3 NPs), and explored their effects on the proliferation, cytokine secretion and uptake of CD8+T cells.Methods We prepared Bio-PA NPs by the dialysis method, conjugated CD3 antibodies to the surface of NPs to make Bio-PA-CD3 NPs, and measured the diameter and Zeta potential of the NPs. We evaluated the effects of the NPs on the proliferation of CD8+T cells and the secretion of cytokines by CCK-8 assay and ELISA, respectively, and quantitatively analyzed the cellular uptakes of the Bio-PA-CD3 NPs by the flow cytometry.Results The Bio-PA-CD3 NPs exhibited regular spherical shapes of even size and with no adhesion. The content of CD3 antibodies on the surface of the NPs decreased with the increased degree of biotin substitution. The CD3 contents of the Bio-PA-CD3 NPs with biotin substitution degrees of 1.6%, 5.4% and 6.3% were (36.1±4.4), (21.4±4.3) and (10.3±4.7) μg/mg, respectively. Compared with Bio-PA NPs, Bio-PA-CD3 NPs at a certain concentration significantly enhanced the proliferation of CD8+T cells in vitro and promoted the secretion of IFN-γ, TNF-β and IL-2 cytokines. The Bio-PA-CD3 NPs manifested a higher cellular uptake with the increased content of CD3 antibodies.Conclusion The Bio-PA-CD3 NPs we prepared could be a promising agent to enhance the immune effect of T cells.

Association between human papillomavirus infection and adverse pregnancy outcome / 预防医学

Objective To investigate the infection rate of human papillomavirus(HPV)among pregnant women, and to explore the effect of HPV infection on adverse pregnancy outcome. Methods A total of 1 679 pregnant women in hospital were collected for the research. The flow-through hybridization and genechip(HybriMax)method was used to detect the infection of HPV. Univariate analysis was used to analyze the factors affecting HPV infection in pregnant women. The binary logistic analysis was used to analyze risk factors affecting adverse pregnancy outcome. Results HPV infection rate was 31.39%(527/1 679), including 14.23%(239/1 679)of HR-HPV, 15.54%(261/1 679)of LR-HPV and 1.61%(27/1 679)of mixed of HR-HPV and LR-HPV. Univariate analysis showed that there was significant difference in initial sex age, education level and smoking history between infection group and non-infection group, with statistical difference(P<0.05). The incidence rate of adverse pregnancy outcomes in infection group(31.50%) was significant higher than that of non-infection group(9.81%), with statistical difference(P<0.01). The incidence rate of premature rupture of fetal membranes, newborn respiratory papillomatosis and other adverse pregnancy outcomes among HR-HPV group, LR-HPV group and mixed group had no significant difference (P>0.05). Binary logistic regression analysis showed HR-HPV infection(OR=4.194, 95% CI: 3.099-5.675), LR-HPV infection(OR=1.771, 95%CI: 1.288-2.434)and mixed type infection(OR= 3.350, 95%CI: 1.630-7.735)were the risk factors affecting adverse pregnancy outcome(P<0.01), however, age and times of gestation had no statistical significance in the binary logistic analysis(P>0.05). Conclusion HPV infection was the risk factors for adverse pregnancy outcome, which indicated that screening work in pre-pregnancy and pregnancy, and persisting in early prevention, early detection and early treatment could reduce the incidence rate of adverse pregnancy outcome.

Differential regulation of CCR5 expression on T lymphocytes in healthy donors after mobilization with rhG-CSF and its correlation with aGVHD / 中国实验血液学杂志

This study was to investigate the differential regulation of CCR5 expression on T cells in healthy donors after mobilization with recombinant human granulocyte colony-stimulating factor (rhG-CSF) and analyze its correlation with acute graft-versus-host disease (aGVHD) so as to understand the possible mechanisms underlying rhG-CSF-induced immune tolerance. Sixty-eight related healthy donor and their corresponding recipient for allogeneic hematopoietic stem cell transplantation (allo-HSCT) were enrolled in this study. The expression of CCR5 on CD4(+) and CD8(+) T cells in the peripheral blood (PB) before and after mobilization were detected by using flow cytometry (FCM) respectively. According to the changes of CCR5 expression on CD4(+) and CD8(+) T cells, the Sixty-two evaluable donors were divided into the downregulated and unchanged/upregulated (non-downregulated) groups, and the incidence of grades II to IV aGVHD in two groups were compared. The results showed that the mean value of CCR5 expression on CD4(+) and CD8(+) T cells in PB was not different significantly after mobilization (P > 0.05). Apparent inconsistency was showed among different individuals. Thirty-four (50%) donors displayed downregulation of CCR5 expression, while 34 (50%) donors manifested unchanged or upregulated CCR5 expression on CD4(+) T cells. CCR5 expression on CD8(+) T cells was downregulated in 42 (61.8%), unchanged or upregulated in 26 (38.3%) donors. The cumulative incidence of grades II to IV aGVHD in the downregulated and non-downregulated groups for CD4(+) T cells were 16.1% and 41.9% (P = 0.032), and recipients with CCR5 downregulation on CD8(+) T cells showed an increased tendency of developing aGVHD (37.8% vs 16.0%, P = 0.065). In conclusion, rhG-CSF mobilization could lead to differential regulation of CCR5 expression on T cells, which might influence the migration of T cells in vivo, decrease T cell trafficking towards GVHD target organs, and thus reduce the incidence of aGVHD after transplantation.

Incidence and risk factors of hemorrhagic cystitis after hematopoietic stem cell transplantation / 中国实验血液学杂志

The aim of this study was to analyze the risk factors of hemorrhagic cystitis (HC) after hematopoietic stem cell transplantation (HSCT). The data of 188 transplant patients treated from July 2003 to August 2009 in Peking University First Hospital were studied. The patients were followed up to 180 days after HSCT. Clinical records of the total 188 cases and the 150 cases underwent allogeneic HSCT out of 188 cases were analyzed by using a Cox regression model respectively. The results indicated as follows: (1) 51 of 188 patients developed HC (27.12%). Univariate analysis showed that sex (male RR = 1.673, p = 0.076), allogeneic HSCT (RR = 1.848, p = 0.061) were related to HC, and allogeneic HSCT (RR = 4.508, p = 0.037) was the independent risk factor for HC by multivariate analysis. (2) HC occurred in 32.67% (49/150) patients who underwent allogeneic HSCT, with 42 cases of grade II-IV HC (28.00%). For the incidence of grade II-IV HC, univariate analysis revealed mismatched related donor/matched unrelated donor (RR 2.444, p = 0.018), CMV viruria (RR 2.059, p = 0.021) and CMV positive in serum and urine at the same time (RR 2.497, p = 0.003) were risk factors. The following factors, as conditioning with Fludarabine (Flu) (RR 0.504, p = 0.059) and TBI (RR 0.185, p = 0.095), were associated with a decreased tendency of II-IV HC at age of 26 - 40 (compared with age ≤ 25 or ≥ 41, RR 0.454, p = 0.056). Some others, as conditioning with CTX (RR2.015, p = 0.063), the application of ATG (RR 2.343, p = 0.054) and CMV viremia (RR 2.123, p = 0.088), were associated with an increased tendency of II-IV HC by univariate analysis. Multivariate analysis demonstrated that CMV positive in serum and urine at the same time (RR 2.269, p = 0.008), conditioning without Flu (RR = 2.106, p = 0.040) were the independent risk factor for grade II-IV HC. And the application of ATG (RR = 2.154, p = 0.083) was related to the tendency of higher incidence of grade II-IV HC. It is concluded that the incidence of HC is high in patients underwent allogeneic HSCT. CMV positive in serum and urine at the same time, while conditioning without Flu are the independent risk factors of grade II-IV HC. Application of ATG is related to the increased trend of grade II-IV HC.

Occult HBV infection in patients with anti-HBc positive alone / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>This study was designed to explore the incidence rate of occult HBV infection in patients with anti-HBc positive alone and analyze the possible reasons of occult infection.</p><p><b>METHODS</b>Sera of 183 patients carrying anti-HBc alone(A < or = 0.1) were collected and real-time PCR was used to select samples with HBV DNA positive. HBV pre-S/S amplification products were obtained by PCR, and clonal sequencing were then used for these samples with HBV DNA positive.</p><p><b>RESULTS</b>DNA quantitative results of three samples were greater than 10(3) copies/ml in 183 samples, with a fraction of 1.6%. Pre-S/S sequencing results of two samples from these three samples were obtained. Point mutations within "a" determinant with Q129R/P mutations and co-existence of the mutant type and wild type were found in the two samples.</p><p><b>CONCLUSIONS</b>Occult HBV infection existed in samples with anti-HBc alone. Factors contributing to the loss of HBsAg detection by immunoassays include S gene mutations and low levels of circulating antigen which are below the assay limit of detection. Occult HBV infection not only can lead to a false clinical diagnosis, but also can result in hematological pollution due to such occult infection of blood donors.</p>

Alteration of chemokines after allogeneic hematopoietic stem cell transplantation and its clinical significance / 中国实验血液学杂志

The objective of this study was to analyze the relationship between some chemokines and the pathogenesis of GVHD and to find some biomarkers with diagnostic significance through observing and comparing the changes of some chemokine levels in samples from patients with or without aGVHD after allogeneic hematopoietic stem cell transplantation (allo-HSCT). 26 plasma samples were obtained from 26 patients undergoing allo-HSCT at various time points after transplantation, of which 13 samples from patients with aGVHD were served as investigating group and 13 samples from patients without GVHD after Allo-HSCT were used as control group. The patient characteristics between the two groups were compared, the levels of 40 chemokines in these samples were detected by ELISA, the changes of chemokine levels in samples of 2 groups were analyzed by means of significance analysis microarray (SAM), clustering method and c-test. The results showed that there were significant differences in levels of 6 chemokines including HCC-1, MIF, IP-10, ITAC, TARC and NAP-2 between 2 groups, in which the level of MIF in plasma samples after HSCT was the highest, the difference of TARC level between 2 groups was over 10-fold. It is concluded that the level changes of chemokines mentioned above can be used as a indicator of GVHD presence, but their pathogenetic role in occurrence of aGVHD remains to be determined.

Efficacy analysis of sequential treatment with chemotherapy, ATRA and As(2)O(3) for acute promyelocytic leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the efficacy and treatment outcome of different induction regimens, and different post-remission therapies for adult acute promyelocytic leukemia (APL).</p><p><b>METHODS</b>The outcome of 73 patients with newly diagnosed APL were retrospectively analyzed. According to the induction regimens, the patients were divided into three groups: chemotherapy-only (14 cases group I), all-trans retinoic acid (ATRA) or combined with chemotherapy (33 cases group II), and ATRA combined with arsenic trioxide (As(2)O(3)) (26 cases group III). The complete remission (CR) rate and the time to CR (TTC) were analyzed. After CR, the patients were divided into 2 groups for post-remission therapies: one with sequential treatment of chemotherapy/ATRA/As(2)O(3) and the other with alternative treatment of chemotherapy/ATRA. The overall survival (OS), disease free survival (DFS) and relapse rate were compared between these two groups. Patients induced CR with both ATRA and As(2)O(3), and then sequentially treated with chemotherapy/ATRA/As(2)O(3) (group A), and those induced CR with ATRA or As(2)O(3) alone and then with non-chemotherapy/ATRA/As(2)O(3) sequentially (group B) were also analyzed and compared for CR, OS and DFS.</p><p><b>RESULTS</b>(1) For induction treatment, the CR rate in ATRA and As(2)O(3) combination group was 100%, in ATRA combined with chemotherapy group was 78.8%, and in chemotherapy-only group was 57.1% (P = 0.030). The median TTC in ATRA with As(2)O(3) combination group was 26 (13 - 40) days being the shortest among the three groups. (2) For the post-remission treatment, 3-year OS rates in group I and group II were (95.7 ± 4.3)% and (68.6 ± 11.2)% (P < 0.05), and 3-year DFS rates were (79.0 ± 9.5)%, and (32.9 ± 15.5)%, respectively (P < 0.01). The relapse rate was 14.8% in group I, and 50.0% in group II (P = 0.011). (3) The CR, 3-year OS and DFS rates in group A were all 100%. The CR rate in ATRA or As(2)O(3) alone induced group was 72.9%, and 3-year OS was (72.3 ± 9.1)% (P < 0.05).</p><p><b>CONCLUSIONS</b>For adult APL induction with ATRA and As(2)O(3) combination can obtain a higher CR rate, and shorter TTC. The post-remission treatment with sequential chemotherapy, ATRA and As(2)O(3) results in a lower relapse rate, and significantly improves OS and DFS. The ATRA and As(2)O(3) combination induction with the sequential post-remission therapy is the best strategy for APL treatment.</p>

Analysis of the articles published in Chinese Journal of Applied Physiology between 2000 and 2006 / 中国应用生理学杂志

<p><b>AIM</b>To analyze the characteristics of publications of Chinese Journal of Applied Physiology between 2000 and 2006, evaluate the academic level and the popularity of the issues, and supplying an evidence for the journal reform.</p><p><b>METHODS</b>With CNKI and manpower search, by use of literature metrology, a comprehensive analysis of the publications of Chinese Journal of Applied Physiology between 2000 and 2006 was made.</p><p><b>RESULTS</b>The number of articles published form 2000 to 2006 in Chinese Journal of Applied Physiology was 968, The average number of each issue is 34.57, the average page of each article is 3.11, in the columns, the article about original articles was top of rank (66.22% of the total). In the quotation, the quotation increase year by year (100% in 2004-2006), the number of English quotation is very more (76.52% in average). In the time lag, the longest is 510 days, the shortest if 60 days, the average is 196.51 days. In the fund support, the level is increase by fund support, the article number by fund support is increase too, It is 97 in 2005. In the authors' professional positions and academic degrees, the authors' level is more and more higher. In the authors column, Beijing's author is the top of rank, has 162 persons (16.74% of the total).</p><p><b>CONCLUSION</b>The Chinese Journal of Applied Physiology has published high quality articles. It is the one of the most important information resource for the physiological research and the most important medical journal.</p>

Correlation between interleukin-31 expression on peripheral blood mononuclear cells and disease severity of pediatric patients with atopic dermatitis / 中华皮肤科杂志

Objective To evaluate the significance of human interleukin-31(IL-31)in the pathogenesis of atopic dermatitis and its correlation with pruritus in patients with atopic dermatitis(AD).Methods Twenty-two children with mild to severe atopic dermatitis and 22 age-matched healthy controls were included in this study.Patients and controls were randomly and equally assigned into stimulation and non-stimulation groups.Venous blood samples were obtained from all participants,peripheral blood mononuclear cells were isolated from these samples and cultured with(stimulation groups)or without(non-stimulation groups)staphylococcal enterotoxin B(SEB)for 24 hours.Then,the mRNA expression of IL-31 on PBMCs was assessed via real-time reverse transcription-PCR.ELISA was used to detect the total serum IgE level in these objects.The severity of AD in patients was rated according to scoring atopic dermatitis(SCORAD).The relationship between the mRNA expression of IL-31 and the level of serum total IgE.severity of atopic dermatitis,and degree of pruritus.was evaluated.Results The expression of IL-31 mRNA on non-stimulated PBMCs from patients was 23.2 folds as high as that from the healthy controls(P＜0.01).The stimulation with SEB upregulated the mRNA expression of IL-31 on PBMCs.and the increase on PBMCs from patients was 20.44 times of that from the controls.The total serum IgE level was 260.05 IU/mL(5.9-1131.01 IU/mL)and 17.7 IU/mL(5-140.7 IU/mL)in the Patients and controls respectively(P＜0.01).There was no significant correlation between the mRNA expression of IL-31 and disease severity or total serum IgE level(r=0.07.0.22respectively.both P＞0.05)in patients witll AD.Condusions IL.3 1 is involved in t11e pathogenesis of AD,which is unlikely to be IgE-dependent.SEB can induce the rapid expression of IL-31 on PBMCs of healthy human,and is an important modulator for the production of IL-31.